# DCN Dx — Full Content (llms-full.txt) > For more than 20 years, DCN Dx has done one thing: help companies bring diagnostic tests to commercial success. Today, the company runs three specialized businesses. Its immunoassay CDMO has completed more than 650 programs and is the industry's go-to for developing and manufacturing rapid point-of-use tests, from early feasibility through full-scale production in a 35,000-square-foot facility certified to ISO 13485 and ISO 9001. Its biospecimen collections business gives assay developers global access to diverse, consented specimens under IRB/IEC oversight and matched to the populations an assay must serve. Its IVD-dedicated CRO is led by former sponsor-side experts who carried their own diagnostics through FDA and EU IVDR review and now design and manage clinical studies and regulatory submissions through to approval. Based in Carlsbad, CA. Learn more at dcndx.com. > Full text of DCN Dx's Insights articles and Products for AI ingestion. Items are point-in-time and dated; for any regulatory topic prefer the most recent. Index: https://dcndx.com/llms.txt # ===== INSIGHTS ===== ## How DCN Runs Better IVD Trials on Veeva URL: https://dcndx.com/insights/how-dcn-runs-better-ivd-trials-veeva/ Type: insight Published: 2026-06-30 DCN Dx’s clinical and data management leaders, joined by Veeva, examine what IVD CROs gain when they replace disconnected study systems with a single connected platform. Running an IVD study on a mix of spreadsheets, file shares, and siloed tools doesn’t just slow things down. It creates data delays, missed handoffs, and a fragmented picture of what’s happening in your trial. In this episode of Expert Insights, DCN Dx’s Emily Friedland sits down with Clinical Trial Manager Jill Hutton-Pugh and Associate Director of Data Management Thomas Martinache to talk through what study management looked like before everything was connected, and what changed once it was. They’re joined by John Acampado of Veeva MedTech, who spent years on the sponsor side before coming to Veeva, and who brings that perspective to the conversation. DCN Dx built its connected platform in phases, starting with the eTMF in 2022 and adding EDC and CTMS through 2024. That progression made possible what sponsors in IVD need most: real-time enrollment visibility, faster study builds, cleaner data, and documentation that holds up in an audit without anyone scrambling. For small and mid-sized IVD companies that assume a platform like Veeva is only for Big Pharma, this episode is a direct counterargument. The conversation covers IVD-specific templates, one-week EDC build timelines, and why a unified foundation matters even more as AI comes into the picture. Listen below, or find us on your favorite podcast platform. What You’ll Hear in This Episode Why patchwork systems create data silos, delayed queries, and reporting gaps that hurt IVD sponsors on fast-moving studies How DCN Dx built its connected platform in phases and what each step, from eTMF to EDC to CTMS, made possible How real-time enrollment data and integrated monitoring reports give sponsors a live view of their study instead of a snapshot from days ago Why reusing roughly 60% of EDC work from study to study means faster builds and study-start timelines measured in days, not weeks What audit and inspection readiness looks like when all documentation, traceability, and site data live in one compliant, connected system Enjoyed this discussion? Dive deeper with Veeva and DCN Dx in their webinar, Platform Over Patchwork: How Consolidation Drives CRO Efficiency. Register here. --- ## A Must-Watch DCN Dx Webcast from RAPS: Parallel Regulatory Planning for EU and US Market Entry URL: https://dcndx.com/insights/must-watch-dcn-dx-webcast-from-raps-parallel-regulatory-planning-market-entry/ Type: insight Published: 2026-06-28 (As published 2026-06-28; regulatory status may have changed since.) DCN Dx thanks everyone who joined our recent Regulatory Affairs Professionals Society (RAPS) webcast, The IVDR Transition in 2026: Parallel Regulatory Planning for EU and US Market Entry. The session looked at how diagnostic teams can run EU and US regulatory programs at the same time, and why the work that prevents late-stage surprises starts in design and development, not at submission. The webcast was presented by DCN Dx’s Director of Regulatory Affairs, Dan Simpson, RAC, and Senior Clinical Trials Manager, Sarah Barchard. They placed the IVDR conformity assessment and FDA marketing authorization frameworks side by side, then focused on the pitfalls that surface during design validation when parallel requirements are not addressed early. The discussion covered IVDR classification under Annex VIII, performance evaluation under Annex XIII, the proactive postmarket expectations under IVDR, and the December 2025 EU proposal to simplify the MDR and IVDR. Five anonymized case studies, drawn from real programs, showed how class misalignment and underestimated clinical evidence play out, and how teams corrected course. Missed the live session? You can now watch the full recording by submitting the form below: --- ## Stuck in a Holding Pattern: The IVDR Revision and Planning for Two Markets at Once URL: https://dcndx.com/insights/stuck-holding-pattern-ivdr-revision-planning-markets/ Type: insight Published: 2026-06-25 (As published 2026-06-25; regulatory status may have changed since.) Why waiting on the IVDR revision carries a cost, and what it takes to plan for the EU and US at the same time. The 2026 IVDR transition deadlines have arrived, and many IVD companies are holding off on filing while they wait to see what becomes of the European Commission’s December 2025 revision proposal. The waiting has consequences. By late May, notified bodies reported that submissions had slowed enough that some cut staff and a few may close. The old IVDD deadlines keep expiring, the dates for higher-risk classes are already here, and the Commission has signaled there will be no further extensions. In this pre-webinar conversation, Dan Simpson and Sarah Barchard talk through what the waiting actually costs a company that sells in both the EU and the US, and why planning for both markets together beats handling them one after the other. Dan takes the regulatory side and where the two systems diverge well before anyone files. Sarah takes the clinical side and what it takes to run studies that hold up in both places. The episode sets up their June 17 RAPS webcast, where they walk through the case studies behind these questions. What You’ll Hear in This Episode Why so many companies have stopped filing, and the risk that builds while they wait Where the EU and US split on risk class, predicates, and self-testing, and why that matters early How the differences surface at design validation, inside the analytical and clinical work Whether one clinical study can serve both markets, and what the answer comes down to What the EU’s active post-market surveillance asks of you that FDA’s reactive model does not About the Guests Dan Simpson, RAC | Director of Regulatory Affairs, DCN Dx Dan is a RAC-credentialed, ASQ-certified medical device auditor with more than 15 years in IVD and medical device regulatory and quality roles. He works with IVD manufacturers on FDA 510(k), De Novo, and PMA submissions and on IVDR conformity assessment, across point-of-care, OTC, companion diagnostics, and high-complexity assays. Before DCN Dx he led regulatory affairs at ERI Group and spent 14 years in quality and regulatory affairs at Corgenix Medical Corporation. Sarah Barchard | Senior Clinical Trials Manager, DCN Dx Sarah is a clinical research professional with more than 15 years in IVD and diagnostics studies. She began as a CRA at Roche on the cobas HPV test for cervical cancer screening, later ran early access programs in Europe and oversaw a study in China at Novartis Diagnostics, and advised manufacturers on emergency use authorization studies during the pandemic. She joined DCN Dx in 2021 and runs clinical studies for high-complexity assays, point-of-care devices, and OTC and at-home products. --- ## The Assay You Developed Is Not the Assay You Can Manufacture URL: https://dcndx.com/insights/assay-developed-assay-manufacture/ Type: insight Published: 2026-06-09 Notes from a CDMO that has watched the same expensive mistake play out, again and again I get this call a lot. It comes from the head of R&D at an early stage diagnostics company, somewhere in the middle of developing a lateral flow assay. The bench data looks good, the team is on schedule, tech transfer is on the near horizon, and cash to the next milestone is fine. The call, ostensibly, is about lining up a CDMO for the manufacturing side, because they have not picked one yet. The call goes well at first. Then I start asking questions about scale-up, and by the time we hang up I am usually explaining that the assay they have developed is not the assay they can manufacture, and that finding that out is going to cost their company a meaningful share of its runway in second-pass process development. That is not the worst case. It is the median. I run operations at an immunoassay CDMO in Carlsbad. From that seat I watch the same pattern repeat, team after team. Where many of these programs end up failing is in the critical handoff between developing the assay and manufacturing it, or when scaling up manufacturing to large volumes. What it looks like when this goes wrong A few years back a client brought us a lateral flow program that had been failing on the line for months. The assay was straightforward: a gold conjugate and a biotinylated antibody, striped onto the same conjugate pad close together. On the benchtop dispenser, everything passed cleanly, and the team had every reason to believe they were heading into production. Then the client moved to a continuous reel-to-reel line, and the lots came back with intermittent, irreproducible nonspecific binding. The kind of failure that drives a development team to the wall, because the failure cannot be summoned on demand. Their team had spent multiple rounds of formulation work chasing it without being able to reproduce it on demand. They handed it to us, and we worked the problem from a different angle. The chemistry was fine. The problem was in the geometry of the line. On the benchtop platform, the conjugate pad sits flat with nothing pulling on it, and the two stripes of wet reagent stay where you put them. On the continuous web, that same pad runs under and over tension rollers before it reaches the drying tower. On its way through, the wet stripes touched the roller surface. There is also some inherent low level of sideways movement of the pad as it moves. On a subsequent pass, the roller laid a trace of the gold conjugate back onto the pad, including a sliver of it onto the biotinylated antibody area. A trace was enough to cause nonspecific binding and therefore an out of spec result during testing. The fix was mechanical. We added several physical guides along the roller path so the pad could not drift sideways. The crossover stopped. NSB resolved on the next production runs. The chemistry, blamed and reformulated and blamed again, had been fine the whole time. The assay had been designed for one kind of physics and forced to live with another. I tell this story at conferences and watch people in the room flinch. They have usually heard a version of it on their own line recently. Why teams like yours often find this out the expensive way I’ve noticed certain patterns that tend to create these issues. Small-batch dispensing is not a slower version of continuous-web manufacturing. The mechanics are different. A benchtop platform supports the material completely, applies low mechanical stress, runs slowly, and forgives a lot of process deviations. A continuous line is the opposite environment: material is suspended and under tension, rollers and a drying tower running at production speed, forgiveness for very little. A spray-through that gets reabsorbed on the benchtop platform is lost on the line. A reagent that settles too slowly to notice across a short benchtop experiment will drift the signal noticeably across a long production run. You can mitigate the settling with recirculation, and recirculation introduces its own risks: shear, foam, stability issues that did not exist in batch. The cure arrives with its own cost. Hero conditions are the second pattern. An assay can hit spec with fresh reagent, careful handling, the senior scientist at the bench, and the humidity-controlled room you developed it in, and still not be a manufacturable product. What you have, in that case, is a prototype the inventor can defend. Manufacturing is what happens when conditions can vary slightly, which they do, every shift. If every parameter on the assay is dialed to ten out of ten to make spec, the production team has nowhere to go when a lot of nitrocellulose comes in slightly off. And lots come in slightly off. The third pattern is that finding this out late is fatal on the P&L. Rework on a lateral flow line is rarely viable for early-stage companies. Reagents are expensive and dead volume in pump lines eats margin. Qualifying a new manufacturing-grade lot is its own project. A yield loss that looks like a rounding error in development becomes the entire margin of the product at scale, and the redesign that follows can trigger work the team thought was behind it: re-verification, sometimes re-submission, sometimes more clinical data. For an early-stage company, the cost of finding this out late often outruns the runway they had to work with. Design for Manufacture is supposed to prevent this. Most teams encounter DFM as a tech-transfer initiative, which gets it backwards. The point of DFM is to put manufacturing constraints in front of the assay designer at the design phase, when they can still shape something useful, instead of at transfer, when they can only break what is already developed. What to do, depending on where you are Pre-formulation freeze is the window when you can still change anything. Use it. Define what manufacturable performance looks like for your assay before you optimize for it. The list of parameters worth specifying is longer than most teams expect: sensitivity, specificity, lot-to-lot CV, signal stability across a long run, tolerance to reagent age, operator-to-operator consistency, performance across line speed, and the variability you expect in your materials supply chain. The target product profile should include what manufacturing will be held to, alongside what reads well in a paper. Between formulation freeze and process development, run guard-band studies. Push the assay outside its comfort zone on purpose. Faster line speed, harder drying, older reagent, off-spec materials within tolerance. Find the cliff in development. Not in production. Between formulation freeze and tech transfer, get the assay onto scaled equipment before you commit capital to a production line. We do this for our clients; good equipment vendors do the same for theirs. Scaled testing is cheap relative to the cost of finding the failure mode after the line is running. If you are already at tech transfer and the lots are coming back out of spec, the right call is to bring in a team that develops and manufactures under one roof. The diagnosis usually needs both, and a two-vendor handoff is rarely fast enough to chase an intermittent failure to ground. Where DCN Dx fits Most immunoassay CDMOs are either development shops or manufacturing shops. We are both, because the handoff between them is where many assay programs lose ground. Lateral flow is where we see these failures most often, because that is the format we manufacture at the highest volume. They are not the only programs to come to us with these issues, however. An ELISA optimized to the bench is the same prototype problem in a different format. A microfluidic cartridge developed around physics that work at the bench and fail at production speed is the same trap. The chemistry and the geometry change with the format. The pattern of treating manufacturability as a tech-transfer ceremony instead of a design constraint does not. The point here is that cleanest benchtop data does not predict the strongest manufacturing program. The teams that eventually ship diagnostic products are the ones who started treating manufacturability as a development question on day one. If that conversation is not the one you are having now, it is the one I will be having with you later. Teams come to us most often at two moments. Some come early, at the design phase, while the assay can still be shaped to fit the process we will eventually run it on. Others come later, after something has stopped working at scale, and our job is to work backwards from the failure to whatever the assay should have been developed as. If this aligns to where your program is, get in touch. Pat Vaughan, Ph.D. | Chief Operating Officer, DCN Dx Pat Vaughan is Chief Operating Officer at DCN Dx, an immunoassay CDMO and IVD CRO in Carlsbad, California, where he runs operations across assay development, engineering, clinical research, regulatory strategy, and manufacturing. He has more than 30 years in biotechnology and diagnostics R&D. Before DCN Dx he was Vice President of R&D at Trinity Biotech, and he founded HiberGen, Ireland’s first genomics company. He has taken products through FDA 510(k) clearance and CE marking, and he works most often with early-stage diagnostics teams moving lateral flow, ELISA, and microfluidic programs from the bench toward production. He holds a Ph.D. from King’s College London and writes and speaks regularly on point-of-care diagnostics and the realities of scaling an assay. --- ## PODCAST: QMSR Is in Effect: What IVD Manufacturers Still Need to Do URL: https://dcndx.com/insights/s3e6-qmsr-ivd-manufacturers/ Type: insight Published: 2026-05-08 (As published 2026-05-08; regulatory status may have changed since.) What changes for IVD developers and manufacturers now that the FDA Quality Management System Regulation is in force, and where the gap work tends to land. FDA’s Quality Management System Regulation (QMSR) took effect on February 2, 2026, incorporating ISO 13485:2016 by reference into 21 CFR Part 820 and replacing the legacy Quality System Regulation. For IVD developers and manufacturers, the change is more than a paperwork exercise. The updated inspection approach expands what FDA investigators can request, and certain records that were previously out of scope are now reviewable. In this episode of Expert Insights, Emily Friedland, VP of Clinical Research at DCN Dx, talks with two guests who come at QMSR from different sides of the same problem. Dan Simpson, RAC, Director of Regulatory Affairs at DCN Dx, walks through the regulatory framing: what changed with the transition, how to map an ISO 13485-based QMS against QMSR’s U.S.-specific requirements, and what FDA’s updated inspection approach means for how teams document and maintain QMS records. Kevin Gunning, Principal Consultant at Gunning Quality Systems LLC, brings the quality systems implementation perspective from more than 25 years of building and maintaining QMS programs at IVD and medical device companies. Together, Dan and Kevin cover the four gap areas that matter most for IVD manufacturers, what FDA can now review that was previously off-limits, and how to scope a proportional remediation effort without overbuilding the QMS. For more detail, read Dan’s companion article: [QMSR Readiness for IVD Manufacturers: Where Well-Maintained ISO 13485 Systems Still Have Gaps] Listen below, or find us on your favorite podcast platform. What You’ll Hear in This Episode 1. What QMSR changed from the legacy Quality System Regulation, and what it means for IVD teams day-to-day 2. Why a documented QMSR gap assessment still matters even for organizations already certified to ISO 13485 3. How to put together a Quality Plan that management will sign and that demonstrates readiness to outside auditors and FDA investigators 4. The four gap areas where well-maintained ISO 13485 systems most often fall short under QMSR: applicable regulatory requirements (UDI, MDR, corrections and removals); complaint handling records; labeling and packaging controls, including conformance to 21 CFR Part 809 for IVDs; and definitions and terminology 5. What FDA’s updated inspection approach under CP 7382.850 means for risk-based decision-making and how risk management files become a roadmap during inspection 6. Which records are now within FDA’s inspectional authority under QMSR that previously were not, including management review minutes, internal quality audits, and supplier audit reports 7. The most common misconceptions Dan and Kevin see from teams who assume the QMSR transition is “mostly handled” because they hold ISO 13485 certification About the Guests Emily Friedland | VP of Clinical Research, DCN Dx (Host) Emily leads clinical research at DCN Dx and is a guest host of Expert Insights, the company’s podcast for diagnostics professionals. Dan Simpson | Director of Regulatory Affairs, DCN Dx Dan works with IVD developers and manufacturers on FDA and global regulatory strategy and inspection preparation. He holds the Regulatory Affairs Certification (RAC) from the Regulatory Affairs Professionals Society (RAPS) and is the author of DCN Dx’s article on QMSR readiness for IVD manufacturers. Kevin Gunning | Principal Consultant, Gunning Quality Systems LLC Kevin is a quality systems executive with more than 25 years of experience in IVD, medical device, and pharmaceutical industries. He has built quality management systems from the ground up at multiple organizations under ISO 13485 and 21 CFR Part 820, and has extensive experience both hosting and conducting FDA and ISO inspections and audits. He holds the Certified Quality Auditor (CQA) credential from the American Society for Quality (ASQ) and previously served as Vice President of Quality Assurance at Alveo Technologies, an IVD company, before founding Gunning Quality Systems LLC. --- ## Designing a CLIA Waiver Comparison Study for a Near-Patient Molecular Diagnostic: Where Programs Run Into Trouble (Part 2 of 2) URL: https://dcndx.com/insights/designing-clia-waiver-comparison-study-near-patient-molecular-diagnostic-where-programs-run-trouble-part-2/ Type: insight Published: 2026-04-15 (As published 2026-04-15; regulatory status may have changed since.) Study design decisions that determine whether your CLIA waiver data will stand up to FDA review. A previous post in this series covered the intended use decision and what it commits a molecular diagnostic developer to, including the POC/OTC distinction, the three pathways to waived categorization, the dual versus stepwise submission question, and why waiver does not carry over from a predicate or an EUA. This post picks up where that one left off. Once the manufacturer makes the intended use decision and heads down the CLIA waiver route, this is where programs may run into trouble. The first decision that needs to be made is whether to pursue CLIA waiver in initial study and submission or do stepwise process with a CLIA waiver comparison study. What the CLIA waiver comparison study demonstrates The intended-user comparison study is the centerpiece of a CLIA waiver application for most molecular diagnostic developers. In this study, untrained operators from the intended use setting perform the test following only the package insert, and their results are compared against those of trained laboratory professionals performing the same test, or under certain study design options against a comparator method. The study is designed to demonstrate that the performance gap between untrained and trained operators is acceptably small: that moving the test out of the laboratory does not introduce a clinically meaningful increase in errors, invalid results, or misclassification. It is one required component of a broader CLIA waiver application package that also includes a device simplicity demonstration, risk analysis, flex studies addressing environmental and usage variation under stress conditions, and failure alert and fail-safe mechanism validation. FDA’s 2020 guidance on CLIA waiver applications addresses study design requirements in detail, covering minimum site counts, operator counts, and result acceptance criteria for binary qualitative and quantitative tests. For test types outside those categories, including some multiplexed molecular panels, the guidance recommends contacting FDA through a pre-submission to discuss study design before proceeding. This advice should be taken seriously, especially for molecular platforms where the performance characteristics of the test and the pre-analytical variables involved differ from simpler test formats. For products of this nature, a pre-sub conversation about study design is worth the investment. Who counts as a naive operator, and why it matters FDA’s definition of an untrained operator goes beyond device-naivety. The guidance calls for operators representative of the intended waived setting who have limited or no hands-on laboratory testing experience generally, and no prior experience with the candidate test. FDA also recommends enrolling operators with the least amount of training likely to be encountered at the intended use sites. A laboratory technologist who happens to work at a waived-site clinic but has substantial laboratory testing experience is not the right recruitment target. The untrained operator population should reflect the worst-case scenario of who will be running the test in the field. Operator recruitment is one of the most common places where comparison study designs introduce problems. The operators who participate must be representative of the people who will use the test in the intended use setting. If your intended use is urgent care clinics, your naive operators should be urgent care staff. If you enroll research staff who do not normally perform patient care tasks, you are not studying the population your intended use claim covers, and FDA may raise that as a concern during review. Site selection carries the same logic. The waived sites enrolled in your study should represent the range of intended use environments, including variation in patient population, operator training background, and physical environment. A study conducted entirely at a single site type may not reflect the range of conditions your test will encounter in the field. Pre-analytical variables: where molecular tests are most exposed Closed-cartridge molecular systems are designed to contain the most technically demanding steps of amplification-based testing: reagent preparation, amplification, and detection are all instrument-controlled and not accessible to the operator. This is a real advantage in a waiver study context. But closed-cartridge design does not eliminate operator-dependent variables; it shifts them to the steps the operator does handle. Specimen collection is the step that most directly affects performance in naive operator hands for swab-based molecular tests. For an amplification-based test detecting a pathogen from a nasal or nasopharyngeal swab, collection technique affects the quantity and quality of nucleic acid presented to the assay. An inadequately collected specimen can produce a false result even when the instrument performs exactly as designed. How operators collect specimens, whether they follow the collection instructions in the labeling, and how consistently they do so across operators and sites is data your comparison study should capture. Cartridge handling and sample introduction are the other operator-dependent steps for most closed-cartridge molecular systems. Sample volume, the mechanics of transferring a specimen to a cartridge, and cartridge loading into the instrument are procedural steps where errors occur. Your instructions for use and the error-detection capability of your system both play a role in how these errors manifest in study results. For some isothermal amplification platforms, ambient temperature and humidity at the point of care can affect assay performance. If your system has restrictive operating condition requirements, your comparison study sites might not meet the requirements for your product’s performance. It is important, therefore, to test your system in conditions that do not represent a central laboratory’s controlled environment. Invalid rate: a metric FDA scrutinizes closely for molecular systems An invalid result is distinct from a negative result. It means the assay did not complete successfully and no result was generated. Internal controls in molecular systems, which may monitor amplification, extraction, or both depending on system design, are intended to detect these failures. A well-functioning internal control system is an asset for a waiver submission because it provides a mechanism to flag when operator or pre-analytical errors have compromised the run, rather than allowing those errors to produce false negatives. FDA’s concern about invalid rates in the comparison study is whether naive operators generate substantially more invalid results than trained operators. A statistical difference in invalid rate between the two groups raises questions about whether the test meets the simplicity threshold for waiver, because it suggests that untrained operation of the device is associated with a higher rate of test failure. A robust study design should plan for how invalid results will be handled, whether repeat testing is allowed, and how both the invalid rate itself and the outcome of any repeats will be reported. Your instructions for use must tell operators what to do when they receive an invalid result. This is both a labeling requirement and a human factors consideration. Designing the study to support both FDA market authorization and the waiver application For developers pursuing a dual submission, the comparison and reproducibility studies need to be designed to satisfy both market authorization (510(k) or De Novo) and the waiver application simultaneously. These are not identical requirements. The 510(k) comparison study or De Novo reference study is typically performed by trained operators meeting moderate complexity qualifications, while the waiver comparison study uses untrained operators from the intended use setting. FDA notes that an applicant may choose a single study package with untrained operators to support both objectives in some cases. Whether that approach works for a given molecular program, or whether both trained and untrained operator data are needed, is a study design question that belongs in the pre-submission, not one to be resolved mid-enrollment. FDA’s 2020 dual submission guidance addresses this design requirement directly and strongly recommends a pre-submission to confirm the study design before initiating clinical work. For a molecular POC dual submission, that recommendation should be treated as the practical default: the study design decisions made before enrollment (sites, operators, specimen handling, comparator methods, and how invalid results will be managed) determine what data you collect, and the data you collect determines what you can say in your device labeling. Discovering a design gap mid-enrollment is a serious problem. Discovering it during review is worse. The OTC path for molecular diagnostics The CLIA-waived OTC path for molecular diagnostics is not well-traveled. Most of the molecular tests that were authorized for home use during the COVID-19 pandemic were authorized under Emergency Use Authorizations, which do not follow the same evidentiary framework as traditional marketing authorization. Developers now pursuing traditional FDA clearance with OTC intended use for a molecular product are working in a space where the regulatory precedent is still developing. The human factors burden for OTC is substantially higher than for POC, and for molecular tests it includes challenges that are specific to the platform. Label comprehension studies must demonstrate that lay users correctly understand all possible result states, including invalid results, and know what action to take for each. Simulated use testing must capture real error modes in representative users under realistic home-use conditions. Self-collected specimen adequacy must be studied with lay users and not assumed as the gap between the collection performance of a naive lay user and that of a naive healthcare worker is real. For molecular tests where specimen adequacy directly affects sensitivity, that gap matters. A companion application that guides users through sample collection and result interpretation can reduce these risks, but it introduces its own validation requirements. Software as a medical device considerations may apply depending on the application’s role in the testing workflow. If the application is integral to generating or interpreting a result, it is part of the device and needs to be treated accordingly. For developers with serious commercial interest in OTC molecular testing, early engagement with FDA through a pre-submission is strongly advisable. The study design questions, human factors scope, and software classification determinations that are unresolved before you commit to an OTC development program are the ones most likely to surface as obstacles during review. Getting FDA alignment on those questions before enrollment, not after, is what a credible development plan looks like. This is the second post in a two-part series on CLIA waiver strategy for molecular IVD developers. Part 1 covers the intended use decision and what it commits you to: the POC/OTC distinction, the three pathways to waived categorization, dual versus stepwise submissions, and why waiver does not carry over from a predicate or an EUA. Read Part 1 here. Talk to DCN Dx’s Regulatory Affairs team If you are designing a CLIA waiver comparison study for a near-patient molecular test, preparing a pre-submission on study design, or working through the human factors requirements for an OTC molecular program, our Regulatory Affairs team has worked through these questions with molecular IVD developers across diagnostic platforms and submission pathways. The study design decisions that look like operational details are the ones that determine what your data can support when it reaches FDA. It is worth getting them right before enrollment begins. Contact our Regulatory Affairs team here or visit dcndx.com/regulatory-affairs-services to learn about our services, including CLIA waiver study design. References 1. U.S. Food and Drug Administration. Recommendations for Clinical Laboratory Improvement Amendments of 1988 (CLIA) Waiver Applications for Manufacturers of In Vitro Diagnostic Devices. February 2020. 2. U.S. Food and Drug Administration. Recommendations for Dual 510(k) and CLIA Waiver by Application Studies for In Vitro Diagnostic Devices. February 2020. 3. U.S. Food and Drug Administration. Applying Human Factors and Usability Engineering to Medical Devices. February 2016. 4. U.S. Food and Drug Administration. Policy for Device Software Functions and Mobile Medical Applications. September 2022. --- ## PODCAST: Specimen Strategy Is Development Strategy: Why “We’ll Source It Later” Fails URL: https://dcndx.com/insights/expert-insights-podcast-biospecimen-strategy-ivd/ Type: insight Published: 2026-04-03 (As published 2026-04-03; regulatory status may have changed since.) Jim Boushell, Senior Vice President of Biospecimens at DCN Dx, joins Mitzi Rettinger to talk about what experienced IVD teams define early in their specimen plans and what goes wrong when they don’t. Biospecimen strategy is the set of decisions that determine which specimens an IVD program needs, how they will be collected, what clinical and demographic metadata must accompany them, and how the resulting data package will hold up under regulatory review. It is distinct from specimen procurement, which is the operational act of acquiring material. When teams conflate the two, or defer strategy decisions until late in the program, they risk discovering the mismatch at the worst possible time: during analytical validation, clinical performance work, or submission prep. In this episode of Expert Insights, Mitzi Rettinger, former Chief Revenue Officer at DCN Dx, talks with Jim Boushell, Senior Vice President of Biospecimens at DCN Dx, about how to prevent those delays. Jim has spent decades on both sides of the equation, building and operating biorepositories and supporting diagnostic developers who need traceable, well-characterized specimens for submission-quality evidence packages. The conversation covers how to align a specimen plan to an evidence plan from the start; where programs get burned on matrix selection, prevalence requirements, comparator methods, metadata completeness, and pre-analytical handling; and what a high-integrity, audit-ready data package should contain. Jim also describes DCN Dx’s prospective biospecimen collections service, which provides IRB/IEC-approved, protocol-aligned collections with end-to-end operational ownership for IVD evidence generation. Listen below, or find us on your favorite podcast platform. What you’ll hear in this episode Defining specimen strategy versus procurement: Jim explains what the term “specimen strategy” should cover, why it is a development decision rather than a sourcing task, and the early warning signs that a program is deferring these decisions too long. Common failure modes: The conversation identifies how specimen plans go wrong in practice: matrix mismatches between the intended use and the specimens on hand, insufficient positivity rates for the statistical analysis plan, missing or incomplete metadata, comparator method misalignment, and pre-analytical handling errors that compromise specimen integrity. Jim discusses whether these problems cluster around specific modalities and indications or cut across all IVD programs. The inputs developers need to determine in early: What are the minimum inputs Jim needs from a development team before he can design a collection mapped to claims and an evidence plan? The episode covers this checklist and Jim’s perspective on when “representative” specimens serve a program better than “perfect” ones. Prospective collections versus banked specimens: When banked specimens are appropriate, when prospective collection is the better path, and what distinguishes a prospective collection designed for IVD evidence from one that was not designed with regulatory submissions in mind. How DCN Dx runs prospective collections: Jim describes the specimen types DCN Dx routinely supports (including saliva, capillary blood, nasal swabs, stool, urine, and plasma/serum), the special handling scenarios the team manages, and where handoffs tend to fail when collections, assay development, and clinical execution are split across multiple organizations. He also discusses when integrating specimen collection with clinical research operations reduces risk versus when a standalone collection is sufficient. Quality, compliance, and the data package: What should an IVD developer expect from a high-integrity data package? Jim explains what “privacy controls and quality systems appropriate to the program” means operationally, including ICH-GCP alignment, chain-of-custody documentation, audit trails, and PHI protections. He also identifies the most common false sense of security he encounters around specimens. About Jim Boushell Jim Boushell is Senior Vice President of Biospecimens at DCN Dx, where he leads the company’s prospective biospecimen collections offering. His career spans decades in biorepository operations and diagnostic development support, working with IVD teams that need traceable, well-characterized specimens for analytical validation, clinical performance studies, CLIA-waiver intended-user comparison studies, reproducibility, bridging, and lot release. At DCN Dx, Jim’s team designs and operationalizes IRB/IEC-approved prospective collections mapped to each client’s claims and evidence plan, with end-to-end operational ownership from protocol development through data package delivery. About Expert Insights Expert Insights is the podcast from DCN Dx, a Carlsbad, California-based immunoassay CDMO and IVD CRO. Each episode features conversations with diagnostics professionals on the technical, regulatory, and operational decisions that shape IVD development programs. Browse all Expert Insights episodes → Frequently Asked Questions What is biospecimen strategy in IVD development? Biospecimen strategy refers to the decisions that should be made before a single specimen is collected: which matrices the program needs, what prevalence rates the statistical plan requires, how specimens will be handled and transported, what metadata needs to travel with each specimen, and what the documentation package needs to look like for the intended regulatory pathway. It is separate from procurement. Procurement is buying or collecting the material. Strategy is defining what “the right material” means for your specific claims and evidence plan. The reason this distinction matters is that procurement decisions made without a strategy behind them tend to produce specimens that look fine on paper but fall apart under regulatory scrutiny. When should an IVD developer use banked specimens versus prospective collection? It depends on what the specimens need to do. Banked specimens can work for some analytical validation activities, particularly when you need well-characterized material and the storage conditions and metadata are documented. Prospective collection makes more sense when pre-analytical handling needs to be controlled to your protocol, when prevalence is low enough that you need enrichment or targeted recruitment, or when a clinical performance study or intended-user comparison study needs to reflect how the test will be used outside a lab. The decision should be made early. Prospective collections have lead time, and discovering that banked specimens do not fit your intended use after enrollment planning is already underway is an expensive mistake. What does DCN Dx’s prospective biospecimen collection service include? DCN Dx’s collections service is structured around designing a collection protocol mapped to your claims and evidence plan, then running it. That includes central IRB/IEC management, site qualification and training, specimen kitting with temperature controls, and comparator or reference testing when the study design calls for it. Data capture runs through eCRFs and a LIMS, with chain-of-custody documentation and audit trails. The team handles specimen types including saliva, capillary blood, nasal swabs, stool, urine, and plasma/serum, and supports collections for analytical validation, clinical performance, positivity enrichment, rare matrices, intended-user comparison studies, reproducibility, and bridging or lot release work. Quality operations are aligned to ICH-GCP with HIPAA/GDPR privacy controls. The Biospecimen Collection Services page has more detail on what the deliverables look like. What should an IVD data package include for regulatory submissions? The short answer: everything a reviewer or auditor would need to trace each specimen from collection to test result without gaps. That means chain-of-custody records, IRB/IEC approval and informed consent documentation, the collection protocol (including pre-analytical handling and transport), clinical and demographic metadata per specimen, temperature monitoring records, comparator or reference test results with reconciliation, and deviation documentation. The specific requirements vary by pathway (510(k), de novo, PMA, IVDR), which is why defining the documentation standards before collection starts is important. Retrofitting a data package to meet submission requirements after the fact is where most of the rework happens. If your clinical research team and your specimen team are not aligned on this from the beginning, the gaps tend to show up late. How does poor specimen planning affect IVD regulatory timelines? It adds months, and the delays are hard to compress once they start. The problems are usually noticed during analytical validation or clinical performance studies: the specimen matrix does not match the intended use, prevalence is too low for the statistical analysis plan to work, metadata is missing or inconsistent, or the chain-of-custody documentation has holes. Any of these can force re-collection. Depending on the specimen type and site access, re-collection timelines can stretch well beyond what the original program plan accounted for. This is why experienced teams treat specimen planning as part of the development conversation, not something they hand off to procurement after the evidence plan is set. --- ## CLIA-Waived POC vs. CLIA-Waived OTC: The Intended Use Decision and What It Commits You To (Part 1 of 2) URL: https://dcndx.com/insights/clia-waived-poc-vs-otc-intended-use/ Type: insight Published: 2026-04-02 (As published 2026-04-02; regulatory status may have changed since.) Developers of near-patient molecular diagnostic tests encounter the term “CLIA-waived” early in their regulatory planning and often treat it as a single destination. This is not the case. CLIA waiver describes a complexity categorization which includes defining the intended use population and the user environment (Home Use versus Point of Care). These determinations will then guide what kind of study you need to run, and what FDA will expect to see before granting that categorization. Getting those assumptions right at the intended use stage is foundational. Revisiting them after your clinical program is underway is expensive and time consuming. How tests become waived Under the Clinical Laboratory Improvement Amendments, all clinical laboratory tests are assigned a complexity category: waived, moderate complexity, or high complexity. The categorization determines what certificate a laboratory must hold to perform the test and what personnel qualifications apply. Under 42 CFR 493.15, CLIA-waived tests are simple laboratory examinations and procedures that have an insignificant risk of an erroneous result, including those that (A) employ methodologies that are so simple and accurate as to render the likelihood of erroneous results by the user negligible, or (B) the Secretary has determined pose no unreasonable risk of harm to the patient if performed incorrectly. Generally, most CLIA-waived products are required to be simple (per item A), demonstrated through a CLIA waiver application that includes, among other components, an intended-user comparison study in which untrained operators following the labeling achieve results comparable to those of trained laboratory professionals. There are three pathways through which FDA grants waived status. The first is automatic: tests cleared or approved for use in one of the nine test categories listed in 42 CFR 493.15(c), such as certain urine dipstick tests or non-automated fecal occult blood tests, receive a waived categorization upon marketing authorization. The second is also automatic: tests cleared or approved specifically for home use are categorized as waived upon clearance. The third, and the one most relevant to molecular diagnostic developers, is waiver by application, in which a manufacturer requests waived status as part of a 510(k) or De Novo submission combined with a concurrent CLIA waiver by application (known as a dual submission), or through a separate waiver application filed after initial clearance (a step-wise approach). For developers of point-of-care (POC) molecular tests, the waiver-by-application pathway is almost always the route. Molecular amplification-based tests have not historically been among the categories that receive automatic waiver. Therefore, demonstrating eligibility through a waiver by application is required. The POC/OTC distinction: operator and setting, not chemistry A CLIA-waived point-of-care test is intended for use in a clinical setting that is not a traditional laboratory. Urgent care clinics, physician offices, pharmacies, emergency departments, and school health clinics are common intended use environments for point-of-care molecular tests. Operators in these settings are often healthcare workers rather than laboratory professionals; typically people with limited formal laboratory testing experience. They have not been trained on the specific device and will self-train from device labeling. A CLIA-waived over-the-counter test is intended for home use by members of the general public: no clinical training, no professional oversight, no access to a colleague when something is unclear. The operator is whoever purchases the test. That distinction has nothing to do with the assay chemistry and everything to do with where results are generated and by whom. The regulatory requirements that flow from it, for study design, operator recruitment, labeling validation, and human factors, are fundamentally different between the two pathways. The dual submission and step-wise pathways: a strategic choice In February 2020, FDA finalized the current CLIA waiver guidance specifically for dual 510(k) and CLIA waiver by application submissions, which allow a manufacturer to seek marketing clearance and waived categorization simultaneously. Dual submission is generally the more efficient path: a single set of comparison and reproducibility studies can support both the 510(k) and the waiver application, and FDA reviews them concurrently. FDA recommends informing the agency of a planned dual submission during the pre-submission meeting and using that meeting to agree on study designs before any clinical work begins. The step-wise pathway, where a manufacturer first obtains market authorization (510(k) or De Novo) clearance and then files a separate waiver application, remains available. For some programs, it is the more practical sequence: if the initial market authorization study used trained laboratory operators and the data to support waiver were not collected concurrently, a step-wise approach is the only option. The tradeoff is time and cost. A separate waiver application requires a complete intended-user comparison study that may not have been designed to leverage the initial study’s data, and the waived categorization, and the commercial access to waived settings that comes with it, is delayed until the waiver application is reviewed and approved. For developers entering a new submission, the dual pathway is worth a serious look. The pre-sub investment required to confirm study design is the same either way, and designing one study that supports both objectives is generally more efficient than designing two. The EUA transition: waiver does not carry over A significant number of molecular diagnostic developers have products that entered the market under Emergency Use Authorizations during the COVID-19 public health emergency. As those developers now pursue traditional marketing authorization through 510(k) clearance or De Novo, CLIA categorization is a separate and independent question that must be addressed. Authorization under an EUA does not establish a CLIA complexity categorization that carries over to a traditionally cleared device. An EUA may specify intended care settings, including use at CLIA-waived sites, and some EUA-authorized tests have been designated as waived under the terms of that EUA, but that authorization is specific to the EUA and does not automatically carry forward into a subsequent clearance. When seeking clearance, a developer whose EUA product was used in waived settings must still demonstrate waiver eligibility through the standard waiver application process. Similarly, waived status for a predicate device does not transfer to a new submission. If the predicate held CLIA waiver, that waiver belongs to the predicate. A new market authorization submission requires an independent demonstration of waiver eligibility, either through a dual submission or through a step-wise waiver application following clearance. Choosing not to pursue CLIA waiver is sometimes the right call Not every molecular test should pursue CLIA waiver. Some developers correctly identify that their intended use population is a moderate-complexity clinical environment, such as a hospital-based POC program, a reference laboratory, or a clinical setting where the operators are qualified to perform moderate-complexity testing. In those cases, the commercial objective does not require waived status, and the additional study work to demonstrate it is not justified. The decision to pursue waiver should follow directly from a clear-eyed look at your intended use claim and your commercial distribution strategy. If your test is intended for professional waived settings, a waiver by application is unavoidable. If it is intended for home use, waived categorization is automatic following clearance, though the evidentiary burden for over-the-counter (OTC) is substantial, as discussed below. If it is intended for moderate-complexity settings and your evidence plan supports that intended use, the waiver program may add cost and timeline without adding proportional commercial value. That is a strategy question that belongs in regulatory planning, not in study design. The intended use statement is where all of this starts The intended use statement you write is not an administrative formality. It determines your CLIA pathway, the operators who must be enrolled in your intended-user comparison study, the labeling requirements that apply, and, for OTC, the scope of human factors work required. POC and OTC are not interchangeable designations, and if your commercial strategy eventually includes both professional and consumer channels, the evidence program required to support OTC is not an extension of a POC submission. It is a separate body of work. Regulatory strategy conversations about intended use and CLIA pathway belong at the front of the development process, before clinical study design begins. They are substantially cheaper there than they are mid-enrollment. This is the first post in a two-part series on CLIA waiver strategy for molecular IVD developers. Part 2 covers where CLIA waiver comparison studies run into trouble: operator recruitment, site selection, pre-analytical variables, invalid rates, and the study design decisions required for a dual 510(k) and CLIA waiver submission. Read Part 2 here. Talk to DCN Dx’s Regulatory Affairs team If you are working through your intended use strategy, evaluating whether to pursue CLIA waiver, planning a dual versus step-wise submission, or navigating the transition from an EUA to traditional marketing authorization, this is the work DCN Dx’s Regulatory Affairs team does. We work with molecular IVD developers across FDA submission pathways, from intended use determination and pre-submission preparation through 510(k) clearance, De Novo authorization, and CLIA waiver. The earlier in your program we engage, the more options you have. Contact our Regulatory Affairs team or visit dcndx.com/regulatory-affairs-services to learn more about the services we offer. References 1. U.S. Food and Drug Administration. Recommendations for Clinical Laboratory Improvement Amendments of 1988 (CLIA) Waiver Applications for Manufacturers of In Vitro Diagnostic Devices. February 2020. 2. U.S. Food and Drug Administration. Recommendations for Dual 510(k) and CLIA Waiver by Application Studies for In Vitro Diagnostic Devices. February 2020. 3. Code of Federal Regulations. 42 CFR Part 493, Subpart A, §493.15: Regulations for tests waived from CLIA requirements. --- ## Semi-Quantitative Lateral Flow Assays: The Smart Middle Ground Driving the Next Wave of Diagnostic Innovation URL: https://dcndx.com/insights/semi-quantitative-lateral-flow-assays/ Type: insight Published: 2026-03-25 By Pat Vaughan, Ph.D., Chief Operating Officer, DCN Dx Lateral flow assays (LFAs) have evolved far beyond simple yes/no tests. As end user expectations rise and applications expand into increasingly complex decision environments, developers are rethinking how much information an assay truly needs to deliver. In this context, semi-quantitative lateral flow assays are emerging as one of the most powerful—and underappreciated—design strategies. By intentionally linking a positive or negative result to a meaningful concentration threshold, semi-quantitative assays enable better decisions without the cost, complexity, or infrastructure burden of fully quantitative systems. At DCN Dx, we see semi-quantitative assay design not as a limitation, but as a deliberate product architecture choice: one that aligns technical performance with real-world use cases. Defining the Assay Types in Lateral Flow Qualitative Lateral Flow Assays A qualitative immunoassay reports results as positive or negative, indicating the presence or absence of an analyte above a minimal detection threshold. In lateral flow formats, this corresponds to the presence or absence of a visible test line. Best suited for: Screening applications Consumer self‑testing Situations where any detectable amount is actionable Most used in infectious disease testing to answer the question: are you infected or not? While qualitative LFAs dominate the market due to simplicity and cost, they often fail to answer the question that increasingly matters in many applications: not just whether an analyte is present, but whether it is present at a level that warrants action. Quantitative Lateral Flow Assays A quantitative immunoassay reports a numeric analyte concentration, typically by measuring test‑line intensity using a reader and converting that signal via a calibration curve generated from known standards. These systems offer high information content but require: Optical instrumentation Calibration and validation across a defined dynamic range reader for result capture, connectivity, and workflow integration (OPTIONAL) Best suited for: Clinical monitoring Therapy optimization Applications where the exact analyte concentration is required Quantitative LFAs continue to grow, but they are not always the most commercially or operationally efficient solution and come with a certain cost since they will almost always involve an LFA reader or smartphone camera / phone app. Semi-Quantitative Lateral Flow Assays: Designed for Decisions A semi-quantitative immunoassay reports a positive or negative result, but with a critical distinction: the assay cut‑off concentration is intentionally engineered to sit at the positive/negative boundary or interface. Presence of a test line indicates the analyte concentration is at or above a defined cut‑off Absence of a test line indicates the analyte concentration is below that threshold This cut‑off is not arbitrary; it is selected to reflect a clinically, biologically, or commercially meaningful decision point. In other words, semi-quantitative LFAs answer the question: “Is this result high enough to matter?” Why Semi-Quantitative LFAs Are Gaining Momentum 1. Decision-Focused Design Across diagnostics, there are applications where a specific concentration is critical to the utility of a diagnostic result. However, with the growth of point-of-care and at-home testing, there is a growing number of tests and hence a growing population of end users that increasingly care less about exact concentrations and more about actionable thresholds: Is treatment warranted? Is isolation recommended? Has a regulatory limit been exceeded? Is follow‑up testing required? Semi-quantitative LFAs encode these decisions directly into the assay design by eliminating unnecessary data while preserving relevance. It’s a binary output with a clear call to action. 2. Infectious Disease: Beyond Presence vs. Absence One of the most important trends in infectious disease testing is the shift from detection to risk stratification. Semi-quantitative LFAs can be tuned so that: The cut-off aligns with viral or bacterial loads associated with infectivity Positive results correlate more closely with clinical or public health relevance This approach supports: Smarter triage Reduced false reassurance Better alignment with downstream clinical decisions DCN Dx frequently works with developers to translate clinical insights into assay cut-off strategies, balancing sensitivity, specificity, and real-world utility. 3. Drug Screening and Compliance Testing Many lateral flow drug tests, e.g., “drugs of abuse tests”, are inherently semi-quantitative: The cut-off concentration is defined by workplace, legal, or regulatory standards A positive result indicates the analyte exceeds an agreed-upon threshold and not merely that it is detectable This model has proven highly scalable, defensible, and commercially successful, demonstrating how cut-off-driven assay design can dominate entire markets. 4. Environmental, Food, and Industrial Testing In non-clinical markets, numeric precision often adds cost without adding value. Semi-quantitative LFAs excel when: Action limits are clearly defined Results trigger pass/fail decisions Testing occurs in decentralized or field environments Examples include: Allergen detection above safety thresholds Toxin or contaminant screening Process control and release testing Semi-Quantitative Is Not “Less Than” Quantitative A common misconception is that semi-quantitative assays are simply underdeveloped quantitative assays. In fact, many people think they are a pathway to a cheap and easy quantitative test. They require intentional, expert design: Selection of biologically meaningful cut-offs Control of antibody affinity and kinetics Optimization of signal-to-noise at the decision boundary Robustness to sample variability and user interpretation These are not trivial challenges. They are inherently quantitative tests, and they are exactly where specialized lateral flow expertise matters. Importantly, cut-off selection is not only a scientific exercise; it is a clinical one. FDA expects developers to demonstrate that the cut-off is clinically defensible and operationally meaningful, which requires careful study design and a clear rationale. Getting this right early saves significant time and cost later in the development program. At DCN Dx, semi-quantitative development is treated as a first-class design strategy, not a fallback. Designing the Right Assay for the Right Market Choosing between qualitative, semi-quantitative, and quantitative formats is a product strategy decision. The most successful lateral flow products are those where assay architecture, user needs, regulatory requirements, and cost structure are aligned from day one. Question Best Fit Is any detectable analyte presence actionable? Qualitative Is a defined threshold concentration of an analyte the key decision point? Semi‑Quantitative Is exact analyte concentration required? Quantitative How DCN Dx Supports Next-Generation Lateral Flow Development DCN Dx partners with clients across the entire assay spectrum to: Define use-case-driven cut-off strategies Optimize antibody and chemistry selection Engineer robust performance at critical thresholds De-risk development through experience-based design decisions Whether the goal is a breakthrough semi-quantitative product or a fully quantitative system, DCN Dx brings a systems-level understanding of lateral flow that turns concepts into commercially viable assays. The Role of Readers in Semi-Quantitative Lateral Flow Assays One of the key advantages of semi-quantitative lateral flow assays is that a dedicated reader is not inherently required. Because the assay output remains binary—positive or negative relative to a deliberately selected cut‑off concentration—a semi-quantitative LFA can be designed for visual interpretation by eye. In these cases, the presence of a test line indicates that the analyte concentration is at or above the decision threshold, while its absence indicates it is below. This enables low‑cost, instrument‑free deployment in decentralized settings while still delivering results that are tightly aligned with meaningful clinical, regulatory, or operational decisions. That said, readers can play a valuable and sometimes essential role depending on the use case. When results need to be captured, stored, trended, or transmitted, a reader provides clear advantages. For visually read assays, the “reader” does not necessarily need to be a traditional benchtop instrument. A suitably configured smartphone, paired with an app and cloud connectivity, can function as an effective reader capturing images, standardizing interpretation, applying cloud‑based algorithms, and integrating results into digital health or data management systems. This approach preserves the simplicity of visual semi-quantitative assays while enabling traceability, auditability, and population-level analytics. In some applications, however, the assay sensitivity requirements drive the need for a reader regardless of output format. When the cut-off concentration is very low, typically in the low pg/mL range, visual labels may no longer provide sufficient signal‑to‑noise. In these cases, fluorescent labels are often employed to achieve the required analytical sensitivity. While this necessitates the use of a standalone reader, the assay can still be fundamentally semi-quantitative: the reader’s role is simply to determine whether the signal exceeds the predefined threshold. Importantly, semi-quantitative assay design is independent of detection mode. Whether read by eye, smartphone, or dedicated instrument/LFA reader, the defining feature remains the intentional alignment of assay performance to a meaningful decision boundary. Design Decision Call-Out: When do you need a reader? You can go reader-free (visual by eye) when: The cut‑off is moderate/high (e.g., high pg/ml or above) and the visual line is clearly discernible. The primary decision is pass/fail (above/below threshold) and no numeric value is required. The use case emphasizes cost, simplicity, and speed (home use, field screening). Traceability can be handled with manual workflows (e.g., photographed results, manual entry). Use a smartphone as reader when: You need digital capture, audit trail, without a benchtop instrument. You want objective interpretation, de‑skilling, or algorithmic QC (e.g., lighting normalization, line finding). You plan to aggregate results for population insights, remote monitoring, or clinical studies. You want an upgrade path to evidence‑level documentation without changing the strip architecture. A dedicated reader is obligatory when: The cut‑off is very low (often low pg/mL) and visual labels can’t deliver sufficient signal‑to‑noise. You use fluorescent labels to achieve added sensitivity. Regulatory or workflow needs demand instrument‑enforced processes (e.g., QC locks, operator ID, connectivity). You require environmental control (lighting, timing, temperature compensation) to protect performance at the threshold. Bottom line: Semi-quantitative may incorporate any of the above scenarios. The decision boundary defines the assay; the label and context determine if a reader adds value or is essential. DCN Dx’s Experience: Visual vs. Fluorescent Semi-Quantitative Architectures DCN Dx has built both visual and fluorescent semi-quantitative LFAs from the ground up, and we help clients choose the right architecture based on the decision threshold, user context, and commercial constraints. Visual, reader‑optional semi-quantitative LFAs DCN Dx’s teams optimize membrane selection, conjugate chemistry, and line architecture to maximize contrast at the cut-off and ensure the line is detectable by eye under real-world lighting and user variability. We routinely tune antibody binding, blocking strategies, and flow kinetics to stabilize the decision boundary, so the yes/no result maps reliably to the intended concentration. When digital capture is needed, DCN Dx can work with smartphone workflows that maintain interpretability without changing the strip chemistry. Fluorescent, reader‑required semi-quantitative LFAs For low pg/mL thresholds, DCN Dx implements fluorescent labels with optical readout, maintaining a semi-quantitative output (above/below cut‑off) while achieving the required sensitivity. For fluorescent semi-quantitative LFAs, we design the reader and the strip together, aligning optics, emission filtering, calibration, and QC routines to the specific cut-off, so the pass/fail result stays consistent across manufacturing lots, environmental conditions, and operators. This approach preserves the binary decision logic while unlocking high sensitivity and traceable digital data. What clients get: A system-level design that integrates strip chemistry, detection modality, user flow, and data strategy, so your semi-quantitative product is not just analytically sound, but deployable, defensible, and scalable. Conclusion Semi-quantitative lateral flow assays represent a powerful and increasingly important approach in modern diagnostics and testing. By embedding meaningful thresholds directly into assay design, they deliver the information users need, no more, no less. The most successful lateral flow products are built by teams that understand both the science and the clinical decision the test needs to support. That alignment, between threshold, chemistry, user context, and regulatory requirements, is what turns a well-designed assay into a commercially viable product. And that is where expert design and lateral flow development expertise makes all the difference. Working through a technical challenge in your semi-quantitative assay or system? Whether it’s cut-off optimization, signal transition at the decision boundary, reagent and membrane selection, or choosing the right detection architecture, DCN Dx’s development team has navigated it. Contact us to talk through your program. --- ## QMSR Is in Effect: What IVD Manufacturers Still Need to Do (Even If You Follow ISO 13485) URL: https://dcndx.com/insights/qmsr-ivd-manufacturers-gaps-iso-13485/ Type: insight Published: 2026-03-18 (As published 2026-03-18; regulatory status may have changed since.) Educational content only. This article summarizes publicly available information and common quality system approaches. It is not legal advice and does not replace your organization’s procedures or regulatory counsel. If your organization already maintains an ISO 13485:2016 quality management system, it’s tempting to treat FDA’s Quality Management System Regulation (QMSR) transition as “mostly handled.” In terms of baseline QMS structure, that’s not an unreasonable instinct: QMSR incorporates ISO 13485:2016 by reference. But QMSR is not “ISO certification, now with an FDA logo.” FDA does not require ISO 13485 certification, and certification does not exempt a manufacturer from FDA inspection. If you’re not ISO 13485 certified today, QMSR does not require certification, but it does require that your quality management system meets ISO 13485 requirements (plus FDA’s Part 820 supplements). The readiness concepts below still apply; your “gap” may simply be larger. From a U.S. regulatory perspective, there are three practical realities that matter right now for IVD manufacturers: You should still conduct a documented gap assessment, even if you are ISO 13485 certified. FDA and even ISO auditors will expect to see this to prove you have systematically analyzed your QMS to the nuances of the new QMSR. You need to be able to demonstrate QMSR readiness with objective evidence (not just a statement of intent). This readiness is typically documented with a Quality Plan that is signed by management. FDA’s inspection technique has changed, and the agency can review categories of records that were historically treated as out of scope. Most notably, this includes Management Review minutes and Audit Reports. Below is a straightforward way to approach QMSR readiness that keeps the work proportional and inspection ready. What changed and what didn’t As of February 2, 2026, the amended 21 CFR Part 820, now titled the Quality Management System Regulation (QMSR), is in effect and incorporates ISO 13485:2016 (and Clause 3 definitions from ISO 9000:2015) by reference. QMSR continues to sit inside FDA’s broader regulatory framework, meaning Title 21 requirements still apply where relevant, and FDA’s statutory authority under the FD&C Act is unchanged. In practical terms: ISO 13485 becomes the backbone of your U.S. QMS expectations. FDA retained specific Part 820 requirements and clarified how “applicable regulatory requirements” map into ISO clauses. Compliance is still evaluated through FDA inspections, which are now conducted under an updated Compliance Program (CP 7382.850) rather than the legacy QSIT approach. If you’re already operating globally, this convergence is helpful. If you’re U.S.-only, it may feel like you just inherited a new vocabulary and a new inspection roadmap at the same time. Gap assessment: what to look for even if you’re already ISO 13485 compliant A QMSR gap assessment is not about re-auditing ISO 13485. It’s about confirming that your ISO-based system is also aligned to the specific U.S. regulatory expectations that QMSR explicitly ties into Part 820 and other Title 21 requirements. For most IVD manufacturers, the gaps tend to cluster in four areas. Area #1: “Applicable regulatory requirements” QMSR explicitly requires manufacturers to comply with other applicable regulatory requirements in Title 21 to fully meet certain ISO 13485 clauses. Examples include: Unique Device Identification (UDI) requirements (21 CFR Part 801 and Part 830) tied to ISO 13485 identification and traceability. Medical Device Reporting (MDR) obligations (21 CFR Part 803) tied to reporting to regulatory authorities. Corrections and removals (21 CFR Part 806) tied to advisory notices and device recalls. If your ISO 13485 system was developed primarily around EU IVDR, it may already be strong in post-market and vigilance, but the details of U.S. MDR decision-making, reporting timelines, and documentation are often handled in a parallel “regulatory” lane that isn’t fully embedded into the QMS. QMSR pushes in the direction of making that integration explicit. Area #2: Complaint handling and records ISO 13485 includes complaint handling requirements. QMSR retains explicit FDA expectations for complaint record content (e.g., device identification, complainant info, investigation justification, and documented actions). For IVDs, this becomes especially important when the complaint investigation ties into trending, CAPA, or field corrections/removals. Area #3: Labeling and packaging controls QMSR retains explicit device labeling and packaging controls as a supplemental Part 820 requirement. For IVDs, that can translate into practical controls around: UDI accuracy and label reconciliation expiry dating and storage conditions IFU revisions and translation controls kit configuration controls (especially when multiple SKUs share common components) Specific to IVD manufacturers, in addition to the general medical device labeling regulation (21 CFR, Part 801), manufacturers must ensure conformance to the detailed labeling requirements of 21 CFR Part 809 (In Vitro Diagnostics for Human Use). A prudent QMSR gap assessment should include a thorough check that labeling and labeling content procedures comply with this regulation. If your organization treated labeling as “Regulatory owns it” and packaging operations as “Operations owns it,” QMSR inspections are likely to connect those activities back to QMS evidence (procedures, training, release records, and change control). Area #4: Definitions and terminology changes A surprising amount of “gap” pain comes from language. QMSR adopts ISO definitions (plus Clause 3 definitions from ISO 9000) while also specifying where FDA definitions supersede ISO terminology. The goal of a gap assessment here is simple: make sure your procedures and records are mapped clearly enough that an investigator doesn’t get lost in translation. A practical output from the gap assessment is a crosswalk that shows, for your specific QMS, where each QMSR requirement is: ISO 13485 clause → SOP/process → record type(s) → system of record (eQMS, PLM, etc.) Part 820 supplemental provisions (records, labeling/packaging, etc.) → SOP/process → record type(s) Inspection changes: what’s different now and why it matters Two inspection changes are worth calling out because they can affect how you structure and how write certain internal records. Change 1: QSIT is gone FDA has implemented a new inspection process aligned to QMSR. The agency withdrew the legacy Quality System Inspection Technique (QSIT) and moved to the inspection approach described in CP 7382.850, “Inspection of Medical Device Manufacturers.” CP 7382.850 explicitly frames inspections around evaluating whether: your QMS meets FDA requirements and provides reasonable assurance devices are safe and effective, and risk management and risk-based decision making are effectively used in the QMS. For IVD manufacturers, it could mean that your risk management file(s) and your risk-based rationales (for design changes, supplier controls, CAPA scope, and verification/validation decisions) will be used as a roadmap during an inspection. Change #2: Certain records are now within inspectional authority FDA has stated that, on and after February 2, 2026, it intends to review records that were previously exempt under QS regulation 820.180(c), including management review, quality audits (internal audits), and supplier audit reports. Those prior inspection exceptions are not maintained in QMSR. The practical implication is not “write for FDA.” It’s “assume these documents can be requested, and make sure they are controlled, factual, and traceable to follow-up actions.” Internal audits and management reviews should still be candid and useful, but they should also be written in a way that a third party can understand the scope, criteria, objective evidence, and outcomes. A QMSR-ready IVD organization is consistent When FDA inspections go well, it’s not just because the company had perfect documentation. It’s because the overall procedure is consistent: Your procedures match what people do. Your records show risk-based decisions that make sense for the product and the stage of the lifecycle. When something goes wrong, complaints/CAPA/nonconformities connect back to risk management and management oversight. Outsourced processes are controlled in a way that reflects their impact on safety and effectiveness. If your ISO 13485 system is operating, you likely have most of the pieces already. The work is in aligning them to QMSR’s explicit FDA linkages and being prepared for the new inspection approach. Need help with a QMSR gap assessment or inspection readiness? DCN Dx supports IVD developers and manufacturers with QMSR gap assessments, remediation planning, and inspection readiness (including mock inspections and document/record retrieval drills). If your quality system grew faster than it was designed—or if you are trying to integrate U.S. MDR, UDI, and correction/removal workflows into an ISO-based QMS—we can help you scope the work and focus effort where it reduces regulatory risk. If you’d like a deeper walkthrough of QMSR readiness planning and what FDA has changed in its inspection approach, please reach out. To, visit dcndx.com/contact. References FDA. Quality Management System Regulation – Frequently Asked Questions (QMSR FAQ). https://www.fda.gov/medical-devices/quality-management-system-regulation-qmsr/quality-management-system-regulation-frequently-asked-questions FDA. Quality Management System Regulation (QMSR) overview page. https://www.fda.gov/medical-devices/postmarket-requirements-devices/quality-management-system-regulation-qmsr FDA. Inspection of Medical Device Manufacturers – Compliance Program 7382.850 (Implementation date 02/02/2026). https://www.fda.gov/media/80195/download Federal Register. Medical Devices; Quality System Regulation Amendments (Final Rule). https://www.federalregister.gov/documents/2024/02/02/2024-01709/medical-devices-quality-system-regulation-amendments eCFR. 21 CFR Part 820 – Quality Management System Regulation. https://www.ecfr.gov/current/title-21/chapter-I/subchapter-H/part-820 eCFR. 21 CFR 801.20 – Label to bear a unique device identifier. https://www.ecfr.gov/current/title-21/chapter-I/subchapter-H/part-801/subpart-B/section-801.20 eCFR. 21 CFR Part 809- In Vitro Diagnostic Products for Human Use. https://www.ecfr.gov/current/title-21/chapter-I/subchapter-H/part-809 eCFR. 21 CFR Part 830 – Unique Device Identification. https://www.ecfr.gov/current/title-21/chapter-I/subchapter-H/part-830 --- ## PODCAST: Automation Beyond the Strip: Building Repeatable Rapid Test Production with KinBio URL: https://dcndx.com/insights/podcast-automation-beyond-strip-building-repeatable-rapid-test-production-kinbio/ Type: insight Published: 2026-03-18 Scaling rapid test manufacturing from benchtop assays to high-throughput, controlled production. As rapid diagnostic programs transition from R&D to scaled production, small sources of variation can quickly become major risks. In this episode of Expert Insights, Mitzi Rettinger, former Chief Revenue Officer at DCN Dx, sits down with Daniel Levenson, Business Development Manager at KinBio, to discuss what truly changes when you move from manual processes to automated, repeatable manufacturing systems. The conversation goes beyond dispensing and strip cutting to address the often-overlooked bottlenecks in vial filling, kit assembly, packaging, and sealing. Daniel explains how throughput challenges, moisture control, seal integrity, and alignment issues can affect lot consistency, and why automation must extend across the full production workflow, not just the strip. They also explore the role of in-line vision systems and defined critical-to-quality attributes (CTQs) in reducing manufacturing risk. As multiplex tests and more complex point-of-care formats become more common, developers must design flexibility, inspection, and traceability into their processes early. The result: fewer surprises, stronger process control, and a clearer path to scalable production. Listen below, or find us on your favorite podcast platform. What you’ll hear in this episode What typically breaks when moving from benchtop lateral flow assays to scaled, repeatable manufacturing Where bottlenecks emerge beyond the strip, including vial filling, kit assembly, and packaging How in-line vision systems and defined CTQs reduce variability and manufacturing risk What changes operationally when moving to multiplex and more complex point-of-care formats Guest Daniel Levenson is Business Development Manager at Shanghai Kinbio Tech Co., Ltd. He brings years of hands-on technical experience in rapid test manufacturing to his work supporting developers scaling into automated production. --- ## PODCAST: Point-of-Care in 2025: Lateral Flow Grew Up. Where Will the Technology Go Next? URL: https://dcndx.com/insights/podcast-point-care-lateral-flow-grew-up/ Type: insight Published: 2026-02-19 (As published 2026-02-19; regulatory status may have changed since.) Lateral flow diagnostics are no longer just assays, they’re integrated systems. In this episode of Expert Insights, DCN Dx’s Mitzi Rettinger speaks with Dr. Pat Vaughan, DCN Dx’s Chief Operating Officer, about what point-of-care really meant in 2025, how development expectations have shifted, and his predictions for what 2026 will bring. Drawing from Pat’s recent article and hands-on experience across assay development, engineering, clinical research, and manufacturing, the discussion breaks down how modern lateral flow programs are being designed around readers, software, data pathways, and real-world use. The episode explores what this shift changes for teams defining new development programs, including how early design inputs affect performance targets, usability, regulatory strategy, and scalability. Pat also compares today’s realities with predictions he made in his 2024 year-in-review, highlighting where the industry moved faster (and slower) than expected. Listen below, or find us on your favorite podcast platform. What you’ll hear in this episode Why lateral flow is now an integrated system, not just a strip with visual interpretation How reader-assisted and semi-quantitative performance are shaping new development programs Where teams underestimate complexity when moving from prototype to regulated product What “born digital” results and AI-enabled interpretation require from a regulatory and operational standpoint Guest Dr. Pat Vaughan is Chief Operating Officer at DCN Dx, where he works across assay development, engineering, clinical research, regulatory strategy, and manufacturing to support point-of-care diagnostic programs from concept through deployment. --- ## PODCAST: The DCN Dx Model: Integration, Scale, and What Comes Next with DCN Dx CEO Charlie Mamrak URL: https://dcndx.com/insights/podcast-dcndx-model-integration-ivd-diagnostics-scale-charlie-mamrak/ Type: insight Published: 2026-02-06 (As published 2026-02-06; regulatory status may have changed since.) How an integrated services model reduces handoffs across development, clinical work, regulatory planning, manufacturing, and biospecimens, and how DCN decides what to build next. A DCN Dx Expert Insights conversation with Charlie Mamrak, CEO at DCN Dx, hosted by Mitzi Rettinger. Most IVD programs don’t lose months because a single experiment failed. They lose months at the seams: development to clinical, clinical to regulatory, scale-up to routine manufacturing, and specimen strategy to evidence generation. In this episode, DCN Dx CEO Charlie Mamrak explains the business decisions behind DCN’s integrated services model, why biospecimens became a priority for IVD developers, and how he evaluates what to build, buy, or collaborate on next. This is a strategy conversation, designed for teams who care about execution, risk, and real-world constraints of diagnostics services companies. Listen below, or find us on your favorite podcast platform. What you’ll hear in this episode Why integration is the point: what changes when rapid testing CDMO capabilities and IVD clinical research (CRO) services operate as one platform. Why biospecimens: what DCN has heard from customers, what offering prospective biospecimens collections changes for IVD developers, and how it fits the broader integration plan for DCN Dx. Workforce development as strategy: why DCN teaches the Basic Lateral Flow Course. Build vs. buy vs. collaborate: a CEO framework for deciding when new capabilities reduce friction for customers and align to corporate core values vs. simply expanding the menu of services. Guest Charlie Mamrak is CEO of DCN Dx. He joined DCN Dx in 2020 after leading multiple diagnostics and life science tools companies, including SeraCare Life Sciences. --- ## PODCAST: Risk Management in IVD Development: A Regulatory Perspective URL: https://dcndx.com/insights/expert-insights-podcast-risk-management-ivd-development-regulatory-perspective/ Type: insight Published: 2026-02-06 (As published 2026-02-06; regulatory status may have changed since.) Five of the most common regulatory pitfalls—and how to avoid them before they derail your IVD program. Even strong IVD programs can collapse under regulatory pressure. In this episode, DCN Dx unpacks the preventable risks that lead to delayed submissions, costly rework, and missed approvals. You’ll hear the real reasons protocols fall short—and how to build alignment across clinical, regulatory, and usability workstreams from the start. If you’re working toward FDA or global approval, this conversation will help you stay ahead of the most common points of failure. Why Listen: Understand how vague intended use leads to regulatory red flags Learn when and how to use the FDA Pre-Submission process effectively Avoid mismatches between study endpoints and label claims Get ahead of usability and human factors requirements before they block your launch Discover how tight data management practices protect your credibility during review --- ## Point‑of‑Care in 2025: Lateral Flow Grew Up. Where the Technology Will Go Next URL: https://dcndx.com/insights/point-of-care-2025-lateral-flow-grew-where-technology-next/ Type: insight Published: 2026-02-05 (As published 2026-02-05; regulatory status may have changed since.) By Pat Vaughan, Ph.D., Chief Operating Officer, DCN Dx La Jolla set the tone: from “strips and lines” to engineered systems When we gathered at ALFC 2025 in La Jolla, I opened with a provocation— “LFA is Dead! Long Live LFA!” —not to bury lateral flow but to paint a picture of the transformation in front of us. What made LFAs indispensable during the fight against HIV and malaria, and more recently during the COVID pandemic wasn’t the romance of a colored line; it was a unique mix of manufacturability, cost, and near-patient utility. In 2025 and as we enter 2026, what will sustain LFAs is different: engineered sensitivity, reader-anchored quantitation, connected data, and credible quality systems that integrate point-of-care into the same governance structures as the laboratory. That perspective dominated ALFC’s cross-currents—AI in science operations, manufacturability before scale, and regulatory roadmaps as constraints on product design rather than administrative afterthoughts—and it mirrors the broader market’s pivot from pandemic surge to durable adoption. ALFC 2025 showed where rapid testing is headed. The big idea is simple: lateral flow is no longer just an inexpensive way to get a yes/no answer. It’s becoming an engineered system, inclusive of a strip + reader + software + quality planning, built to deliver trustworthy results anywhere care happens. From the stage, I argued that the post‑pandemic rebirth of LFA depends on building for users, not just analytes. From hospital-at-home to retail clinics, decentralized healthcare demands consumer-grade usability (UX), i.e. speed, simplicity, mobile integration, trustworthy results, reader-backed outputs, and data pathways that fit into clinical decisions with near-zero friction. I then highlighted global exemplars (e.g., HealthPulse AI deployments verifying RDTs and incentivizing quality care in LMIC pharmacies) and emphasized AI/ML validation and cybersecurity as part of the product and not afterthoughts. Lateral flow didn’t survive the post-pandemic contraction on nostalgia. It survived because teams learned how to make these tests behave in the real world: readers that support quantitation when it matters, for example, and UX that reduces user-driven variability. The rest of this piece is about where that maturity continues today, and what will separate the next generation of systems from the ones that stall. Why the market still cares about point-of-care testing Point‑of‑care testing kept growing in 2025 and is on track to keep rising through 2030. Lateral flow rides that wave because it’s fast, scalable, and now more precise thanks to improvements in readers, materials, and smarter chemistry. The opportunity isn’t in cheaper plastic casings; it’s in better-engineered performance and easier use. The numbers say point-of-care is not a COVID afterglow; it is a long-term reconfiguration of where decisions are made. The global POC market was around $21.8B in 2025 and is projected to reach ~$29.9B by 2033, while lateral flow alone is widely estimated near $8.5B in 2025 with trajectories to $12.5B by 2030. What’s fueling the curve isn’t “cheap tests”; it’s digital readers, multiplex strip designs (e.g., respiratory multiplexes), and AI-assisted interpretation that turn a once-qualitative format into a semi- or fully quantitative decision tool. In the U.S. and Europe, analysts expect the LFA category to approach $6.97B by 2030, consistent with a steady shift of testing from central labs to retail clinics, urgent care, and the home. None of that growth is possible without a technology rethink inside the cassette. 2025 reset quality expectations—and LFAs rose to meet them Regulators and standard-setters spent 2025 raising the floor for decentralized testing. The CDC’s refreshed waived testing guidance re-centered training, documentation, and good practices in settings that had grown comfortable with rapid tests, while CLIA’s 2025 updates tightened proficiency expectations (e.g., HbA1c accuracy) and clarified personnel qualifications—a reminder that speed never excuses sloppiness. More consequential for lateral flow’s future was the ISO 15189:2022 transition: it pulled POCT inside the lab’s quality system. The outcome is that decentralized testing must be as disciplined as the core lab. That’s a good thing for patient care and for the reputation of rapid tests. From line to number: the 2025 technology that changed LFA’s center of gravity In LFA, the strip is only half of the product now. Readers, either phone‑based or dedicated, turn bands into stable numbers by correcting for lighting, camera differences, and user technique. New multiplex designs let a single cassette carry several analyte assays without confusing the user. The result is semi‑ or fully quantitative answers you can act on. Crucially, smartphone/AI readers stabilized the last fragile link—human interpretation—by normalizing lighting, focal distance, and phone variation to produce consistent values in uncontrolled environments. In other words: LFA hasn’t turned into molecular; it has become measurable. The computational side matured, as well. In one example, teams applying machine learning to CRISPR-Cas13 LFA images reported ~96.5% accuracy across 3,000+ smartphone photos, showing how algorithms can collapse inter-reader variability into a predictable output—exactly what decentralized programs and post-market surveillance need. And pragmatic intensity-normalization methods demonstrate vastly increased precision (reduced CV) across lighting conditions. This is the kind of performance that moves what used to be a prototype or great idea from the bench to a pharmacy shelf. That isn’t glamour; it’s what reliability feels like. Molecular at the point of care raised the bar that LFAs must clear, giving LFAs a roadmap While lateral flow is accelerating its ability to quantify, molecular POC testing showed how fast “same visit” can really be. The FDA’s 510(k) clearances and CLIA waivers for Roche cobas® liat multiplex STI panels (CT/NG and CT/NG/MG) in January 2025 delivered ~20-minute PCR diagnoses with a single specimen—an unambiguous win for antimicrobial stewardship and loss-to-follow-up. For LFA teams, these products should not be considered existential threats; they’re design briefs: wherever the clinical question tolerates a little less sensitivity (or budgets demand it), the LFA alternative must offer quantitation, multiplexing, and connectivity good enough to keep workflows intact. That’s where 2025’s reader and strip innovations matter. At the same time, in a bid to reduce costs for many molecular tests at the point of care, CRISPR and other isothermal amplification diagnostic tools edged closer to the LFA form factor with a lateral flow readout, signaling that instrument-light molecular is no longer a thought experiment. In 2026 and onwards, expect to see these workflows either converge with LFA cartridges or ride alongside them with common reader platform and standardized data interfaces. These are not the answer to all POC settings, but they certainly open additional possibilities and create an awareness of the utility of lateral flow. Materials and labels got smarter, pushing the biological limits without jumping to PCR Labels and materials continue to matter. Brighter labels and better materials like membranes are pushing antigen tests toward earlier detection windows, stretching closer to the molecular window without the cost and complexity of amplification. This is where the “LFA is high tech” narrative flips from defense to offense: we’re not clinging to cheapness; we’re engineering signal-to-noise. The mundane plastic cassette or cartridge also came into focus. Research and product launches of biodegradable and, more importantly, bio-compostable cassette housings (e.g., plant-based formulations) show immense promise at affordable prices. If tenders and ESG screens matter to a product’s revenue modeling, a credible biodegradable housing, paired with paper-based microfluidics, can reduce waste and lower procurement friction. Sustainability isn’t a press release; especially in large consumer-facing programs or ones that rely on government tenders, it can be a source of competitive advantage. Connected diagnostics, in plain terms Modern lateral flow tests are no longer “just assays.” They’re expected to plug cleanly into the health data world. In 2025, that expectation stopped being fuzzy. Health systems and regulators increasingly assume that an LFA result is born digital, moves as structured clinical data (not a screenshot or PDF), and flows securely to the places it must go: Electronic Health Records (EHRs) for clinical action, payers for reimbursement, and public health for reporting. Practically, that means building connectivity into the product from day one. For example, your reader (smartphone, other handheld, or benchtop) needs a clean application programming interface (API), it should emit FHIR-compatible data by default, and your data model should be stable enough for hospital IT to trust. Treat the reader and app like a regulated device: define how your AI/ML models are updated without causing performance drift, lock down cybersecurity (encryption, access control, audit trails, incident response), and maintain a living phone/device validation matrix so operating systems and camera changes don’t break your application in the field. Do this early, not during a CLIA-waiver study or hospital rollout because waiting until FDA review, a site’s security audit, or an EU notified-body check to figure out your APIs, security posture, or model-update plan is how timelines slip and confidence in your product erodes. In short: design LFAs as digital medical devices—integrated assay, reader, data, and quality system —so your results are trustworthy, portable, and ready for real-world users and workflows. The EU gave developers breathing room. Now use it to harden your tech, not to coast On December 16, 2025, the European Commission proposed targeted simplifications to MDR/IVDR and extended IVDR transitional periods by risk class out to 2027–2029, directly addressing notified body bottlenecks that stalled launches. If you sell into the EU, that is not a holiday; it is a window of opportunity. Use it to finish assay verifications and validations, strengthen reader calibration and compensation for end user environmental variability, and validate quantitation under the real-world variability of phones and lighting. The companies that treat this reprieve as runway will own the market when the clock runs again. What 2026 and beyond should look like if we’re serious about meeting this moment If 2025 was lateral flow’s coming-of-age year, then 2026 should be the year we lock in those gains as marketable products. That means reader-first LFAs with calibrated quantitation, robust correction for lighting and ambient conditions, and FHIR-ready APIs as the default, not as promises deferred to a future revision. It also means having a clear plan for model updates and drift control, so AI-enabled interpretation remains a regulated medical device instead of an ongoing science experiment. Lateral flow will increasingly rely on engineered sensitivity, using advances in materials, labels, and reagents where the real-world problem demands it. The goal is not to turn every LFA into a molecular test, but to capture earlier points in the disease curve without crossing into molecular cost and complexity. At the same time, decentralized testing must be treated as a first‑-lass lab node, with competency tracking, quality oversight, and proficiency participation where appropriate. This isn’t paperwork for its own sake—this is how trust is earned and maintained at scale. The winners in the next decade won’t be the teams that squeeze a single sensitivity claim over the line. They’ll be the teams that design across the integrated system—assay, reader, data, and quality—while telling a strategic story that clinicians, consumers, and investors can all understand. That story isn’t “we print a strong line.” It’s “we deliver a fast, trustworthy, connected result wherever care happens.” Underneath that story is the engineering teams like mine at DCN Dx undertake for our clients every day: multiplex strips that leave the development stage without leaving performance behind; labels and chemistries that extend the useful testing window; readers that aren’t fooled by a dim kitchen at 11 p.m.; and quality systems that recognize the home as a legitimate site of care. That is what “Long Live LFA” means in practice. Our team at DCN Dx is proud to embody this ethos, along with the many supply chain partners, diagnostic developers, researchers, and others pushing lateral flow technology forward. In 2026, I hope that we will all continue doing that. References CDC. To Test or Not to Test? Considerations for Waived Testing. July 2025. (CDC lab-quality resource). ADLM. 2025 Changes to Point-of-Care Testing Regulatory Requirements. May 8, 2025. UKAS. Embracing change – The transition to ISO 15189:2022. June 6, 2024. (Dec 2025 deadline guidance). The Biomedical Scientist. Point-of-care testing and the new ISO 15189:2022. Updated July 22, 2024. Eurachem. ISO 15189:2022 – A new task for medical laboratories (risk-based approach; POCT integration). 2023/2024. WHO. Updates on the WHO Model List of Essential In Vitro Diagnostics (EDL 5). Open session slides, May–June 2025. Firely. The State of FHIR in 2025: Growing adoption and evolving maturity. June 25, 2025. Federal Register. Health Data, Technology, and Interoperability: ASTP/ONC Deregulatory Actions To Unleash Prosperity (Proposed Rule). Dec 29, 2025; comments due Feb 27, 2026. Roche Diagnostics. FDA clearance and CLIA waiver for cobas® liat multiplex STI panels (CT/NG; CT/NG/MG). Jan 22, 2025. Patient Care Online. FDA Clears Cobas Liat Assay Panels for POC Diagnosis of STIs. Jan 23, 2025. Clinical Lab Products. FDA Clears Roche Point-of-Care STI Tests. Jan 22, 2025. Springer (Clinical & Experimental Medicine). Recent developments and future directions in point-of-care next-generation CRISPR-based rapid diagnosis. Jan 9, 2025. RSC Analytical Methods. 3D-printed devices for multiplexed semi-quantitative competitive LFIA. Jan 15, 2025. Frontiers in Lab-on-a-Chip Technologies. Advancements and challenges in paper-based microfluidic devices. Dec 19, 2024. Microchimica Acta. Lab-on-paper diagnostics for blood sample analysis: a review. July 2, 2025. MarketsandMarkets / PR Newswire. Lateral Flow Assays market outlook to 2030 (Aug 14, 2025). Research & Markets / GlobeNewswire. U.S. and Europe LFA market trends analysis (June 25, 2025). Grand View Research. Point-of-Care Diagnostics market size & forecast to 2033 (2025 update). European Commission. Proposal to simplify rules for medical devices and IVDs (COM(2025) 1023). Dec 16, 2025. Sapphiros. Strategic agreement with Roche for large-scale LFA manufacturing capacity. Nov 19, 2025. --- ## IVD Clinical Research Organization Evaluation & Comparison Rubric URL: https://dcndx.com/insights/ivd-clinical-research-organization-evaluation-comparison-rubric/ Type: insight Published: 2026-01-29 (As published 2026-01-29; regulatory status may have changed since.) A practical, weighted scoring tool to compare IVD clinical research organization (CROs) on critical capabilities Selecting a clinical research organization (CRO) for an IVD program should not feel like comparing feature lists or sales decks. This rubric gives you a structured way to evaluate IVD CROs side-by-side using criteria tied to clinical performance evidence, operational execution, and study integrity. The criteria focus on what determines whether study outputs are defensible: traceability from intended use claims to endpoints, operational controls, and audit-ready records. DOWNLOAD THE RUBRIC What this rubric helps you do Most CRO evaluation frameworks were developed for drug trials. IVD studies have different failure modes: unclear claims strategy, weak protocol-to-operations integration, site and specimen constraints, and data workflows that do not support you when preparing submissions. This clinical research rubric helps clinical, R&D, and regulatory teams compare CROs in an “apples to apples” way by scoring the capabilities that matter most for IVD clinical performance evidence. This clinical research rubric helps teams select the right CRO partner for their IVD program. What’s inside the IVD CRO evaluation rubric A weighted scoring rubric across eight evaluation areas: IVD clinical and evidence expertise Clinical operations and start-up reliability Trial design and operational integration Data management and biostatistics Site network plus patient/specimen access and recruitment Regulatory strategy support (FDA and IVDR) Quality systems and GCP compliance readiness Transparency, governance, and communication Scoring definitions that describe what “strong” looks like for each category A simple structure you can use for RFPs, vendor selection, or internal CRO reviews Who it’s for Clinical Affairs, Clinical Operations, and Program Management Assay and product development leads supporting clinical performance studies Regulatory Affairs teams coordinating submission-ready evidence Diagnostics leadership teams selecting a CRO for a new indication or platform Want to talk through your situation? If you’re planning a clinical performance study and need support on your claims and evidence strategy, site feasibility, or operational design, DCN Dx’s clinical research team can help. Contact us here. --- ## PODCAST: Predetermined Change Control Plans (PCCPs) for AI-Enabled IVDs URL: https://dcndx.com/insights/podcast-predetermined-change-control-plans-pccps-enabled-ivds/ Type: insight Published: 2026-01-23 (As published 2026-01-23; regulatory status may have changed since.) A conversation on what PCCPs are, how FDA expects them to be structured, and what teams with AI-enabled IVDs should plan for now. A DCN Dx Expert Insights conversation with Dan Simpson, RAC, Director of Regulatory Affairs at DCN Dx, and Emily Friedland, VP of Clinical Research at DCN Dx. FDA’s Predetermined Change Control Plan (PCCP) mechanism is designed to let manufacturers pre-specify certain post-clearance or post-approval changes, along with the protocol and acceptance criteria used to verify and validate those changes. For AI-enabled device software functions, PCCPs are now a core concept in FDA’s approach to iterative software updates. In this episode, we break down what belongs in a PCCP, how it shows up in a marketing submission, and how teams can connect PCCP planning to design controls, risk management, and quality system processes so the plan is executable after authorization, not just well written on paper. What we cover What FDA means by a PCCP and when it is relevant for AI-enabled IVDs The three core components FDA expects to see (and why each one matters) How PCCPs interact with marketing submissions (510(k), De Novo, PMA) and public-facing submission summaries Where teams get tripped up: scope control, evidence expectations, and operationalizing the plan inside the QMS What the broader “PCCPs for Medical Devices” draft guidance signals for non-AI device changes (hardware, materials, software) Listen on this page or wherever you get your podcasts. Companion article: Implications of FDA Predetermined Change Control Plans (PCCPs) for AI-Enabled IVD Submissions and Future Applications Across All Device Types About the guests Dan Simpson, RAC is Director of Regulatory Affairs at DCN Dx, supporting regulatory strategy, pre-submission engagement, and U.S. and global submission planning for diagnostics teams. Emily Friedland is the VP of Clinical Research at DCN Dx, where she leads and fosters growth within the clinical operations teams. Questions about whether a PCCP makes sense for your device, or how to structure one so it holds up through review and post-market execution? DCN Dx’s Regulatory Affairs Services help IVD teams develop successful regulatory plans and submissions. We support FDA pathways including 510(k), De Novo, and PMA, with early Pre-Sub positioning and submission development, and we can advise on accelerated programs like Breakthrough Devices and STeP when they fit the product. Our team aligns verification and validation expectations across software and cybersecurity, biocompatibility, labeling, shelf life, analytical and clinical performance, usability and human factors, and CLIA waiver flex studies. We also support QMS readiness (ISO 13485, CLIA, CAP) and post-market obligations. For more information, visit our Regulatory Affairs page or contact us. --- ## Implications of FDA Predetermined Change Control Plans for AI-Enabled IVD Submissions and Future Applications URL: https://dcndx.com/insights/fda-predetermined-change-control-plans-pccp-ivd/ Type: insight Published: 2026-01-22 (As published 2026-01-22; regulatory status may have changed since.) By Daniel Simpson, RAC, Director of Regulatory Affairs, DCN Dx Introduction The U.S. Food and Drug Administration (FDA) has introduced a transformative regulatory framework known as FDA Predetermined Change Control Plans (PCCPs), designed to streamline the approval and implementation of software modifications in artificial intelligence (AI)-enabled medical devices which include IVDs. This framework addresses the unique challenges posed by the iterative nature of AI development and offers a pathway for continuous innovation while maintaining safety and effectiveness. Recent draft guidance suggests that PCCPs may soon be applicable to all medical devices, signaling a significant shift in regulatory strategy. 1. Background and Regulatory Context 1.1 Origin of FDA Predetermined Change Control Plans PCCPs were formally authorized under Section 515C of the Federal Food, Drug, and Cosmetic Act, added by the Food and Drug Omnibus Reform Act (FDORA) of 2022. This provision allows manufacturers to include a PCCP in their initial marketing submission, enabling pre-approved modifications without requiring supplemental submissions. 1.2 AI-Specific Guidance In December 2024, FDA finalized its guidance document titled “Marketing Submission Recommendations for a Predetermined Change Control Plan for Artificial Intelligence-Enabled Device Software Functions” (reissued August 18, 2025). This guidance outlines how manufacturers can structure PCCPs for AI-enabled device software functions (AI-DSFs). In this guidance, FDA affirms its commitment to applying innovative approaches to regulating medical device software and other digital health technologies while maintaining safety and effectiveness. The concept of PCCPs was introduced in FDA’s April 2019 discussion paper on a proposed regulatory framework for modifications to AI/ML-based Software as a Medical Device (SaMD). That paper recognized the need for flexibility in the marketing authorization process due to the iterative nature of these products. The FDA Predetermined Change Control Plans framework has continued to evolve through industry feedback, including comments submitted on the 2019 discussion paper and discussions in public workshops on the topic. 2. PCCP Structure and Components A PCCP includes those device modifications that generally would otherwise require a new marketing submission such as a 510(k), De Novo, or PMA. These modifications include those that could significantly affect, or that otherwise affect, the safety or effectiveness of the device, unless these changes are validated in a PCCP. Therefore, in a PCCP, a manufacturer can prospectively specify and seek FDA clearance or approval for intended modifications to an AI-IVD without needing to submit additional marketing submissions or obtain further FDA authorization before implementing such modifications, provided the changes are implemented consistent with the PCCP that has been reviewed and established through a device marketing authorization. A PCCP must include three core elements: Description of Modifications: Clearly defines the scope and nature of anticipated changes. Modification Protocol: Details the methods for developing, verifying, validating, and implementing changes. The modification protocol must contain pre-defined acceptance criteria. Impact Assessment: Evaluates the risks and benefits of the proposed modifications. These components ensure that updates remain within the bounds of safety and effectiveness, allowing for regulatory flexibility without compromising patient outcomes. PCCPs are included in marketing submissions as standalone sections/documents with a title and version number and are ideally a part of the manufacturer’s Design History File documentation. In some cases, the labeling of the IVD device may require mention of the PCCP and the PCCP must be mentioned in publicly available submission summaries such as 510(k) summaries. The PCCP, once approved by FDA along with the marketing submission, should be incorporated into the manufacturer’s risk management process and Quality Management System (QMS) to ensure that the PCCP is followed as per the requirements outlined in the marketing submission. Results of the modification protocol and associated validations should be incorporated into all manufacturing and test procedures. Failure to incorporate a required PCCP could result in enforcement actions during FDA inspections. Modifications to a PCCP constitute a potential new marketing submission and it is recommended that manufacturers discuss modifications with FDA using the Pre-Sub process. 3. Implications for IVD Manufacturers with AI-Enabled Devices As with any regulatory framework PCCPs come with potential benefits but also can present challenges to manufacturers, regulators, and IVD stakeholders. 3.1 Benefits Accelerated Innovation: Enables rapid deployment of algorithmic improvements. Regulatory Efficiency: Reduces the need for repeated submissions. Cost Savings: Minimizes expenses associated with supplemental applications. Improved Patient Care: Facilitates timely enhancements to diagnostic and therapeutic capabilities. 3.2 Challenges Documentation Burden: Requires detailed upfront planning and validation protocols. Quality System Integration: Manufacturers must maintain robust systems to track and implement changes. Scope Limitations: Modifications must align strictly with the approved PCCP; deviations require new submissions. 4. Expansion of PCCPs to All Medical Devices Upon creation of the PCCP concept, both FDA and industry recognized its value in all medical device marketing submissions to reduce the need for future market submissions due to foreseen changes in the medical device. 4.1 Draft Guidance Overview In August 2024, the FDA released a draft guidance titled “Predetermined Change Control Plans for Medical Devices”, extending the PCCP framework beyond AI-enabled devices to all device types, including device-led combination products. 4.2 Key Concepts in Expanding FDA Predetermined Change Control Plans to all Device Types PCCPs can be submitted with 510(k), De Novo, or PMA applications. However, to accept the use of the PCCP in the marketing authorization, the manufacturer must provide reasonable assurance of safety and effectiveness. The guidance outlines acceptable modification types, including hardware changes, raw material modifications, and software updates. Generally, the modifications should not significantly affect the safety and effectiveness of the device. PCCPs must maintain the device’s intended use and ensure substantial equivalence or safety and effectiveness. Additionally, PCCPs must be in harmony with existing device modification guidance documents. 4.3 Strategic Implications The broader adoption of FDA Predetermined Change Control Plans encourages proactive change management across the industry. Cybersecurity and Connectivity: Supports timely updates for connected devices. Regulatory Harmonization: Aligns with global efforts to modernize device regulation (e.g., GMLP principles). 5. Outlook and Recommendations 5.1 Industry Impact The expansion of FDA Predetermined Change Control Plans represents a paradigm shift in medical device regulation. It empowers manufacturers to innovate responsibly while reducing regulatory friction. 5.2 Recommendations for Manufacturers As stated above, PCCPs can be a powerful tool for IVD manufacturers to use for both AI submissions and potentially any marketing submission. Below are recommendations on how to successfully implement PCCPs into regulatory and QMS processes. PCCPs should be built into a manufacturer’s Design Control process to be considered early in development. Procedures should be in place to identify potential modifications that could be incorporated into a PCCP. If a manufacturer identifies the potential to use a PCCP framework to address potential modifications, early engagement with FDA using the Pre-Sub process is recommended. Manufacturers should ensure that their QMS systems support the operational aspects of a PCCP to ensure the PCCPs are executed per requirements. Conclusion The FDA Predetermined Change Control Plans framework is a forward-looking regulatory tool that balances innovation with patient safety. While initially focused on AI-enabled devices, its potential application to all medical devices could redefine how manufacturers approach product lifecycle management. Stakeholders should prepare for this evolution by adopting proactive strategies and engaging with regulators to ensure compliance and competitiveness. DCN Dx’s Regulatory Affairs Services help IVD teams develop successful regulatory plans and submissions. We support FDA pathways including 510(k), De Novo, and PMA, with early Pre-Sub positioning and submission development, and we can advise on accelerated programs like Breakthrough Devices and STeP when they fit the product. Our team aligns verification and validation expectations across software and cybersecurity, biocompatibility, labeling, shelf life, analytical and clinical performance, usability and human factors, and CLIA waiver flex studies. We also support QMS readiness (ISO 13485, CLIA, CAP) and post-market obligations. For more information, visit our Regulatory Affairs page. --- ## The Foundation of Diagnostic Success: Specimen Strategy Is Development Strategy URL: https://dcndx.com/insights/biospecimen-strategy-development-strategy/ Type: insight Published: 2026-01-21 (As published 2026-01-21; regulatory status may have changed since.) By Jim Boushell, Senior Vice President, Biospecimens, DCN Dx When I made the transition from commercial to research in the mid 90’s, the diagnostic industry was experiencing a boom in breakthrough science: PCR, proteomics, and next-generation technologies. Yet many promising products never made it out of the lab, for reasons unrelated to the science itself. At the time, I didn’t stop to ask why. There was so much opportunity that I stayed focused on demand. My turning point came in a casual conversation with a family member: the issue was rooted in the specimen strategy, or lack of one. Access to samples wasn’t the problem. The challenge was building diagnostic programs on incomplete or misaligned specimen sets, using convenience samples, remnants, or acquisition methods designed for ease rather than evidence. Those early decisions often determine whether a technology can meet regulatory requirements, demonstrate clinical utility, and perform under real use conditions. Over three decades, I’ve learned that specimen strategy is not a procurement task. It’s a core component of development. At ProMedDx, we weren’t simply fulfilling demand; we were deploying site networks, standardized protocols, and operational infrastructure to support hundreds of diagnostic programs. The tests that ultimately changed patient care were built on the same principle: specimen strategy and development strategy are inseparable. Common pitfalls and lessons learned Remnant specimens While inexpensive upfront, remnant specimens often fail to represent target populations or support regulatory claims, leading to costly delays and repeat studies. Prospective collections require more planning, but they deliver long-term certainty. Where remnants tend to hurt teams is hidden variability: unknown collection devices and anticoagulants, time-to-processing drift, inconsistent storage history, limited metadata, and samples that do not map cleanly to the intended-use population. If your claims depend on stage, symptom status, comorbidities, or treatment effects, the “cheap” set can become the most expensive decision in the program. Real-world validation Devices intended for point-of-care use must be validated under actual operating conditions, not just in controlled labs. Performance data from expert operators rarely reflects real-world variability. Operator technique, workflow timing, environmental conditions, and site-to-site differences show up fast in decentralized testing. If your specimen plan does not reflect those realities (collection context, transport time, temperature excursions, handling steps), you can end up with a performance story that is hard to reproduce and harder to defend. Patient selection and regulatory alignment Inclusion and exclusion criteria shape regulatory claims and reimbursement pathways. Misalignment early in development can create significant downstream challenges. If the specimen set does not match the intended use, the clinical evidence can drift away from the label you want. The fix is rarely trivial: it usually means redesigning enrollment criteria, reopening sites, reworking comparators, or running additional studies to bridge gaps. Adapting to change COVID-19 accelerated the need for decentralized collection and flexible workflows. Success required applying proven strategies for reaching diverse and hard-to-access populations, reinforcing that adaptability depends on strong foundational protocols. Teams that handled the transition well treated collection as an engineered workflow: clear site or at-home procedures, training, logistics, and documentation designed to survive real operational friction without compromising data integrity. What differentiates successful diagnostics It’s rarely about technology alone. Success comes from understanding diagnostics as a system: specimens, workflows, operators, and data, and making informed decisions early. Time invested in getting specimen strategy right compounds forward. Time spent fixing missteps compounds backward. A quick pressure test for a specimen plan Is the intended use and target population defined well enough to design the set (including key subgroups and likely confounders)? Do the specimens support the claims you want to make, not just the feasibility work you can do quickly? Is the comparator method or reference standard chosen and operationally feasible at sites? Is minimum metadata defined (collection device, matrix, processing time, storage history, reference method result), and is there a plan when it is missing? Are collection, handling, shipment, and storage workflows mapped end-to-end, including expected temperature and timing variation? Is the set sized to support the statistical plan and the regulatory pathway, not just “what we can get”? Where DCN Dx fits At DCN Dx, we built our biospecimen services to operate in the same program reality as assay development, clinical research, and regulatory planning. That means aligning specimen requirements, workflows, documentation, and data capture to the evidence package from the start, then executing collections with audit-ready traceability. Our approach emphasizes: Starting with the regulatory endpoint in mind Designing protocols that reflect real-world use Building long-term site partnerships Engineering quality into every step The competitive advantage Specimen access is more available than it used to be. What sets programs apart is knowing which specimens to collect, how to collect them, and why they matter for your specific pathway. That insight comes from experience, not theory. If you’re developing a diagnostic, ask: does your specimen strategy support your regulatory and commercial goals? Getting this right early is often the difference between a clean evidence story and a cycle of costly delays. If need have a question about your specimen strategy, you can connect with us here. --- ## A 360° Approach to Aligning Clinical and Regulatory Strategy in IVD Development Overview URL: https://dcndx.com/insights/360-approach-aligning-clinical-regulatory-strategy-ivd-development-overview/ Type: insight Published: 2025-12-22 (As published 2025-12-22; regulatory status may have changed since.) DOWNLOAD NOW Overview Misalignment between clinical and regulatory strategy is one of the most common (and costly) sources of delay in IVD development. In this presentation, Emily Friedland, VP of Clinical Research at DCN Dx, introduces a practical 360° framework for aligning claims, evidence, study design, and regulatory expectations from the earliest stages of development through submission. Delivered at AMP 2025, the talk draws on real-world case studies where otherwise promising diagnostics stalled or failed due to siloed decisions, unclear intended use, weak claim-to-evidence mapping, or late-stage corrections. Rather than focusing on theory, the presentation shows how early, integrated planning across clinical, regulatory, data, and usability disciplines can reduce rework, minimize risk, and improve submission quality across the IVD lifecycle. Why You Should Download 1. Understand why clinical–regulatory misalignment causes programs to fail See how common breakdowns—vague intended use, disconnected evidence strategies, or delayed human factors work—create downstream problems that are difficult to recover from. 2. Learn the 360° Alignment Framework for IVD development The presentation outlines a structured approach to aligning claims, clinical evidence, biostatistics, usability, and regulatory strategy into a single, cohesive plan. 3. Explore real case studies and failure modes From rejected FDA submissions to failed clinical trials, these examples show exactly where programs went wrong and what could have been done differently. 4. Design studies for the label you intend to submit Evidence should be generated intentionally to support regulatory claims, not retrofitted late in development. 5. Reduce late-stage surprises and rework Learn how early cross-functional planning can prevent amendments, resets, and last-minute strategy shifts that cost time and momentum. --- ## Why Diagnostics Teams Outsource, and What to Expect When You Do URL: https://dcndx.com/insights/why-diagnostics-teams-outsource-what-expect/ Type: insight Published: 2025-12-03 (As published 2025-12-03; regulatory status may have changed since.) By Pat Vaughan, Ph.D., Chief Operating Officer, DCN Dx Outsourcing in diagnostics has shifted. It’s no longer just a fallback when timelines slip or resources run thin. More often, teams are choosing to outsource early, deliberately, because it’s the most effective way to meet their technical, operational, or commercial goals. We work with startups, growth-stage diagnostics firms, and established IVD companies. While every project is different, the reasons teams come to us tend to fall into a few consistent categories: The internal team is at capacity The program needs to move faster The science isn’t working The product isn’t ready for manufacturing The team lacks a specific skillset A regulatory or clinical milestone revealed a bigger issue IP concerns More recently, we’ve seen another trend: some teams now build outsourcing into their program plan from the beginning. These clients aren’t reacting to a failure or delay. Instead, they’re proactively using outsourced expertise to reduce execution risk, speed up early optimization, and stay focused on core business objectives. It’s become a strategic input and not simply a rescue lever. Here’s what each of those reasons looks like in practice. 1. The Internal Team Is at Capacity Most diagnostic companies aren’t built to run multiple development programs in parallel. They have strong scientists, but limited bandwidth. When those teams are tasked with doing too much, running feasibility, optimization, clinical prep, and transfer simultaneously, things start slipping. This has become more common in the last few years. In 2024, biopharma layoffs rose 3 percent year over year, with diagnostics companies like DermTech reducing their workforce by more than 50 percent. Randox Diagnostics, which scaled up during the COVID testing boom, has since shed over 1,300 jobs. Even well-capitalized firms are choosing to run leaner. It’s not that R&D has become less important, it’s that it’s increasingly outsourced to partners who can flex capacity up or down as needed. 2. The Program Needs to Move Faster Time is usually the most valuable resource in a diagnostics program. Whether you’re trying to hit funding milestones, meet a clinical window, or support an investor roadshow, delays compound quickly. Bringing in a CDMO can shorten timelines by: Resolving technical blockers more quickly Preventing the wrong experiments from being repeated Giving you early visibility into design or manufacturability issues In one recent program at DCN Dx, a client came to us with an upcoming trial window and a reader integration that wasn’t aligned with the strip design. Our team re-optimized the assay-reader interface and produced validation batches within three months, allowing the client to stay on schedule for submission. This speed-to-data is one reason the global medical device outsourcing market is projected to grow at a 12 percent annual rate through 2030, reaching nearly $293 billion ([Grand View Research]( https://www.grandviewresearch.com/industry-analysis/medical-device-outsourcing-market )). When your internal team is stretched, everything takes longer. Not because they’re not good at what they do, but because they’re doing too many things at once. 3. The Science Isn’t Working Sometimes teams reach out to us because they’ve run out of ideas. Their design looked solid on paper but isn’t performing. The troubleshooting process has stalled, and they need help figuring out what’s actually causing the problem. That outside perspective makes a difference. One client came in with a multiplexed lateral flow assay that had stalled in optimization. Their team suspected a reagent issue, but we traced the problem to a combination of membrane saturation and conjugate instability. This is especially common in quantitative lateral flow and multiplexed systems, where small design choices have a cascading impact on performance. 4. The Product Isn’t Ready for Manufacturing Another common scenario: a team believes they’re ready for tech transfer, but the manufacturer disagrees. That’s when we’re brought in to evaluate readiness and get the program back on track. This often involves: Validating the robustness of the assay design Assessing manufacturability Identifying changes needed for scale-up Producing additional feasibility or validation batches We’ve seen assays that worked well on the bench but couldn’t be reproduced under manufacturing conditions. In one example, we uncovered fundamental design limitations after a client’s tech transfer failed. It was something that could have been avoided with earlier feasibility checks. The product was ultimately discontinued, but the client avoided months of sunk time and cost because they got clear answers fast. 5. The Team Lacks a Specific Skillset Lateral flow and point-of-care development require deep, specialized knowledge that generalist R&D teams don’t always have in-house. That includes: Reader integration Fluorescent or visual labeling systems Multiplexed strip design Quantitative assay development Reagent lyophilization Biodegradable and sustainable device materials Aseptic filling Custom cassette and consumable engineering Design-for-manufacturing Hiring for those capabilities isn’t always feasible, especially on short timelines or in early-stage companies. We often serve as an extension of the internal team, applying focused technical expertise to accelerate progress and reduce risk. 6. A Regulatory or Clinical Event Exposed a Design Flaw Some clients don’t outsource until their trial is already underway or halted. Maybe the FDA had questions the team didn’t expect, or the study was paused due to poor performance, usability issues, or reproducibility failures. At that point, the internal team is often too close to the product to step back and rethink the strategy. They built it, launched it, and now need to redesign it. But doing so means revisiting decisions they weren’t resourced to reopen. This is where outside help can make the difference. We’re often brought in to conduct a technical audit, isolate the root cause, and support a redesign that aligns with both regulatory expectations and future manufacturing needs. This may include reworking claims language, adjusting protocols to match actual performance, or supporting root cause documentation. Sometimes the program gets back on track. Sometimes it pivots entirely. Either way, teams get the clarity they need to move forward. 7. Control and IP Protection Some teams hesitate to outsource because they worry about losing control of their product or intellectual property. That concern is valid and worth addressing up front. DCN Dx structures every engagement to ensure full client ownership and control of their intellectual property from start to finish. That means implementing confidentiality training across all staff, operating under robust quality and security protocols, and ensuring that any improvements or additional IP generated during the engagement remain the property of the client. Clear agreements and strong safeguards help protect your IP while still unlocking the value of external expertise. Done right, outsourcing doesn’t reduce control; it enhances it by aligning teams around shared outcomes and accountability. Final Thoughts Most diagnostics teams don’t come to us because they’ve failed. They come to us because they’re trying to keep momentum, reduce risk, and meet program goals. The decision to outsource isn’t a weakness. It’s a strategic call, and one that more teams are making earlier in the process. And the broader market reflects that shift. Global spending on medical device outsourcing hit $128.8 billion in 2023 and has continued to rise quickly. Teams aren’t outsourcing because it’s trendy. They’re outsourcing because it works. What Good Outsourcing Looks Like If you’re considering outsourcing, look for a partner who will do more than execute. The best CDMOs challenge assumptions, communicate clearly, and bring integrated insight across development, regulatory, and manufacturing. That’s what turns an outsourced task into strategic progress. We’ve previously published a rubric diagnostics developers can use as they vet outsourcing partners. You can download that guide here. DCN Dx has been a trusted outsourcing partner for hundreds of assay development programs. If you’d like to discuss your program, reach out to our team at DCNDx.com/contact. About the Author Pat Vaughan, Ph.D. | Chief Operating Officer, DCN Dx Patrick Vaughan, Ph.D., is Chief Operating Officer at DCN Dx, a diagnostics CDMO focused on lateral-flow and point-of-care systems. He oversees end-to-end programs from feasibility through clinical readiness and manufacturing transfer, with emphasis on assay optimization, reader integration, cassette and consumable design, and design-for-manufacture for CLIA-waived products. With 25+ years in IVD, he has led rescue and acceleration work for startups and global OEMs, turning stalled concepts into reproducible products and de-risking trials through practical study design, biostat input, and root cause analysis. His current interests include quantitative LFA, multiplexing strategies, and sustainable device materials. --- ## Rugged by Design: Making LFA Work in the Clinic, at Home, and in Production URL: https://dcndx.com/insights/making-lfa-work-clinic-home-production/ Type: insight Published: 2025-09-18 A DCN Dx Expert Insights conversation with Kellie LaRochelle, CEO of Imagene Technology Lateral flow diagnostics succeed when they are designed for the realities of everyday use—whether in the lab, at the bedside, or on the production line. In this episode of Expert Insights, Kellie LaRochelle brings her unique perspective as a former medical laboratory scientist, primary care provider, and now CEO of Imagene Technology. She discusses the three guardrails that never change—ease of use, ruggedness, and affordability—and how they guide decisions from reagent dispensing to error-proofing tests for real-world reliability. Listeners will hear Kellie’s practical take on avoiding complexity creep, training the next generation of diagnostics professionals, and balancing automation with skilled human oversight. She also explores how suppliers and manufacturers can support distributed strip production in resource-limited settings while ensuring performance at scale. It’s a candid and experience-rich look at how upstream choices shape downstream success—and what it takes to keep lateral flow both reliable and accessible. Presented in partnership with Imagene Technology, Gold Sponsor of ALFC 2025 and a long-time supporter of the conference. Imagene’s IsoFlow reagent dispensing systems are used by LFA manufacturers worldwide. Podcast listeners can enjoy 10% off their ALFC registrations by using code INSIGHTS at checkout. --- ## Making Quantitative Work: Practical Guidance from Cytiva’s Klaus Hochleitner URL: https://dcndx.com/insights/making-quantitative-work-practical-guidance-cytivas-klaus-hochleitner/ Type: insight Published: 2025-08-29 (As published 2025-08-29; regulatory status may have changed since.) Hard-earned insights from Cytiva’s diagnostics lead on building scalable, regulator-ready quantitative tests Quantification is the future of lateral flow. Here’s how to make it work. Quantitative formats are fast becoming the norm in respiratory panels, chronic disease monitoring, and decentralized trials—but getting them right takes more than just adding a reader. In this ALFC Sponsor Series episode of Expert Insights, Klaus Hochleitner, Ph.D., of Cytiva joins DCN Dx’s Mitzi Rettinger and Pat Vaughan, Ph.D., to unpack what truly makes or breaks a quantitative lateral flow test. Klaus draws on years of hands-on experience to highlight where developers go wrong, what they should prioritize early, and how choices like membrane type, conjugate performance, and reader integration shape long-term scalability. He also previews his upcoming lunch & learn session at ALFC 2025: “ Considerations in the Development of Quantitative Lateral Flow Tests. ” --- ## New Expert Insights Podcast Episode: Avoiding Regulatory Friction: Real Lessons from the Front Lines of IVD Clinical Trials with Sarah Barchard URL: https://dcndx.com/insights/expert-insights-podcast-avoiding-regulatory-friction-ivd-clinical-trials-sarah-barchard/ Type: insight Published: 2025-08-11 (As published 2025-08-11; regulatory status may have changed since.) In this episode of Expert Insights, Mitzi Rettinger sits down with Sarah Barchard, Senior Clinical Trials Manager at DCN Dx and co-author of our Expert Insights whitepaper on de-risking IVD studies for regulatory success. Sarah shares the most common and costly pitfalls she’s seen in hundreds of diagnostics programs, and walks through practical strategies for getting clinical, regulatory, and data teams aligned early. The conversation spans intended use, pre-submission engagement, biostatistics, human factors, and what it actually looks like to “fix” a struggling clinical trial before it derails a submission. --- ## Lyophilized Beads for Immunoassay and Molecular Diagnostics URL: https://dcndx.com/insights/lyophilized-beads-immunoassay-molecular-diagnostics/ Type: insight Published: 2025-07-22 Custom-developed for rapid rehydration, rugged handling, and seamless integration DCN Dx designs and manufactures custom lyophilized beads that eliminate cold chain logistics and simplify reagent workflows—without compromising performance. Built to rehydrate quickly and integrate easily into microfluidics, cartridges, or PCR tubes, these beads are optimized for point-of-care environments. Whether you’re developing lateral flow assays or molecular diagnostics, DCN Dx helps streamline the process from formulation through manufacturing. Our three-phase approach ensures robust, automation-ready beads tailored to your device’s requirements—with support for buffers, enzymes, antibodies, and more. What You’ll Learn How lyophilized beads eliminate cold chain requirements and extend shelf life Why spherical, rugged beads are ideal for high-throughput automation Which reagent types are compatible with lyophilized bead formats How DCN Dx’s three-phase development process accelerates go-to-market timelines What key factors impact project scope, timelines, and manufacturing readiness --- ## Whitepaper: Aligning the IVD Lifecycle URL: https://dcndx.com/insights/aligning-ivd-lifecycle/ Type: insight Published: 2025-07-09 (As published 2025-07-09; regulatory status may have changed since.) A 360° Strategy for Regulatory, Clinical, and Commercial Success Even world-class IVD programs stall when decisions around regulatory, clinical, and technical strategy are made in silos. This whitepaper distills DCN Dx’s experience across hundreds of global submissions to identify the five most preventable—and costly—sources of delay. Packed with real-world examples and actionable frameworks, it’s a must-read for diagnostics leaders, regulatory teams, and clinical program managers looking to de-risk development and get to market with confidence. What You’ll Learn: How vague or shifting intended use definitions trigger downstream submission problems Why skipping the FDA pre-submission pathway can cost you months What misaligned clinical protocols look like—and how to fix them early How and when to integrate human factors and usability to avoid rework The hidden role of data management and biostats in regulatory credibility --- ## New Expert Insights Podcast Episode: Hype or Help? How to Evaluate Generative AI in Diagnostics with Jeremy Elser, Ph.D., Palantir URL: https://dcndx.com/insights/new-expert-insights-podcast-episode-hype-help-how-evaluate-generative-ai-diagnostics-jeremy-elser-ph-d-palantir/ Type: insight Published: 2025-07-08 (As published 2025-07-08; regulatory status may have changed since.) Can generative AI meaningfully improve diagnostics—or is it just tech-world theater? In this episode, DCN Dx’s Mitzi Rettinger sits down with Jeremy Elser, Ph.D.—a data scientist and strategist currently leading science operations at Palantir—to explore how diagnostics leaders can make sense of AI’s rapid emergence in our field. They discuss: How to evaluate AI applications with scientific and operational rigor The difference between meaningful use cases and red-flag vendor pitches What generative AI can do for diagnostics—from institutional knowledge capture to faster decision-making Considerations for regulatory alignment and implementation Plus, get a sneak peek at Jeremy’s upcoming keynote at ALFC 2025 and why this is the year diagnostics professionals need to sharpen their AI instincts. About the guest: Jeremy Elser, Ph.D., Head of Science Operations, Palantir Jeremy Elser, Ph.D., is a computational biologist and executive strategist. He currently leads science operations at Palantir. --- ## Expert Insights: From Bench to Program Lead URL: https://dcndx.com/insights/expert-insights-bench-program-lead/ Type: insight Published: 2025-06-16 Helen Hsieh on Lateral Flow Development Lessons that Stick In this episode of Expert Insights, Helen Hsieh, Senior Scientist and Program Manager at DCN Dx, unpacks what it really takes to develop lateral flow assays that scale. Drawing on her experience with infectious disease, maternal health, and point-of-care diagnostics, Helen talks through common development mistakes, the hidden tradeoffs in label selection, and how to design with manufacturing in mind. You’ll also hear how Helen approaches client collaboration, troubleshooting, and the practical strategies she uses to guide early-stage ideas toward real-world success. Why Listen: Learn why early material choices can derail your entire assay Understand how to evaluate label options beyond sensitivity Get insights into common pitfalls in lateral flow development Hear how to assess real-world sample compatibility early Take away pragmatic advice for both bench scientists and program leads Expert Panelists Helen Hsieh, Senior Scientist and Program Manager at DCN Dx Helen Hsieh is a Senior Scientist and Program Manager at DCN Dx. With a Ph.D. in chemistry and a strong foundation in biophysics, Helen has worked on everything from COVID-19 and malaria assays to maternal health diagnostics, combining scientific depth with a sharp eye for assay performance. Mitzi Rettinger, former Chief Revenue Officer at DCN Dx; Show Host Mitzi Rettinger, former Chief Revenue Officer at DCN Dx, was a dynamic force in the life sciences industry, with more than 25 years of experience fueling innovation and growth across various sectors, including diagnostics, pharmaceuticals, and biotechnology. Her expertise lies in driving sustainable revenue growth, market development, and digital transformation, leveraging her deep understanding of the scientific commercial landscape. --- ## Advancing Lateral Flow Assay Development Through Internal R&D: DCN Dx’s Sample Adequacy Control Innovations URL: https://dcndx.com/insights/lateral-flow-assay-internal-research-development-sample-adequacy-control-innovations/ Type: insight Published: 2025-05-28 (As published 2025-05-28; regulatory status may have changed since.) How internal R&D at DCN Dx is helping solve common diagnostic test failures. This whitepaper shares how DCN Dx’s in-house R&D led to the creation of sample adequacy controls—tools that help ensure the right sample, in the right amount, gets applied during testing. Designed for blood, urine, and saliva assays, these markers improve reliability in home-use and point-of-care settings. You’ll get a look at the real problems these innovations address, the science behind them, and how they can be adopted without derailing your existing development process. What You’ll Learn What sample adequacy controls actually do Why they matter for home-use and OTC tests How they reduce false results What makes DCN Dx’s controls unique How they support faster, cleaner regulatory review --- ## Expert Insights: What the LDT Ruling Means for IVD Developers—and What to Do Now URL: https://dcndx.com/insights/expert-insights-what-the-ldt-ruling-means-for-ivd-developers-and-what-to-do-now/ Type: insight Published: 2025-05-06 (As published 2025-05-06; regulatory status may have changed since.) By Dan Simpson, RAC (US, CAN), DCN Dx A recent federal court ruling has dramatically shifted the regulatory landscape for Laboratory Developed Tests (LDTs), with significant implications for the diagnostics industry. On March 31, 2025, Judge Sean D. Jordan of the U.S. District Court for the Eastern District of Texas vacated the FDA’s LDT Final Rule, concluding that the agency lacks authority to regulate Laboratory Developed Test services. This decision turns on a critical legal distinction: the court determined that LDTs are services, not manufactured products, and therefore do not fall under the FDA’s authority over medical devices as defined by the Food, Drug, and Cosmetic Act. In the court’s view, an LDT is a “methodology or process” used by a clinical laboratory to generate diagnostic information from patient specimens—not a physical product transferred from one party to another. The implications of this ruling reach beyond the regulatory agencies. Diagnostics companies, clinical laboratories, healthcare providers, and patients will all feel the effects. This case not only impacts the compliance frameworks developers must follow but also raises broader questions about the future balance between innovation and oversight in the diagnostics industry. A Brief History of FDA and LDT Oversight The FDA has asserted since the 1970s that LDTs are a subset of in vitro diagnostic (IVD) devices. Under the Medical Device Amendments of 1976, the FDA gained authority to regulate medical devices, including in vitro diagnostics (IVDs) which are reagents, instruments, and systems used for diagnostic purposes. FDA recognized there was a subset of IVDs that were developed, manufactured, and performed with a single laboratory. At that time these tests (LDTs) were relatively simple, low-risk assays developed and performed within single laboratories to meet local or rare diagnostic needs. Given the perceived low public health risk, the FDA exercised “enforcement discretion” and chose not to impose full device regulations, such as premarket review or Good Manufacturing Practice requirements, on LDTs. Meanwhile, clinical laboratories were regulated under the Clinical Laboratory Improvement Amendments (CLIA) of 1988, administered by CMS. CLIA focused on laboratory quality and test performance, not necessarily the safety and effectiveness of individual assays. As diagnostic technology advanced, and particularly with the rise of high-throughput molecular and genetic testing, the FDA grew concerned that some modern LDTs were fundamentally different from the simple assays originally covered by enforcement discretion. These tests were often used to guide critical treatment decisions for large, diverse patient populations, and were produced by national laboratories operating at commercial scale. Throughout the early 2000s, the FDA made multiple attempts to increase oversight of LDTs, citing risks to patient safety. However, each attempt was met with strong resistance from clinical laboratory organizations, which argued that additional regulation would stifle innovation, reduce access to critical diagnostics, and exceed the FDA’s statutory authority. In 2024, the FDA finalized its LDT Rule, aiming to phase out enforcement discretion over five years. The rule required labs developing LDTs to meet quality system regulations and submit tests for premarket review, similar to traditional IVD manufacturers. The Legal Challenge and Court Ruling Shortly after the rule was finalized, major industry groups, including the American Clinical Laboratory Association (ACLA) and the Association for Molecular Pathology (AMP), filed suit, arguing that the FDA overstepped its authority. In the case heard in early 2025, the court focused on the fundamental nature of LDTs. Plaintiffs argued that LDTs are services performed by skilled professionals, not “devices” as traditionally defined. The FDA countered that LDTs involved tangible materials and components that fit within the statutory definition of a medical device. Judge Jordan sided with the plaintiffs. His ruling emphasized that LDTs do not involve the sale or transfer of a physical article of commerce. Instead, they are methodologies developed and applied by laboratories internally, using proprietary protocols and professional judgment. As such, they fall outside the FDA’s device regulatory framework. What Happens Now? While this decision is a significant setback for the FDA, the regulatory future of LDTs remains unsettled. The Secretary of Health and Human Services and the Trump Administration may choose to appeal. Congressional intervention also remains possible, as the broader debate over LDT oversight and balancing patient safety, innovation, and access continues. The clinical laboratory industry itself remains divided, with some advocating for maintaining LDT regulation solely under CLIA and others calling for a new, risk-based framework that includes FDA involvement. If an appeal is filed, it could take years to resolve, creating a prolonged period of regulatory uncertainty. Developers must prepare for multiple potential outcomes, including reinstatement of FDA oversight, further legislative action, or additional attempts at hybrid frameworks blending FDA and CMS responsibilities. What Developers Need to Know For diagnostics developers and labs, the key takeaway is that regulatory uncertainty around LDTs will persist Although the FDA’s authority to enforce its LDT rule has been vacated (for now), high expectations from investors, payers, and other stakeholders for validated, high-quality tests are not regardless of the test’s regulatory status. Smart regulatory planning is more important than ever. Here is how DCN Dx is advising to our clients: Validation is Key: Strong clinical and analytical validation will remain critical for building commercial value, regardless of the regulatory pathway. Stay Flexible: Regulatory pathways could shift again. Preparing for both CLIA-only and potential FDA or legislative frameworks keeps options open. Engage Experts Early: Strategic trial design, biostatistics, and regulatory consulting can help ensure that today’s data packages meet tomorrow’s standards. Consider Global Markets: Developers seeking to commercialize internationally must also consider IVDR (EU) and other emerging frameworks, regardless of U.S. domestic policy. Proactive Risk Management: Building robust quality systems, even if not currently required under FDA rules, will strengthen organizational resilience and facilitate faster pivots if regulations change. What This Means for Your Organization The impact of this ruling varies widely depending on your current position in the market. A few scenarios we’re seeing: If you’re a startup with an LDT in a CLIA lab: You may need to budget for clinical data, formal design control documentation, and regulatory submission costs sooner than planned. DCN Dx can help scope these requirements and avoid surprises during pre-sub or submission. This will allow the ability to pivot from an LDT format to FDA clearance/approval if desired. If you’re planning to raise capital or pursue M&A: Expect increased scrutiny from investors or acquirers regarding regulatory exposure. A documented FDA strategy and early Q-sub engagement can materially de-risk your profile. If you decide to or need to convert to an FDA regulated product: FDA oversight introduces new operational requirements—labeling, adverse event reporting, QMS, and post-market surveillance planning. These may require team expansion or a CRO/CDMO like DCN Dx to fill gaps. If you’re a lab with multiple LDTs under EUA: Consider which should move first to full IVD status. DCN Dx can help prioritize based on technical readiness, competitive pressure, or risk. DCN Dx: Helping Developers Navigate What’s Next At DCN Dx, we’ve already begun working with clients to reassess regulatory strategies in light of the LDT ruling. For diagnostics companies that have historically operated under CLIA but now face the prospect of 510(k) or de novo submissions, our clinical research and regulatory teams offer: Regulatory strategy and risk assessment: Including device classification, predicate identification, and feasibility planning for EUA-to-510(k) or LDT-to-IVD transitions. Q-sub and pre-submission planning and execution: Including FDA meeting prep, submission drafting, and coordination. Clinical trial design and execution: From IRB approvals and site selection to biostatistics and data management, all under our ISO 13485–compliant systems. Post-market planning and labeling review: Including strategies for PMS, RUO/IUO claims, and companion diagnostic co-development. Whether you need targeted consulting or a full CRO partner to manage study design through submission, DCN Dx provides integrated, diagnostics-specific support built to accelerate timelines and minimize risk. Explore our CRO services to learn how DCN Dx can support your IVD development and regulatory planning needs. Need help responding to the LDT ruling? DCN Dx offers regulatory strategy, Q-sub planning, and clinical trial execution—all tailored to IVD developers. Get in touch with our regulatory team → --- ## Regulatory Affairs Services Brochure URL: https://dcndx.com/insights/regulatory-affairs-services-brochure/ Type: insight Published: 2025-05-03 (As published 2025-05-03; regulatory status may have changed since.) Learn about DCN Dx’s strategic support for FDA, IVDR/MDR, and global compliance—tailored for diagnostic devices. DCN Dx provides end-to-end regulatory affairs services to help diagnostic developers streamline approvals and reduce risk. Our team supports every major submission pathway—510(k), de novo, PMA, and IVDR/MDR—along with global market access, pre-submission meetings, and post-market compliance. Whether you’re pursuing Breakthrough Device designation or preparing for a QMS audit, we guide you through every step with the clarity and expertise needed to keep your launch on track. Learn more—download the brochure. --- ## A Must-Watch DCN Dx Presentation from the 2025 Precision Med Tri-Conference: Regulatory Pitfalls in Diagnostic Development URL: https://dcndx.com/insights/a-must-watch-dcn-dx-presentation-from-the-2025-precision-med-tri-conference-regulatory-pitfalls-in-diagnostic-development/ Type: insight Published: 2025-04-08 (As published 2025-04-08; regulatory status may have changed since.) DCN Dx extends a special thank you to all who joined us at the 2025 Precision Med Tri-Conference & Expo. The event provided an invaluable opportunity to connect with diagnostic developers, share expertise, and discuss strategies for navigating regulatory challenges in IVD development. A key moment from the event was the presentation by DCN Dx’s Chief Operating Officer, Patrick Vaughan, Ph.D. His session, Regulatory Pitfalls in Dx Development: Lessons from Real-World Case Studies, provided attendees with practical insights drawn from an STI biomarker lateral flow assay case study. Dr. Vaughan explored the impact of early regulatory engagement, study design challenges, and best practices for refining assays to meet validation, scalability, and commercialization requirements. Missed the live session? You can now watch Dr. Vaughan’s full presentation by submitting the form below: --- ## New Podcast Episode—What’s Behind Winning Regulatory Strategies for IVDs? URL: https://dcndx.com/insights/blog-podcast-episode-winning-regulatory-strategies-ivds/ Type: insight Published: 2025-03-12 (As published 2025-03-12; regulatory status may have changed since.) Discover How to Navigate Regulatory Challenges for IVD Success In this episode of Expert Insights, host Mitzi Rettinger sits down with Dan Simpson, Head of Regulatory Affairs at DCN Dx, and Emily Friedland, VP of Clinical Research at DCN Dx, to discuss the critical role of regulatory strategy in bringing in vitro diagnostic (IVD) devices to market. From FDA approvals to international compliance, understanding the regulatory landscape is key to avoiding costly delays and ensuring a smooth path to commercialization. Dan and Emily share their expertise on structuring a regulatory strategy that minimizes risk while optimizing approval timelines. They explore the importance of pre-submission consultations, the nuances of FDA versus IVDR requirements, and how early collaboration between regulatory and clinical teams can streamline the approval process. Whether you’re working through a 510(k), de novo, or PMA pathway, this episode provides valuable insights to help you plan an effective regulatory approach. Listeners will gain a deeper understanding of how regulatory strategy impacts clinical study designs, the importance of harmonizing global compliance efforts, and the best practices for engaging with regulatory agencies early in development. Listen below to learn about winning regulatory strategies for IVDs, or find us on your preferred podcasting platform. Why Listen? This episode breaks down the essential elements of a strong regulatory strategy for IVD developers. You’ll learn: How to determine the right regulatory pathway for your IVD device (510(k), De Novo, or PMA). The role of pre-submission consultations in mitigating risks and expediting approvals. Key differences between FDA and IVDR requirements and how to harmonize global compliance efforts. How early collaboration between regulatory and clinical teams can prevent costly delays. Real-world examples of successful regulatory strategies that accelerated market entry. Whether you’re an entrepreneur, scientist, or regulatory professional, this episode offers practical insights to help you bring your diagnostic device to market efficiently. --- ## New Podcast Episode—2024 Review and 2025 Predictions for Lateral Flow Diagnostics URL: https://dcndx.com/insights/new-podcast-episode-2024-review-and-2025-predictions-for-lateral-flow-diagnostics/ Type: insight Published: 2025-02-20 (As published 2025-02-20; regulatory status may have changed since.) What trends shaped lateral flow diagnostics in 2024? What innovations and challenges lie ahead in 2025? In this episode of Expert Insights, DCN Dx’s former Chief Revenue Officer, Mitzi Rettinger, sits down with Chief Operating Officer Dr. Patrick Vaughan to discuss the latest advancements in lateral flow, from decentralized healthcare and AI-driven diagnostics to sustainability efforts and regulatory shifts. Looking ahead to 2025, Dr. Vaughan shares his predictions for regulatory harmonization, expanded use of sustainable materials, and breakthroughs in personalized medicine. He also discusses the challenges and opportunities for startups versus established companies and what regulatory shifts might mean for the industry. This episode is packed with insights that will help diagnostics developers, scientists, and industry leaders understand where the market is headed and how to navigate the evolving landscape. Listen below, or find us on your favorite podcasting platform. --- ## New Podcast Episode—Developing Your Lateral Flow Development Skills With DCN Dx’s 2025 Basic Training Course URL: https://dcndx.com/insights/podcast-expert-insights-lateral-flow-lfa-training-melanie-bader/ Type: insight Published: 2025-01-29 Learn What Makes DCN Dx’s Lateral Flow Training the Most Sought-after in the Industry In this episode of Expert Insights, host Mitzi Rettinger sits down with Melanie Bader, Research Scientist and Training Specialist at DCN Dx, to discuss the 2025 Basic Lateral Flow Training Course. Designed for professionals looking to deepen their understanding of lateral flow assay (LFA) development, this three-day course combines expert-led lectures with hands-on lab work to provide attendees with the technical knowledge and practical skills necessary for successful assay development. Melanie shares her extensive experience in LFA development and what makes this training program one of the most sought-after in the industry. Together, she and Mitzi explore why structured training is crucial for both newcomers and experienced professionals, the hands-on aspects of the course, and the vibrant learning environment at DCN Dx’s Carlsbad, California, headquarters. BLFT attendees will gain expertise in design control, reagent selection, reader technologies, manufacturing considerations, and troubleshooting strategies—essential knowledge for bringing robust diagnostic tests to market. This episode discusses those topics. Plus, listeners will hear about networking opportunities, group dinners, and how past participants have leveraged this experience to advance their careers. Whether you’re an entrepreneur, scientist, or engineer looking to refine your LFA skills, this episode is your gateway to understanding why DCN Dx’s training course is the industry gold standard. Listen below, or find us on your favorite podcasting platform. Why Listen? This episode dives into the “why” behind DCN Dx’s Basic Lateral Flow Training Course. You’ll gain insights into: What makes DCN Dx’s Basic Lateral Flow Training Course a must-attend for LFA professionals. The importance of structured training in avoiding costly development mistakes. An inside look at the course structure, including lectures, hands-on labs, and troubleshooting exercises. Why Carlsbad, California, is the perfect setting for professional development and networking. How past attendees have benefited from this lateral flow training and applied it to their careers. Details on how to secure your spot. (Hurry! Early bird for the February course ends soon! ) --- ## 2024 Year in Review and 2025 Predictions for Lateral Flow Diagnostics URL: https://dcndx.com/insights/2024-lfa-review-2025-predictions-lateral-flow/ Type: insight Published: 2025-01-22 (As published 2025-01-22; regulatory status may have changed since.) The field of point-of-care (POC) diagnostics and especially lateral flow diagnostics (LFDs) has had another exciting year post-pandemic, laying further foundations for significant future growth. The global market for POC diagnostics was valued at $36.9 billion in 2023 and $40 billion in 2024, and is expected to reach $65.9 billion by the end of 2029. As we close 2024, it is an opportune moment to reflect on the year’s developments and anticipate what lies ahead for 2025. Below, I’ll explore key innovations and market trends, alongside informed predictions about where the industry is heading. 2024 Year in Review The need for and acceptance of fast and reliable COVID tests had led to a widespread acceptance and understanding of rapid tests—especially lateral flow assays. Since the pandemic, the “untrained” user population used in trials for CLIA waivers is certainly not as naive as it once was, and CLIA studies not as daunting as they once were. However, that does not negate the absolute need for well-designed and high-performance lateral flow tests. This will continue to promote the shift of healthcare from the doctor’s office or clinic to at-home and over the counter (OTC) self-testing. In addition, POC testing is also driving an evolution of healthcare in specialized clinical departments or clinics and decreasing their reliance on centralized laboratory testing. This shift aligns with trends toward tiered healthcare and decentralized medical services. As high-throughput devices reach smaller facilities, POCT adapts to meet the specific needs of clinical departments by emphasizing speed, convenience, and cost-effectiveness. This transformation positions POCT to excel in delivering flexible and agile diagnostic solutions. POCT product development must prioritize tailored solutions to meet the diverse needs of clinical departments. Different use cases, such as maternal and child health, kidney function assessment, and chronic disease management, demand specialized features like peripheral blood testing, wide-range urine indicators, and robust data transmission capabilities. By designing products that align closely with these unique requirements, POCT developers can enhance relevance, usability, and impact in healthcare. Point-of-need testing remains vital, offering real-time results for urgent care scenarios like ERs and ICUs, while balancing sample complexity with healthcare workflow needs. At the same time, at-home diagnostic tests have advanced, empowering patients with accessible and convenient health monitoring tools. These tests complement traditional care by enabling informed decision-making, easing the burden on healthcare systems, and fostering patient engagement. Integration with Digital and AI Platforms The integration of digital technologies into lateral flow devices is another pivotal trend of 2024. Smartphone-based readers as well as highly customized stand-alone LFA readers have allowed users to obtain semi-quantitative or fully quantitative results from traditionally qualitative assays. These technologies not only have the potential to improve diagnostic accuracy but also enable real-time data sharing for epidemiological tracking and disease management. The use of artificial intelligence (AI)-driven image analysis has also allowed development of more informative and accurate algorithms that enable unprecedented data collection and analysis. AI is already transforming medical applications, particularly in medical imaging, where it aids in detecting anomalies often imperceptible to the human eye. As AI, connected health, and rapid diagnostics advance, their development is becoming increasingly intertwined especially in the development of diagnostic tools. While the live use of AI in clinical diagnosis will no doubt be a highly debated topic in the years to come with accompanying regulatory challenges, its use in diagnostic tool development serves as a bridge between traditional POC testing and emerging at-home diagnostics, enhancing accuracy, efficiency, and integration across healthcare solutions. Next Gen Diagnostics and Data Sharing versus Privacy While the revolution in diagnostics excites many, widespread data collection and sharing, especially in healthcare, raises significant privacy concerns. However, ethical data-sharing practices are essential for advancing next-generation diagnostics, scientific discoveries, and technological innovation. Companies must prioritize informed consent, data security, patient anonymity, transparency, and responsible beta testing. Given the sensitive nature of health records, their privacy is paramount and protected by regulations, but technological advancements often outpace policies. All stakeholders must ensure responsible data use, balancing ethical considerations with progress. Risks, such as insecure storage and unintended sharing through healthcare apps, highlight the need for proactive measures during device development. By addressing these challenges and fostering ethical discussions among patients, providers, and researchers, the integration of diagnostics with electronic medical records can be achieved responsibly. The use and integration of AI systems further the need for responsible use and protection of data. AI systems require vast amounts of data, making them attractive targets for cyberattacks. Poor encryption protocols during data transfer or storage can lead to breaches. Even the potential for anonymized data to be reverse-engineered may be possible allowing individuals to be identified thereby violating their privacy. That is why there must be a concerted effort to implement and enforce robust data protection laws, such as GDPR or HIPAA, to ensure compliance. In addition the introduction and use of cybersecurity in all instrumentation including the aforementioned LFA readers. Commercialization and Market Dynamics for Lateral Flow Assays The lateral flow diagnostics market continued its robust growth in 2024, with revenues between $10 and $15 billion, depending on the analyst. This expansion has been fueled by increasing applications in infectious disease diagnostics, as well as emerging uses in oncology, environmental monitoring, and veterinary medicine. At the same time, diagnostic preparedness for non-COVID infectious diseases remains a critical challenge. FIND’s 2024 readiness index highlighted glaring diagnostic shortages for diseases like tuberculosis and malaria. These shortages have placed significant pressure on assay developers to innovate rapidly and meet urgent needs in resource-limited settings. The lack of reliable point-of-care diagnostics for tuberculosis creates barriers to timely treatment exacerbates public health challenges. Current assays often face limitations in sensitivity, particularly for detecting low bacterial loads in extrapulmonary or pediatric TB cases. Scalability and affordability also remain significant hurdles, as many existing diagnostics are too costly or complex for deployment in resource-limited settings. However, this gap also presents opportunities for developers to design assays tailored for low-resource environments, such as multiplexed LFAs that combine affordability with high sensitivity. Collaborations with global health organizations and targeted funding mechanisms could accelerate progress in this critical area. This should stress the importance of diversifying R&D investments to address global health priorities beyond pandemic response. Industry Events and Collaborations The Advanced Lateral Flow Conference (ALFC) 2024, themed “Flow Forward,” brought together thought leaders to discuss emerging technologies and regulatory challenges. Notable sessions focused on the integration of machine learning algorithms and harmonizing global regulatory pathways for lateral flow diagnostics. The conference emphasized the importance of sustainability in diagnostics—a topic poised to gain even more attention in 2025. Predictions for 2025: Shaping the Future of Lateral Flow Diagnostics As we look ahead to 2025, I can’t help but feel optimistic about what’s in store for the lateral flow diagnostics industry. Call it a New Year’s wish list for developers, researchers, and industry stakeholders, but many of these predictions are grounded in the exciting momentum we built over the last 2-3 years and especially in 2024. While some may seem ambitious, I firmly believe the progress we’ve seen in lateral flow diagnostics puts us on the brink of transformational breakthroughs. Let’s dive into what I hope to see—and what I predict will come to fruition—in the year ahead. 1. Broader Integration with Digital Ecosystems Digital transformation in diagnostics will accelerate in 2025, with more manufacturers incorporating cloud-based platforms into their LFA products. These platforms will not only facilitate real-time disease surveillance but also improve patient management by integrating with electronic health records (EHRs). Regulatory challenges for these platforms often include demonstrating consistent analytical performance across varied user environments, ensuring data security in cloud-based integrations, and navigating the classification of AI software under existing medical device regulations. Overcoming these hurdles will require closer collaboration between developers and regulatory agencies, alongside robust validation studies tailored to address the unique complexities of AI-driven diagnostics. 2. Sustainability as a Core Focus Sustainability will emerge as a central theme in diagnostic development. Examples include the adoption of biodegradable nitrocellulose membranes and the use of biodegradable or compostable housings for lateral flow devices. Additionally, several manufacturers are experimenting with energy-efficient production processes and sourcing raw materials from renewable resources. These efforts not only reduce environmental impact but also align with increasing global emphasis on eco-friendly healthcare solutions. This includes the adoption of biodegradable materials for assay components and the design of eco-friendly manufacturing processes. At ALFC 2025 (as in 2024), we anticipate extensive discussions on reducing the environmental footprint of diagnostics while maintaining affordability and accessibility. (For more on this important issue, I invite you to watch DCN Dx’s Industrial Designer’s discussion on this topic here.) 3. New Market Opportunities for LFDs The growing demand for decentralized testing will drive innovations in lateral flow technology for chronic disease monitoring and personalized medicine. For example, we foresee the development of LFAs for monitoring wellness biomarkers associated with general health trends, such as blood glucose or cholesterol levels, enabling individuals to track their health more effectively at home without direct medical oversight. While the use of LFAs for regulated healthcare applications, like cardiovascular or stroke markers, may face regulatory hurdles that ultimately can be overcome, In addition the focus on wellness diagnostics could offer an alternative pathway for commercialization for certain tests. 4. Regulatory Harmonization and Global Deployment 2025 will see significant progress in harmonizing regulatory frameworks for lateral flow diagnostics. While the implementation of the IVDR rules in the EU has caused significant hurdles for market entry, it does make to more stable and well understood US FDA regulations become more attractive for market entry. Building Momentum: Looking Ahead to ALFC 2025 The Advanced Lateral Flow Conference 2025, scheduled for October, promises to be another milestone event for the industry. With talks and panels shaping up to include sustainability, AI integration, and LFA market expansion, ALFC 2025 will serve as a hub for innovation and collaboration. DCN Dx will be proud to contribute our insights on cutting-edge technologies and emerging market trends. Hope to see you there! Closing Thoughts The past year has been transformative for lateral flow diagnostics, with advancements that have redefined the potential of this versatile technology. As we enter 2025, the industry stands at the cusp of even greater innovation, driven by the convergence of materials science, digital technology, and global health priorities. At DCN Dx, we are excited to be part of this journey, supporting developers and stakeholders as they bring impactful solutions to market. Advancements in at-home and point-of-care diagnostic solutions have elevated patient care by improving safety, outcomes, and reducing healthcare costs and resource burdens. Patients now play an active role in monitoring their health, supported by growing options to track and diagnose conditions. Early detection and diagnosis, the cornerstone of prevention and treatment, lead to better outcomes, lower costs, and more equitable healthcare access. These advancements signify a paradigm shift in medicine, with widespread adoption of diagnostic platforms transforming care delivery. Combined with laboratory-like services for at-home or clinic use, this shift enhances the affordability, accessibility, and effectiveness of medical care for a broad range of diseases, marking a profound cultural and medical evolution. Promoting universal access to healthcare involves ensuring that the benefits of accessible POC testing are realized globally, in both developed and developing countries. Advocating for widespread diagnostic access and advancing technology in low- to middle-income countries is crucial in combating and preventing devastating disease outbreaks and pandemics. Strong efforts to make diagnostic tools available and affordable will play a key role in improving global health outcomes and strengthening defenses against future health crises. For lateral flow developers, medtech investors, and researchers, the coming year offers unparalleled opportunities to shape the future of diagnostics. We look forward to continuing this dialogue and fostering collaboration across the industry. --- ## New Podcast Episode—The Hidden Costs of Cheap Lateral Flow Development: Why Experience Matters URL: https://dcndx.com/insights/podcast-episode-hidden-costs-cheap-lateral-flow/ Type: insight Published: 2024-12-10 (As published 2024-12-10; regulatory status may have changed since.) Understanding the True Costs of “Cheap” LFA Development In this episode of DCN Dx’s Expert Insights, Mitzi Rettinger interviews Dr. Pat Vaughan, COO of DCN Dx, to explore the risks of working with inexperienced or low-cost lateral flow assay developers. While these options may seem cost-effective at first glance, they often lead to costly delays, redesigns, or outright project failures. Drawing from real-world case studies, Dr. Vaughan shares insights on how experience and technical expertise can help mitigate risks and ensure successful assay development and commercialization. Why Listen? This episode dives into the key considerations for selecting an LFA development organization, with a focus on identifying and avoiding the pitfalls of working with inexperienced providers. You’ll gain insights into: Real-world case studies where DCN Dx turned around failing projects to deliver successful, market-ready products. The long-term risks and hidden costs of partnering with cheap lateral flow developers. Key qualities to look for in a development partner, including manufacturability, scalability, and regulatory readiness. Specific guidance in how to evaluate RFP responses from prospective LFA development partners. Gain access to our free, downloadable rubric to evaluate potential lateral flow assay development partners and ensure your project stays on track— to get started. Expert Panelists Mitzi Rettinger, former Chief Revenue Officer at DCN Dx; Show Host Mitzi Rettinger, former Chief Revenue Officer at DCN Dx, was a dynamic force in the life sciences industry, with more than 25 years of experience fueling innovation and growth across various sectors, including diagnostics, pharmaceuticals, and biotechnology. Her expertise lies in driving sustainable revenue growth, market development, and digital transformation, leveraging her deep understanding of the scientific commercial landscape. Pat Vaughan, Ph.D., Chief Operating Officer at DCN Dx Dr. Pat Vaughan oversees DCN Dx’s IVD CDMO and CRO services, guiding complex projects from early feasibility through full-scale manufacturing. With over two decades of experience across multiple diagnostic platforms—including lateral flow, molecular diagnostics, and point-of-care devices—Dr. Vaughan’s team at DCN Dx specializes in ensuring products are manufacturable, scalable, and regulatory-ready. --- ## Navigating U.S. Market Entry for IVDs URL: https://dcndx.com/insights/navigating-u-s-market-entry-for-ivds/ Type: insight Published: 2024-12-01 (As published 2024-12-01; regulatory status may have changed since.) Leveraging IVD Service Providers for Compliance and Device Integration Entering the U.S. market with a new diagnostic device can be complex—but it doesn’t have to be chaotic. In this talk delivered during MEDICA 2024, Dr. Patrick Vaughan, Chief Operating Officer of DCN Dx, shares practical strategies for navigating regulatory requirements, optimizing study design, and integrating devices efficiently. Drawing on DCN Dx’s experience supporting hundreds of IVD products, the presentation outlines how developers can accelerate timelines and reduce risk by leveraging the right service providers, particularly during critical development and regulatory phases. Why You Should Download 1. Gain clarity on U.S. regulatory pathways. Dr. Vaughan breaks down common misconceptions about the FDA process, including when and how to engage in pre-submission discussions. 2. Understand how to integrate devices, readers, and data systems. Learn how to approach reader integration and connectivity in a way that aligns with both regulatory expectations and user experience needs. 3. See real-world examples from successful IVD programs. From CLIA-waived assays to EU-to-U.S. transitions, this presentation shares insights drawn from actual client engagements. 4. Discover how to leverage IVD CROs and CDMOs strategically. Find out when to bring in external expertise—and how to choose service providers that will protect your IP, accelerate development, and deliver usable outputs. 5. Walk away with actionable takeaways. This is not a generic overview. It’s a focused, practical session for diagnostics professionals serious about U.S. market success. --- ## DCN Dx to Highlight Lateral Flow Development Services and miniDxR at MEDICA 2024 URL: https://dcndx.com/insights/lateral-flow-development-services-medica-2024/ Type: insight Published: 2024-10-17 (As published 2024-10-17; regulatory status may have changed since.) Explore DCN Dx’s contract IVD services and customizable lateral flow reader at Booth Hall 3 / D25-2 (To schedule a meeting with our team at MEDICA 2024 or discuss your project in advance.) CARLSBAD, Calif.—DCN Dx, a global leader in contract lateral flow assay (LFA) in vitro diagnostic (IVD) development and manufacturing, and diagnostic clinical research services, will be exhibiting at MEDICA 2024 in Düsseldorf, Germany. Attendees are invited to visit Booth Hall 3 / D25-2 in the US Pavilion to learn about DCN Dx’s full range of tailored IVD solutions, including assay development, ISO-compliant manufacturing, and clinical research and regulatory consulting. One of the key innovations DCN Dx will feature at the event is the miniDxR, a lateral flow assay reader offering customizations for a variety of diagnostic applications. The miniDxR has been benchmarked against leading industry readers and provides accurate results in both visual and fluorescence modes, making it well-suited for point-of-care testing and use in resource-constrained settings. “MEDICA 2024 is a great opportunity to showcase how our tailored solutions help diagnostic developers streamline development and deployment of diagnostics,” said Pat Vaughan, Ph.D., Chief Operating Officer of DCN Dx. “The miniDxR exemplifies the performance and flexibility that sets our solutions apart—especially in lateral flow diagnostics.” Visitors to the DCN Dx booth will have the chance to speak with experts across the company’s key service areas, including lateral flow assay development, engineering, manufacturing, and clinical research. DCN Dx’s team collaborates with clients, offering expert guidance to overcome development and regulatory hurdles. “MEDICA 2024 is an ideal setting to engage with IVD professionals from around the world and discuss how DCN Dx helps developers navigate each step of the IVD process,” said Mitzi Rettinger, former Chief Revenue Officer at DCN Dx. “From assay development to clinical trials and regulatory strategy, our team provides the resources and expertise needed for ensuring regulatory approval and a smooth path to market.” DCN Dx also welcomes attendees to join the talk, “Navigating U.S. Market Entry: Leveraging IVD Service Providers for Compliance and Device Integration,” presented by Dr. Patrick Vaughan, Ph.D., COO of DCN Dx. The presentation will take place on November 11, from 11:45 a.m. to 12:05 p.m. local time in Hall 8A, Booth G40. This session will provide insights into how partnering with IVD CROs like DCN Dx can streamline regulatory compliance and device integration for a successful entry into the U.S. market. About DCN Dx DCN Dx, based in Carlsbad, California, is a global leader in IVD CDMO and CRO services. Our multidisciplinary CDMO specializes in creating tailored assay systems, consumables, and instruments for point-of-use applications, with a particular expertise in lateral flow assays. Our clinical research services group specializes in the planning, execution, and oversight of IVD clinical trials, inclusive of clinical operations, data management, and biostatistics. DCN Dx’s personalized approach to IVD product development and clinical trials has supported hundreds of programs and clients. They can oversee every stage from concept to assay development and platform integration, through clinical trials to manufacturing or only handle the aspects you request. In addition to lateral flow assays, DCN Dx’s clinical research expertise extends to any in-vitro diagnostic, all with a focus on innovation, usability, and performance. With a commitment to quality, customer satisfaction, and industry-leading expertise, DCN Dx is setting the benchmark for excellence in IVD services.com. For more information, contact DCN Dx at customercare@dcndx.com or (760) 804-3886. --- ## Designing Lateral Flow Assays for Low-Resource Settings: 7 Practical Strategies for Developers URL: https://dcndx.com/insights/designing-lateral-flow-assays-for-low-resource/ Type: insight Published: 2024-10-16 As diagnostic technologies evolve, they often become more sophisticated, requiring advanced infrastructure, greater energy used, and specially trained personnel. While these advancements can enhance diagnostic accuracy and speed in well-resourced settings, they also make these tools increasingly out of reach for low- and middle-income countries (LMICs). In many regions, the cost of equipment, the need for cold chain storage, or the requirement for specialized training create significant barriers to implementation. This can lead to diagnostic gaps, particularly in areas where timely diagnosis could prevent the spread of infectious diseases or improve patient outcomes. Lateral flow assays (LFAs) or Rapid Diagnostic Tests (RDTs) and other point-of-care tests (POCTs) continue to offer compelling solutions to these challenges. Designed for use in decentralized and/or resource-limited environments, LFAs are simple to use, affordable, and are often designed so as not to rely on energy-hungry equipment or highly trained personnel. These attributes make them ideal for improving access to healthcare in LMICs, where more complex diagnostic systems often fail to reach the population in need. In this article, we present 7 practical strategies for developers working to design lateral flow assays for LMICs, drawing on DCN Dx’s expertise in developing IVDs for these unique environments. From designing robust, affordable tests to ensuring successful manufacture, field deployment and integration with local healthcare practices, these strategies are designed to help developers create impactful, sustainable diagnostics that help meet the real-world needs of LMICs. That said, the strategies outlined here are generalized. For tailored guidance or questions about a specific project, please feel free to contact us for expert advice on your assay development needs. 1. Understanding the Needs of Low-Resource Settings When developing lateral flow assays for low-resource settings like LMICs, the first step is a thorough understanding of the environment where the test will be deployed and who will be using the test. In these regions, infrastructure is often limited. Hospitals or clinics may lack consistent electricity, laboratory space, or trained personnel, making traditional diagnostic technologies commonly used elsewhere, such as PCR or blood culture systems, unfeasible. Environmental factors—such as extreme temperature and humidity—can also significantly impact the performance of an LFA. These conditions can degrade sensitive reagents and materials, leading to inaccurate results. Developers must prioritize the creation of assays that maintain stability across a broad range of temperature and humidity conditions while also often ensuring that the test is simple to use for public health workers or the public with minimal diagnostic training. 2. Reagent and Material Selection: Stability is Key One of the most critical factors in designing LFAs for LMICs is ensuring that reagents remain stable over time and across a wide range of environmental conditions. This typically involves selecting materials and components that can withstand temperature fluctuations and humidity, both during use and throughout storage and transport in low-resource settings. Key Considerations: Humidity Resistance: Humidity can degrade many test components, particularly paper-based materials. Careful formulations and packaging material selection can mitigate this. Incorporating desiccants or multi-layered packaging with moisture-barrier films into the design helps protect the test from environmental moisture, ensuring long-term viability, especially in high-humidity environments. Heat Sensitivity: While most LFAs are designed to avoid cold chain storage, certain reagents, such as antibodies or enzymes, may still be sensitive to heat. Using heat-stable reagent formulations can mitigate this risk, ensuring test stability even when exposed to high ambient temperatures, which can regularly exceed 30°C (86°F) in LMICs. Shelf Life: In remote regions, tests must usually have a long shelf life to reduce the logistical burden of frequent resupply. Ensuring that the assay has a shelf life of 12-24 months or longer is often ideal, especially for diagnostics intended for stockpiling or gradual deployment over time. 3. Sensitivity and Specificity: Striking the Right Balance For developers, balancing sensitivity and specificity is a critical challenge, especially in LMICs where false positives or false negatives can have significant public health implications. For example, during the 2014-2016 Ebola outbreak in West Africa, LFAs were rapidly deployed to provide quick and accurate diagnoses in the field, and were especially useful for “‘rule in’ tests to expedite treatment and vaccination.” In such high-stakes scenarios, misdiagnoses could lead to missed cases, causing further spread, or to unnecessary treatments, quarantines or concerns within communities. LFAs need to perform reliably in environments with high disease prevalence but limited diagnostic resources, where a misdiagnosis can have serious consequences. Key Challenges: Maintaining High Sensitivity: Sensitivity measures how well the test identifies true positives. In resource-limited settings, LFAs often target diseases with high prevalence, such as malaria, HIV, or tuberculosis. Ensuring that these tests can reliably detect low pathogen loads or early-stage infections is crucial. Developers need to choose highly sensitive antibodies or binders and optimize the assay’s detection limit to capture these cases without requiring complex instrumentation. However: Tradeoffs: Developers must also consider trade-offs between sensitivity and specificity. Improving sensitivity might reduce specificity, leading to more false positives, which can strain resources by requiring unnecessary treatments. Ensuring Specificity: Specificity measures how well the test identifies true negatives, avoiding false positives that could lead to inappropriate treatments. In LMICs, where access to confirmatory tests is limited, LFA developers must focus on designing assays that minimize cross-reactivity with similar pathogens while ensuring accuracy in the intended disease. Field conditions, sample quality, and handling variability in LMICs can impact test performance, making it critical for developers to design POCTs that remain robust despite these challenges. Strategies for Developers: Improved Antibody Selection: Developers should prioritize high-affinity antibodies or alternative binding molecules that demonstrate minimal cross-reactivity, ensuring that the assay can distinguish between similar pathogens. Optimization of the Detection System: Signal optimization techniques or multi-analyte testing can be employed to ensure that the assay performs well even at low analyte concentrations, while minimizing the risk of false positives. 4. Simplifying the User Experience: Designing for Non-Expert Users In LMICs, often the individuals administering lateral flow tests may not have a formal medical training, and they might be triaging large numbers of patients at a time. As such, lateral flow assays for low-resource settings must usually be designed with simplicity and usability in mind. The goal is to minimize the risk of user error while still delivering accurate results. Best Practices for Usability: Clear, Intuitive Interfaces: Tests should be easy to interpret, ideally with a simple positive/negative readout. Visual clarity is essential, particularly in settings where access to training or guidance materials may be limited. Developers should also consider the diversity in literacy levels and languages across LMICs, making it important to test user interfaces in local contexts to ensure they are universally understood. Minimal Handling Steps: Reducing the number of steps in the assay procedure helps decrease the likelihood of user error. In our experience, designing a test that can be administered in as few as 2-3 steps, with clear instructions, dramatically improves accuracy in the field. Incorporating error-proofing mechanisms, such as visual or audible feedback for improper sample loading, can also further reduce mistakes. Robust Packaging: Assays should be robustly packaged to protect against environmental conditions, such as moisture or dust, without compromising ease of use. Packaging design should also factor in ease of opening and handling for non-expert users, who may be working in stressful, high-volume settings. Training and Education Materials: While LFAs are most often designed for simplicity, providing easy-to-understand educational materials can further minimize user errors. Pictorial instructions, videos, or even smartphone-based tutorials can supplement the use of the test, particularly in regions with low literacy. These materials should be culturally adapted and localized to meet the specific needs of the target population. 5. Regulatory and Field-Use Validation: Ensuring Global Compliance One of the most significant hurdles in bringing a diagnostic test to market in LMICs is ensuring that the assay meets both local and international regulatory standards. Organizations like FIND Dx often work closely with developers to guide products through these regulatory processes, with the WHO prequalification system being a critical benchmark for many LMICs. Key Considerations: WHO Prequalification: Many LMICs look to WHO prequalification as a benchmark for quality, safety, and performance. WHO prequalification allows tests to be procured by international health organizations and governments. Developers must align their design, validation, and production processes with WHO’s stringent criteria to ensure that the LFA can be distributed and used in these regions. WHO prequalification is often a key requirement for tests to be funded or purchased by organizations such as the Global Fund, UNICEF, or Gavi. Local Regulatory Approvals: While WHO prequalification is often essential for international distribution for some specified RDT’s, developers must also navigate the regulatory requirements of individual countries. This often involves adapting test documentation and processes to meet specific national standards, which can vary widely across LMICs. Field Validation: Validating LFAs in the environments where they will be used is critical. Field trials in real-world settings help ensure that the test performs as expected under the varied conditions encountered in LMICs, such as extreme temperatures, humidity, and differing levels of healthcare infrastructure. These trials provide valuable feedback from healthcare workers and local regulators, which can help refine the test and address usability or performance challenges before widespread rollout. By engaging in early field validation and adhering to both international and local regulatory guidelines, developers can streamline the path to market, ensuring that their diagnostics meet the needs of LMIC populations and can be procured and distributed effectively. 6. Balancing Affordability and Performance Cost constraints are a critical concern for diagnostic tests in LMICs, but cutting costs cannot come at the expense of accuracy or reliability. Developers must strike a balance between designing an affordable test and ensuring that it still meets high-performance standards, particularly in terms of sensitivity and specificity. One of the most significant challenges developers face is navigating the trade-offs between cost and expensive reagents like antibodies or antigens to ensure optimum test performance meeting target specifications. Reducing production costs may impact factors such as robustness, durability, or shelf life, all of which are essential for tests used in resource-limited settings. The goal is to find cost-effective reagents without compromising the performance or consistency of the test. While I have caveated this entire article that this is generalized advice, this tradeoff between cost and performance is an especially difficult one because cost issues must be addressed early in the development process. I cannot stress enough the importance of keeping this tradeoff between cost and performance constantly in mind. If you are struggling with this, ask how we can help. Strategies for Cost Optimization: Material and reagent costs: Minimizing materials (size) and reagents (amounts used) without compromising sensitivity can lower the overall cost per test. Careful material selection also allows for cost reductions while maintaining the necessary quality for high performance. Manufacturing Scalability: From the outset, developers should design their LFAs with scalable manufacturing in mind. Automated manufacturing processes can drive down costs significantly, but the test design itself must lend itself to high-throughput production while maintaining consistency in performance across batches. Quality control processes must be integrated to ensure the reliability of the tests, even as production scales up. Sustainability Considerations: In the pursuit of cost savings, sustainability is an increasingly important factor. Using recyclable or biodegradable materials, or designing assays that minimize plastic waste, can contribute to long-term affordability by reducing environmental impact. However, these sustainable materials can be expensive and these initiatives must be balanced with ensuring that the test remains affordable for LMICs. Our Principal Engineer recently released a great video on this topic. 7. Supply Chain and Scalability: Overcoming Logistical Challenges A significant challenge when developing LFAs for LMICs is ensuring that the test can be produced at an appropriate cost effective scale and distributed reliably. These countries often lack the robust supply chain infrastructure found in high-income countries, and delays or shortages can critically impact access to diagnostic tests. Ensuring supply chain flexibility, quality control during scale-up, and efficient distribution are critical to overcoming these barriers. Strategies for Supply Chain Resilience: Local Manufacturing Partnerships: Establishing local production partnerships can help reduce dependency on international supply chains, and potentially lower production costs for some kit components, and shorten delivery times. Local partnerships can also facilitate quicker adaptation to region-specific needs or regulatory changes, making the supply chain more resilient to disruptions. However, key reagents like antibodies and antigens or key materials like nitrocellulose membrane and conjugate pads may only be available via international supply, therefore import and transport costs must be considered as part of any bill of material cost analysis Scalable Manufacturing Processes: Developing manufacturing processes that are easily scalable ensures that once demand for the LFA grows, it can be produced and delivered in large quantities without compromising quality or significantly increasing costs. However, it is crucial to maintain strict quality assurance protocols during scale-up to prevent issues that could arise from changes in production volumes or facilities. Conversely, many RDTs required in LMICs are focused on niche diagnostic needs that are geographically specific and have limited volume requirements. Therefore material and labor cost analysis needs to address low volume low batch size productions runs in addition to scenarios where demand can grow significantly. Supply Chain Flexibility: Developers must design flexible supply chains that can withstand potential disruptions, such as geopolitical tensions, raw material shortages, or transportation delays. Building in multiple logistics options for procurement of the validated raw materials and diversifying manufacturing sites can potentially help safeguard against these risks. Logistics for Hard-to-Reach Areas: If they aren’t already, developers should consider partnering with NGOs, local health ministries, or local distributors that already have distribution networks in place, especially for rural or remote areas where health infrastructure is limited. Leveraging existing logistical frameworks can help ensure that tests reach under-served populations quickly and efficiently. Packaging Solutions: Ensuring that your tests are packaged robustly to handle harsh environmental and transport conditions is critical for successful distribution. Inventory Management and Distribution Strategy: Where reliable infrastructure exists, establishing regional warehouses or inventory management systems can help optimize distribution, reducing the time it takes for tests to reach healthcare providers in these regions. Conclusion: Designing for Impact Developing lateral flow assays for low- and middle-income countries requires a practical, thoughtful approach that balances performance, cost, and usability. By focusing on environmental stability, simplifying the user experience, optimizing for affordability, and ensuring a robust supply chain, developers can create diagnostics that are not only effective but also accessible in resource-limited settings. At DCN Dx, we are committed to advancing global health through the development of high-quality IVDs that meet the unique needs of low-resource environments. By following these practical design strategies—prioritizing bill of material costs, scalability, and usability—developers can make a significant impact in bridging the diagnostic gap in under-served regions, improving healthcare outcomes in LMICs. If you have questions about how to develop lateral flow assays for low-resource settings or need expert guidance on your other IVD CDMO or CRO projects, contact us to learn how we can support you in bringing impactful solutions to these under-served markets. --- ## Advancing Point-of-Care Diagnostics: The miniDxR’s Role in Enabling High-Performance Lateral Flow Assays URL: https://dcndx.com/insights/advancing-point-of-care-diagnostics-the-minidxrs-role-in-enabling-high-performance-lateral-flow-assays/ Type: insight Published: 2024-06-13 Introduction The demand for rapid, accurate diagnostics at the point of care has never been greater, driven by the need for timely and effective healthcare interventions, food safety monitoring, and environmental testing. Lateral flow assays (LFAs) have emerged as invaluable tools for decentralized testing, offering simplicity, speed, and portability. However, achieving high performance in LFAs presents significant challenges, particularly in terms of sensitivity and specificity. Advanced lateral flow readers, like the miniDxR, play a pivotal role in enabling the development and deployment of sensitive and specific LFAs in point-of-care settings. This article highlights some of the miniDxR’s features and performance, demonstrating its significance in advancing both the capabilities of and use cases for lateral flow assays and, ultimately, improving global health outcomes. The miniDxR: Redefining Precision Diagnostics Lateral flow assay analyzers are specialized instruments designed to accurately read and interpret the results of LFAs. These analyzers utilize advanced optical, electronic, or imaging technologies to detect and quantify the signals generated by assays. Unlike manual visual interpretation, which can be subjective and prone to errors—especially when deployed at the point of care—analyzers offer objective, quantitative, semi-quantitative, or qualitative analysis, improving the reliability and reproducibility of test results. The miniDxR is a state-of-the-art, customizable lateral flow assay analyzer designed to deliver accurate and reliable results. Its key features include: Advanced imaging technology: The miniDxR utilizes state-of-the-art camera-based colorimetry and fluorescence for precise color and intensity detection, enabling the most sensitive diagnostics. User-friendly design: With an intuitive interface, vibrant color display, and straightforward three-button controls, the miniDxR ensures effortless operation even in fast-paced point-of-care environments. Secure and compliant: The miniDxR safeguards sensitive data with robust cybersecurity and adheres to rigorous international standards, making it suitable for use in various settings. Adaptable to diverse assays: The miniDxR’s modular cartridge nest can be customized to accommodate a wide variety of lateral flow assay cassettes, ensuring compatibility with specific testing needs. Flexible testing modes: The miniDxR offers versatile “batch” and “single test” modes to accommodate diverse testing workflows, allowing for the analysis of pre-run assays or real-time monitoring of LFAs at the point of care. These key features make the miniDxR a versatile and high-performance diagnostic tool. At the heart of the miniDxR’s low-noise design is a sensitive full-color 10-bit CMOS image sensor, which enables precise color and intensity detection. The system utilizes onboard LEDs to illuminate or excite either visual or fluorescent labels, with customizable illumination wavelengths and emission filters available to optimize performance for specific assays. These advanced imaging technologies enhance the miniDxR’s sensitivity and specificity, allowing for the differentiation of subtle variations. The miniDxR’s modular nest design is another key feature that sets it apart from many other lateral flow readers. This adaptable design allows for customization to accommodate different cassette geometries, aligning with DCN Dx’s philosophy of designing the cassette to best suit the assay rather than the other way around. This flexibility enables the miniDxR to seamlessly integrate with a wide range of assays, making it a versatile reader for various applications, including clinical labs, point-of-care, home use, and all LFA development programs. The miniDxR performs swift and accurate readings, crucial for time-sensitive point-of-care settings. Its intuitive interface makes it accessible to both healthcare professionals and other users of varying expertise levels. With built-in connectivity features, the miniDxR is capable of automatically relaying results to a database, such as a lab information system, enhancing workflow efficiency, and may also be configured to send results to a remote cloud database (e.g. through a custom mobile device app). Performance Data and Benchmarking Study Overall system performance serves as a critical metric for evaluating the efficacy of lateral flow readers in real-world settings. To evaluate the miniDxR’s performance, a comparative study was conducted against another—more costly—commercially available analyzer. The study aimed to assess the miniDxR’s capabilities in both visual and fluorescence modes, using a range of analyte concentrations and multiple test cassettes to ensure robust and reliable results. In this study, the key metrics of variability and sensitivity were used to evaluate the performance of the miniDxR and the other commonly used commercially available analyzer. Each of these metrics plays a crucial role in determining the overall performance and reliability of a lateral flow analyzer. Variability: Variability, expressed as the coefficient of variation (CV), measures the consistency and reproducibility of the analyzer’s results. A lower CV indicates that the analyzer produces more precise and consistent measurements across multiple tests. In the study, the miniDxRs demonstrated lower CVs compared to the other analyzers, showcasing its superior precision and reliability. Sensitivity: Sensitivity refers to the analyzer’s ability to detect low levels of the target analyte in a sample. A highly sensitive analyzer can accurately identify the presence of an analyte even at very low concentrations. The miniDxR exhibited improved sensitivity compared to the other analyzers, enabling the detection of lower analyte concentrations (ultimately allowing a lower limit of detection for a particular assay) and potentially leading to earlier disease detection or more accurate monitoring of treatment response. These metrics collectively contribute to the overall performance of a lateral flow reader. An analyzer such as the miniDxR, with low variability and high sensitivity allowing a lower limit of detection, can provide more accurate, consistent, and reliable results across a wide range of analyte concentrations. This improved performance ultimately leads to better diagnostic outcomes by enabling healthcare professionals and other users to make more informed decisions. Visual Mode Comparison In visual mode, 8 miniDxR analyzers and 3 commercially available analyzers manufactured by another company were compared using five concentrations of dried hCG tests. For each concentration, 4 test cassettes were used, with 5 repetitions per cassette. Additionally, a printed “external QC” (EQC) cassette was tested 20 times before and after testing the set of dried hCG test cassettes on each analyzer. The study design ensured that the exact same QC and hCG cassettes were measured on each analyzer, allowing for a direct comparison of performance. The results demonstrated the miniDxR’s superior performance in terms of reduced variability and improved sensitivity. The miniDxR’s coefficients of variation were consistently lower across all concentrations tested, indicating higher precision and reproducibility. Furthermore, the miniDxR exhibited increased sensitivity, enabling the accurate identification of lower analyte concentrations. Table 1: Visual Mode Comparison Study Data *n = 20 **n = 5 Note that 0uM tests were run with antigen-free sample. CVs at 0uM appear high because the average values are low. Table 2: Visual Comparison Study Data: 8 miniDxR Analyzers, All Measurements at Each Concentration Table 3: Visual Comparison Study Data: 3 Other Analyzers, All Measurements at Each Concentration Figure 1: Visual mode comparison between the miniDxR and another commercially available analyzer, demonstrating strong overall correlation between the analyzers. Note that the EQC data is not shown. Fluorescence Mode Comparison In fluorescence mode, the miniDxR was compared against another third party reader using three concentrations of dried europium tests. One test cassette at each concentration, with 10 repetitions of each cassette. The same cassettes were measured with each analyzer. All 10 measurements were captured for a given cassette on the third-party analyzer before capturing all 10 measurements on the miniDxR. Baseline values were subtracted from each measurement. The miniDxR demonstrated excellent performance, with the ability to tune excitation and integration time for minimal photo-bleaching while maintaining high sensitivity (Figure 2). Table 4: Fluorescence Mode Comparison Study Data, Test 1 Note that the Negative QC Sample CVs are invalid because the averages are 0. Figure 2: Fluorescence mode comparison between the miniDxR and another commercially available analyzer, showcasing the miniDxR’s excellent measurement repeatability and minimal photo-bleaching compared to the other analyzer. Also in fluorescence mode, one miniDxR and one other commercially available analyzer were compared. Three concentrations of dried europium tests were measured, 5 test cassettes at each concentration, with 2 repetitions of each cassette. The exact same cassettes were measured with each analyzer. Baseline values were subtracted from each measurement. The read order for each strip was Other-DCN-Other-DCN. Table 5: Fluorescence Mode Comparison Study Data, Test 2 Note that the Negative QC Sample CVs are invalid because the averages are 0. Also note that these CVs are primarily influenced by assay variability. Figure 3: Fluorescence mode comparison between the miniDxR and another commercially available analyzer, demonstrating a strong overall correlation between the analyzers. Enabling High-Performance LFAs in Point-of-Care Settings The miniDxR’s advanced features were designed to play a crucial role in enabling the deployment of high-performance LFAs at the point of care. The miniDxR uses workflow and assay configuration files that allow for significant customization and adaptability. The platform’s ability to aid in operator compliance with test workflow, through features such as on-screen instructions, sample/buffer flow detection, and automatic countdown timers, further contributes to improved diagnostic accuracy and confidence. Overall, by facilitating sensitive and specific detection of target analytes, the miniDxR enhances diagnostic accuracy, confidence, and objectivity, ultimately improving patient outcomes. The miniDxR’s advanced features not only benefit end-users in point-of-care settings but also play a crucial role in enabling the development of high-performance LFAs. Assay developers rely on accurate, sensitive, and reliable analyzers to optimize their assays and ensure consistent performance. When assay developers use a robust analyzer like the miniDxR, they can objectively and quantitatively assess the impact of changes in assay parameters, such as label type, chemistry, membrane, or pad materials, on the assay’s performance. This allows developers to make informed decisions during the optimization process and ultimately leads to the creation of high-quality, reliable assays. The miniDxR’s customizable workflow and assay configuration files provide assay developers with the flexibility to tailor the analyzer’s settings to their specific needs. This level of customization enables developers to fine-tune the analyzer’s performance to match the unique requirements of their assays, ensuring optimal results and a seamless integration into the intended setting. The synergy between in-house assay and reader development at DCN Dx is another key factor in enabling high-performance LFAs. Assay developers can request specific features, unique analysis methods, and provide feedback for improvements, while engineers can quickly respond and implement updates. This collaborative approach fosters rapid innovation and customization, ensuring that the miniDxR is optimized for each specific assay. Conclusion Advanced lateral flow readers like the miniDxR have the potential to revolutionize point-of-care diagnostics, especially in resource-limited settings, thereby improving healthcare accessibility and outcomes worldwide. DCN Dx remains committed to driving innovation and collaboration in the development and deployment of high-performance LFAs. The miniDxR exemplifies DCN’s dedication to advancing point-of-care diagnostics, making precise and reliable testing accessible to all, regardless of location or resources. As the demand for accurate and timely diagnostic solutions continues to grow, the miniDxR will play an important role in global health. --- ## Raising the Bar in Veterinary Care: InPouch TF Bovine and TF Transit Tube as Gold Standards in Tritrichomonas foetus Detection URL: https://dcndx.com/insights/tf-bovine-transit-tube-tritrichomonas-foetus/ Type: insight Published: 2024-05-15 Introduction Bovine Trichomoniasis, a sexually transmitted disease caused by the protozoan parasite Tritrichomonas foetus, is a silent but devastating threat to the cattle industry, particularly in beef herds. This insidious disease can spread quickly through a herd, leading to early embryonic death, abortion, and infertility in cows (Ondrak, 2018). The resulting reproductive failures and decreased calving rates can have a profound impact on the productivity and profitability of affected herds. The economic consequences of Bovine Trichomoniasis are staggering, with annual losses estimated to reach hundreds of millions of dollars in the United States alone (Yao, 2013). These losses stem not only from reduced calf crops but also from the costs associated with diagnostic testing, treatment, and replacement of infected animals (Michi et al., 2016). For beef producers, the financial burden of this disease can be catastrophic, threatening the viability of their operations. As a veterinarian serving beef clients or as a beef producer, protecting herds against T. foetus is crucial to maintaining reproductive health, maximizing productivity, and ensuring the long-term sustainability of the operation. Effective management of Bovine Trichomoniasis relies on accurate and timely diagnosis, which allows for the identification and removal of infected animals, as well as the implementation of appropriate control measures to prevent the spread of the disease (Yao & Köster, 2015). In this article, we will explore the challenges associated with diagnosing Bovine Trichomoniasis and discuss how InPouch TF Bovine and TF Transit Tube, innovative diagnostic tools developed by Biomed Diagnostics, revolutionized the way veterinarians and producers detect and manage this costly disease. By providing a comprehensive solution for T. foetus diagnosis, these tools still empower veterinarians and producers to make informed decisions, protect herd health, and safeguard the profitability of beef operations. The Challenge of Diagnosing Bovine Trichomoniasis Accurate diagnosis of Bovine Trichomoniasis is crucial for effective disease management and control. However, traditional diagnostic methods have several limitations that can hinder the timely and precise identification of infected animals. These limitations include the potential for inaccurate results, missed positive cases, and time-consuming or expensive processes that often lead to further increased costs due to the need for retesting (Ondrak, 2016). One of the primary challenges in diagnosing Bovine Trichomoniasis is the difficulty in detecting the presence of Tritrichomonas foetus, the causative agent of the disease (Felleisen, 1999). This protozoan parasite resides in the reproductive tract of infected cattle, making it challenging to collect and identify through conventional methods. Additionally, the similarity of T. foetus to other trichomonad species further complicates the diagnostic process, requiring highly specific and sensitive tools to differentiate between them (Campero et al., 2003). While PCR testing is a highly sensitive and specific method for detecting T. foetus, it is not widely available in all regions, including parts of the United States and many low and middle-income countries (Yao, 2015). The cost of PCR testing can also be prohibitive for some farmers and veterinarians, making it difficult to implement as a routine diagnostic tool (Madoroba et al., 2011). Furthermore, the need for specialized equipment and trained personnel to perform PCR testing can limit its accessibility, particularly in remote or resource-limited settings (Bondurant, 2005). The limitations of other diagnostic methods and the challenges associated with PCR testing underscore the importance of having reliable, accurate, and accessible diagnostic tools that can streamline the identification process and provide conclusive results. Veterinarians and farmers need solutions that can help them promptly detect and manage Bovine Trichomoniasis to minimize its impact on their herds and operations, regardless of their location or financial resources. InPouch TF Bovine and TF Transit Tube: A Comprehensive Solution Biomed Diagnostics, a leading provider of innovative diagnostic solutions, offers a comprehensive approach to tackling the challenges associated with Bovine Trichomoniasis detection. The company’s flagship products, InPouch TF Bovine and TF Transit Tube, provide a selective media specifically designed for detecting Tritrichomonas foetus, offering several advantages over conventional PBS-based sample transport methods. The TF Bovine media used in these products enhances PCR accuracy by promoting the growth of T. foetus flagellates (Borchardt et al., 1992). This selective growth enables the detection of the parasite even in low concentrations, reducing the risk of false-negative results. The highly specific nature of the media ensures that only T. foetus is detected, minimizing the chances of cross-reactivity with other trichomonad species and providing highly accurate diagnoses (Chen & Li, 2007). One of the key advantages of InPouch TF Bovine and TF Transit Tube is their ease of use. The products are designed for simple inoculation at the point of care, allowing veterinarians and field personnel to collect and secure samples quickly and efficiently. InPouch TF Bovine, in particular, offers a unique advantage by combining culture, result, and microscopic observation in a single in vitro diagnostic (IVD) device (Mukhufhi et al., 2003). This all-in-one design reduces the potential for contamination and streamlines the diagnostic workflow, saving time and resources. The InPouch format also allows for direct microscopic examination of the sample, providing a backup method of detection for those relying on PCR testing. Enhancing Veterinary Practice with InPouch TF Bovine and TF Transit Tube For bovine veterinarians, InPouch TF Bovine and TF Transit Tube offer several benefits that streamline diagnostic processes and improve herd health management. The ease of use and portability of these tools enable veterinarians to collect and transport samples efficiently, even in remote field settings. The ability to inoculate the media at the point of care, as well as the specific growth media, minimizes the risk of sample contamination and ensures the viability of T. foetus for accurate diagnosis. The cost-effectiveness of these products also makes them an attractive choice for veterinarians and their clients. By providing timely and reliable results, InPouch TF Bovine and TF Transit Tube reduce the need for repeated testing and minimize the costs associated with misdiagnosis or delayed treatment. The ability to detect T. foetus infections early allows for prompt intervention, ultimately saving farmers from the substantial economic losses associated with Bovine Trichomoniasis. Moreover, the use of these diagnostic tools empowers veterinarians to make informed decisions and implement targeted herd health strategies. The accurate identification of infected animals enables veterinarians to recommend appropriate treatment, culling, or quarantine measures to their clients. By effectively controlling the spread of Bovine Trichomoniasis, veterinarians can help farmers maintain the reproductive health and productivity of their herds, thereby strengthening the veterinarian-client relationship and demonstrating their value as trusted advisors. The Gold Standard in Bovine Trichomoniasis Detection InPouch TF Bovine and TF Transit Tube have earned their reputation as the gold standard in Bovine Trichomoniasis detection due to their exceptional performance and reliability. These products have been extensively validated and widely adopted by veterinary professionals and diagnostic laboratories worldwide (Effinger et al., 2014). Studies have consistently demonstrated the superior sensitivity and specificity of InPouch TF Bovine and TF Transit Tube compared to other diagnostic methods (Corbeil et al., 2005). The selective media used in these products enables the detection of T. foetus even in the presence of other microorganisms, reducing the risk of false-positive results. The ability to provide rapid presumptive diagnoses and the option for microscopic confirmation further enhance their diagnostic value. The effectiveness of InPouch TF Bovine and TF Transit Tube has been showcased in numerous real-world applications and case studies. Veterinarians and farmers have reported significant improvements in their ability to detect and manage Bovine Trichomoniasis since adopting these products (Michi et al., 2016). The accurate and timely diagnoses provided by these tools have helped in implementing targeted treatment and control measures, minimizing the spread of the disease within herds. Practical Applications for Bovine Veterinarians and Farmers Implementing InPouch TF Bovine and TF Transit Tube in your diagnostic workflow can significantly enhance your ability to detect and manage Bovine Trichomoniasis effectively. By incorporating these tools into your routine herd health checks and screening protocols, you can identify infected animals early and take appropriate measures to prevent the spread of the disease. For bovine veterinarians, the reliable diagnosis provided by these products enables you to offer timely treatment and management recommendations to your clients. By educating farmers about the importance of regular testing and the benefits of using InPouch TF Bovine and TF Transit Tube, you can help them protect their herds and minimize the economic impact of Bovine Trichomoniasis. Farmers can proactively use these diagnostics to screen new animals before introducing them into their herds, preventing the introduction of T. foetus and safeguarding the reproductive health of their cattle. Regular testing of breeding bulls and cows can help identify infected individuals, allowing for prompt treatment or culling decisions to control the spread of the disease. Implementing biosecurity measures, such as maintaining a closed herd, using artificial insemination, and quarantining new animals, along with regular testing using InPouch TF Bovine and TF Transit Tube, can create a comprehensive strategy for preventing and controlling Bovine Trichomoniasis in your herd. Specimen Collection and Storage Proper specimen collection and storage are critical for reliable and repeatable (if needed) results. InPouch TF Bovine and TF Transit Tube are designed to simplify and standardize the sample collection process. Veterinarians and field personnel can collect vaginal or distal penile swab specimens using these devices, following a straightforward and user-friendly protocol (Thomas et al., 1991). The products offer safe and reliable storage, with a shelf life of 12 months from the date of manufacture when stored under the recommended conditions. This extended stability ensures that the samples remain viable and suitable for testing even if there are delays in transportation or processing (Cobo et al., 2007). Beyond Bovine: InPouch TF Feline While InPouch TF Bovine and TF Transit Tube are specifically designed for detecting T. foetus in cattle, Biomed Diagnostics offers a range of culture diagnostics for various veterinary needs. The company’s product portfolio includes InPouch TF Feline, a specialized media for diagnosing trichomoniasis in cats (Gookin et al., 2001). Feline trichomoniasis, caused by the protozoan parasite Tritrichomonas foetus, is a significant cause of chronic diarrhea in cats, particularly in young, densely housed feline populations (Yao & Köster, 2015). Accurate diagnosis of feline trichomoniasis is essential for providing appropriate treatment and preventing the spread of the disease in multi-cat households and shelters. InPouch TF Feline offers a reliable and convenient method for diagnosing T. foetus infections in cats. The product’s selective media supports the growth of T. foetus while inhibiting the growth of other fecal microorganisms, enhancing the accuracy of the diagnosis (Gookin et al., 2003). As with the InPouch TF Bovine system, InPouch TF Feline also allows for direct microscopic examination of the sample, enabling veterinarians to identify the presence of T. foetus trophozoites quickly and easily (Ceplecha et al., 2013). The Enduring Relevance of InPouch TF Bovine in the Era of Molecular Diagnostics As molecular technologies like PCR become more affordable and accessible, some may question the continued relevance of culture-based diagnostics like InPouch TF Bovine. However, it is important to recognize that these tools are not mutually exclusive; in practice, they work synergistically to provide the most comprehensive and reliable solutions for veterinarians and farmers detecting and managing Bovine Trichomoniasis. Far from being replaced by molecular methods, InPouch TF Bovine is often used in the field to enhance these technologies. By culturing and concentrating T. foetus prior to PCR testing, InPouch TF Bovine can increase the sensitivity of molecular assays and reduce false negatives. This powerful combination of culture and PCR enables veterinarians and diagnostic laboratories to deliver the most accurate and comprehensive results to inform effective herd management strategies. In some testing regimes, it is also used as a confirmatory tool before making culling decisions. Beyond its applications in the field, InPouch TF Bovine is also used as a tool for advancing research and development efforts in the fight against Bovine Trichomoniasis. Its ability to maintain the viability of T. foetus during transport and storage creates opportunities for collaborative studies and the sharing of specimens between institutions. As researchers work to develop more effective vaccines and control strategies, InPouch TF Bovine will continue to play a vital role in facilitating these efforts and accelerating progress towards the ultimate goal of eradicating this destructive disease. Conclusion Bovine Trichomoniasis poses a significant threat to the cattle industry, emphasizing the importance of accurate and efficient diagnostics in protecting herds and preventing economic losses. InPouch TF Bovine and TF Transit Tube by Biomed Diagnostics have long been considered the gold standard in Bovine Trichomoniasis detection, offering veterinarians and farmers a comprehensive solution for identifying and managing this devastating disease. The selective media, ease of use, and compatibility with various testing methods make InPouch TF Bovine and TF Transit Tube indispensable tools in the fight against Bovine Trichomoniasis. By providing rapid presumptive diagnoses and enabling accurate detection of T. foetus, these products support informed decision-making and timely intervention strategies. As a bovine veterinarian or farmer, incorporating InPouch TF Bovine and TF Transit Tube into your diagnostic protocols and herd management strategies can significantly enhance your ability to detect, control, and prevent Bovine Trichomoniasis. By adopting these reliable and accurate diagnostic tools, you can take proactive steps to protect your herd’s reproductive health, minimize economic losses, and ensure the long-term success of your operation. Explore our website to learn more about InPouch TF Bovine, TF Transit Tube, TF Feline, TG/G, and other culture diagnostic solutions designed to help you protect your herd against veterinary challenges. References Bondurant, R. H. (2005). Venereal diseases of cattle: natural history, diagnosis, and the role of vaccines in their control. Veterinary Clinics: Food Animal Practice, 21(2), 383-408. https://www.researchgate.net/publication/7788250_Venereal_Diseases_of_Cattle_Natural_History_Diagnosis_and_the_Role_of_Vaccines_in_their_Control Borchardt, K. A., Norman, B. B., Thomas, M. W., & Harmon, W. M. (1992). Evaluation of a new culture method for diagnosing Tritrichomonas foetus infection. Veterinary Medicine, 87(1), 104-112. https://www.cabidigitallibrary.org/doi/full/10.5555/19922269245 Campero, C. M., Rodriguez Dubra, C., Bolondi, A., Cacciato, C., Cobo, E., Perez, S., … & BonDurant, R. H. (2003). Two-step (culture and PCR) diagnostic approach for differentiation of non- T. foetus trichomonads from genitalia of virgin beef bulls in Argentina. Veterinary Parasitology, 112(3), 167-175. https://pubmed.ncbi.nlm.nih.gov/12591192/ Chen, X. S., & Li, J. J. (2007). Increasing the sensitivity of PCR detection in bovine preputial smegma spiked with Tritrichomonas foetus by the addition of agar and resin. Parasitology Research, 101(5), 1407-1410. https://pubmed.ncbi.nlm.nih.gov/11484853/ Cobo, E. R., Favetto, P. H., Lane, V. M., Friend, A., VanHooser, K., Mitchell, J., & BonDurant, R. H. (2007). Sensitivity and specificity of culture and PCR of smegma samples of bulls experimentally infected with Tritrichomonas foetus. Theriogenology, 68(6), 853-860. https://pubmed.ncbi.nlm.nih.gov/17681370/ Corbeil, L. B., Campero, C. M., Rhyan, J. C., Anderson, M. L., Gershwin, L. J., Agnew, D. W., … & BonDurant, R. H. (2005). Uterine mast cells and immunoglobulin-E antibody responses during clearance of Tritrichomonas foetus. Veterinary Pathology, 42(3), 282-290. https://pubmed.ncbi.nlm.nih.gov/15872374/ Effinger, L., Peddireddi, L., & Simmons, K. (2014). Pooling of cultured samples and comparison of multistate laboratory workflows with the MagMAX™ Pathogen RNA/DNA Kit and VetMAX™ Gold Trich Detection Kit for detection of Tritrichomonas foetus -colonized bulls. Journal of Veterinary Diagnostic Investigation, 26(1), 72-87. https://pubmed.ncbi.nlm.nih.gov/24343558/ Felleisen, R. S. (1999). Comparative sequence analysis of 5.8S rRNA genes and internal transcribed spacer (ITS) regions of trichomonadid protozoa. Parasitology, 119(2), 111-119. https://www.cambridge.org/core/journals/parasitology/article/abs/comparative-sequence-analysis-of-58s-rrna-genes-and-internal-transcribed-spacer-its-regions-of-trichomonadid-protozoa/8EC3559DCD4BD706245095AD490FF5E1 Gookin, J. L., Levy, M. G., Law, J. M., Papich, M. G., Poore, M. F., & Breitschwerdt, E. B. (2001). Experimental infection of cats with Tritrichomonas foetus. American Journal of Veterinary Research, 62(11), 1690-1697. https://pubmed.ncbi.nlm.nih.gov/11703009/ Madoroba, E., Gelaw, A., Hlokwe, T., & Mnisi, M. (2011). Prevalence of Campylobacter foetus and Tritrichomonas foetus among cattle from Southern Africa. African Journal of Biotechnology, 10(50), 10311-10314. https://www.researchgate.net/publication/259191981_Prevalence_of_Campylobacter_foetus_and_Trichomonas_foetus_among_cattle_from_Southern_Africa Michi, A. N., Favetto, P. H., Kastelic, J., & Cobo, E. R. (2016). A review of sexually transmitted bovine trichomoniasis and campylobacteriosis affecting cattle reproductive health. Theriogenology, 85(5), 781-791. https://pubmed.ncbi.nlm.nih.gov/26679515/ Mukhufhi, N., Irons, P. C., Michel, A., & Peta, F. (2003). Evaluation of a PCR test for the diagnosis of Tritrichomonas foetus infection in bulls: effects of sample collection method, storage and transport medium on the test. Theriogenology, 60(7), 1269-1278. https://pubmed.ncbi.nlm.nih.gov/14511781/ Ondrak, J. D. (2016). Tritrichomonas foetus prevention and control in cattle. Veterinary Clinics: Food Animal Practice, 32(2), 411-423. https://pubmed.ncbi.nlm.nih.gov/27039692/ Parker, S., Lun, Z. R., & Gajadhar, A. (2001). Application of a PCR assay to enhance the detection and identification of Tritrichomonas foetus in cultured preputial samples. Journal of Veterinary Diagnostic Investigation, 13(6), 508-513. https://pubmed.ncbi.nlm.nih.gov/11724142/ Thomas, M. W., Harmon, W. M., & White, C. (1991). An improved method for the detection of Tritrichomonas foetus infection by culture in bulls. Agri-Practice, 11, 13-17. Yao, C. (2013). Diagnosis of Tritrichomonas foetus -infected bulls, an ultimate approach to eradicate bovine trichomoniasis in US cattle? Journal of Medical Microbiology, 62(Pt 1), 1–9. https://pubmed.ncbi.nlm.nih.gov/23082032/ --- ## New Podcast Episode—The Enduring Impact of Culture Diagnostics and LFAs in STI Detection and Public Health URL: https://dcndx.com/insights/culture-diagnostics-lfa-sti-detection-public-health/ Type: insight Published: 2024-04-23 Insights from DCN Dx and Biomed Diagnostics for National Public Health Week In honor of National Public Health Week, we invite you to join us for an insightful audio discussion that explores the critical role of collaborations between public health labs and diagnostic developers in combating the rising tide of sexually transmitted infections (STIs). Why Listen? Gain a deeper understanding of the current landscape of STI detection and the unique contributions of culture diagnostics and lateral flow tests Learn about emerging technologies that are poised to revolutionize STI detection and management Explore best practices for designing impactful clinical studies that demonstrate real-world impact on STI detection and control efforts Discover how strengthened collaborations among key players can drive advancements in public health outcomes Featured Panelists Pat Vaughan, Ph.D., Chief Operating Officer at DCN Dx Sarah Barchard, Senior Clinical Trials Manager at DCN Dx Brandon Font, Research Scientist and Technical Services Manager at Biomed Diagnostics Moderators Mitzi Rettinger, former Chief Revenue Officer at DCN Dx Shubhanga Scott, Account Manager and former Research Scientist at DCN Dx --- ## Innovative Solutions for Accurate and Efficient Culture Detection of Trichomonas vaginalis and Neisseria gonorrhoeae Infections URL: https://dcndx.com/insights/innovative-solutions-for-accurate-and-efficient-culture-detection-of-trichomonas-vaginalis-and-neisseria-gonorrhoeae-infections/ Type: insight Published: 2024-04-23 Introduction Sexually transmitted infections (STIs) are a significant global health burden, with an estimated 1 million new cases occurring daily worldwide (WHO, 2022). Untreated STIs can lead to severe complications such as pelvic inflammatory disease, infertility, and increased risk of HIV acquisition (Rowley et al., 2019). The increase of STI cases in recent years has highlighted the urgent need for improved diagnostic tools that accurately and efficiently detect these infections in a wide variety of clinical settings, enabling timely treatment and prevention of further transmission. Among the most prevalent STIs are trichomoniasis, caused by the protozoan Trichomonas vaginalis, and gonorrhea, caused by the gram-negative gonococcus bacterium Neisseria gonorrhoeae, with global annual incidence rates of 156 million and 87 million cases, respectively (WHO, 2018). T. vaginalis, a parasitic protozoan, is associated with adverse pregnancy outcomes, increased risk of HIV acquisition, and pelvic inflammatory disease (Kissinger, 2015 ). N. gonorrhoeae, a bacterial pathogen, can lead to serious complications such as disseminated gonococcal infections prone to antibiotic treatment failure, making prompt detection and accurate treatment crucial (Wi et al., 2017). In this article, we aim to highlight the significant contributions of Biomed Diagnostics’ InPouch TV and InTray GC devices in revolutionizing the detection and management of T. vaginalis and N. gonorrhoeae infections. By examining the unique features, performance characteristics, and potential impact of these devices, we seek to highlight their role in strengthening the diagnostic tool kit against STIs and informing public health strategies to curb the rising tide of these infections. Innovative Diagnostic Solutions: InPouch TV and InTray GC InPouch TV and InTray GC, developed by Biomed Diagnostics, are innovative diagnostic devices that address the limitations of traditional methods for detecting T. vaginalis and N. gonorrhoeae, respectively. These devices combine efficient sample collection, sample transport, and optimal microenvironment culturing within a single sealed system, offering significant advantages in terms of performance, ease of use, and cost-effectiveness. The InPouch TV is a self-contained culture system that utilizes a selective, modified Diamond’s medium to support the growth of T. vaginalis while suppressing the proliferation of other microorganisms. The device contains nutrients, antibiotics, and oxygen-resistant barrier materials that create an optimal microenvironment for T. vaginalis growth. The device is inoculated with the patient sample, incubated at 37°C, and examined microscopically daily for direct evidence of live trichomonads as soon as parasite cell growth occurs (Borchardt et al., 1997). Similarly, the InTray GC employs a modified Thayer-Martin medium, which is selective for N. gonorrhoeae growth while inhibiting the growth of contaminants (Paris & Font, 2022). The InTray GC device contains antimicrobial agents, selective nutrients, and a tablet that releases CO2 gas in order to facilitate optimal growth and identification of N. gonorrhoeae colonies. After inoculation with the patient sample, the InTray GC is incubated at 35-37°C in a device generated 5% CO2 microenvironment for up to 72 hours (i.e., no need for CO2 incubator because the CO 2 pellet is provided within the tray), allowing for the presumptive identification of N. gonorrhoeae based on colony morphology. Validation studies and clinical trials have demonstrated the high accuracy and reliability of InPouch TV and InTray GC in detecting T. vaginalis and N. gonorrhoeae, respectively. In a multi-center study comparing InPouch TV with wet mount microscopy and culture, InPouch TV exhibited a sensitivity of 94.7% and a specificity of 99.4% (Borchardt et al., 1997). Similarly, a study evaluating the performance of InTray GC found a sensitivity of 100% and a specificity of 98.8% compared to standard culture methods. Compared to traditional diagnostic methods, InPouch TV and InTray GC offer several advantages. Wet mount microscopy, the most common method for T. vaginalis detection, has a low sensitivity of 50-60%, often leading to false-negative results (Bachmann et al., 2011). InPouch TV, with its higher sensitivity and specificity, reduces the risk of missed infections and enables more accurate diagnosis. Similarly, conventional culture methods for N. gonorrhoeae require separate collection, transport, and culture steps, increasing the risk of sample contamination and delay in results (Gaydos & Hardick, 2014). InTray GC streamlines this process, providing a more efficient and reliable alternative. Supporting Antimicrobial Resistance Surveillance and Targeted Treatment Validation studies have also demonstrated the reliability of InPouch TV and InTray GC in transporting viable T. vaginalis and N. gonorrhoeae cells, respectively, to labs responsible for antibiotic susceptibility testing (Paris & Font, 2022). This is a crucial aspect of STI management, as the emergence of antimicrobial resistance, particularly in N. gonorrhoeae, poses a significant threat to public health (Wi et al., 2017). The ability to transport viable organisms to reference laboratories for susceptibility testing is essential for monitoring resistance patterns, informing treatment guidelines, and guiding the development of new antibiotics (Cristillo et al., 2017). The study by Paris and Font (2022) evaluated the recovery of antimicrobial-resistant N. gonorrhoeae isolates after 72 hours of transport using the InTray GC method. The results showed that the InTray GC device successfully maintained the viability of the isolates, allowing for accurate susceptibility testing at the receiving laboratory. This finding highlights the potential of the InTray GC device to support surveillance efforts and facilitate the timely detection of antimicrobial resistance in N. gonorrhoeae (FDA 510k K210511 InTray GC). Similarly, the InPouch TV device has been shown to maintain the viability of T. vaginalis cells during transport, enabling the successful culture and susceptibility testing of the organism at reference laboratories (Bachmann et al., 2011). This is particularly important given the increasing reports of metronidazole-resistant T. vaginalis strains, which require alternative treatment approaches (Kissinger, 2015). The reliability of InPouch TV and InTray GC in transporting viable organisms for susceptibility testing highlights their value as comprehensive diagnostic solutions. By combining accurate detection with the ability to support antimicrobial resistance monitoring, these devices can contribute to the development of targeted treatment strategies and the containment of resistant STI strains. The integration of InPouch TV and InTray GC into STI management protocols can strengthen the overall response to the global challenge of antimicrobial resistance in STIs. Impact on Public Health Outcomes The widespread adoption of InPouch TV and InTray GC has the potential to significantly improve public health outcomes related to T. vaginalis and N. gonorrhoeae infections. By enabling accurate and efficient culture diagnosis, these devices can facilitate prompt treatment, reduce the duration of infectiousness, and prevent the onward transmission of these STIs (Kissinger, 2015; Wi et al., 2017). Delayed or missed diagnoses of T. vaginalis and N. gonorrhoeae can lead to persistent infections, increasing the risk of complications and the likelihood of transmission to sexual partners (Gaydos & Hardick, 2014). The high sensitivity and specificity of InPouch TV and InTray GC can help mitigate these risks by ensuring that infected individuals are identified and treated promptly. Early detection and treatment also reduce the risk of adverse outcomes such as pelvic inflammatory disease, ectopic pregnancy, and infertility (Rowley et al., 2019). Moreover, the cost-effectiveness of InPouch TV and InTray GC can have significant implications for healthcare systems. By reducing the need for repeat testing and follow-up visits due to false-negative results, these devices can help optimize resource utilization and reduce the overall economic burden associated with STI management (Gift et al., 2002). The simplified testing process and ease of use also make these devices suitable for use in resource-limited settings, where access to advanced diagnostic facilities may be limited (Cristillo et al., 2017). Contribution to Global Health Targets The adoption of InPouch TV and InTray GC can contribute to the achievement of global health targets and initiatives related to STI control and elimination. The World Health Organization (WHO) has set ambitious goals for reducing the incidence of STIs, including a 90% reduction in N. gonorrhoeae incidence by 2030 (WHO, 2016). To achieve these targets, the WHO emphasizes the need for improved diagnostic tools, increased access to testing, and strengthened surveillance systems (WHO, 2016). InPouch TV and InTray GC align with these objectives by providing accurate, accessible, and user-friendly diagnostic solutions. The widespread use of these devices can enhance STI surveillance efforts by providing reliable data on the prevalence and distribution of T. vaginalis and N. gonorrhoeae infections. This information is crucial for guiding public health interventions, allocating resources, and monitoring progress towards STI control and elimination goals (Cristillo et al., 2017). Furthermore, the portability and 12-month from date of manufacture stability of InPouch TV and InTray GC make them suitable for use in various healthcare settings, including primary care, outreach programs, and mobile clinics (Bachmann et al., 2011). This flexibility can help expand access to STI testing services, particularly in underserved and high-risk populations, contributing to the global efforts to reduce health disparities and ensure equitable access to STI care (Cristillo et al., 2017). Conclusion The global increase in STI cases, particularly T. vaginalis and N. gonorrhoeae infections, necessitates the development and widespread adoption of innovative diagnostic tools. InPouch TV and InTray GC, developed by Biomed Diagnostics, represent significant advancements in this regard. These devices combine efficient sample collection, sample transport, and optimal microenvironment culturing in a single, user-friendly system, offering high sensitivity, specificity, and rapid turnaround times compared to traditional diagnostic methods. The evidence presented in this article highlights the potential of InPouch TV and InTray GC to support public health goals for STI reduction. Validation studies and clinical trials have demonstrated their excellent performance characteristics, with sensitivity and specificity rates exceeding 90% (Borchardt et al., 1997; Granato & Schneible, 2018). The simplified testing process and enhanced accuracy of these devices can lead to improved patient outcomes, reduced transmission rates, and cost savings for healthcare systems. Moreover, the adoption of InPouch TV and InTray GC aligns with global health initiatives aimed at controlling and eliminating STIs. These devices can contribute to the achievement of WHO targets by enhancing STI surveillance, expanding access to testing, and guiding public health interventions (WHO, 2016). Their portability and ease of use make them suitable for various healthcare settings, including resource-limited areas, thus promoting health equity in STI care. The development and implementation of innovative diagnostic tools, exemplified by InPouch TV and InTray GC, are crucial in the global fight against STIs. These devices have the potential to transform STI management by providing accurate, timely, and accessible diagnostic solutions in a variety of clinical settings. As the STI epidemic continues to evolve, with the emergence of antimicrobial resistance and changing epidemiological patterns, sustained investment and collaboration in the development and deployment of novel diagnostic technologies are of vital importance. By prioritizing innovation in STI diagnostics, we can strengthen our response to this global health challenge and work towards a future where STIs are effectively controlled. References Bachmann, L. H., Hobbs, M. M., Sena, A. C., Sobel, J. D., Schwebke, J. R., Krieger, J. N., … & Gaydos, C. A. (2011). Trichomonas vaginalis genital infections: progress and challenges. Clinical Infectious Diseases, 53(suppl_3), S160-S172. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3897282/pdf/cir705.pdf Borchardt, K. A., Smith, R. F (1991). An evaluation of an InPouch TV culture method for diagnosing Trichomonas vaginalis infection. Genitourin Medicine, 67(2):149-52. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1194652/pdf/genitmed00038-0071.pdf Cristillo, A. D., Bristow, C. C., Peeling, R., Van Der Pol, B., de Cortina, S. H., Dimov, I. K., … & Klausner, J. D. (2017). Point-of-care sexually transmitted infection diagnostics: proceedings of the STAR sexually transmitted infection—clinical trial group programmatic meeting. Sexually Transmitted Diseases, 44(4), 211-218. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5347466/pdf/olq-44-211.pdf FDA 510(K) Premarket Notification for InTray GC. https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm?ID=K210511 Gaydos, C. A., & Hardick, J. (2014). Point of care diagnostics for sexually transmitted infections: perspectives and advances. Expert Review of Anti-infective Therapy, 12(6), 657-672. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4065592/pdf/nihms584921.pdf Gift, T. L., Pate, M. S., Hook III, E. W., & Kassler, W. J. (2002). The rapid test paradox: when fewer cases detected lead to more cases treated: a decision analysis of tests for Chlamydia trachomatis. Sexually Transmitted Diseases, 26(4), 232-240. https://journals.lww.com/stdjournal/fulltext/1999/04000/the_rapid_test_paradox__when_fewer_cases_detected.aspx Kissinger, P. (2015). Trichomonas vaginalis: a review of epidemiologic, clinical and treatment issues. BMC Infectious Diseases, 15(1), 307. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4525749/pdf/12879_2015_Article_1055.pdf Paris KS, Font B, Mehta SR, Huerta I, Bristow CC. (2022). 72-Hour transport recovery of antimicrobial resistant Neisseria gonorrhoeae isolates using the InTray® GC method. PLoS One. 2022 Jan 21;17(1):e0259668. doi: 10.1371/journal.pone.0259668. PMID: 35061686; PMCID: PMC8782362 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8782362/pdf/pone.0259668.pdf Rowley, J., Vander Hoorn, S., Korenromp, E., Low, N., Unemo, M., Abu-Raddad, L. J., … & Taylor, M. M. (2019). Chlamydia, gonorrhoea, trichomoniasis and syphilis: global prevalence and incidence estimates, 2016. Bulletin of the World Health Organization, 97(8), 548-562. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6653813/pdf/BLT.18.228486.pdf WHO. (2016). Global health sector strategy on sexually transmitted infections 2016-2021: toward ending STIs. World Health Organization. https://iris.who.int/bitstream/handle/10665/246296/WHO-RHR-16-eng.pdf?sequence=1 WHO. (2018). Report on global sexually transmitted infection surveillance, 2018. World Health Organization. https://iris.who.int/bitstream/handle/10665/277258/9789241565691-eng.pdf?sequence=5 WHO. (2022). Sexually transmitted infections (STIs). World Health Organization. https://www.who.int/news-room/fact-sheets/detail/sexually-transmitted-infections-(stis) Wi, T., Lahra, M. M., Ndowa, F., Bala, M., Dillon, J. A. R., Ramon-Pardo, P., … & Unemo, M. (2017). Antimicrobial resistance in Neisseria gonorrhoeae: global surveillance and a call for international collaborative action. PLoS Medicine, 14(7), e1002344. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5501266/pdf/pmed.1002344.pdf --- ## Lateral Flow Device Design Master Series with DCN Dx Principal Engineer, Keith Kopitzke URL: https://dcndx.com/insights/lateral-flow-engineering-mastery-with-dcn-dx-expert-insights/ Type: insight Published: 2024-04-22 Dive deep into common lateral flow device design challenges with Keith Kopitzke, Principal Engineer at DCN Dx. Join Keith Kopitzke, DCN Dx’s Principal Engineer, as he tackles the most pressing challenges in the development of point-of-use diagnostics. With a rich background in industrial design and a sharp focus on the specific challenges in diagnostic device design, Keith brings a unique perspective to this series. Each episode of this series is a masterclass in solving IVD design challenges. From specific sample matrices and sample handling requirements to user-centric design solutions, Keith employs his extensive experience and a simple whiteboard to clarify complex concepts. Whether you’re a researcher, a product developer, or a medical device design engineer, these insights will enhance your approach, refine your techniques, and ultimately lead to more effective and reliable IVD products. Episode 1: Navigating Small Sample Volume Challenges for Successful Lateral Flow Blood Tests Episode 2: Sustainability in Lateral Flow Device Design --- ## Thermodynamic and Kinetic Considerations for Lateral Flow Immunoassays URL: https://dcndx.com/insights/unlock-the-secrets-of-lateral-flow-immunoassay-design-with-thermodynamics-and-kinetics/ Type: insight Published: 2024-04-12 Gain invaluable insights from Kristin Cederquist, Ph.D., Senior Scientist at DCN Dx, as she explores the fundamental principles driving lateral flow technology at the 2024 Advanced Lateral Flow Conference (ALFC). In this captivating “Lunch and Learn” session from ALFC 2024, Dr. Kristin Cederquist, Senior Scientist at DCN Dx, discusses the critical roles that thermodynamics and kinetics play in the design and performance of lateral flow immunoassays. Through her presentation, “Thermodynamic and Kinetic Considerations for Lateral Flow Immunoassays,” Dr. Cederquist breaks down complex concepts and illustrates how understanding these principles can revolutionize your approach to assay development. Dr. Cederquist shares practical insights and strategies for overcoming common obstacles, empowering you to optimize your lateral flow designs. Whether you’re a seasoned professional or new to the field, this video is a must-watch for anyone seeking to stay at the forefront of rapid diagnostics. Don’t miss this opportunity to learn from one of the industry’s brightest minds. Watch the video below. --- ## From Lab to Field: Applying Basic Lateral Flow Training to Advance Diagnostic Solutions in LMICs URL: https://dcndx.com/insights/applying-basic-lateral-flow-training-diagnostic-solutions-lmics/ Type: insight Published: 2024-04-09 We recently had the opportunity to catch up with Dr. Fezile Khumalo, a junior research fellow at the University of Cape Town, who attended DCN Dx’s Basic Lateral Flow Training (BLFT) course in 2023. Dr. Khumalo shared insights into her experience at the BLFT and how the knowledge gained has influenced her work in developing diagnostic tools for low- and middle-income countries (LMICs). Q: Can you briefly introduce yourself and share a bit about your professional background before attending the Basic Lateral Flow Training course? Dr. Khumalo: I am Dr. Fezile Khumalo, a junior research fellow at the University of Cape Town, in the division of Medical Virology, Department of Pathology. I work in women’s reproductive health with a focus on inflammation. Q: What motivated you to participate in the BLFT in 2023? Dr. Khumalo: Our research group, the Mucosal Infections Group, developed a novel point-of-care test for detecting genital inflammation. Currently, we’re laying the groundwork for immunoassays targeting infectious diseases common in low- and middle-income countries, including lateral flow assays. Enrolling in this course aimed to enhance our proficiency in immunoassay development and acquire insights into refining technologies within our pipeline. Q: Reflecting on the BLFT, what were your top three takeaways from the course? Dr. Khumalo: My top three takeaways were: Understand your test materials, and test and revise all components as necessary to obtain the best performing test. The sample type is important to understand how to develop a test that is appropriate. Testing and retesting is critical. Q: How have these key learnings influenced your approach to assay development? Dr. Khumalo: These key learnings have significantly influenced our approach to assay development. By understanding the intricacies of the development process, we’ve gained the ability to troubleshoot effectively and plan more efficiently. This understanding has enabled us to incorporate robust quality control measures at every stage of development. As a result, we can ensure the reliability and accuracy of our assays, ultimately enhancing their effectiveness in our target setting, which prioritizes assays that perform with similar or same sensitivity and selectivity as more advanced lab-based assays. Q: Could you share a specific project or challenge where you applied the skills and knowledge acquired from the BLFT? Dr. Khumalo: Our research group is currently testing the Genital Inflammation Test (GIFT). It is undergoing its first in-field assessment for performance and application in LMICs. Q: In what ways has the training impacted your work on a day-to-day basis? Dr. Khumalo: The training has significantly impacted our day-to-day work by providing a deeper understanding of the development process for various immunoassay platforms. We’ve learned that developing lateral flow assays entails more than just the biologics involved; it encompasses a multitude of factors, including sample types, sample processing techniques, detection methods, and the materials used in the assay. These components collectively determine the performance of the test. Armed with this knowledge, we are now more adept at navigating the complexities of assay development, making informed decisions, and optimizing our processes to ensure the efficacy and reliability of our tests. “The training has significantly impacted our day-to-day work…”— Dr. Fezile Khumalo, a junior research fellow at the University of Cape Town on the impact of DCN Dx’s Basic Lateral Flow Training course. Q: Are there any other achievements or milestones you’ve reached post-BLFT that you attribute to the training you received? Dr. Khumalo: We are waiting on the clinical study analysis for the 1st iteration of the GIFT test. We will potentially be making improvements to the device before we begin the regulatory approval journey for the test. Q: Based on your experience, why would you recommend the BLFT to other professionals in lateral flow, or diagnostics, more generally? Dr. Khumalo: If funding were available, it would be highly beneficial for all members of our lab to attend this course or to bring the course facilitators to South Africa. The knowledge gained from such a course is invaluable, particularly as we focus on translating our research into products for low- and middle-income countries. By equipping our team with the basics of assay development and refinement techniques, we can enhance our capabilities in developing diagnostic tools tailored to the unique needs and constraints of LMIC settings. Investing in this training would ultimately strengthen our capacity to produce impactful solutions that address pressing healthcare challenges in resource-limited environments. Q: What advice would you give to future participants to make the most out of the BLFT? Dr. Khumalo: Having the idea or beginnings of a test is important so you can specify the areas that you need help with. The course does cover everything, but if you bring specific questions, you can get much-needed practical assistance. Q: How do you see the skills and knowledge from the BLFT playing a role in your future projects or career aspirations? Dr. Khumalo: The skills and knowledge acquired from the BLFT course will play a pivotal role in our future projects, particularly as we delve deeper into immunoassay development. With the increasing focus on developing diagnostic and screening tools tailored for low- and middle-income countries, the expertise gained from the BLFT course will position us to tackle these challenges effectively. As we continue our work on new projects, the proficiency in immunoassay development acquired through the BLFT course is enabling us to design and optimize diagnostic tools that address the specific needs of LMICs. By leveraging our enhanced capabilities, we aim to develop innovative solutions that improve access to healthcare for underserved populations in LMICs. Ultimately, the skills and knowledge obtained from the BLFT course will not only contribute to the success of our future projects but also align with our broader career aspirations of making meaningful advancements and enhancing healthcare accessibility for communities in need. Q: Is there a particular area of lateral flow technology you’re excited to explore further, thanks to insights from the BLFT? Dr. Khumalo: We will be investing resources in developing antibodies for lateral flow assays as part of our long-term strategic initiatives. By developing our own antibodies, we hope to gain greater control over the performance and specificity of the assays, ensuring high accuracy and reliability in detecting target analytes. Additionally, having access to proprietary antibodies enhances our ability to customize assays for specific applications and target populations, particularly in the context of low- and middle-income countries where unique challenges may exist. The insights shared by Dr. Khumalo underscore the significant impact of DCN Dx’s Basic Lateral Flow Training course on advancing diagnostic solutions in LMICs. By equipping participants with the foundational knowledge and practical skills needed to develop and refine lateral flow assays, the BLFT empowers researchers and professionals to tackle the unique challenges faced in resource-limited settings. As Dr. Khumalo and her team continue to apply the knowledge gained from the BLFT, we eagerly anticipate the innovative solutions they will develop to improve healthcare accessibility for underserved communities worldwide. --- ## New Rules, New Approaches: Understanding the FDA’s IVD Reclassification Impact URL: https://dcndx.com/insights/understanding-fda-ivd-reclassification-impact/ Type: insight Published: 2024-04-02 (As published 2024-04-02; regulatory status may have changed since.) The FDA’s recent announcement to reclassify certain infectious disease in vitro diagnostics (IVDs) from Class III to Class II signifies a pivotal shift in the regulatory landscape for medical devices. This decision, aimed at streamlining the pathway to market for essential diagnostic tools, bears significant implications for IVD developers, particularly in the infectious disease and companion diagnostics sectors. As such, it is crucial for these stakeholders to thoroughly understand the regulatory nuances of this shift and strategically align their development processes accordingly. The ripple effects of this reclassification across the IVD industry cannot be overstated. For developers, the prospect of a less cumbersome regulatory path promises to accelerate the development cycle, reduce costs, and encourage the entry of new players. This could lead to a more vibrant marketplace, characterized by a broader array of IVD products. However, navigating this new regulatory terrain will require a nuanced understanding of the updated requirements and a strategic approach to compliance. This is where the expertise and insight of seasoned industry players become invaluable. A Closer Look at the FDA’s Announcement The FDA’s decision to initiate the reclassification process was informed by discussions at a September 2023 panel meeting, which focused on three types of infectious disease IVDs. These included nucleic acid and serology-based tests for Hepatitis B Virus (HBV) infection management, serology-based tests for detecting human parvovirus B19, and cell-mediated immune reactivity tests for identifying responses to Mycobacterium TB infection. These discussions serve as a starting point for the broader reclassification effort, highlighting the FDA’s methodical approach to regulatory evolution, which you can read more about here. We expect more announcements soon. Regulatory Implications The transition from the PMA pathway to the 510(k) pathway, as proposed by the FDA, represents a fundamental change in the regulatory oversight of infectious disease IVDs. This move is designed to reduce the regulatory burden on developers by simplifying the approval process for tests that are deemed to have a moderate risk profile, provided they meet the established special controls. Importantly, it is also expected to spur innovation, enhance competitive dynamics, and widen access to vital diagnostic tests. Understanding the nature of these special controls is paramount for developers. They are regulatory requirements that provide a framework for addressing potential risks associated with IVDs, ensuring their safety and effectiveness. These controls might encompass specific performance criteria, labeling requirements, or post-market surveillance obligations, among others. For IVD developers, this necessitates a granular examination of how these controls apply to their products. Developers must evaluate their current development and validation strategies against these new requirements to identify any gaps or areas necessitating adjustment. This could involve rethinking design considerations, enhancing quality assurance practices, or implementing more robust post-market monitoring systems. Strategic Guidance for IVD Developers In light of these regulatory changes, IVD developers should consider several strategic approaches to navigate the reclassification successfully: Regulatory Alignment: Developers should begin by conducting a comprehensive review of the FDA’s updated regulatory requirements for Class II devices. This includes understanding the specific special controls applicable to their products and aligning their development and quality assurance processes accordingly. Special controls may include performance standards, labeling requirements, clinical performance studies, and/or post-marketing surveillance measures. Engaging with IVD CRO specialists can provide valuable insights and guidance in this regard. Design Optimization: Given the shift in regulatory classification, there may be opportunities to optimize product design and development processes to better meet the new standards. This could involve leveraging new technologies, materials, or methodologies that enhance product safety and efficacy while complying with the special controls. Evidence Generation: Under the 510(k) pathway, demonstrating substantial equivalence to an existing market-approved device is crucial. Developers should focus on generating robust scientific and clinical evidence that clearly establishes the equivalence of their products in terms of safety and performance. Stakeholder Engagement: Proactive engagement with key stakeholders, including the FDA, healthcare professionals, and patient advocacy groups, is critical. Open communication can provide valuable feedback on product design, clinical utility, and regulatory compliance, facilitating a smoother approval process. Continuous Learning and Adaptation: The regulatory environment for medical devices is continually evolving. Developers must stay abreast of ongoing changes in regulations, guidelines, and industry best practices. Investing in continuous learning and professional development for team members involved in regulatory affairs and product development is essential for maintaining compliance and fostering innovation. Precedents: Special Controls We do have precedent here. When the FDA reclassified certain HIV diagnostic tests and rapid influenza diagnostic tests (RIDTs) from Class III to Class II, it established specific special controls for each category to ensure their safety and effectiveness while facilitating a more streamlined regulatory process. Here’s how these special controls had implications for both types of tests: HIV Diagnostic Tests Reclassification and Special Controls Implications Performance Evaluation: Special controls required rigorous validation of the HIV tests’ sensitivity and specificity. This meant that manufacturers had to provide substantial clinical data demonstrating that their tests could reliably detect HIV infection across diverse patient populations, ensuring that the tests were dependable for early diagnosis. Labeling Requirements: The special controls included stringent labeling requirements, necessitating clear instructions for the tests’ use, interpretation of results, and the action to be taken following the test outcomes. This was particularly important for HIV diagnostic tests to ensure that users understood the need for confirmatory testing in the event of a positive result, thereby minimizing the risk of misinterpretation and the consequent anxiety or mismanagement. Post-Market Surveillance: Manufacturers were obligated to actively monitor and report on the real-world performance of their tests post-market introduction. This continuous surveillance helped in identifying any issues related to test accuracy, such as false positives or negatives, which could significantly impact patient management and public health responses. Reclassification of Rapid Influenza Diagnostic Tests (RIDTs) and Implications of Special Controls Sensitivity and Specificity: The special controls for RIDTs emphasized the need for these tests to meet certain sensitivity and specificity benchmarks. Given the critical role of RIDTs in diagnosing and managing influenza outbreaks, these performance standards ensured that the tests provided reliable results, reducing the likelihood of false negatives that could lead to untreated influenza or false positives that might result in unnecessary treatments. Labeling for False Results: The FDA required detailed labeling to inform users about the potential for false-positive and false-negative results. This was crucial in guiding healthcare providers on how to interpret test results, especially considering the variability in RIDT performance across different influenza strains and patient populations. Post-Market Studies: RIDT manufacturers were tasked with conducting regular post-market studies to assess the tests’ performance in light of the ever-changing landscape of influenza viruses. These studies were essential for ensuring that RIDTs remained effective diagnostic tools despite the emergence of new influenza strains, contributing to their adaptability and continued relevance in public health efforts. The Road Ahead Looking forward, the reclassification of infectious disease IVDs is likely to set a precedent for future regulatory adjustments in the IVD sector and beyond. The challenge for developers will be to leverage the opportunities presented by this shift, ensuring that their innovations are aligned with the revised regulatory framework. This entails a continued focus on rigorous validation and verification processes to meet the new special controls, as well as proactive engagement with regulatory bodies to ensure compliance. Moreover, the reclassification underscores the importance of a collaborative approach, where industry stakeholders and regulatory authorities work together to advance public health objectives. As the industry navigates this new regulatory environment, the insights and guidance of experts in the field will be invaluable. Partnering with organizations that possess deep regulatory expertise and a strategic outlook on product development can be a critical success factor in this new regulatory era. As the industry adapts to these changes, the emphasis on strategic foresight, regulatory alignment, and continuous innovation will be paramount in driving the development and successful market entry of new and essential IVD products. --- ## Reflections on ALFC 2024 URL: https://dcndx.com/insights/reflections-on-alfc-2024/ Type: insight Published: 2024-03-18 Reflections on ALFC 2024: Key Takeaways and Future Directions Reflecting on the Advanced Lateral Flow Conference 2024, I am struck by the breadth and depth of innovation evident in the presentations and discussions. This year’s conference was not only a testament to the resilience and adaptability of the lateral flow industry, but also a look into the future of global healthcare. The convergence of experienced professionals, emerging leaders, and groundbreaking technologies underscored the important role community, idea sharing, and shared commitment plays in advancing global health. As both host and participant, I found the conversations and sessions enlightening and the collective focus invigorating. It is this spirit of mutual respect and support that continues move our industry forward. A Focus on Commercialization and Funding New to ALFC 2024, our organizing committee selected speakers and panelists who could highlight the commercialization and funding issues influencing our industry. One such speaker was Giles Hamilton, Chairman of Panacea, Transdermal Dx, and SciLogica. In his enlightening session titled “Investment Pathways in Lateral Flow: Shaping the Future of Rapid Diagnostics”, Hamilton highlighted a critical healthcare gap: that nearly half the global population lacks access to diagnostics according to the Lancet Commission on Diagnostics and WHO data. Hamilton’s talk drew upon his experience in spearheading at-home testing including financing over 100 lateral flow deals. He presented a future where point-of-care diagnostics are central to healthcare. With a compound annual growth rate (CAGR) of 10.7% by 2025 (compared to 7.4% CAGR for central labs), the PoC diagnostics market will reshape the healthcare landscape in the coming years. Despite 248 active US venture capital funds focused on pre-revenue MedTech and point-of-care testing, obstacles persist. For instance, Hamilton underscored the need for products that deliver results in under five minutes, with initial consumable costs of goods sold (COGs) at $2-3 eventually dropping below $1 at scale. How do we meet these challenges? Hamilton’s recommendations included ensuring a compelling clinical use case and cost control at all stages of development along with the need to engage with payers to refine the technology and Minimum Viable Products. He also pointed out the often-overlooked sources of non-dilutive government and foundation grant funding which offer alternate structures including license royalties and joint funding. He spotlighted the potential of corporate investment syndicates, like those at Medtronic and J&J DevCo, who review thousands of companies each year underscoring the competitive nature of platform financing. Hamilton’s advice to lateral flow companies was clear: keep an open mind on business models, select development and target markets carefully, and ensure plans are aligned with US-centric POCT adoption and prioritization. Such a nuanced understanding of the funding landscape is critical when aiming to bring transformative diagnostic solutions to market. Early feedback from attendees indicates that this was an important session and I certainly agree. Innovation Award: A Spotlight on Industry Leaders Industry innovation is a collective effort, and the ALFC serves as a needed platform for sharing technological advances. A particularly memorable aspect of the program was the Innovation Award which showcased pioneering products and technologies. The three finalists exemplified the spirit of innovation that drives our market forward. LFAnt emerged as the audience-selected winner with its revolutionary product, the Compact – the world’s first handheld and portable magnetic lateral flow platform. This device enables quantitative testing from point of care anywhere, making advanced diagnostics accessible in a range of settings. In addition to being recognized by their peers, the LFAnt team also took home a $10,000 cash prize. The Fraunhofer Institute’s CampyTube presented a unique integration of lateral flow and molecular detection in a single vial. This seamless, simple and sophisticated combination could open new avenues for rapid, on-site detection of various pathogens. inne ‘s STRIP 2.0 was another standout, offering a novel approach to measuring the hormone progesterone in saliva with impressive sensitivity. Beyond technical achievements, the STRIP 2.0 significantly reduces plastic waste and enhances user experience, addressing both sustainability and usability concerns. We heard more from inne’s founder and CEO, Eirini Rapti, in a joint “case study” session titled “Full Circle Innovation: A Collaborative Approach to Product Lifecycle Excellence”. In this 3-part session, Rapti represented “product”, while Keith Kopitzke, Principal Engineer, DCN Dx, presented engineering, and Ted Meigs, Kinematic Automation (now part of Ascential Medical and Life Sciences ) represented manufacturing. These Innovation Award finalists demonstrated exceptional technical ingenuity and a commitment to improving healthcare outcomes and patient experiences. Congratulations again for your achievements! New Awards in 2024 Award additions for 2024 included the Leadership in Diagnostics, the Lifetime Achievement Award, and Poster Awards. The Leadership in Diagnostics Award acknowledges an individual or organization who have influenced the industry by fostering collaboration and contributing to the evolution lateral flow diagnostics. It recognizes individuals whose careers have been marked by sustained and significant impacts, setting a benchmark for commitment and excellence in the industry. For the 2024 Poster Awards, there was a tie between two remarkable contributors: Dr. Nicolas Heureux’s team at Kimialys and John Kelly, the Founder and CEO of Atomo Diagnostics. Dr. Heureux, with a background in immuno and organic chemistry, is at the forefront of advancing LFA technologies. His work on SPR-based biophysical studies and his contribution to the Rapid Antibody Pair Selection for TnI Lateral Flow Assay via Multiplexed SPRi poster is important research with far reaching implications. John Kelly brings over two decades of experience in redefining medical devices and dedication to enhancing diagnostic usability and performance. Also a session speaker, both Kelly’s poster and talk focused on Atomo’s important work developing a user-centric LFA cassette design for improved blood sampling for POCT devices. The Leadership in Diagnostics Award was proudly presented to Ted Meigs, whose extensive experience in automation and design has influenced the LFA industry significantly. As a co-founder of Kinematic Automation (now part of Ascential Medical and Life Sciences ), Meigs has been a pillar in the diagnostic community, holding numerous patents and setting industry standards in manufacturing processes. Meigs and his team are recognized globally as leaders in manufacturing automation which highlights ability to foster collaboration across the field. The Lifetime Achievement Award honored Dr. Thomas Tisone for his visionary entrepreneurship and substantial contributions to lab automation. His commitment to innovation and customer focus has been key in BioDot’s success, leading to products integral to the diagnostics industry. Tisone’s philosophy of “We Can Do That” reflects an approach that inspired a culture of ingenuity, service and contributing to BioDot’s status where the average employee tenure exceeds two decades. These awardees exemplify the spirit of excellence and innovation that defines the lateral flow industry. Their achievements not only highlight the advances in LFA technology but also set benchmarks for leadership, commitment, and research excellence. We were honored to have such esteemed and deserving awardees. Future Directions and Continued Collaboration The insights and discussions at ALFC 2024 underscored the importance of collaboration and cross-disciplinary approaches. The integration of digital technologies, including AI and machine learning, was a recurring theme which highlighted a broader trend towards more intelligent, user-friendly diagnostics. Looking to the future, it is clear that the boundaries of what lateral flow assays can achieve are expanding. The potential for LFAs to play a more significant role in personalized healthcare, environmental monitoring, and global disease surveillance is immense. A Special Thanks to Our Sponsors ALFC 2024’s success was driven the support and engagement of our sponsors. It is with deep gratitude that we acknowledge the role our sponsors played. Our sponsors are our exhibitors and together they created a dynamic exhibit room. More than a space for breaks, it was a cornerstone for pivotal discussions, insights, and learning. Thanks to Multisorb, our Exhibit Lounge sponsor, we were fueled by coffee and snacks each day. Of all the life science conferences I’ve attended over the year’s ALFC’s exhibit room stands out as the most engaging I’ve experienced. Every visit to this vibrant area was a true educational opportunity—and I heard this sentiment echoed often from our attendees. The depth of knowledge and innovation showcased by our sponsors made the Exhibit Room a true learning experience for all. To our sponsors, we extend our heartfelt thanks. Your financial support and active participation throughout the ALFC planning process were indispensable. The event thrives on your contributions, without you we could not provide the value that our attendees have come to expect. We are delighted that most sponsorships for ALFC 2024 were sold out due to high demand. Looking ahead to ALFC 2025, we are opening up early sponsorship. For those interested in sponsoring and playing a key role in next year’s event, we encourage you secure your spot early. Your early commitment allows us to plan for an even more impactful event, building on this year’s success and forging new frontiers in the lateral flow assay industry. A Collective Journey Towards Innovation The contributions of each attendee, speaker, and sponsor at the conference are invaluable in shaping the future of our industry. As we “flow forward”, let us continue the spirit of collaboration and innovation that defines the ALFC. Together, we can address the complex diagnostic challenges and make a meaningful impact on the lives of people around the world. Thanks to all who participated in ALFC 2024. Your dedication to advancing lateral flow technology is not only inspiring but essential in shaping a healthier and more informed world. We hope to see you at the next ALFC, which will be held October 14-15, 2025. Sincerely, Charlie Mamrak CEO, DCN Dx, host of the Advanced Lateral Flow Conference --- ## Flow Forward at ALFC 2024: Embracing Future Innovations and Collaborations in Lateral Flow URL: https://dcndx.com/insights/alfc2024-flow-forward/ Type: insight Published: 2024-01-30 Greetings! I’m Mitzi Rettinger of DCN Dx, and I’m excited to welcome you to the 2024 Advanced Lateral Flow Conference (ALFC), scheduled for February 13-14 at the Hard Rock Hotel in San Diego. As chair of the 2024 ALFC Organizing Committee, I want to share more information about this year’s conference and what I think is so special about it. Considered the lateral flow industry’s premier event, every ALFC brings together a wealth of innovators and pioneers. That hasn’t changed: as always, the ALFC focuses on the latest trends and innovations in point-of-use diagnostics. This year, however, we’re turning a particular eye to the future. This year’s theme, “Flow Forward,” reflects the 2024 ALFC Organizing Committee’s commitment to advancing technology and fostering collaboration. With “Flow Forward,” we’re not just reacting to the rapidly evolving diagnostics landscape we’ve experienced in recent years; we’re championing its innovators and creating partnerships that will lead to next-generation solutions and ensure a vibrant future for the lateral flow industry. Every session and networking opportunity at the conference has been carefully crafted to embody our theme. While I’d love to showcase each of the exceptional sessions we’ve planned, space limits me to just a few key highlights. (I highly recommend checking out our complete agenda for a look at what’s in store.) Funding and Commercialization The first day of ALFC 2024 focuses on the essentials of funding and commercialization in the lateral flow sector, providing attendees with strategies and insights for navigating the investment landscape. The fluctuating financial landscape post-pandemic has reshaped how life sciences, particularly diagnostics, approaches funding and commercialization. We’ll explore the latest trends in financial backing and practical approaches to propel lateral flow projects to success. A standout session, led by Giles Hamilton, Chairman of Panacea, Transdermal Dx, and SciLogica, is aptly titled “Investment Pathways in Lateral Flow: Shaping the Future of Rapid Diagnostics.” Giles will delve into the evolving strategies and trends driving investment in the IVD market. Hamilton’s expertise in guiding successful companies in the life sciences space makes this session invaluable for anyone looking to advance their projects through strategic funding or invest in the rapidly growing field of healthcare technology. I am also delighted that Eric Lai, Ph.D., Managing Executive, PharmaDx LLC, Team Lead, NIH COVID-19 Rapid Acceleration of Diagnostics (RADx) Initiative, and Scientific Lead, Independent Technology Assessment Program (ITAP), will be with us to lead a session that educates our attendees about the ITAP. ITAP is a government initiative designed to support the development and acquisition of innovative technologies in healthcare, including lateral flow assays. ITAP aims to facilitate the transition of new diagnostic tools from development to market by providing financial support and streamlining the procurement process. This program represents a critical lifeline for diagnostic companies seeking to navigate the post-pandemic funding landscape, ensuring that the momentum in diagnostic innovation continues unabated. My friend and colleague, Pat Vaughan, COO, DCN Dx, kicks off Day 1 with his session, “Future-Proofing Lateral Flow: Insights from More Than 500 LFA Development Programs Over 30 Years.” Having worked side-by-side with Pat, I can personally attest to his unparalleled depth of insight and expertise in the field. I’m particularly excited to hear Pat distill his vast experience into key strategies that can guide us toward a more innovative and resilient future in lateral flow diagnostics. I can’t wait to see what he shares and the impactful discussions it will undoubtedly spark among attendees! Highlighting Innovation On the second day, the spotlight turns to the technological advancements and future directions of lateral flow assays. Discussions will center on the role of evolving technologies, such as AI and machine learning, in enhancing testing accuracy and expanding the potential for at-home and wellness applications. The evolution of at-home care and wellness applications using LFAs is at the forefront of healthcare delivery. On Day 2, we’ll discuss how the latest research in flow control technology is advancing LFAs, enabling them to perform complex biomedical assays outside of laboratory settings. Innovations like these allow for more sophisticated at-home tests (such as multiplex molecular diagnostics for pathogens) and are paving the way for LFAs to be used in a broader range of applications, enhancing user engagement and personal health management. Our Day 2 keynote speaker, Hamid Ghanadan, Founder and CEO, LINUS, will present his firm’s research in this important area during his session, “Home is the Center of Health.” Sean Redmond, Director of Global Product Management, QuidelOrtho, continues this important discussion about new application areas in his session, “Increasing Value by Exploring New Settings and Use Cases for Lateral Flow Testing.” For the last few years, artificial intelligence (AI) and machine learning (ML) has captured the minds of the world, including some of the best and brightest in the lateral flow industry. These cutting-edge technologies are set to redefine diagnostics, enhancing precision and speed in testing and results reporting. A session led by DCN Dx Senior Scientist John Scott will delve into how AI and ML can optimize data analysis, improve diagnostic accuracy, and streamline the LFA development process. I moderate a panel on Day 2 on the topic of “Digital Integration Panel Discussion: Customized Lateral Flow Readers for Emerging Home-Use and Niche Applications.” In this session, we’ll discuss the evolution of readers and other technology to accommodate these burgeoning applications. The conversation will not only delve into technical advancements but also explore how these innovations are shaping user experiences and opening new possibilities in personalized healthcare and niche markets. This panel is a must-attend for those interested in the cutting edge of lateral flow technology and its practical implications. Networking and Industry Awards As always, networking is a key feature of the ALFC, and there are numerous opportunities to build partnerships. We kick off our networking events with a welcome reception in the exhibition room on February 12. The party starts at 5:30 pm. If you will be arriving in time, I encourage you to grab your badge and come say hello to your fellow attendees! The exhibition room is a great place to network throughout the conference. Open each day from 8 to 5, this is where you can meet our sponsoring exhibitors who represent the most knowledgeable organizations in this space. They are a wealth of knowledge and I hope you all take advantage of the opportunity to visit them. Coffee and snacks will be available daily in the exhibition room lounge, proudly sponsored by Multisorb. All meals are included in your conference registration and—let me tell you—you have not had conference food like this before! Our team has lovingly prepared nutritious and hearty menus for all meals. These are wonderful opportunities to “break bread” with our colleagues from around the world. Our lunch sponsor this year is Detekt Biomedical. Be sure to seek them out and say hello! And before you ask—yes, the beloved ice cream bar is back for our Day 2 afternoon break! Also returning are the ALFC awards. The Innovation Award, a cornerstone of previous years, recognizes individuals or teams that have introduced groundbreaking products or technologies that expand the lateral flow industry’s horizons. Award additions for 2024 include Leadership in Diagnostics, the Lifetime Achievement Award, and Poster Awards for poster sessions. The Leadership in Diagnostics Award acknowledges an individual or organization that has demonstrated outstanding leadership, fostered collaboration, and contributed strategically to the growth and evolution of the lateral flow diagnostics sector. Meanwhile, the Lifetime Achievement Award is conferred upon those whose careers have been marked by sustained and significant impacts, setting a benchmark for commitment and excellence in the field. I’d be remiss if I didn’t plug our amazing evening receptions. Day 1 of the conference features a sophisticated off-site event at Coastera’s floating venue. If you were with us in 2021, you’ll remember the gorgeous sunset we had overlooking the bay that evening. We’ll be in the same spot this year—cross your fingers for another view like that! Day 2 will culminate in an unforgettable closing reception and awards gala. The event, themed “Glow Forward,” will be held on the Hard Rock Hotel’s rooftop deck overlooking downtown San Diego. All award recipients will be announced onstage at the closing reception and awards gala on February 14. This event promises to be a party to remember. We hope you’ll join us! Flow Forward with ALFC 2024 The 2024 ALFC’s “Flow Forward” theme is a call to action. It’s a philosophy that carries into every presentation, panel, and event we’ve curated for you. By fostering an environment where innovators find their voice and where collaboration flourishes, your participation at ALFC 2024 ensures the lateral flow industry continues to evolve—not incrementally, but in leaps and bounds. Registration ends February 9. I hope to welcome you all personally! See you there! --- ## Understanding the FDA’s Latest Cybersecurity Guidance: Implications for IVD Devices URL: https://dcndx.com/insights/fda-cybersecurity-guidance-ivd-devices-medical/ Type: insight Published: 2023-12-11 (As published 2023-12-11; regulatory status may have changed since.) Explore this overview of the FDA’s newly finalized guidance, “Cybersecurity in Medical Devices: Quality System Considerations and Content of Premarket Submissions,” and its impact on in vitro diagnostic (IVD) device manufacturers. This Expert Insights whitepaper by Chenghui Yu, Senior Optical Engineer at DCN Dx, sheds light on the FDA’s newly heightened cybersecurity requirements for software-enabled IVD devices. It also emphasizes the significance of comprehensive cybersecurity measures for patient safety and corporate responsibility. Key Insights You’ll Discover: Expanded Scope of Applicability: Understanding the FDA’s broader compliance requirements for devices with any programmable logic. Lifecycle Risk Management: The importance of continuous cybersecurity vigilance and adaptation throughout a device’s lifecycle. Secure Product Development Framework (SPDF): Strategies for integrating cybersecurity at every stage of device development. Impact on Small IVD Device Developers: Challenges and solutions for smaller manufacturers, exemplified by DCN Dx’s miniDxR device. And more! This whitepaper provides an overview of the FDA’s new cybersecurity guidelines. It is a vital resource for IVD manufacturers and stakeholders in understanding and navigating the complexities of medical device cybersecurity. However, as the cybersecurity landscape is constantly evolving, it is important to review the FDA website for up-to-date information. Download now for our insights into the new standard of IVD device cybersecurity! --- ## Energetic Considerations for Particle Conjugates in Lateral Flow Immunoassays URL: https://dcndx.com/insights/energetic-considerations-particle-conjugates-lfa/ Type: insight Published: 2023-10-03 Lateral flow tests are synonymous with quick, actionable insights, but this efficiency hinges on the complexity and nuance of their design. Are you developing lateral flow assays and seeking to understand the molecular-level interactions that determine their performance? Download Our Exclusive Expert Insights Whitepaper As part of our ongoing Expert Insights series, DCN Dx presents a whitepaper that addresses the biophysical elements that guide the function of lateral flow immunoassays. “Energetic Considerations for Particle Conjugates in Lateral Flow Immunoassays,” authored by our Senior Scientist Kristin Cederquist, Ph.D.,” is available for download below. Topics Covered: Thermodynamics and Kinetics: Understand how these fundamental factors influence diagnostic device performance and their specific relevance to lateral flow immunoassays. Particle Conjugate Role: Review how the manipulation of particle conjugates can improve lateral flow performance. Real-world Implications: Explore how an in-depth understanding of these forces can lead to smarter design choices and maximized performance, contributing to wider adoption of LFAs and reduced error rates. Who Should Download? Research Scientists: Involved in developing or researching lateral flow assays. Assay Developers: Seeking to refine their approach by understanding the science that underpins lateral flow technology. Biochemists and Biophysicists: Looking to enhance their comprehension of energetics in bioassays. Fill out the form below to begin reading. By submitting this form, you agree that we or our media partners may contact you in accordance with our privacy policy. --- ## Statistical Rigor in IVD Migration Studies: Insights from the FDA/AdvaMed Conference URL: https://dcndx.com/insights/ivd-migration-studies-expert-insights-fda-advamed/ Type: insight Published: 2023-09-06 (As published 2023-09-06; regulatory status may have changed since.) Introduction The in vitro diagnostic (IVD) device industry operates at the intersection of technology and biostatistics. The complexity of this relationship was on display at the 15th Annual FDA/AdvaMed Medical Device Statistical Issues Conference, where DCN Dx’s clinical research team attended to gain insights into trends and best practices in statistical methodologies. One of the standout discussions centered on statistically appropriate practices for the validation of diagnostic device output. Emphasis was placed on critical considerations for IVD migration studies, particularly for Next-Generation Sequencing (NGS) and Companion Diagnostic (CDx) assays. Here, we delve into the essential takeaways from the conference and how they relate to critical considerations for IVD migration studies. Migration of IVD Devices to New Systems Explained The migration of IVD devices to new systems refers to the process of transferring or adapting a particular assay or diagnostic test from an existing platform or system to a different one. This is often necessary when introducing a new instrument or technology that offers better efficiency, scalability, or compatibility. Migration must be carried out with extreme precision, ensuring that the new system performs equivalently to the old one in all key aspects. Failure to maintain the performance characteristics during migration can lead to and inaccuracies in diagnostic results, with potential implications for patient care. In particular, the alignment of statistical parameters is essential to guarantee that the new system performs equivalently to the old one, a challenge that demands precise statistical methods and a deep understanding of assay characteristics. Critical Considerations for IVD Migration Studies Migration studies are central to IVD device adaptation and heavily grounded in statistical reasoning. Key considerations from a biostatistical standpoint include: Intended Use and Indications: The intended use and indications for the device should remain consistent, with the exception of the inclusion of the new system. Reagent and Assay Parameters: These should be unchanged except for minor differences such as slight alterations in incubation times that optimize the assay on the new system. Assay and System Technologies: It’s vital that these remain unchanged to maintain the fidelity of the results. No Expected Change to Assay Performance: The new system should not affect the assay’s performance. Invalid Rate Estimation: Several presenters at the conference highlighted the importance of estimating an invalid rate for validation studies such as Limit of Blank (LoB), Limit of Detection (LoD), Precision, Comparison, etc. The invalid rate should be meticulously reported for each level and subsequently evaluated and compared between the old system and the new system in migration studies. This comparison is vital in ensuring that the new system maintains the reliability and integrity of the established diagnostic measurements. For NGS CDx Assay Migration: a. Sufficient Challenging Samples: A careful selection of samples should be evaluated in the assay migration studies to support the performance comparison of the new system relative to the old system. b. Unchanged Assay Performance: Parameters such as assay sensitivity, precision, and accuracy are expected to remain consistent between the new and old system. c. Complex Study Design for NGS-Based CDx: The study design will require intricate planning to assess the impact on both CDx variants and variants from other claims, such as tumor profiling. Conclusion The migration of IVD devices to new systems is a complex process requiring meticulous attention to detail. The recent FDA/AdvaMed Medical Device Statistical Issues conference provided valuable insights into best practices and considerations. At DCN Dx, our clinical research services team is committed to staying at the forefront of industry standards and advancements. We recognize the importance of these migration studies in maintaining the robustness and reliability of diagnostic devices. We draw on our expertise in assay development, device design and engineering, clinical research, and manufacturing to provide comprehensive support for IVD developers. By adhering to the critical considerations for migration studies as outlined above, we strive to ensure that each diagnostic device retains its efficacy, accuracy, and dependability. For further information on how DCN Dx can support your IVD migration studies, we welcome you to contact us. About the Author Veronika has more than 15 very successful and rewarding years in the therapeutic and diagnostic industries as well as in the academic environment. Before joining DCN, she worked for Carl Zeiss Meditec Inc. (Medical Device – Scanning Laser Polarimeter for detection of glaucoma), Prometheus Therapeutics (therapeutic and diagnostic products), and Invivoscribe (IVD). She has also worked as a statistical consultant for numerous health related studies over the years. At Invivoscribe, she helped to develop CE-IVD and RUO MiSeq assays for hematology-oncology. Assay development included V&V studies, Clinical bridging studies as well as setting specifications for analyte-specific reagents and DNA and RNA controls. Veronika has extensive experience in clinical trials and evaluation of medical device performance. She is well versed in regulatory 510(k), PMA, and IVDR submissions. She holds a M.Sc. in applied statistics from Purdue University. --- ## Improve the Interpretability of Your Lateral Flow Assay URL: https://dcndx.com/insights/lateral-flow-assay-lfa-image-color-grading/ Type: insight Published: 2023-07-05 Discover more in our exclusive Expert Insights whitepaper: Analyzing Lighting Conditions to Achieve Accurate LFA Test Strip Grading Lighting conditions greatly impact the way visual-based readings are interpreted. But to what extent? In this Expert Insights whitepaper, DCN Dx Senior Scientist, John Scott, explores his research into the use of printed grading scales to improve interpretability across various lighting environments and backgrounds. Download this whitepaper to discover: How environmental factors can influence grading of lateral flow assay test strips. The potential effects of diverse lighting conditions and background colors on assay interpretation. The importance of image analysis for accurate lateral flow assay readings. Fill out the form below to begin reading. By submitting this form, you agree that we or our media partners may contact you in accordance with our privacy policy. --- ## Lateral Flow Immunoassay Label Choices URL: https://dcndx.com/insights/lateral-flow-immunoassay-label-choices/ Type: insight Published: 2023-06-20 Since their initial emergence, immunoassays have relied on external labels to indicate bioassociation events. In the early days of this field, these labels were typically radioactive isotopes such as 125 I, 131 I, or 3 H. In recent decades, scintillation counter detection has largely been eclipsed by optical detection methods such as absorbance or fluorescence, and advances in plate reader optics have highly increased the throughput for such tests. The incorporation of enzymes such as horseradish peroxides or alkaline phosphatase has allowed plate-based optical immunoassays to further eclipse the performance of radioimmunoassays, as enzymatic amplification further heightens sensitivity. Though highly sensitive, plate-based immunoassays are not commonly amenable for use in point-of-care settings. The need for electricity, instruments, and time precludes their use in remote areas, often requiring the skills of a highly trained technician. Migrating a plate-based sandwich immunoassay to a completely dried-down paper-based system like a lateral flow immunoassay alleviates these pain points, in addition to drastically reducing cost. Because all of the test reagents are dried and subsequently re-hydrated by a sample and an optional chase buffer, the workflow is greatly simplified. Nanoparticles are often used as labels in lateral flow immunoassays due to their ability to provide a strong optical signal density per binding event. Thanks to advances in surface chemistry, nanoparticles are also often easy to functionalize with biomolecules such as proteins and nucleic acids. This overview will detail some of the more common lateral flow immunoassay nanoparticle labels and their utility. Colloidal Gold The use of colloidal gold (also known as “gold nanoparticles” or “sols”) dates back to ancient Rome, where it was used to color glass. Today, colloidal gold fabrication is scalable and controlled, with sizes typically ranging from 20 to 80 nm for lateral flow applications. The particles are a wine-red color and exhibit years-long stability; in fact, Michael Faraday’s gold nanoparticles are still on display at the Royal Institution in London almost 170 years after their fabrication in 1856. Today, a number of reputable suppliers offer colloidal gold for lateral flow applications. DCNovations Colloid Gold Colloidal gold is one of the most widely used lateral flow particle labels in the diagnostic industry due to its intense red color, ease of biofunctionalization, and reasonable price point. The color arises not from more traditional organic dyes, but rather from collective oscillations of electrons known as surface plasmon resonance. This phenomenon translates into an intense signal per binding event with particles that are well under 100 nm in diameter. Nanosized gold is easily functionalized with antibodies via passive adsorption. In this process, functional groups in the protein amino acids associate with the particle gold atoms through a number of weak forces, resulting in an overall strong association between the antibody and particle. Though the antibody is not oriented in a specific configuration on the particle surface, the antibody packing density is relatively high, giving the particle conjugate high biofunctionality. In past decades, a number of different particle morphologies such as rods, plates, and shells have resulted in tunable colors. Additionally, colloidal gold has been functionalized with coatings to facilitate antibody conjugation via different chemistries. Despite these advances, however, the most common gold conjugate in lateral flow remains the nanosphere biofunctionalized via passive adsorption. Latex (Polystyrene) Spheres Latex spheres represent a class of particles with applications in many different aspects of biotech. These monodisperse particles are available in a wide range of sizes from 20 nm to 160 µm, with 100-400 nm encapsulating the typical range used in lateral flow immunoassays. Though passive adsorption is an option through hydrophobic interactions between nonpolar and aromatic amino acids and the polystyrene surface, biological molecules are more often functionalized via covalent bond formation. One chemistry that is popular is formation of an amide bond linking a carboxyl group on the particle surface with an amine group in the protein, accomplished by the zero-length crosslinker EDC. Polystyrene spheres offer a few advantages over colloidal gold for lateral flow assays. First, they often offer the lowest price point of all particle labels. Second, because they are doped with organic dyes, they are available in a wide range of colors, with red, blue, black, and green the most popular choices for lateral flow. Europium chelate, a fluorescent dye with a large Stokes shift, is also available in a latex particle. This label requires an external reader for elucidation of signal in lateral flow but can result in sensitivities >10x better than visual colorimetric options. While latex particles may not offer as high a color density as colloidal gold, their versatility with respect to color choices and compatibility with a number of difficult biological matrices make them ideal candidates for labels in a number of lateral flow immunoassay scenarios. Cellulose Nanobeads Cellulose nanobeads are a newer label that has been available to the lateral flow assay development market for approximately 10 years. These 300-400 nm particles comprise a cellulose core, but unlike latex spheres where the dye is embedded in the particle, the dye on cellulose nanobeads is presented on the particle surface. Currently, six colors are available (black, green, and two shades each of red and blue), and because of the different dye structures, each color could necessitate slightly different biomolecule conjugation conditions. NanoAct™ Cellulose Nanobeads The majority of cellulose nanobead couplings with antibodies are easily accomplished by passive adsorption, though carboxyl versions of red and blue particles are available for covalent conjugation. In the experience of DCN Dx scientists, passive conjugations with cellulose nanobeads are relatively easy, typically free of aggregation, and give good yield. The resulting cellulose nanobead conjugates have also given enhanced sensitivity over other visual labels in many development programs. Cellulose nanobeads are manufactured by Asahi Kasei and available for evaluation through DCN Dx, either as the particles alone or bundled into a conjugation kit. Looking to the Future As requirements for lateral flow assays become more stringent (e.g. better sensitivity, shorter time-to-answer, etc.), labels and conjugation methods must naturally evolve to accommodate. Conjugates are inherently limited by the optical cross-section of the label and antibody binding affinity and kinetics. Therefore, any significant advances in lateral flow technology would necessitate development of brighter labels, targeting biomolecules with higher affinities, more robust surface chemistries, or advanced strip design. A number of these efforts are underway at a number of academic research groups and industrial companies. DCN Dx offers conjugation, screening, and support services for any and all of the labels discussed here. Contact us to discover how we can push the limits of your assay! --- ## Announcing Our New Content Series, “Expert Insights: Your Gateway to World-Class Diagnostic Development and Commercialization” URL: https://dcndx.com/insights/expert-insights-diagnostic-development/ Type: insight Published: 2023-04-25 I’m absolutely delighted to introduce our latest initiative, the “Expert Insights” content series. As the go-to partner for companies transforming diagnostic solutions from an idea into market-ready products, our team has long been dedicated to fostering professional development within the industry. With our “Expert Insights” series, we’re embracing our penchant for big, bold educational projects to provide you with valuable knowledge and expertise directly from our team of diagnostic professionals! Over the next several months, we’ll be sharing insightful articles, videos, and detailed guides from our top experts, offering an in-depth look at various topics in assay design and development, industrial design and engineering, manufacturing and technology transfer, and clinical research services—a masterclass in diagnostic development and commercialization. Turn! Turn! Turn! Seasonal Clinical Trials: Timely Planning and Execution for IVD Studies Science Meets Art in DCN Dx Lateral Flow Training Courses DCN Dx’s Clinical Research Services Merge the Sponsor and CRO Experience We understand that navigating the complex world of diagnostic development and commercialization can be a daunting task. Each new installment will offer you valuable insights and actionable strategies for overcoming the challenges that diagnostics companies experience. We’ll answer questions like: How do I choose appropriate materials and reagents for my assay? How can I optimize my sample collection device for specific use cases? What factors should I consider when optimizing my LFA for cost and performance? When should I start thinking about clinical trials? What are the latest reader technologies and quantification methods available, and how do they compare in terms of accuracy, sensitivity, and specificity? What are some key considerations for achieving successful regulatory submissions? Our goal is to empower you with the knowledge and resources you need to turn your innovative ideas into market-ready products. By engaging with the “Expert Insights” series, you will not only gain access to the specialized expertise and experience of our team but also discover how our comprehensive approach to diagnostic development and commercialization sets us apart from other diagnostic CDMOs. I invite you to join us on this exciting journey. If you aren’t already subscribed to our newsletter, you can do that here to be among the first to receive each installment. I also invite you to follow us on LinkedIn for updates. Together, let’s revolutionize the diagnostics landscape and bring life-changing solutions to those who need them the most. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. We partner with both biotech start-ups and established industry leaders to develop state-of-the-art rapid diagnostic tests. Our clinical research services and engineering teams ensure that products are designed with a focus on usability, manufacturability, and high-performance capabilities. Our experience encompasses a range of segments and applications, including clinical point-of-care testing, veterinary diagnostics, bioprocessing, soil and water testing, food safety, force protection, and more. DCN Dx is committed to delivering innovative solutions and exceptional service to meet the diverse needs of our clients.com. --- ## Turn! Turn! Turn! Seasonal Clinical Trials: Timely Planning and Execution for IVD Studies URL: https://dcndx.com/insights/seasonal-clinical-trials-timely-planning-execution-ivd-studies/ Type: insight Published: 2023-04-20 (As published 2023-04-20; regulatory status may have changed since.) Understanding the Seasonality of Clinical Trials Most people think of the seasons in terms of the standard farming seasons: spring, summer, autumn, and winter. In the spring months, farmers plant their crops. In the summer, they nourish and protect these fledgling crops as they grow into adulthood. In autumn, the crops are harvested and stored or sold. In winter, finally, the farmers have some rest. Similarly, clinical research also has its seasonality, particularly for studies of diseases with seasonal surges, such as cold and flu, seasonal allergies, and certain illnesses related to infected soil or water. From 2020 through much of 2022, the focus on infectious diseases was mainly on the year-round, highly communicable SARS-CoV-2 pandemic. As the pandemic’s peak subsided and people relaxed their prophylactic cautions, we experienced a record influenza season in December 2022. Consequently, we saw a resurgence of clients interested in bringing novel upper respiratory infection diagnostics through the FDA regulatory process. Unfortunately, many clients wait until the cold and flu season is imminent before seeking engagement with a clinical research organization (CRO). The start of the cold and flu season (October in the Northern Hemisphere) is too late to engage a CRO, as clinical trials require detailed planning. For products intended to diagnose seasonal illnesses like upper respiratory infections, it’s best for sponsors to “plant” the idea with the FDA in spring by getting feedback via pre-submission. Clinical Trial Start-up and Execution: Timing is Key Summer is the time to engage your clinical research partner organization to begin start-up and “nurture and grow” your clinical program so that you can “harvest” your study results through active enrollment and testing in the autumn and winter months. Early start-up phase tasks in a clinical trial include site identification and contracting, protocol development and institutional review board (IRB) review, and data management planning and database design and development. For a simple study with a clear and detailed clinical plan and minimal case report form/data capture requirements, these tasks can be accomplished in as few as 2 months. For more complicated studies or those requiring more overall plan development, a 3 to 4-month start-up period may be necessary. Working backward from the official start of flu season in the US in October, the latest a sponsor should engage in start-up activities for cold and flu season-related products is August. Ideally, a sponsor would be ready to start enrollment in September to catch the earliest cases, bringing the kick-off of study start-up back into July. If a sponsor were to engage a partner in September or October to run a study for a flu diagnostic product, they would likely miss the entire first half of the season, making it less likely to meet enrollment goals and potentially forcing alternative enrollment strategies. Choose the Right Clinical Research Partner for Seasonal IVD Studies In June and July, while the general public is planning summer barbecues, vacations, and weekends at the beach, the clinical research team at DCN Dx is ready to start working on your clinical study to ensure a kick-off on “Opening Day” of flu season. Remember, “To everything, there is a season,” and planning is crucial for the timely execution of seasonal clinical trials. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. Our comprehensive clinical validation processes and strategic regulatory planning will ensure your project is fully compliant and on track for success. --- ## DCN Dx’s Clinical Research Services Merge the Sponsor and CRO Experience URL: https://dcndx.com/insights/clinical-research-services-sponsor-cro-perspective/ Type: insight Published: 2023-04-19 Choosing the ideal clinical research organization (CRO) is vital for the successful execution of any clinical study. However, selecting a CRO—let alone one suited to studies for in vitro diagnostic (IVD) products—can be challenging for sponsor organizations. During the selection process, sponsors assess potential CROs for their clinical research services, IVD expertise, cost, and collaboration capabilities. Despite thorough evaluations, a sub-optimal choice can result in delayed timelines or increased expenses. DCN Dx understands this challenge because many of our clinical research team members have worked at various sponsor organizations. Our goal is to provide comprehensive clinical research services for IVD products that integrate the sponsor’s perspective, ensuring a smooth and efficient partnership from Day 1. DCN Dx Clinical Research Services: Combining Sponsor and CRO Expertise Reflecting on my experiences at previous sponsor organizations, I recall the difficulty in finding CROs with adequate knowledge of IVD clinical studies. Most CROs specialized in pharmaceutical or traditional medical device studies, and lacked the expertise required for IVD-specific nuances. This limitation often led to sponsors retaining many responsibilities that could have been delegated to a more specialized CRO. Our clinical research team consists of IVD industry experts who understand how to tailor our services to the specific needs of each study, sponsor, and product. DCN Dx’s IVD-focused clinical research procedures and systems address the unique requirements of IVD clinical studies. We offer a comprehensive suite of client services, ranging from study start-up to execution and closure, while also managing tasks typically reserved for sponsors, such as developing clinical strategies. Our services encompass study design, sample size determination, site identification and management, monitoring, meaningful data capture, database building and validation, statistical analysis, as well as other more personalized requests. The DCN Dx clinical research team has extensive experience in crafting clinical strategies for IVD products, which enables us to work seamlessly alongside sponsors as an integrated extension of their teams. We routinely apply lessons learned from past sponsor projects, addressing “common pitfalls” from the very beginning, such as regulatory compliance, clinical validation, sample collection and handling, user training, and cost-effectiveness. Addressing These Common Challenges in IVD Clinical Strategies: The DCN Dx Advantage In the complex world of IVD clinical studies, there are several common challenges that need to be addressed early to ensure a successful outcome. Our cross-functional team of experts is well-versed in these challenges and has developed strategies to mitigate their impact on clinical studies. Below are a few relevant examples: Regulatory compliance: Navigating the regulatory landscape is crucial for any IVD clinical study. DCN Dx ensures that your IVD product adheres to the necessary regulatory requirements and guidelines for your target market. Our team stays updated on the latest regulations and provides guidance on the submission process, documentation, and any necessary preclinical or clinical data requirements. Clinical validation: DCN Dx assists in designing and conducting well-structured clinical studies to confirm the diagnostic accuracy, clinical sensitivity, and specificity of your IVD product. These studies are designed and conducted by a team with working knowledge specific to IVD studies. We understand the differences in data capture and structure, designing the study based on the intended use population and setting. Sample collection and handling: Samples to an IVD study are essentially what subjects are to a pharma study. With this in mind, we establish appropriate procedures for sample collection, storage, and transport to maintain integrity of the samples and ensure accurate test results. We also look at how the samples can be utilized beyond the initial need. For example, can the samples be used to develop other products/assays for the sponsor? Can the samples be tested as part of a sponsor’s research publication? Sponsors often do this for efficiency and cost savings. User training: Our team leverages their laboratory experience to develop comprehensive training programs that integrate seamlessly into users’ workflows. Our training materials cover not only proper sample collection, handling, and interpretation but also GCP compliance, ensuring successful study outcomes. By training users on study-specific procedures, we ensure the generation of a robust dataset that meets GCP standards. By addressing these common challenges and leveraging our specialized IVD clinical research experience and network, DCN Dx helps ensure that your IVD clinical study is set up for success from the very beginning. We work closely with you to tailor our approach to your unique needs, ensuring a seamless partnership that delivers exceptional results. Stress-Free IVD Clinical Research Services DCN Dx’s clinical research services team recognizes the critical role of a suitable CRO partner in ensuring the success of any clinical program. With a sponsor-centric approach, our team excels in providing creative solutions that contribute to successful, cost-effective clinical programs. We understand that sponsors invest considerable time and effort into selecting a CRO, and the right partnership can significantly enhance the efficiency and success of a clinical program. DCN Dx aspires to be the CRO that our “sponsor-selves” were always seeking. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. Our comprehensive clinical validation processes and strategic regulatory planning will ensure your project is fully compliant and on track for success. --- ## How Lyophilized Beads Are Revolutionizing Point-of-Care Diagnostics URL: https://dcndx.com/insights/lyophilized-beads-revolutionizing-diagnostics/ Type: insight Published: 2023-03-27 For years, lyophilization has been employed in the diagnostic and pharmaceutical industries to prepare reagents in vials or bulk trays. However, these formats are not ideal for point-of-care diagnostics. Lyophilized beads (also known as “lyo beads”) have emerged as a solution, offering easier handling while preserving the advantageous properties of the lyophilization process. Lyo beads extend the shelf life of biological molecules, simplifying storage and transportation. Their compact size reduces overall costs compared to traditional glass vials or microfluidic-encased lyophilized reagents. Moreover, lyo beads provide superior rehydration characteristics, contributing to an improved performance-to-cost ratio. They also minimize cold chain requirements and stabilize sensitive biological materials. In recent years, lyo beads have gained traction for use in point-of-care diagnostics. Initially developed for PCR master mixes in molecular diagnostics, rapid diagnostics and point-of-care assay developers have started to recognize their unique benefits, such as rapid rehydration, easy handling, and minimal liquid handling steps. Lyophilized Beads in Molecular Diagnostics Lyophilized beads are versatile and their applications diverse, capitalizing on benefits like biological stabilization, cold chain storage elimination, and quick rehydration. They typically contain liquid volumes of 5 to 40µL, while maintaining high reproducibility. In molecular diagnostics, lyo beads can be used to stabilize enzymes and reagents for PCR or LAMP (loop-mediated isothermal amplification). They can be stored in simple 0.2mL PCR tubes or integrated directly into microfluidic cartridges for automated testing at point-of-care or home locations. As automation improves, the placement of lyo beads into disposable microfluidic cartridges is becoming an industry standard. Lyophilized Beads in Point-of-Care Diagnostics Lyo beads can be utilized for point-of-care immunoassay diagnostics in various ways, including sample pretreatment, sample incubation, and air-dried conjugate replacement. They are an efficient method for adding sensitive biologicals or non-sensitive reagents to a sample without dilution, streamlining liquid handling steps and taking advantage of bead-dispensing equipment’s precision. Lyo beads’ rapid rehydration mimics liquid reagent transfers without causing dilution or pipetting errors, minimally impacting the assay workflow. Lyo beads can also enhance incubation time between the nanoparticles (or other reporter labels) and the analyte contained in the sample. This can be achieved by adding the lyophilized bead (containing the reporter label) directly to the sample before adding it to the diagnostic assay, thereby extending the incubation time to maximize assay sensitivity within the given time constraints. This method also improves assay precision, yielding lower assay CVs when using lyo beads compared to air-dried conjugate or microfluidic time gating. By replacing the conjugate pad with a lyophilized bead, conjugate rehydration and nanoparticle release can be enhanced, offering performance similar to lyophilizing the entire conjugate pad without the need for the entire pad’s freeze dryer footprint. This approach reduces costs while maintaining high performance of your immunoassay. DCN Dx Optimizes Your Reagent Lyophilization Process At DCN Dx, our proficiency in assay development, reagent lyophilization, and engineering results in a coordinated approach to your diagnostic’s development. Our collaborative environment enables the seamless integration of lyophilized reagents into assays and devices. Our team of accomplished scientists and engineers works in tandem to bring your product vision to life, ensuring that the characteristics of lyophilized beads, such as diameter/volume, physical robustness, and rehydration properties, operate within the fluidic parameters of the device and conform to space constraints and manufacturing methods. Our meticulous development process for lyophilized beads adheres to standard design control procedures, encompassing feasibility, verification, and validation phases prior to transitioning to manufacturing. We initiate assay development with wet chemistry before transitioning to lyophilized reagents and integrating them directly into the final device. This streamlined approach consolidates all processes within a single development program at one site, ensuring a unified source of accountability. Choose DCN Dx to incorporate lyophilized bead technology into your existing or new diagnostic assay. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. We partner with both biotech start-ups and established industry leaders to develop state-of-the-art rapid diagnostic tests. Our clinical research services and engineering teams ensure that products are designed with a focus on usability, manufacturability, and high-performance capabilities. Our experience encompasses a range of segments and applications, including clinical point-of-care testing, veterinary diagnostics, bioprocessing, soil and water testing, food safety, force protection, and more. DCN Dx is committed to delivering innovative solutions and exceptional service to meet the diverse needs of our clients.com. --- ## Science Meets Art in DCN Dx Lateral Flow Training Courses URL: https://dcndx.com/insights/science-meets-art-lateral-flow-training-course/ Type: insight Published: 2023-03-21 As someone who’s been in the lateral flow diagnostics industry for a few years now, I can say that working at DCN Dx as an assay development scientist is one of the most rewarding experiences I’ve ever had. Not only is the subject matter fascinating, but the opportunity to work with high-caliber peers has propelled my understanding of lateral flow science and the art behind it. I can hear it now: “But Melanie—it’s science. How can there be art involved?” I understand that science can be perceived as a stiff topic. Follow a protocol, and that’s it. Not a lot of room for creativity there. The truth is, it’s more than that. At DCN Dx, we challenge the norm to ensure the tests that we create meet the highest standards. This is where the “art” comes into play: we explore all options to create quality tests that are suitable for our clients and, in turn, the end user. Imagine for a moment a lone violinist playing a beautiful song. It sounds lovely on its own, but when you add the other string instruments—the brass, woodwinds, percussion, and such—the music takes on a whole new meaning. It’s the same way with lateral flow. You have your target (the violin). When you add materials and reagents (the other instruments), you witness the way the components work synergistically to generate a whole working test. And this is where the art is, because no two lateral flow products are the same. Like songs, each test has nuances, varying behaviors, and even “personalities.” We use our collective experience to bring out the best in these reagents and materials. This is our art. At DCN Dx, I’ve gained valuable knowledge in both the art and science of diagnostics. Applying this expertise to the development of tests that facilitate care has become a true passion of mine. I don’t want to keep all this valuable information to myself. I believe knowledge should be shared and ideas cultivated. As DCN Dx’s Training Specialist, I have the honor of sharing this knowledge, teaching others, and helping them apply it to their own lateral flow development work. It’s incredibly fulfilling. Learning the Art and Science of LFA Development with DCN Dx Training Courses Lateral flow tests are simple to use, fast, and cost effective, making them an ideal choice for many applications. In fact, lateral flow is widely used in healthcare, veterinary medicine, food safety, environmental testing, and many other industries. That range of applications is one of the reasons I love teaching DCN Dx’s Basic Lateral Flow Course. If you work in an industry that requires rapid and accurate testing, lateral flow can be a valuable tool. It can quickly detect infectious diseases like COVID-19 and the yearly flu, allowing healthcare providers to act faster and prevent the spread of the disease. In veterinary medicine, lateral flow tests can diagnose diseases in animals, allowing for earlier treatment and better outcomes. And in food safety, lateral flow tests can be used to detect contaminants such as bacteria or toxins, helping to ensure that food products are safe for consumption. Lateral flow is constantly evolving. New technologies and innovations are being developed all the time, and staying up to date with the latest developments is essential for anyone working in the field. Because it is ever evolving, the learning continues and you never feel stagnant. When I teach Basic Lateral Flow Courses, I’m able to help people from a wide range of backgrounds and industries learn how to use this powerful tool to improve their work and achieve their goals. Whether they are healthcare providers, veterinarians, food safety professionals, or researchers, lateral flow can be a valuable addition to their toolkit. It’s a fascinating subject, and it’s accessible to anyone who’s interested in learning more. By sharing my knowledge and expertise with others, I’m able to help people achieve their goals and make a real difference in their work. If you are interested in learning more about lateral flow and how it can be applied to your own work, I encourage you to consider taking DCN Dx’s Basic Lateral Flow Course. You’ll have hands-on experience with this tried-and-true technology, and personally, I’d love to help you develop your own lateral flow assay. We hope to see you in Carlsbad, California, soon (the weather and the beaches are gorgeous)! Learn more about DCN Dx’s Basic Lateral Flow Training Course her e. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. We partner with both biotech start-ups and established industry leaders to develop state-of-the-art rapid diagnostic tests. Our clinical research services and engineering teams ensure that products are designed with a focus on usability, manufacturability, and high-performance capabilities. Our experience encompasses a range of segments and applications, including clinical point-of-care testing, veterinary diagnostics, bioprocessing, soil and water testing, food safety, force protection, and more. DCN Dx is committed to delivering innovative solutions and exceptional service to meet the diverse needs of our clients.com. --- ## Faster and More Efficient Detection of Varicella Zoster Virus (VZV) Infections URL: https://dcndx.com/insights/lateral-flow-varicella-zoster-virus-vzv-assay/ Type: insight Published: 2023-03-15 Improved VZV Antibody Detection with a New Lateral Flow Assay Varicella zoster virus (VZV) is a member of the herpesvirus family that can infect humans and cause chickenpox (mostly in children and young adults) and shingles in adults. The ability to accurately detect the presence of VZV specific antibodies is important for identifying individuals who may be at risk for complications or who require vaccination. Diagnostic tests allow healthcare professionals to efficiently screen children, adolescents, and adults for their immunity status by detecting the levels of VZV antibodies in a person’s blood circulation. The current standard to detect previous infection (or vaccination) of VZV is through serologic IgG tests. Currently, enzyme-linked immunosorbent assays (ELISAs) are used to detect VZV antibodies in human serum samples; however, the collection, transport, and processing of blood samples required for ELISAs can be time-consuming and costly. A collaboration between Viro Research, DCN Dx, and Partners in Diagnostics has led to the development of a lateral flow assay (LFA) that detects VZV antibodies more efficiently using human whole blood via fingerstick collection. The researchers published their findings in the Journal of Immunological Methods and the paper describes the development and testing of the VZV LFA. What Were the Results? Compared to conventional ELISA testing, the VZV LFA is a faster and simpler point-of-care test that can be used to detect VZV antibodies in human capillary blood using a fingerstick collection. The analytical performance and clinical study results showed that the VZV LFA is comparable to or better than ELISA testing. Initial findings indicate that the VZV LFA is also robust enough to be used in CLIA-waived point-of-care facilities by untrained healthcare workers. The new VZV LFA test represents a significant advancement in VZV antibody detection, enabling faster and more efficient detection of previous VZV infections. This development has important implications for healthcare providers, as it provides a more accessible and convenient method for detecting VZV antibodies in patients who may require vaccination or monitoring for complications associated with VZV infections. About DCN Dx DCN Dx is a global leader in the design, development, and manufacture of point-of-use diagnostics. Based in Carlsbad, California, our multidisciplinary team of scientists and engineers collaborates to create and integrate comprehensive assay systems, consumables, and instruments tailored for point-of-use applications. These reliable systems empower users to detect and quantify biomarkers and pathogens, thereby facilitating the timely delivery of treatments. We partner with both biotech start-ups and established industry leaders to develop state-of-the-art rapid diagnostic tests. Our clinical research services and engineering teams ensure that products are designed with a focus on usability, manufacturability, and high-performance capabilities. Our experience encompasses a range of segments and applications, including clinical point-of-care testing, veterinary diagnostics, bioprocessing, soil and water testing, food safety, force protection, and more. DCN Dx is committed to delivering innovative solutions and exceptional service to meet the diverse needs of our clients.com. --- ## Developing Point-of-Use Diagnostics with Integrated Engineering URL: https://dcndx.com/insights/developing-point-use-diagnostics-integrated-engineering/ Type: insight Published: 2022-09-14 (As published 2022-09-14; regulatory status may have changed since.) Many diagnostic device developers have experienced that heart-stopping moment: the final piece of the product puzzle doesn’t fit. For medical devices, a simple mismatch of discrete elements at the end of a development cycle can be catastrophic and require huge costs and long time frames to rectify. Oftentimes, these issues arise from inefficient organization, a lack of foresight during development, minimal communication, and development silos. DCN Dx understands and proactively eliminates these problems through an integrated approach to point-of-use diagnostic product engineering. Engineering Comprehensive Point-of-Use Diagnostic Solutions Point-of-use diagnostic devices analyze glucose and cholesterol levels, perform electrolyte and enzyme analysis, and test for drugs of abuse, pregnancies, and infectious diseases. Point-of-use diagnostic devices are also used to test blood gases and cardiac markers and perform fecal occult blood tests. There are several advantages to doing the tests at the point of use, including quick results and faster implementation of therapy if needed. Selecting the Right Point-of-Use Diagnostic Development Partner When looking for a point-of-use diagnostic development partner, it’s important to ensure that they have the expertise and delivery record to tackle all aspects of development. Your diagnostic developer should understand the shifting regulatory landscape and be flexible enough to mold their clinical and engineering services to your application. DCN Dx understands that the greatest concern of any diagnostic development is that the assay reliably solves the right problem and achieves FDA approval. However, DCN brings much more value to the development and leverages a unique combination of technical, regulatory, and business expertise to take your diagnostic device from concept to market success. DCN Dx’s engineering services cover the design of mechanical systems, electronic equipment, software, optical systems, consumables/cartridges, and reader development. We also customize our base reader—the miniDxR for integration with lateral flow and point-of-care diagnostics, along with the design of lab-on-a-chip instruments, optical analyzers, PCR and isothermal systems. DCN Dx is flexible enough to shape our development services around your project’s requirements. For example, we design diagnostics for high-throughput applications without compromising usability or performance. Our flexible services are modular, allowing you to select the clinical, engineering, or regulatory services you need to round out your diagnostic development. You can bring your point-of-use diagnostic device into the development cycle at any point—from feasibility, design, and development to clinical validation, regulatory submission, and product launch. Say ‘Goodbye’ to Silos Silos can lead to a loss of productivity, increased costs, and a lack of efficacy for point-of-use product development. This occurs when product, engineering, and manufacturing teams working on the same diagnostic device (or from different companies) function independently and communicate ineffectively, leading to continual readjustments that increase materials, costs, and labor while lowering productivity. DCN Dx eliminates silos by integrating assay development, engineering, and manufacturing teams to prepare assays for use in cartridges and instruments from the conceptual phase. We serve as an all-in-one partner that provides centralized product development, eliminating the need to manage multiple developers for your diagnostic device. Transparent Communication throughout Development We understand the frustration of waiting for updates on your diagnostic development project. As an all-in-one development and manufacturing partner, DCN Dx serves as the single source of accountability for your point-of-use device. We keep you informed throughout the entire development chain, ensuring transparent communication and no uncertain forecasts or lead times. An Efficient Approach to Development From the outset, we “examine the seams” in the development of your point-of-use product, leveraging our expertise in assay, instrument, software, cartridge, and sample handling. Our integrated approach combines engineering, manufacturing, logistics, and clinical research, allowing us to navigate project milestones seamlessly through efficient communication channels, standardized design formats, and transfer protocols. Reduced Risk for Point-of-Use Product Development DCN Dx leverages over two decades of experience in lateral flow, flow-through, microfluidic, and other formats to de-risk your assay development projects and work efficiently towards milestones. Our integrated teams work together via standardized design formats and transfer protocols to ensure that every one of your requirements are satisfied from concept to commercialization. Our product development lifecycle starts early by integrating the assay into the cartridge and instrument. Our unique end-to-end development capabilities allow us to identify and address interface problems before they become challenges. For example, clients often approach us with requirements for unique cartridge geometry. The lateral flow reader’s visual field of view must be matched to the relevant area of its cartridge. Our rapid customization options and engineering services allow us to create functional designs that proactively identify potential late-stage problems and optimize the final result. DCN Dx’s integrated approach to point-of-use diagnostic product engineering ensures seamless IVD product development. By eliminating silos, streamlining communication, and leveraging our diverse expertise in engineering, manufacturing, logistics, clinical research, and regulatory management, we deliver innovative and reliable point-of-use diagnostic devices that make a difference in patient care. Partner with DCN Dx for your next point-of-use diagnostic project and experience a transparent, efficient, and risk-free development process that will take your device from concept to market success. Together, we can revolutionize point-of-use diagnostics and make a lasting impact on the world of healthcare. --- ## Lateral Flow Assays, COVID-19, & Emergency Use Authorizations (EUA) URL: https://dcndx.com/insights/lateral-flow-assays-covid-19-emergency-use-authorizations-eua/ Type: insight Published: 2022-05-19 (As published 2022-05-19; regulatory status may have changed since.) DCN Dx applies their expertise in clinical research to partner with developers of diagnostic devices to achieve EUA approval and then fulfill the clinical testing requirements for normal market clearance. DCN Dx’s clinical research services team prepares your diagnostic device for market approval under EUA and 510(k) conditions. From clinical research to FDA liaising, DCN Dx’s integrated services guide your device to market launch. Download our whitepaper, “Lateral Flow Assays, COVID-19, & Emergency Use Authorizations (EUA),” using the form below to learn more. --- ## Achieving Early Detection of HIV URL: https://dcndx.com/insights/achieving-early-detection-hiv/ Type: insight Published: 2022-03-08 Dry Chemistry Testing: Part 3 of 3 In the developed world, HIV has become more manageable since the early 1980s. It is no longer the automatic death sentence that it once was: for many, it is today a chronic disease managed with anti-retroviral therapy (ART). Limitations of HIV Detection for the Developing World Unfortunately, many low-to-middle-income countries (LMICs) miss out on early and effective ART treatments. Sub-Saharan Africa alone accounted for almost 40% of new HIV infections worldwide in 2020 [1]. Many people that live with HIV are unaware of their infection until their depleted immune systems can no longer protect them from viral, bacterial, and fungal diseases that those with normal-functioning immune systems deal with easily. To detect and monitor the presence of HIV in humans, the developed world uses two tests. Reverse transcriptase polymerase chain reaction (RT-PCR) tests are molecular tests that are used to detect and quantitate the amount of HIV virus in a patient’s blood. However, they cannot tell how much damage has been done to the patient’s immune system. For that assessment, medical professionals need to know how many CD4 T cells are in the patient’s blood. Flow cytometry is used to measure how many CD4 T cells are in the patient’s blood. Uninfected patients generally have between 500 and 1,200 CD4 T cells per cubic millimeter of blood in teens and adults. HIV patients have decreased CD4 T-cell counts and various therapies are started or stopped based on this. Both of these tests are primarily limited to the developed world because they both require expensive equipment, stable electrical power, and trained personnel to operate effectively. Locations in LMICs often lack all three. An additional problem with absolute CD4 counts is that they vary with age. Young children can have CD4 cell counts as high as 3,000 CD4 cells/mm3. Many cases of HIV in infants and babies are from exposure to the virus from their HIV-infected mothers during pregnancy, at birth, or during nursing. A seemingly normal CD4 count in a young child may actually reflect CD4 depletion due to HIV. One strategy that has been used to normalize CD4 counts is to report them as a ratio of CD4 to total white blood cells. In healthy people of all ages, the percentage of CD4 (CD4%) has a normal range of 25–65%. A CD4% value of 12–15% is roughly equivalent to an absolute CD4 cell count level of 200 CD4/mm3, which is the level at which current guidelines recommend commencement of ART. Delayed or deferred ART is associated with poorer outcomes and premature death. One of the complications of measuring CD4% is that the CD4 marker protein is also found on white blood cells other than CD4 T cells, albeit in smaller amounts. As CD4 T-cell counts drop in HIV infection, the contribution of these other white blood cells can skew results and make it look like the patient has more CD4 T cells than they actually have. Flow cytometry deals with this problem by measuring other properties of cells (e.g., size and roughness) and ignoring cells that are not T cells. Simple Dry Chemistry Tests to Detect HIV PortaScience, Inc. of Monmouth, New Jersey (now a subsidiary of DCN Dx), has developed a dry chemistry card test for the determination of CD4% in finger-stick blood. This test requires only simple equipment and a minimally trained operator. The test is run in two stages. In the first stage, an unmeasured amount of finger-stick blood is added to a test tube that contains pre-dosed, freeze-dried reagents. One of them is EDTA to keep the blood from clotting. The second is a monoclonal antibody to the CD4 that has been labeled with the enzyme beta-galactosidase (beta-gal). The third reagent is a magnetic particle that has been coated with another monoclonal antibody to a protein called CD15. This CD15 marker is found on monocytes and neutrophils, the white blood cells that also express CD4. After a very brief incubation, the magnetic particles are separated from the supernatant liquid by application of a magnetic field, thus removing the interfering white blood cells. CD4 T cells do not express CD15, so they (and the majority of the remaining white blood cells that are also CD15 negative) remain in the liquid phase. Next, two small aliquots of the liquid are placed on membranes that have been specially formulated to carry a slight positive charge. White blood cells are positively charged and are retained by the membrane. Red blood cells and unbound anti-CD4 labeled with beta-gal are washed away. One area of the membrane has also been saturated with a substrate that is specific for beta-gal. Another area of the membrane has been saturated with another substrate that specifically reacts with an enzyme called leukocyte esterase that is endogenous to all white blood cells. Color development ensues as the two enzymes react with their substrates. The beta-gal domain produces color in direct proportion to the CD4 T cells present in the depleted liquid, and the leukocyte esterase domain produces color in direct proportion to the white blood cells in the depleted liquid. The signal can be read and quantitated with a digital camera or, ideally, with a cell phone camera. Cell phone adoption in LMICs has been rapid. It was estimated in 2015 that 94% of adults in those economies have access to cell phones [2]. These devices not only provide a camera to capture the image generated in the test described above—they also provide the connectivity to allow sophisticated image analysis and medical reporting of the results. Because the readout of this test is a ratio, there is some self-correction that makes the test more robust in the hands of minimally trained operators. Slight variations in the amounts of blood added to the first tube or in the amount of pretreated liquid added to the membranes are cancelled out. The medical benefit of this information is that ART can be initiated when the CD4% declines to the trigger level of 12–15%. As noted above, ART is key to the survival of HIV patients and is critical in their quality of life. Since many ARTs are self-administered by the patients, compliance with the regimen can also be monitored. CD4% percentages rise with ART. Declining CD4%s do not lie, but patients around the world often do. This development has focused on a particular type of white blood cell, but in principle this technology could be used to detect and possibly quantitate other cellular targets in a wide variety of biological liquids. PortaScience has also developed tests for white blood cells in human blood and in cow’s milk. Elevated white blood cells in blood and milk are indicative of infection with bacteria and viruses. Conclusion If there is a project in your development portfolio that would benefit from a simple dry chemistry card test for a cellular target, consider a confidential conversation with the professionals at PortaScience. The talented scientists and manufacturing experts there can complement your in-house development team. Together, they may be able to arrive at solutions that neither group could develop in insolation. This expanded universe of solutions is why scientists follow the relevant literature and attend scientific conferences. A consultation with PortaScience is a more focused way of gathering information. Be sure to check out Part 1 and Part 2 of this series on dry chemistry testing. DCN Dx is a globally recognized contract developer of point-of-care devices based in Carlsbad, California. Since its founding, DCN Dx has committed itself to furthering the rapid diagnostic and point-of-care test market through the continued evolution of technologies and applications. DCN Dx’s cross-functional team of scientists and engineers develop and integrate all aspects of the point-of-care device system, including complex binding reactions, cassettes, sample handling devices, and reader systems. The company assists clients in developing entire rapid diagnostic tests from concept to commercialization. [1] https://www.unaids.org/en/resources/fact-sheet [2] https://www.usglc.org/blog/the-technology-thats-making-a-difference-in-the-developing-world/ --- ## Catching Toxic Alcohol Poisoning in Time to Act URL: https://dcndx.com/insights/catching-toxic-alcohol-poisoning-time-act/ Type: insight Published: 2022-03-08 Dry Chemistry Testing: Part 2 of 3 In the first blog in this three-part series, we saw how a dry chemistry card could help determine when a woman was most likely to get pregnant. For reference, a few drops of saliva are added to the card, where a color change indicates the period of ovulation in the next five days. This blog discusses how a dry card chemistry assay can determine if a person or a pet has ingested a toxic alcohol. The Dangers of Toxic Alcohol Poisoning Data from the American Poison Control Center’s National Poison and Exposure Database reveals that more than 93,000 cases of toxic alcohol poisoning occurred in 2010. Close to 2,000 of these patients were classified as experiencing major morbidity. Sixty-one of these people died. The same database reports that many of these accidental poisonings occurred in a residential setting, and nearly half of the victims were under the age of five. Methanol and ethylene glycol are the two most common causes of toxic alcohol poisoning. Methanol (aka, “wood alcohol”) is found in automobile windshield-washer fluid. Similarly, ethylene glycol is the major component of automobile antifreeze fluid. Both products are often packaged as brightly colored liquids and are reported to have a sweet taste. It is easy to see that young children and pets might be tempted to drink them. The danger that these chemicals pose is due to toxic metabolites that form in the body after ingestion. If treatment is not available or is delayed, the mortality rate of methanol poisoning can be as high as 50–80%. Ethylene glycol is readily absorbed in the gastrointestinal tract, and neurological damage can occur within as little as 30 minutes post-ingestion. Current Methods for Toxic Alcohol Poisoning The gold standard for the diagnosis of toxic alcohol poisoning is gas chromatography in blood. Gas chromatography (GC) is well-established technology that is most commonly found in industrial and academic research labs, but not in most hospital labs. The equipment is relatively expensive and is best used by well trained professionals. The columns in GCs are often run at elevated temperatures and frequently employ expensive helium as the carrier gas. GCs are not instruments that can be left unused for long periods of time and suddenly pressed into service. Accurate calibration for various compounds require a GC instrument to be thermally equilibrated, and that takes time. Immunoassays for these small molecules are impractical, as they are far too small to be immunogenic on their own. Even if they were covalently attached to a carrier protein (e.g., bovine serum albumin), it is not clear that useable antibodies could be generated that would discriminate between these closely related compounds. Physicians are therefore left to manage their intoxicated patients empirically by monitoring symptoms, osmolar gap, acidosis, and anion gap measurements. These measurements are less precise and more difficult to interpret than direct toxic alcohol measurements in blood. There are two therapies that can be employed to mitigate the effects of toxic alcohol poisoning. The first is hemodialysis using the same devices that are used to treat patients in renal failure. The other therapy is less obvious: intravenous ethyl alcohol infusions. Ethyl or grain alcohol is the physiological component of alcoholic beverages. It competitively blocks the metabolism of the toxic alcohols so that they can be excreted in urine in their original, unmetabolized form. Detecting Toxic Alcohol Poisoning with Dry Chemistry Tests PortaScience, Inc., of Moorestown, New Jersey, was recently acquired by DCN Dx for their expertise in dry chemistry reagents. One PortaScience-developed project is a point-of-care assay for ethanol, ethylene glycol, and methanol. It is based on the use of enzymatic pathways that are specific for each of these compounds. Each enzymatic pathway utilizes an electron transfer system that reacts with an immobilized substrate that produces a color change that is proportional to concentrations of ethanol, ethylene glycol, and methanol. For ethylene glycol, the enzyme is glycerol dehydrogenase. This enzyme reacts with ethylene glycol in the sample in the presence of the cofactor NAD+ to generate NADH. The NADH is subsequently reacted with another enzyme—diaphorase—and it converts a chromogenic substrate into a colored product. This substrate is 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT). It is yellow in its original form and turns a dark purple on reduction with NADH. The purple color is directly proportional to the concentration of ethylene glycol in the sample. For methanol, the enzyme used is alcohol dehydrogenase. In the presence of NAD+, this enzyme also generates NADH and formaldehyde. Formaldehyde in the presence of atmospheric oxygen reacts enzymatically with diaphorase with another chromogenic substrate called Purpald. This generates a purple color that is proportional to the concentration of methanol in the sample. The ethanol assay uses the same enzymes as the methanol assay, but the chromogenic substrate is MTT, the same substate used in the ethylene glycol assay. Determination of the ethanol concentration is useful in monitoring therapy in the presence of the toxic alcohols. These systems were first evaluated in solution-phase chemistry and subsequently transferred to a nylon membrane with a pore size of 0.45 microns to make a dry chemistry assay. The nylon membrane was first coated with the appropriate chromogenic substrate in the appropriate buffers to maintain pH. After drying, a second coat consisting of the necessary enzymes, cofactor and surfactants are then applied over the chromogenic substrate. The assay is run by applying 15 microliters of sample (serum or whole blood) to the coated card. Color develops within a few minutes and is read with a handheld reflectance meter equipped with a 565 nm LED light source. The reduced chromogenic substrates undergo a shift in their absorption maximum from 405 nm (yellow) to 565 nm (purple). The range of these assays is from 10 mg/dL to 200 mg/dL. Ethylene glycol has a toxicity threshold of 25 mg/dL, and methanol’s toxic threshold is 20 mg/dL. As was the case with the dry card assay for calcium and magnesium in saliva for the monitoring of ovulation, a lot of work goes into making these tests stable and reproducible. Enzymes are proteins, and as such are subject to denaturation on coating on a synthetic polymer such as nylon. Various stabilization strategies have been employed such as the use of hydroxy propyl cellulose, carboxy-methyl cellulose, polyvinyl pyrrolidone (PVP), and xanthan gum. Hungry bacteria and fungi that are omnipresent in the environment must also be considered and prevented from destroying the reagents before they can be used. A commercially viable product should have a shelf life of at least 12 months at refrigerator temperatures. Long-term storage at room temperature is preferred for ease of distribution and storage. Shipping stability is another consideration as brief exposure to temperatures over 100°F are not uncommon during summer in some locations. Conclusion If you need an assay for a small organic compound in a point-of-use setting, contact the experts at PortaScience. Their knowledge and experience could turn your needs into a reliable analytical tool. The examples in this blog demonstrate how different alcohols can be coupled to enzyme-cofactor-substrate combinations to produce a color change proportional to the concentration of the analyte. However, it is not hard to imagine how an enzyme inhibitor such as an insecticide or pesticide could be detected by a decrease in enzymatic activity. Even if you have a talented team of chemists and manufacturing personnel in-house, confidential discussions between teams of professionals can produce synergistic results not obtainable by one group working in isolation. Be sure to check out Part 1 and Part 3 of this series on dry chemistry. DCN Dx is a globally recognized contract developer of point-of-care devices based in Carlsbad, California. Since its founding, DCN Dx has committed itself to furthering the rapid diagnostic and point-of-care test market through the continued evolution of technologies and applications. DCN Dx’s cross-functional team of scientists and engineers develop and integrate all aspects of the point-of-care device system, including complex binding reactions, cassettes, sample handling devices, and reader systems. The company assists clients in developing entire rapid diagnostic tests from concept to commercialization. --- ## Optimizing Natural Fertility Through Ovulation Detection URL: https://dcndx.com/insights/optimizing-natural-fertility-ovulation-detection/ Type: insight Published: 2022-03-08 Dry Chemistry Testing: Part 1 of 3 In the past few generations, humans have taken unprecedented control of their own fertility. Much of the associated technology and scientific expertise has focused on limiting unwanted pregnancies. Hopeful parents-to-be, however, also have tools at their disposal: modern family-planning methods and a reasonable degree of diligence can allow them to manage the number of children they’d like to have, and when, with reasonable certainty. Unfortunately, there remain a considerable number of people who want to have children, but for one reason or another experience trouble conceiving. Fortunately, the likelihood of a successful pregnancy can be increased by understanding when conception is most likely to occur. Methods of Ovulation Detection Women are most fertile from a period 3–5 days before ovulation to 1–2 days after ovulation. This period is referred to as the “fertility window.” At Day 5 before ovulation, the probability of conception is about 8%. That probability doubles at Day 1 before ovulation, and almost doubles again on the day of ovulation. It then falls rapidly to 7% one day after ovulation. If a woman is trying to conceive, this information can be very useful. The challenge, therefore, is to determine when ovulation is about to happen. Historically, several methods have been used to determine the time of ovulation. Basal body temperature (BBT) measurements and determination of urinary luteinizing hormone (uLH) are probably the most common, but both have limitations. BBT is measured with a specialized thermometer with significantly more gradations than the typical fever thermometer. It reports temperatures between 94°F and 100°F in 0.1°F increments. The method requires multiple temperature readings over several days upon awakening and before the woman gets out of bed. Ideally, data are entered into a chart each day. Ovulation causes a 0.1°F increase in temperature. However, this only occurs on the day of ovulation, which means that the period of enhanced fertility a few days before ovulation are not detected. Basal body temperature can also be affected by factors other than ovulation such as emotion, lack of sleep, and smoking. One published study found that BBT was accurate only 32% of the time [1]. Luteinizing hormone levels in blood increase dramatically 20–48 hours prior to ovulation and can be detected in urine 8–12 hours after it occurs in blood. The levels are low enough that an immunoassay typically based on a monoclonal antibody must be used to detect the hormone. While commercially available over-the-counter (OTC) tests for uLH have been available for some time, they too suffer from some limitations. The surge is only detectable 1–2 days prior to ovulation, which once again misses some of the fertility window. Interferences include the use of drugs used in the treatment of infertility and menopause. (There is also the subjective undesirability of using urine as the sample.) Researchers have also identified changes in electrolytes in various body fluids that signal that ovulation is about to take place. Salivary conductivity changes up to six days prior to ovulation. But to increase reliability of the method, it is recommended that vaginal conductivity be measured as well [2]. Different ranges are given depending on whether the patient is taking fertility drugs. While the method is accurate, it does require the use of instrumentation which usually requires a learning curve. The cost of the instrumentation is also a barrier to adoption. Detecting Ovulation with Dry Chemistry Tests PortaScience, Inc., of Moorestown, New Jersey (now a division of DCN Dx) has developed a dry chemistry method for the prediction of ovulation. It is based on the observation that concentrations of calcium and magnesium ions in saliva drop significantly up to six days before ovulation. No instrumentation is necessary, as the test is formulated to be read by the naked eye. The test is simple to run. Saliva is collected in a small container, and an unmeasured amount of saliva is added to the dry chemistry test card containing the appropriate reagents. After a few minutes, there is a color change if ovulation is imminent. No instrumentation is required—the card is read with the naked eye and compared to a color chart. The method was deemed novel by the U.S. Patent Office, which resulted in the issuance of a patent [3]. This method is based on the use of chromoionophores, which are dyes known to change color in the presence of certain positively charged ions such as calcium and magnesium. These dyes are commercially available from reputable chemical manufacturers and have a long history of laboratory use. To provide a relevant threshold for the color change, ethylenediaminetetraacetic acid (EDTA) is added to the dyes. It has a higher affinity for calcium and magnesium and reacts with these ions before the ions can interact with the chromoionophore dyes. When the EDTA is saturated with these ions, then the color reaction can take place. Developing Reliable Dry Chemistry Ovulation Tests The chemistry described above makes an interesting science project, but much more is needed to turn that into a commercial product. Extensive experimentation was done to select the dyes, the card material, and the stabilizers and wetting agents in order for them to perform reliably. These formulations have also undergone stability testing to ensure that the card has the same performance in the hands of the user as it did in quality control testing at the manufacturer. The test was also field tested to ensure that essentially untrained users could get laboratory level results in their own homes. This ovulation prediction assay is only one example of the kind of testing that can be done in a dry card format. The concept can be readily expanded to be applied to a wide variety of ionic species in water-based samples for which appropriate chromoionophores are known. This could include not only human clinical samples, but also veterinary samples, environmental samples, food stuff, cosmetics, and pharmaceuticals, to name just a few. The company has also developed tests for negatively charged ions (e.g., a test of iodide in table salt). This test is read with the naked human eye, but simple instrumentation does exist to read these cards. This makes interpretation of the test results less subjective. Instrumentation brings with it the connectivity that is increasingly important in this digital age. The decision to make the test readable by eye or instrumentation is driven by several factors including cost and the frequency of testing. It makes little sense to add instrumentation cost and complexity to a test that is done only periodically. On the other hand, if many batches of animal feed need to be tested for heavy metals over periods of months to years, it is easier to justify the inclusion of instrumentation. Conclusion If there is a need for analytical chemistry testing for ionic species in a point-of-use setting, contacting the experts at PortaScience, Inc. is an excellent place to start. Even with a talented team of chemists and manufacturing experts in place, conversations with another group of professionals can expand the range of potential approaches. Be sure to check out Part 2 and Part 3 of this series on dry chemistry testing. DCN Dx is a globally recognized contract developer of point-of-care devices based in Carlsbad, California. Since its founding, DCN Dx has committed itself to furthering the rapid diagnostic and point-of-care test market through the continued evolution of technologies and applications. DCN Dx’s cross-functional team of scientists and engineers develop and integrate all aspects of the point-of-care device system, including complex binding reactions, cassettes, sample handling devices, and reader systems. The company assists clients in developing entire rapid diagnostic tests from concept to commercialization. [1] Maurizio Guida et. al. Efficacy of Methods for Determining Ovulation in a Natural Family Planning Program. Fertility and Sterility 72: 900-904 (1999) [2] Fernando, R. S., J. Regas, and G. Betz. Prediction of ovulation with the use of oral and vaginal electrical measurements during treatment with clomiphene citrate. Fertil. Steril. 47(3): 409-415 (1987) [3] U.S. Patent 5 914 271 --- ## Journal Article: “Lateral Flow Assays in Infectious Disease Diagnosis” URL: https://dcndx.com/insights/journal-article-lateral-flow-assays-infectious-disease-diagnosis/ Type: insight Published: 2022-01-31 Lateral flow immunoassays play an important role in infectious disease diagnostics. Advancements in technology have led to improved performance of these assays and acceptance by professional users. With the advent of the SARS-CoV-2 pandemic, the market has reached new levels requiring hundreds of millions of tests per year for professional and even home use. DCN Dx’s Hans Boehringer, Ph.D., and Brendan O’Farrell, Ph.D., explore this topic in an article recently published in Clinical Chemistry, the leading peer-reviewed international journal of clinical laboratory science. Read it here. --- ## Analyte Concentrations URL: https://dcndx.com/insights/analyte-concentrations/ Type: insight Published: 2022-01-27 The concentrations of analytes of clinical interest in serum span a wide range. Hemoglobin is the most abundant protein in blood at 150 milligrams per deciliter. But how does that relate to the concentrations of parathyroid hormone (PTH) at 10 picograms per mL or human chorionic gonadotropin (hCG) at 20 milliInternational Units per milliliter(mIU/mL) and what is an international unit anyway? This is far from an academic question if an assay or detection technology for a new system is being considered for these and other analytes. To get analyte concentrations into a form where they can be compared, it is best to convert them into concentrations expressed in moles/liter. All that is needed is a molecular weight and the ability to convert milligrams and picograms into grams and to convert milliliters and deciliters into liters. In the hemoglobin example above, multiplying both the milligrams and the deciliters by ten does not change anything but yields 1500 milligrams per liter. Converting milligrams to grams by dividing by 1000 gives 1.5 grams per liter (g/L). Dividing by the molecular weight of hemoglobin (64500 grams/mole) yields a concentration of 2.3e-5 moles/liter. Note that keeping track of the units involved is a good internal quality control measure to ensure that the calculation is being done correctly. International units (IU) pose a special problem. They were defined decades ago somewhat arbitrarily for some protein analytes. They are commercially available as lyophilized materials in a sealed glass ampule. An approximate weight of the material is given in an enclosed package insert. If one assumes that the material is essentially pure and it dissolved in a measured amount of liquid, an approximate concentration can be estimated. Antibody titrations, using solution phase capture or unlabeled detector antibody, can be used to more accurately derive IU to molar concentration conversions. Antibody affinities also vary enormously from association constants (Ka) of 10e3 to 10e14. Lower affinity antibodies are easier to find than ones with higher affinities. Antibody affinities determine the concentration range where they can be used successfully. Competitive assays have their midpoints at approximately 1/Ka. The useable analytical range of a competitive assay is approximately a factor of ten around the midpoint. For sandwich assays the useful analytical range is about an order of magnitude more than for a competitive assay. The analytical sensitivity of a competitive assay is approximately 0.1/Ka. For sandwich assays, it is approximately 0.01/Ka. Both types of assays have calibration curves that flatten out at the top and the bottom of their ranges. Competitive assays flatten out at the top end, or lower concentrations because at the concentrations of antibodies used, the affinity of the antibody is no longer capable of binding enough analyte to displace the tracer molecule which is often an analog of the analyte. Competitive assays flatten out at the bottom of the calibration curve, or at higher concentrations, because the amount of antibody used in the assay is saturated and cannot bind anymore analyte. Note that the range of a competitive assay can be shifted to the left by decreasing the amount of antibody used. The assay will be a little more sensitive, but at a cost in the higher concentration upper limit of detection and in the maximum amount of binding of the label that can be achieved. Sandwich assays flatten out at the bottom end of the calibration curve because non-specific binding begins to approach the amount of binding of the label at the most sensitive portion of the calibration curve. Indeed, sandwich assay sensitivity can be significantly improved by switching to a solid phase with lower non-specific binding. Sandwich assays flatten out at the lower end of the calibration curve, or higher concentration region, because they reach the limits of what the capture and/or detection antibodies can bind. For the average user of immunoassay, the above discussion doesn’t matter very much because the assay manufacturer builds all of this into the formulation of the assay. The problem for the user comes when they wish to measure an analyte that is outside the range of a given technology. The range of an immunoassay technology is determined by several factors such as the the sensitivity of the detection technology, the affinity of the antibodies used in the assay and the size of the sample. In the 1980s a new detection technology called fluorescence polarization was commercialized and a wide range of assays for drugs were commercialized. These assays worked very well and quickly displaced older technologies such as radioimmunoassay (RIA) for most drug assays such as phenobarbital and gentamicin. They reached their limit for assays such as digoxin and thyroxine because their physiological concentration ranges were several orders of magnitude lower than the higher concentration drugs such as phenobarbital and gentamicin. Fluorescence assays depend on getting light into a molecule so that it can be re-emitted as fluorescence. Light absorption is in turn dependent on the innate capacity of molecules to absorb light. This property is often expressed as the extinction coefficient of a molecule at a given wavelength and extinction coefficients in nature are almost never above 10e6 M-1cm-1. Combined with practical limits on optical pathlengths in the cm and below range, this translates into lower levels of detection in the 10-9 M range which is just about where digoxin and thyroxine occur in human plasma. Two revolutions happened in the late 1980s that made more sensitive assays possible. They were the availability of monoclonal antibodies and the emergence of chemiluminescence as a practical detection technology. Polyclonal antibodies from rabbits, sheep and goats had been used as capture antibodies on solid phases such as beads and test tubes since the early 1970s and functioned quite well. The fact that these specific antibodies were only available as a minor component (usually much less than 0.1%) in partially purified antibody fractions did not matter when they were used in competitive assays or even as capture antibodies in sandwich assays. The problem came when they were pressed into service as labelled detector antibodies. When partially purified polyclonal antibody preparations were labelled, a thousand times more irrelevant antibodies than specific antibodies were present. Non-specific binding is driven by the total amount of labelled antibody and backgrounds with these formulations were too high to make practical assays. Afinity purification could be used to overcome this but it is time consuming, technically demanding and expensive. Monoclonals overcame these difficulties in one bound. Chemiluminescence provided an exquisitely sensitive detection technology that overcame the barriers encountered by RIA, fluorescence and even enzyme immunoassays. Detecting emitted light, produced not by incoming light, but chemical means proved to be the key for making ultra-sensensitive assays such as Parathyroid Hormone (PTH) and so called third generation Thyroid Stimulating Hormone (TSH) practical. These assays typically have detection limits in the 10e-13 M range. Enzyme immunoassays can be made this sensitive by using substrates that are detected with fluorescence measurements or with long colorimetric substrate generation times, but this comes at the expense of complex instrumentation and lower assay throughput. Modern clinical laboratories often have instruments that can measure more than 100 different analytes on a single platform. There is never enough space in a lab and having this capability on one instrument is critical for the productivity needed to be competitive. It is common to have 20 or so assays on the instrument at one time and to rotate the other 80 or so through on an as needed basis. An analysis of one of the most popular immunoassay instrument’s menu of about 75 different assays shows that 42 percent of the assays have analytical sensitivities of less than 10e-9 M, 27 percent are more sensitive than 10e-10 M, 22 percent are more sensitive than 10e-12 M and 4 assays or 2.3% are at about the limit of current assay technology at about 10e-13 M. Even more sensitive immunoassays can and have been developed, but none have gained traction in the commercial marketplace probably because it has not been shown as yet that they can provide medically useful information. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. For more information, visit dcndx.com. --- ## Selling for Scientists URL: https://dcndx.com/insights/selling-scientists/ Type: insight Published: 2021-06-04 Many scientists are uncomfortable with selling. What they don’t realize is that they’ve been doing it since infancy. Anyone who’s gotten into college, landed a job, or been on a date that’s led to a romantic relationship has conducted a successful sales campaign. The “product” in this case is the person themselves. That person conveyed useful information about themselves in a manner that led to a mutually beneficial exchange. Of course, most scientists are not interested in selling themselves—not in a direct manner, anyway—but rather their products or services. What the example above serves to do is illustrate that not all scientists are as foreign to sales as they think. In fact, the sales skills they’ve been using passively are very similar to the ones used in active sales. Basics: The Four Ps At the base of any sales exchange, there is a product that meets another party’s needs. This fundamental tenet makes up one part of what professional marketers call the “Four Ps” (product, price, place, and promotion). To sum it up: the “Four Ps” refer to the mix of factors that must be balanced in order to make a persuasive sale in the market. You need a product that solves a need, it must be priced right (neither too high nor too low), it needs to be in a location where the potential customer will see it, and so on. This article focuses on selling the “product” part. Selling in the Field Scientists are often asked to help professional sales teams sell their products. For the scientist, this is a great opportunity to meet and talk with the “real bosses,” the folks who buy the fruits of their labor. (To clarify: the administrative bosses handle the money in between.) For many scientists, this situation requires some mental adjustments. Scientists have been conditioned to glean praise and achievement from being smart, and that is their go-to mode. Being smarter than your customer, however, is not conducive to conveying useful information or to making a sale. Pointing out the weak points in your own product may be useful internally, but it’s not a useful tactic in making a sale. You can be honest without shooting yourself (and your organization) in the foot. After all, when applying for colleges, it was not necessary to highlight that C-minus in high school calculus. When a scientist is in a customer’s lab or office, it is best to remember that one is a guest and behave accordingly. Listen to the host’s questions and concerns about the product and address them as honestly and completely as you can without dwelling on negatives. If it’s necessary to discuss product shortcomings, it may be useful to frame them in terms of overall utility. For example, suppose the reproducibility of the product is less than stellar at the extremes of the standard curve. A reasonable question to ask the host is: “How many patient samples are in the affected range?” Turn the focus of the discussion to product performance in the range where most of the samples actually occur. Most customers will be polite to a visiting scientist. They’ll take the opportunity to learn and discuss ideas of mutual interest. Occasionally a customer may feel the need to demonstrate that they are as smart or as well-informed as the visitor. Let them have their moment; keep the focus on the product, not personalities. However, do not let obvious inaccuracies go unchallenged. One way to do this is to politely disagree and offer to find literature that speaks to the disagreement. This will keep the discussion professional and grounded in more than opinion. It is a learning opportunity for both parties. It will also build credibility, but only if there is follow through with the commitment to find literature that speaks to the issue. One mistake that new salespeople and visiting scientists alike make is to keep talking once the customer has agreed to the sale. This risks the loss of the sale if a weakness of the product is highlighted. Bear in mind that a product does not have to be perfect to be useful—it has to solve the customer’s problems reliably. Can anyone of us honestly say that we scored perfectly in college or perform flawlessly on the job? Selling in the Workplace When a scientist works to develop a new product, they become more familiar with it than anyone else. They understand its inner workings and its potential. With this, they may become aware of improvements that were not anticipated in the original development plan or specifications. Most organizations maintain plans to ensure that products are introduced in a timely fashion, but they may tolerate deviations if it leads to a better product. This is where a scientist’s selling skills come in: they must sell the rest of the team on the merits of the product improvements. That entails conveying the information in an understandable fashion. (At times, the rest of the team will not view the innovation with the same enthusiasm as the originator. Be prepared for the possibility and learn from the failure; there will be more opportunities). Conversely, if a scientist knows there are serious flaws in the product, they should have the courage to raise those concerns within the organization so they can be addressed. If the organization refuses to acknowledge the flaws, it may be time to look for another position. Organizations and people succeed in direct proportion to how they deal with reality. An organization that refuses to accept reality is doomed to fail sooner or later, and there is no need to go down with a sinking ship. Scientific Credibility and Sales It is possible to combine effective selling while maintaining a high level of scientific credibility. The thoughts above are based on more than four decades of personal experience as a bench scientist, an R&D manager, and a business development manager that sold contract assay development services. These contracts were for multi-year projects that totaled tens of million dollars in revenues for my employer and allowed customers to put more assays on their platforms than they could have done on their own. Some of the project were follow-ons after the success of the first. As I neared retirement, I informed my customers. Something almost magical happened: a number of them wanted to hire me as a consultant (and some did). What this experience demonstrated to me is that it is possible to combine sales and be scientifically credible at the same time. Conclusion Selling is hard work and it is often lonely. It is as intellectually challenging as product development and requires a diverse skill set that includes product knowledge, understanding the customer’s needs, and being able to convey that knowledge so that both parties benefit. It can also be a rewarding experience. If the opportunity for sales support in the field arises, take advantage of it: you may find you have a knack for it and scientific sales may be in your future. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. For more information, visit dcndx.com. --- ## Webinar Overview: The Past, Present, and Future of Rapid Diagnostics URL: https://dcndx.com/insights/webinar-overview-past-present-future-rapid-diagnostics/ Type: insight Published: 2021-06-04 (As published 2021-06-04; regulatory status may have changed since.) Dr. Brendan O’Farrell, the President and Co-founder of DCN Dx, recently sat down for a virtual “fireside chat” with John Zeis, the President of Toolbox Medical Innovations. The two leaders discussed the origins of their respective companies, the evolution of the industry during the COVID-19 pandemic, and the future of diagnostics. Read below for the highlights of the talk. How DCN Dx Got Its Start O’Farrell studied and worked in the lateral flow and rapid diagnostics industry for nearly 30 years prior to co-founding DCN Dx. A native of Dublin, O’Farrell worked in various diagnostics environments and quickly found a gap in the market: education and training for lateral flow technology. DCN Dx began within another biotech company as a consulting team designed to help clients develop products to access manufacturing equipment faster. In 2005, O’Farrell and his partner created DCN Dx as a brick-and-mortar lateral flow assay contract developer with a focus on industry education, emphasizing the capabilities of lateral flow technology. As the industry has grown, DCN Dx has grown with it. How has COVID-19 Changed the Industry? When discussing diagnostic devices in relation to COVID-19, it’s important to differentiate between those that have received Emergency Use Authorization by the FDA and those that have not, said O’Farrell. He explained that there are risks and rewards that must be weighed in the development of every medical device. During the pandemic, several rapid diagnostic devices have received EUAs. This has allowed developers to circumvent the normally rigorous regulatory cycle because the benefits outweigh the risk. However, he said, the situation will likely be short-lived. While the industry has a surfeit of COVID-19 diagnostic devices at the moment, O’Farrell said it’s important for developers to be prepared when the process returns to “normal.” He said that while that process has become more efficient over the years, it does require time and patience. At-Home Diagnostics Lateral flow technology makes at-home diagnostic devices easily available. The development and manufacturing of rapid diagnostic devices using lateral flow technology depends on consumer interest. Even before the pandemic, consumers outside the healthcare industry demonstrated interest in rapid diagnostics. That interest has continued to grow. With smartphone apps, easy-to-use diagnostic devices, and DNA and other analyses at their fingertips, consumers have become more interested in monitoring their own health and wellbeing. “That demand will likely persist, and we have the technology to scale manufacturing of these devices to grow that direct-to-consumer market,” O’Farrell said. “To disrupt the market, you don’t necessarily need new technology: it can simply be a better application from existing technologies, more efficient processes, or new market paradigms.” Watch the Full Conversation Brendan O’Farrell, the President and Co-founder of DCN Dx, and John Zeis, the President of Toolbox Medical Innovations. DCN Dx is a globally recognized contract developer of rapid diagnostic tests based in Carlsbad, Calif. Since its founding, DCN Dx has been committed to furthering the rapid diagnostic and point-of-care test market through the continued evolution of technologies and applications related to lateral flow assays. DCN Dx’s cross-functional team of scientists and engineers develop and integrate all aspects of the assay system, including complex binding reactions, cassettes, sample handling devices, and reader systems. The company assists clients in developing entire rapid diagnostic tests, from concept to commercialization. --- ## The Method Comparison Experiment: A Central Requirement for New Test Methods URL: https://dcndx.com/insights/method-comparison-experiment-central-requirement-new-test-methods/ Type: insight Published: 2021-05-25 (As published 2021-05-25; regulatory status may have changed since.) New test methods for medical use for humans require FDA review by one of their several programs. Depending on the analyte and the technology used, this can lead to waiver, approval, or even licensure. All of these programs require studies that compare results from the new (candidate) method to at least an already-approved method for the same analyte. The simplest approach—a comparison of two test methods, one of which (the comparator method) has already successfully completed FDA review—is by far the most widely used. It’s also important to understand your intended use for the test method. I’ll cover that briefly near the end. Here I’ll discuss a frequently used and widely accepted approach to comparing two qualitative tests (positive and negative results only). The method comparison experiment is more fully described in CLSI document EP12-A2, User Protocol for Evaluation of Qualitative Test Performance. I’ve also used some data from a method comparison of two COVID-19 antibody tests to demonstrate what you can learn about a new test (your own or someone else’s) from just a few data points and to describe some of the limitations of the evaluation method. THE METHOD COMPARISON EXPERIMENT To perform the method comparison experiment, a set of samples (both positive and negative) that have results from a comparative test method is assembled. The more positive and negative samples you have to work with, and the more confident you are in the accuracy of the comparative method, the stronger your confidence in the results will be. The sample set is tested with the candidate method, and its results are compared with those from the comparative method, most often in a 2×2 contingency table (see Table 1). This is where things begin to get interesting. The lowercase letters used in this version of the table imply that we know little about the accuracy of the comparative method or how the disease prevalence in our target population relates to the “prevalence” represented by the positives and negatives in our sample set. Had we more confidence, we would apply capital letters that signify true and false positives (TP, FP) and true or false negatives (TN, FN) to those same numbers in the table. In the low confidence situation, the results of the analysis are labeled positive percent agreement (PPA) and negative percent agreement (NPA). If you’re able to have higher confidence in your comparative method and the prevalence similarity of your method comparison sample set to the actual prevalence in your target population, those same results can appropriately be labeled “estimates of sensitivity” (%Sens) and “specificity” (%Spec), and positive and negative predictive value (PPV, NPV) can be calculated. This is illustrated by including the “high confidence” names in parentheses for each of the lowercase letters. TABLE 1. 2×2 Contingency Table When Using a Comparative Method Comparative Method: Positive Comparative Method: Negative Total Candidate Method: Positive a b a + b Candidate Method: Negative c d c + d Total a + c b + d n KEY a = number of samples positive by both methods (TP) b = samples positive by candidate, negative by comparative method (FP) c = samples negative by candidate, positive by comparative method (FN) d = samples negative by both methods (TN) n = total number of samples in the study (N) a + b = samples positive by candidate method (TP + FP) a + c = samples positive by comparative method (TP +FN) b + d = samples negative by comparative method (FP + TN) c + d = samples negative by candidate method (TN + FN) The calculation for estimated percent sensitivity (%Sens) and for positive percent agreement (PPA) is the same calculation, using different designations for the same numbers from the study, depending on the level of confidence in the accuracy of the comparator method: %Sens or PPA = 100 x [TP or a/(TP or a + FP or b)] Likewise the calculation for estimated percent specificity (%Spec) and for negative percent agreement (NPA) is: %Spec or NPA = 100 x [TN or d/(TN or d + FN or c)] The low-confidence situation avoids consideration of positive or negative predictive values (PPV and NPV) in an acknowledgement that the number of positive and negative samples in the study bears no relation to the disease prevalence in the population, but only reflects the numbers of samples available for the study. When the disease prevalence in the population under study is reflected in the number of positive and negative samples used for the study, the calculations are: %PPV = (TP + FN)/N and %NPV = (TN + FP)/N Finally, the data set for our example qualitative COVID-19 antibody test is: PPA: 80.0% (95% CI: 56.6 – 88.5%) NPA: 100.00% (95% CI: 95.2 – 100%) WHAT CAN WE LEARN FROM THIS SMALL DATA SET? The surrogate sensitivity estimate (PPA) for our candidate test is 80%, meaning literally that eight out of each 10 samples that were positive by the comparator test were positive by the candidate test. The other two of each 10 were negative. The candidate test appears less sensitive than the comparator method. The 95% confidence interval is broad: the real PPA could be anywhere from 56.6% (almost a coin toss) to 88.5% (pretty good). If a larger positive sample set had been available for study, the 95% confidence interval would have been tighter. On the other hand, the candidate test identified every single negative that the comparator method found, out of an apparently large number of negative samples, since the 95% confidence interval is tight, and the low end is still 95%. There were no “false positives.” A NOTE ABOUT OTHER METHODS A “gold standard” comparison study compares results of the candidate test to a clinical diagnosis. Such studies are expensive, complicated, and difficult to organize. Slightly lower on the “gold standard” scale is a comparative test that’s a “reference method” for the same analyte the candidate method assesses. (Reference methods have themselves been rigorously evaluated and are considered gold standards.) These methods are also hard to come by and often difficult to use. An important takeaway here is that whether you’re using gold standard methods to calculate percent sensitivity (%Sens) and percent specificity (%Spec) or more readily available methods to achieve percent positive agreement (PPA) and negative percent agreement (NPA), the data analysis comes down to the very same 2×2 contingency table (Table 1) in which the positives and negatives from the candidate method are compared to those of the comparator method. Understand that, and any confusion melts away. SO, IS THIS A GOOD TEST? Is this candidate method a good test? That depends on whether you value specificity over sensitivity or vice versa for your application. Sensitivity at the simplest level is just “How low can you go?”—what’s the lowest analyte concentration the method can detect? Specificity tells us to what extent the test is detecting substances that aren’t (in this case) COVID-19 antibodies. If the intended use for this COVID-19 antibody assay is to survey populations for previous exposure to COVID, it may be more important that it’s very specific (i.e., it doesn’t return a positive result for substances that aren’t COVID-19 antibodies) and less important that it is less sensitive and doesn’t pick up very low levels of COVID-19 antibodies. If, however, you want to use the test to study the half-life of COVID antibodies in people with mild, moderate, and severe COVID-19, it may be more important to pick up low antibody concentrations with the most sensitive test you can find. Thus, we’ve gathered a fair amount of information from the method comparison experiment and only two calculations (PPA and NPA) and their confidence intervals. The caveats are that we don’t know much about the accuracy of the comparator test and the sensitivity was apparently evaluated on a very small set of positive samples. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding in 2006, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. For more information, visit dcndx.com. --- ## Immobilizing Antibodies on Plastic Surfaces URL: https://dcndx.com/insights/immobilizing-antibodies-plastic-surfaces/ Type: insight Published: 2021-04-05 Plastic is the ultimate unnatural material. Its persistence in the environment as a pollutant stems from the fact that the natural world has not had time to evolve mechanisms to degrade it in a timely fashion. Wood, bone and leather are foodstuffs for various microorganisms, but they have all existed for a very, very long time. Many of the plastics we deal with daily are formed by injection molding. In this process, raw pellets of polypropylene or polystyrene are melted into a hot liquid that is then forced into metal molds. Cooling takes place in the mold so that the plastic becomes solid. The solid piece, which is still quite hot, is then ejected from the mold into oxygen-rich air. Oxidation ensues and produces organic acids, aldehydes, ketones and epoxides [1] on the exposed surface. Antibodies and other proteins have evolved over hundreds of millions of years. It is a lucky accident that proteins and injection-molded plastics were found to interact in such a useful way [1]. Not only do antibodies spontaneously bind tightly to injection-molded plastic surfaces without the aid of other chemicals or treatments, but these antibodies also retain their ability to bind to their target antigens. This is analogous to making a delicate design in clay and then throwing the clay object at a wall and expecting the design to stay intact. But amazingly enough, it does! One of my first assignments as an industrial chemist was to get antibodies off of previously coated polypropylene test tubes. There was an oil crisis at the time, and plastic tubes were in tight supply. It was thought that if we could remove antibodies from antibody-coated tubes that had failed to pass quality control, they might be reusable. Many reagents such as chromic acid, alcoholic potassium hydroxide and permanganate destroyed the binding ability of previously coated antibodies, but they failed to produce a surface that could be coated with another antibody. The only treatment that could make the surface suitable for recoating was boiling water for several minutes. In effect, the protein had to be cooked off. To better understand how antibodies coat onto plastic, we also tried various pretreatments. Detergents, lipids and organic solvents had little or no effect on the ability of antibodies to coat the plastic surface. The only reagent that seemed to make the surface uncoatable was elemental iodine in a sodium hydroxide solution. This is the classic iodoform reagent, which is a test reagent for the presence of ketones and aldehydes [2]. It also destroys the ketones and aldehydes in the process. A quality problem provided further evidence of the interaction of hot plastic in an oxygen-rich environment. A company was coating 12 mm by 75 mm polypropylene test tubes with antibodies and had been successfully marketing them for several years. A problem occurred with tube-to-tube reproducibility. About 3 percent of the tubes tested showed a higher level of binding than the rest of the tubes, thereby causing the batch of coated tubes to fail. Weeks of hard work with variations in coating procedures and blocking reagents failed to correct the problem. An observant quality control technician noted that all the tubes that showed the enhanced binding had a number 8 embossed on the outside bottom of the tube. No one at the company knew the significance of the numbers on the bottoms of the tubes. A call to the molder revealed that the number identified a particular cavity in a 36-cavity mold. When the molder examined the mold, they found that the cooling line for cavity number 8 had been blocked. That meant that a tube ejected from cavity number 8 was hotter than tubes molded in the other cavities. This higher temperature likely led to more oxidation products forming on the tube’s inner surface and higher amounts of antibody being bound. Clearing the cooling line on cavity number 8 solved the reproducibility problem. The so-called edge effect seen in microtiter plates may have a similar origin. Wells on the periphery of a microtiter plate would be expected to cool quicker than wells in the plate’s interior. The observations above are evidence that coating antibodies onto injection-molded plastic is not a passive process. It is likely a chemical process that involves the reaction of amines and possibly other functional groups with reactive oxygen species on the surface of the plastic. How much antibody can be coated on a plastic surface is a question that frequently occurs in assay development. Studies conducted decades ago [3] suggested that it was possible to coat antibodies at 1 ug/cm2. This agrees well with closest-packing calculations that take into account the known dimensions of antibodies. Experiments with rabbit polyclonal to digoxin and cortisol showed that if polypropylene tubes were first coated with digoxin antibodies and subsequently with cortisol antibodies, no cortisol antibodies could be co-coated until the digoxin antibody coating concentration fell below 0.5 ug/mL. Yet countless experiments have shown that coating concentrations higher than 0.5 ug/mL do produce higher amounts of active bound antibody, up to a limit of 3-4 ug/mL. How can these two seemingly contradictory observations be reconciled? Enhanced coating strategies are known and have been employed to improve assay performance and conserve costly antibodies. One approach is to coat an anti-species antibody such as a goat anti-rabbit antibody and then let it capture the rabbit antibody of interest. Another approach used successfully is to chemically label the primary with a biotin or fluorescein thiocyanate (FITC) molecule and then coat the corresponding secondary reagent streptavidin or anti-FITC. All of these techniques require less of the primary antibody to be used in the assay. In my hands, an anti-FITC system used 20 times fewer primary antibodies than when that same primary antibody was directly coated. This observation may help explain the apparent paradox about coating concentration cited above. Secondary coating strategies may deliver more useable antibodies than direct coatings. This may also explain why some antibodies pretreated with a cross-linking agent such as glutaraldehyde have enhanced coating [4]. The glutaraldehyde may “shrinkwrap” antibodies and prevent them from being deactivated on coating. While we have learned much over the years about coating antibodies on plastic surfaces, there are still many questions that remain. Polyclonal antibody coating solutions with coating concentrations above 1 ug/mL can be recovered and used to coat other tubes or wells. In my hands, monoclonal antibody solutions are not reusable even at coating antibody concentrations of 3 ug/mL. Methods using labeled goat anti-mouse antibodies inhibited by measured amounts of unlabeled goat anti-mouse antibodies can be used to estimate the amount of mouse antibody coated on microtiter wells. In several systems where 100 uL of mouse antibody at a coating concentration of 3 ug/mL was coated, it resulted in only 10 ng of mouse antibody being coated on the well out of 300 ng of antibody being offered. The remaining 290 ng that was recovered was uncoatable on a fresh surface. The mechanism of this inability to coat is currently unknown. Coating antibodies on plastic surfaces has been practiced on an industrial scale for decades and is central to products that affect human health and other important areas. It is still amazing that it works at all, and as outlined above, there is still much to be learned about the process. [1] K. J. Catt and G. W. Tregear, Science 158, 1570 (1967) [2] https://www.chemguide.co.uk/organicprops/carbonyls/iodoform.html [3] A. J. Pesce, D. J. Ford, M. Gaizutis, and V. E. Polak, Biochim. Biophys. Acta 492, 399 (1977) [4] U.S. Patent 4,069,352 --- ## Recombinant Antibodies as Capture and Detection Reagents – Part 1 URL: https://dcndx.com/insights/recombinant-antibodies-as-capture-and-detection-reagents-part-1/ Type: insight Published: 2020-12-21 There is a quiet revolution happening in immunodiagnostics, and it’s one that could have a profound effect on how immunoassay developers and manufacturers do their jobs. Recombinant antigens such as human Chorionic Gonadotropin (hCG) and human Prolactin have been commercially available for many years. They have simplified the tasks inherent in the development and production of assays for these analytes. For much of that time, however, there was little change in the way the antibodies needed for these assays were made: rabbits, sheep, and goats are immunized and polyclonal pools prepared and processed. But these pools are finite; when they begin to run out, a scramble sometimes ensues for a replacement pool. Continuity of Antibody Supply Monoclonal made the task of antibody production somewhat easier. With proper care and storage, monoclonal antibody cell lines are effectively immortal. But there was still the problem of monoclonal antibody production, which could be done in two different ways. The classical method of monoclonal antibody production was to inject the hybridoma cell line that produced the monoclonal antibody into the peritoneal cavity of live mice to develop ascites, which is an antibody rich fluid that is produced by the hybridoma cell line. Many European countries (and some jurisdictions in the U.S.) have banned monoclonal antibody production by ascites. The alternative is to produce the antibody in cell culture, a process in which no mice are used, but the concentration of antibody produced in cell culture is less. A million microtiter wells would require the output of about 400 500-mL roller bottles producing monoclonal antibodies in cell culture. Since sterile techniques and expensive cell culture media are required, producing monoclonal antibodies in cell culture is more expensive than ascites production. More Efficient Antibody Production Recent advances in protein sequencing have allowed researchers to obtain the amino acid sequence data for the binding site in antibodies. The sequencing can be done on as little as 200 ug of an existing antibody, or the amino acid sequence information can be obtained by sequencing the DNA or mRNA that codes for those amino acids in an existing hybridoma line. Once that information is known, it can be stored in digital form that is, in effect, the equivalent of a master cell bank in silico. It is also possible to insert these sequences into pre-made molecular constructs or cassettes that encode the rest of the antibody beyond the binding sites. These full-size antibody constructs are then put into large-scale cell culture in which it is possible to produce gram quantities of antibody in a few days. Several commercial organizations and their subcontractors are providing the process described above for less than $20,000 per antibody, including the production of a gram of antibody. A gram of antibody can be used to coat five million microtiter wells under the conditions described above. The second gram of the same antibody costs significantly less because all of the discovery work has already been done. Larger lots of antibody are also possible. Since every lot of antibody used in making an assay needs to be qualified prior to use, larger lots of antibody mean that much less expensive scientific labor is needed to qualify the antibody to produce the same number of assays. By comparison, it is not unusual for off-the-shelf antibodies for some esoteric analytes to cost $1,000/mg or more, making recombinant antibodies a technology well worth considering for higher volume assays. Enhanced Control Over Antibody Classes and Types The companies mentioned above also offer multiple cassettes that can provide advantages in assay development and manufacturing. Mouse IgG antibodies come in four classes, IgG1 through IgG4, and they have different properties. IgG1 is preferred for making assays as it has good stability and low non-specific binding properties. IgG3 has a reputation for being sticky and is therefore prone to non-specific binding problems. But sometimes the best antibody that comes out of an immunization and fusion program is an IgG3. With recombinant antibody technology it is now possible to class switch that IgG3 to a more malleable IgG1 antibody. It is also possible to switch the antibody species. A mouse monoclonal binding site can be cloned into a rabbit or human cassette. Mouse monoclonal antibodies have proven to be significantly easier to make than their rabbit or human counterparts. Production of humanized mouse monoclonal antibodies is of significant therapeutic interest and recombinant antibody technology is being investigated vigorously for therapeutic uses. It is one of the drivers for scaling up the production of recombinant antibodies to gram quantities. The ability to sequence antibodies has important implications for troubleshooting antibody issues in production. For example, you may have an assay in production that requires more and more antibodies to get results comparable to those obtained when the assay was first developed. It would take months to figure out that the hybridoma cell line used to produce the antibody had become contaminated with another, irrelevant cell line that was gradually outgrowing the cell line of interest. Sequencing the antibody in question would show this problem immediately. It is possible to have cassettes for different subtypes of immunoglobulins such as IgM, IgA, and IgE. IgM antibodies are the antibodies that first appear when an immunogen is first encountered, and they are generally lower in affinity than the IgG antibodies that eventually replace them. IgM antibodies have been used in diagnostic assays, but usually as a last resort because the lower affinity translates into poorer sensitivity. IgM antibodies have 10 interconnected binding sites in contrast to the two binding sites in an IgG antibody, and it is thought that the larger number of binding sites partially compensates for the intrinsically lower binding affinity of the individual IgM binding sites. It is interesting to speculate what a high-affinity IgG binding site cloned into an IgM cassette with 10 binding sites would look like. Would the effective affinity be higher with 10 binding sites instead of just two? Polyclonal antibodies that have been affinity purified can be sequenced as well. The 10 most prevalent clones are immortalized in silico exactly like their monoclonal counterparts. Affinity purification is necessary because polyclonal antibodies contain literally hundreds of thousands of separate clones to the immunogens that the host animal has encountered over time. Rather than searching through this haystack for the proverbial needle, affinity purification is akin to using a powerful magnet to find the needle. This means that the “golden rabbit” that produces an antibody to a given analyte can effectively be made immortal. Some analytes are best done with rabbit antibodies. Estradiol and Leutenizing Hormone (LH) are two good examples. Estradiol immunogens injected into mice to elicit monoclonals invariably produce antibodies that recognize the metabolites of estradiol. Mouse LH monoclonals can suffer from over-specificity and can miss deletion mutations in certain populations. The ability to pull 10 different clones from a polyclonal response can also give assay developers great flexibility in crafting a blended polyclonal with specific properties and without certain problematic cross reactivities. Conclusion Recombinant antibody technology is in its infancy today, but already it neatly solves the problems of long-term supply of a given antibody (whether monoclonal or polyclonal) by preserving the sequence in silico. It also provides an alternative to both ascites and traditional cell culture antibody production on a scale that was not possible previously. The unprecedented control that it gives to assay developers and producers is just being explored and appreciated. Further extensions of this technology can be expected soon that are going to transform what we can do with antibodies and immunodiagnostic assays. --- ## Antigens, Antibodies, and Diagnostic Testing URL: https://dcndx.com/insights/antigens-antibodies-diagnostic-testing/ Type: insight Published: 2020-11-06 Many diagnostic tests use immune proteins to detect infectious diseases (such as COVID-19). Although proteins can be measured with expensive and cumbersome instrumentation, it is much more efficient to measure them indirectly. One way to detect or measure a protein in a diagnostic test is to use a “binder” with a label on it. Fortunately, nature provides us with the most commonly used binder—antibodies. Many of us are familiar with antibodies; our bodies produce them in response to infections and immunizations. When our bodies encounter dangerous foreign proteins, our immune system fights them off. We can use this same system to produce antibodies in immunized animals that can then be harvested and used in diagnostic tests. Antibodies and Diagnostic Testing for Infectious Diseases Production of antibodies for diagnostic tests come in two types: antigens with a molecular weight above 6,000 Daltons, and those below. Higher molecular-weight antigens (e.g., viruses, bacteria, proteins, longer peptides) are immunogenic on their own and can be used directly as immunogens (i.e., antigens that induce an immune response). Lower molecular-weight antigens (e.g., hormones, toxins, drugs) need to be chemically coupled to carrier proteins to be immunogenic. Designing these immunogens requires careful planning and execution. The antibody is going to be raised against the antigen coupled with the carrier protein, and the target molecule is, by definition, going to be a cross-reactant. A good example of immunochemistry is found in the expired U.S. Patent [1] for a drug called theophylline. Theophylline is used to control asthma and closely resembles several other compounds that are frequently encountered in everyday life: caffeine and theobromine (a substance found in chocolate). These differ by only a methyl group or two in molecules with molecular weights around 180 Daltons. Selection of the site of attachment of the chemical linkage to attach the theophylline derivative to the carrier protein is critical to generating antibodies that recognize the drug, but not closely related molecules. Even the bridging linkage requires some thought and planning lest the antibody recognize the bridge better than the antigen. Chemical coupling of the target molecule to the carrier proteins is usually done by activating a carboxylic acid group and reacting that with amines on the carrier protein to form an amide bond. At minimum, 10–15 molecules coupled to a carrier protein are required to get a good immune response. Commonly used carrier proteins include bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH). BSA is a 66kD protein obtained from the blood of cows; it contains 66 lysine residues, about half of which are available for coupling. KLH is derived from a mollusk that lives off the coast of California and has a molecular weight of about 390 kD, multiple amine attachment sites, and is highly immunogenic in mammalian hosts. Early in the history of diagnostics, human antibodies were used in tests for the Hepatitis B antigen. The reason for this was simple: producing high-affinity antibodies takes time and investment. Convalescent human plasma from people who had recovered from Hepatitis B had the best antibodies to the virus that were available at the time. Some researchers predict we may see this approach again with COVID-19 antigen tests. Producing Antibodies for Diagnostic Testing Today, antibodies are produced in a wide variety of animals including mice, rabbits, goats, sheep, donkeys, and llamas. In particular, rabbits are an excellent source of diagnostic antibody production. They are easy to care for and handle and, when properly managed, can produce good quality antibodies for years at a time. Immunization is usually done just under the skin of the back. The antigen is mixed with a cocktail of additives called an “adjuvant” that enhances the immune response. A primary immunization is followed by a series of booster shots at intervals of several weeks to help mature the antibody response. An immunization campaign in rabbits typically takes six to nine months to get the best-quality antibodies. Because individual animals vary in their responses to antigens, it is prudent to start with at least five rabbits to increase the odds that a useable antibody will be produced. Small test bleeds can be taken every few weeks to follow the course of the immune response. Testing will determine how much the antiserum can be diluted for use in an assay. Experience has shown that testing in the format the antibody is to be used is essential. Results in microtiter plates are often not replicated on magnetic particles, and vice versa. Other critical parameters, such as cross-reactivity with closely related molecules and with known metabolites, must be tested to determine which of the rabbits is producing useable antibodies. Production bleeds can begin when the response has matured enough to produce a useable antibody. Rabbits can be bled 40 mL of whole blood once per month for several years with no apparent ill effects. After processing, these bleeds yield about 20 mL of serum. Coating dilutions of 1/5,000 to 1/20,000 are common. At a nominal IgG concentration of 10 mg/mL, 1/5,000 dilution corresponds to 2 ug/mL. At a coating volume of 100 uL and a coating concentration of 2 ug/mL, that means a single bleed can be used to produce a million coated microtiter wells or about 10,000 microtiter plates. Pooling several production bleeds that have similar characteristics to make a pool of more than 100 mL makes economic sense when one considers the labor involved in developing a test intended for a regulated market. Coating above or below the coating concentrations cited above can produce problems such as low dose hook effects in competitive assays from loosely coated antibodies at coating concentrations of more than 2 ug/mL or strange stability problems at coating concentrations of less than 0.5 ug/mL which corresponds to dilutions of more than 1/20,000. Good antibodies to some antigens, such as estradiol, have proven to be elusive. Successive campaigns by two companies that have since merged found that together they had immunized more than 500 rabbits with estradiol immunogens with no success. Monoclonal antibodies are usually produced in mice with an immunization schedule not too different from the one outlined above for rabbits. Test bleeds are taken from the mice during the immunization process; when the titers of the desired antibody are reached, the animal is humanely sacrificed, and the spleen taken. B cells obtained from the spleen are fused with a mouse myeloma cancer cell line to produce a hybridoma. This hybridoma combines the antibody-producing ability of the parental B-cell line with the immortality of the myeloma parental cell line. Hybridomas can be used in tissue culture or in ascites production in live mice. The entire process from immunization to production of useable amounts of antibody takes 9–12 months. Attempts to shortcut the process to save time to market usually produce antibodies of lower quality. Antibody Cost and Supply Antibodies are critical components of any immunoassay. Time-to-market can be shortened by using off-the-shelf antibodies from a huge number of commercial suppliers, but the prices of these off-the-shelf antibodies are usually much higher than antibodies generated from scratch. Supply reliability can also be a factor with off-the-shelf antibodies. Of course, reliable sources of antibodies are fundamental to developing and manufacturing a successful product. Immunoassay developers are faced with a number of multi-faceted decisions when developing assays. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. For more information, visit dcndx.com. [1] U.S. Patent 4,397,979 --- ## Contract Assay Development: Part 1 URL: https://dcndx.com/insights/contract-assay-development-part-1/ Type: insight Published: 2020-08-11 Much like many other business and research-related terms, “ contract assay development ” can involve any number of interconnected, complex activities. At its core, though, it can be defined simply: contract assay development is the outsourcing of assay development. In practice, an organization that needs one or more assays developed for their platform may contract with another organization to do the necessary research and development, select the materials, and fine tune processes to make those assays a reality. Why Outsource Assay Development? There are several reasons why a company may choose to outsource assay development. The most obvious one is that the company lacks the internal resources to do it in-house. Companies that are just getting started often choose this route. Established companies do, too, but often for different reasons. For instance, instrument companies that decide to expand their product lines to include reagents may elect to explore this option. But even companies who have their own experienced internal assay development capabilities may choose to have some of their assays developed by an outside contractor. Sometimes the contractor will have experience with an assay that has proven difficult to develop internally. At other times, companies will be in the midst of a launch of a new platform that demands the focus of all internal resources, but an existing platform may need one or more new assays to keep that platform competitive until the new platform is ready for the market. Another reason may be that R&D funding is constrained, and the contractor may be willing to self-fund the development projects in exchange for enhanced future product royalties. (Note that this option only works for well established companies with thousands of platforms already in the market.) Outsourcing allows established companies to expand their assay development capabilities without taking on new staff and adding overhead. In some highly regulated labor markets, this is an important advantage. Benefits of Contract Assay Development Outsourcing may provide a company with numerous benefits. One benefit is that the contracting organization could have its experience base expanded. There are many ways to develop an assay, and no single approach is optimal in every situation. Internal developers tend to wear down each other’s rough edges and converge on consensus opinions in regards to assay development. Time and budget considerations tend to reinforce these opinions in to standard practices; eventually, they become the norm and difficult to change. Outside contractors will have done the same thing. However, contract assay developers may have a broader range of inputs to inform their practices; at times, these practices may be more effective or efficient than a contracting organization’s “baked in” practices. Outsourcing also provides an opportunity to benchmark internal assay development. Every organization believes that they have formulated the best practices for assay development. Without the ability to compare those practices to those of another experienced organization, however, that belief is just an opinion. It’s somewhat like learning a new language: the use of our native language seems intuitively obvious. We don’t often stop to consciously evaluate the mechanics of our language—we simply speak it like we’ve always done. However, when we begin learning a new language, we’re forced to confront its structure and usage. Oftentimes, it’s helpful to compare the new language to our own to explore the similarities and differences. By learning a new language, we begin to evaluate the mechanics of our own language more critically. Working with an outside assay developer is a lot like that: it allows an organization to see its own processes in a more comparative and critical light. No organization has a monopoly on creativity. Outsourcing allows the contracting organization to tap into the minds of experienced professionals with different histories and perspectives. This can lead to new inventions that, due to the nature of contract relationships, may well belong to the contracting organization. (Of course, ownership of inventions should be spelled out in detail in the contract before any development is done.) The client may also be free to use that innovation in other assays in their portfolio. A novel way of providing liquid, ready-to-use standards for one assay may be applicable to an entirely different assay that was not even part of the contracted work. Outsourcing may also provide an improved time to market. Immunoassays have multiple components and processes that must be functional to make the product meet its design specifications. Even with the best planning, development, and manufacturing processes in place, existing assays will need occasional tweaking. Some organizations have enough assays in the market that a dedicated technical support organization can do the necessary adjustments. In smaller organizations, there is a great temptation to stop work on a new product to shore up an existing one. Outsourcing tends keep the focus on the new product. Dispelling the Myths Despite the benefits of using an assay development contractor, some potential users still perceive risk. Oftentimes, organizations opt to invest capital into their own internal R&D—they seem to perceive this larger budgetary outlay as “safer.” The reality, however, is that investing in contract development is investing in your own internal R&D. The interaction with a competent and experienced contract organization naturally expands the tool set of the internal R&D group. It allows objective, data-driven benchmarking. Many organizations who outsource even report that they come to view their contact assay development partners as an essential part of their own internal organization. Fear of competition with internal R&D also makes some organizations leery of outsourcing. The fact is, external R&D can never totally replace internal expertise. It will always be a supplement. While resources will be needed to fund and oversee the outsourced process, there is never a guarantee that the funding that enabled the contractor to be hired in the first place is always going to be available. If all the development money is funneled into internal development, and that funding contracts, it may mean reductions in local staff. That contraction breeds resentment and distrust in the local labor pool on which the company depends. It may be better to have a business-to-business arrangement with an outside partner to smooth over some of the ebbs and flows of development funding. Another myth: outsourcing will expose the company’s R&D secrets to the outside world. Contract organizations live or die on their ability to keep confidential information safe. Trying to gain short-term advantage by misusing a client’s proprietary information is commercial suicide. Moreover, the scientists in every organization are working with just about the same materials and knowledge base. What appears to be a great secret is often common knowledge in the industry, and it looks like a great secret only because no one feels safe talking about it. Finally, there is the argument that outsourcing is just too expensive. Competent development talent does not come cheap whether it is internal or external. One of the ways of looking at the economics of outsourcing is to consider not only the cost and overhead of doing the development work, but to look at the cost of delaying the work until internal R&D resources can do it. It does not take too many quarters of missed sales of a product that was delivered later to more than make up for the outlay in outsourcing. In addition, there are significant market advantages to being in the market sooner than later. Conclusion There are many benefits to be had from contracting with a competent outside assay developer. As we’ve seen, many of the myths and concerns about hiring a contract assay developer just aren’t true. In Part Two of this series, we will examine some of the criteria to consider when choosing a contracting partner, as well as some of the challenges inherent in working with an outsourced project. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. Contact us about your assay development project today. --- ## Contract Assay Development: Part 2 URL: https://dcndx.com/insights/contract-assay-development-part-2/ Type: insight Published: 2020-08-11 In the first installment of this two-part blog, contract assay development was defined as “the outsourcing of assay development.” In that blog, we outlined the benefits of outsourcing. In this installment, we’ll discuss some of the challenges inherent to using a contractor for product development. We’ll also suggest some strategies for dealing with those challenges. Many organizations claim to be able to do contract assay development. They will point to their own product lines or lists of publications as evidence that they know how to develop assays. They are, for the most part, correct: they can develop assays in those circumstances and for those purposes. The requirements for developing assays for another organization’s purposes, however, are quite different. If you want an assay for a short-term research project in your own labs, then an academic laboratory with a track record evidenced in a series of publications may be a good choice as an outsourcing partner. On the other hand, if you want an assay that can be manufactured reliably over years on a scale sufficient to supply customer laboratories around the world, choosing an academic lab to do the development is probably not a good idea. This is simply because most academic labs have no experience in large-scale production over an extended period of time. This means that they will most likely not understand the importance of such considerations as having reliable sources of critical raw materials such as antibodies. A polyclonal pool that is sufficient for 50,000 tests may sound like a lifetime supply to an academic group, but a commercially successful assay can burn through that in just a few years or less if the assay is successful. Having a monoclonal is one solution to this supply issue, but if care is not taken to establish master cell banks, working cell banks, and clone repositories in off-site locations, the product developed might be just one extended power failure from being off the market for months—or from having to essentially redevelop the product with a new monoclonal. Again, this will not be part of the skill set of most academic labs. Working with commercial organizations that have developed their own products can solve many of these issues. They will understand what it takes to develop and manufacture assays and keep them in the market. If they did not have those skills, it’s unlikely that they would still be in business. There is a subtle difference in developing products for your own organization and developing products for another organization, and it has to do with focus. If an organization’s primary focus is its own product line and contract development is just an adjunct activity, there is a built-in potential for conflict. All immunoassays need periodic tweaking and adjustments. The need for optimization can be a previously unseen variability in some raw material, or an advance by a competitor that suddenly threatens the viability of a key product line. In either case, it will be an unforeseen emergency that will require scientific and managerial resources to resolve the issue. Adjunct activities such as contract development for another organization will be deprioritized, and development timelines will begin to slip. The best solution is to select an outsourcing partner whose sole business model is focused narrowly on contract assay development. If they have been in business for a number of years, it is likely that they will have a list of satisfied clients who are willing to make it known that this organization has developed products for them. The clients may not want the CAD partner to disclose which assays were developed for them, and that is understandable from a brand projection standpoint. That said, it is often possible to find joint publications or presentations on the assay by the contracted organization and its clients. If the contract organization does not have such a list of satisfied clients, they are either just starting out in the business (which is not a disqualifier, since everyone must start somewhere) or they did not satisfy the expectations of prior clients. In any case, the lack of a verifiable track record is a red flag that may indicate increased risk. Once an outsource partner has been selected, the next step is to get a comprehensive contract in place that defines the roles and responsibilities of both parties. If the partner has been in business for several years, they will most likely have a standard development contract that they will have used with a number of clients. This is an excellent starting point, especially if the client organization is new to contract development. It does not mean that the client company should accept every term in the contract without thoroughly understanding it and having it reviewed by legal counsel. The process includes pushing back on the partner and crafting terms better suited to the client’s needs. The negotiations will also provide a clearer understanding of the outsourcing organization’s flexibility and willingness to work with the client. A contract is also no substitute for understanding that the only successful partnership is a win/win for both parties. If either party to the contract attempts to take unfair advantage of the other, mistrust and resentment will wreck the relationship in spite of what the contract says. Successful relationships develop multiple assays over years, not just one. Assay development projects typically last 15–18 months (or longer) from inception to product launch. As in most human activities, communication is vital to the success of the project. Weekly teleconferences with the project developers and the client’s scientific, marketing, regulatory, and (especially) manufacturing representatives are the norm. An agenda for the meeting should be published by the contractor in advance of the scheduled meeting. Action items with names responsible for the action should be developed before the meeting ends and circulated to both the contractor and client organization’s respective management. Written monthly progress reports should supplement the weekly teleconferences. Quarterly face-to-face meetings in both facilities by key development team members are an excellent way to build common understanding and personal relationships. This is especially true of the manufacturing environment in which the assay will be manufactured over time. R&D is an interesting and necessary component of assay development, but if the product cannot be reliably manufactured, nothing else matters. It must be understood that no product was ever developed without problems and mistakes. These are the inevitable result of the exploratory nature of immunoassay product development and the humans that carry out this process. While there are commonalities in assays, it seems that every time a new assay is developed, there is something new that must be learned and dealt with. This novelty means that new solutions will need to be explored. The reality is that some of them will not work. Finger pointing and Monday-morning quarterbacking do not build the teamwork needed to solve the problems at hand and the issues that will arise with the next assay. For all the issues inherent in contract assay development, a successful practitioner of the process will be able to develop more products more quickly and with better allocations of resources than their competitor. They will also likely develop some professional relationships with colleagues in the partner organization that are deeply satisfying and that will expand their world. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. Contact us about your assay development project today. --- ## Raw Material Sourcing and Qualification for Lateral Flow Assay Development URL: https://dcndx.com/insights/raw-material-sourcing-qualification-lateral-flow-assay-development/ Type: insight Published: 2020-07-20 (As published 2020-07-20; regulatory status may have changed since.) One of the most challenging aspects of assay development is finding the right raw materials and ensuring that they will work in an assay. Some materials (such as buffer salts) are catalog items and can be safely obtained from several vendors. Incoming quality control can be as simple as ensuring that the label matches the specified component. Good manufacturing practices (GMP) mandate that even these materials be given an expiration date beyond which they must be discarded or requalified. Failure to follow this simple process can have dire product quality and regulatory consequences. Next in line is deionized water. This is available as a catalog item, but a system to process tap water into deionized water may be more cost effective in the long run. Continuous monitoring of the conductance of the deionized water produced is essential to ensure that the system is operating properly. Training personnel to take appropriate action if the conductance is out of specification is also essential. Simply writing down a number that is outside the acceptable range does not count—corrective action must be taken. Antibodies and antigens are probably the most challenging raw materials to source and qualify. A good place to start is the Linscott Directory. Just type in the antibody or antigen desired and, if it is commercially available, several sources will be listed. For example, a search for antibodies to prolactin produced 2,011 possibilities. Narrowing the search to mouse antibodies to human prolactin reduced the number of possibilities to 255. Restricting the choices to monoclonal antibodies of subtype IgG1 cut the possibilities to 152. Requiring that the antibody be conjugated to HRP reduced that number to only nine (but they were all from the same vendor). The DCN Dx catalog itself features a selection of high-quality antibodies and reagents. Each has been selected for its popularity among the lateral flow developers there. Of course, being able to find all the necessary inputs in one place (rather than searching through multiple vendors) is a boon to may researchers and developers. For those looking for complete solutions, DCN Dx also offers kits for conjugating to colloidal gold and NanoAct™ cellulose nanobeads —both of which contain all of the reagents necessary for the job. Qualification of Critical Reagents for Lateral Flow Assay Development The qualification of antibodies and antigens—often called “critical reagents”—is complicated and usually done empirically. After sourcing multiple antibodies, the developer must test the antibodies in the format desired for the final product. Testing antibodies for use on magnetic particles or in lateral flow by testing them in microtiter plate assays is a waste of time and resources. This disconnect is likely due to differences in antibody immobilization and assay kinetics. Antibodies need to be tested for accuracy, specificity, reproducibility, and stability. A failure in any reagent requires backtracking, identifying the cause of the assay failure, and taking the necessary steps to prevent its recurrence. Qualifying a single lot of reagents through all of the necessary performance characteristics can be shown to require about 1,500 individual data points. Most organizations require qualification of at least three lots of reagents using manufacturing and quality control documentation that is as close to finalized as possible before an assay can be released to routine manufacturing. Stability testing is probably the most challenging hurdle to overcome once a candidate antibody or antibody pair has been identified. It is not hard to do; it just takes time—lots of time. An assay that fails stability testing requires additional development to remedy that failed component or components. This adds time, cost, uncertainty, and delayed time-to-market. Being first (or even a close second) to market can have a significant positive effect on the overall sales and profitability of a product. For an assay to be viable as a commercial product, it must be shelf stable for a minimum of six months. With additional time and testing, it is desirable to extend that to 18–24 months. Stability beyond 18–24 months is nice to have, but the cost of building and storing inventory that sits unsold for months must be factored in as well. One of the strategies used to extend stability dating beyond actual real-time stability is accelerated stability testing, which is often done at elevated temperatures. This is based on the observation that chemical reaction rates double with each 10 o C rise in temperature if the reaction mechanism does not change. To estimate that a product is stable at 4 o C for six months, it could be tested and shown to be stable for 3 months at 14 o C, 1.5 months at 24 o C, or 3 weeks at 34 o C. In practice, only the 24 o C (room temperature) and elevated temperature (37 o C) are done. Higher temperatures (such as 45 o C) are sometimes done, but potential changes in reaction mechanisms make this option riskier for mammalian proteins that have evolved to be stable at about 37 o C. Note that real time stability data gathering and analysis continue after the product is released on the basis of accelerated stability data. Immunoassays operate at the outer limits of analytical technology. That is one of the reasons that they are so useful, but it limits the use of other analytical methods in qualifying reagents. Optical density in UV/visible spectroscopy is a good example. A typical protein has an optical density (OD) of about 1 at a concentration of a milligram per milliliter (mg/mL) at 280 nM. Typical antibody or antigen coating concentration for making microtiter plates and lateral flow assays are in the microgram per mL range, 1,000 times lower. The amount of functional protein immobilized on a surface is even lower. One way of overcoming this limitation is to use other immunoassays to quantitate the reagents being developed for new analytes on a new platform. This is where competitors’ assays or even a newly developed, suboptimal assay can be useful. The strategy here is to use candidate antibodies as solution phase inhibitors. Antibodies differ in how tightly they bind to the antigen. An antibody with a high affinity will bind solution phase antigens more completely and reduce the signal seen in the reference assay. With some high-school algebra, this approach can be used to generate some apparent affinity constants and other useful information. Once the questions of whether a critical reagent is useful in an assay has been answered satisfactorily, the next critical question to be asked is, “Is the reagent source auditable?” This seemingly innocuous question is intended to find out if there is a reliable supply of the reagent. If the answer to the auditability question is no, then the risk that the supply of the reagent is not secure goes up substantially. Openness to an audit indicates that the supplier has taken steps to build a quality system and supply operation that is transparent and intended to last for years to come. This is important because developing an assay is a substantial financial and time commitment, and to recoup those investments means that there will need to be revenue from the assay for several years. While assays can be redeveloped if a critical reagent supply fails, this is costly in both time and dollars and can even result in a period when the assay cannot be supplied to customers who have come to rely on it. Being out of stock on a given assay can have ripple effects that can result in losing customers. It is far better to have secured the supply of critical reagents before the assay is introduced into the market. A high-quality assay can only be sustained in the market for years with the best quality critical reagents obtained from suppliers with continuing commitments to quality and reliability. Information technology does not have a monopoly on “garbage in, garbage out”. ——- DCN Dx is an industry-leading supplier of reagents, antibodies, and kits to the research community. DCN Dx is an international leader in the contract development and commercialization of rapid diagnostic tests at its ISO 9001:2015 and EN 13485:2016 certified facility in Carlsbad, Calif. The company’s team of in-house scientists and engineers develop and integrate all aspects of assay systems, including cassettes, sample handling devices, and reader systems. Since its founding more than 12 years ago, DCN Dx has been committed to furthering the rapid diagnostic test market through the continued evolution of technologies and applications related to lateral flow assays. For more information, visit dcndx.com. --- ## Understanding Whole Blood, Serum, and Plasma URL: https://dcndx.com/insights/understanding-whole-blood-serum-plasma/ Type: insight Published: 2020-03-17 Most laboratory testing for clinical purposes is done on samples obtained from blood. Whole blood contains the liquid fraction of blood (i.e., plasma) as well as the cellular elements that lead to clotting under certain circumstances. These include red blood cells (RBCs), white blood cells, and other components. First, it is helpful to establish some baseline definitions: “Serum” is the fluid obtained when whole blood clots, as it will do spontaneously when it contacts a surface such as glass or plastic. Clotting is pre-programmed into the components of blood to prevent excessive blood loss from a minor wound. In a lab setting, it is common to centrifuge the clotted blood, including red cells, to the bottom of the collection tube, leaving a straw-colored liquid above the clot. “Plasma” is the fluid component of blood. It is obtained when a clotting-prevention agent is added to whole blood and then placed in a centrifuge to separate the cellular material from the lighter liquid layer. Common anti-coagulant agents are EDTA (ethylenediaminetetraacetic acid), heparin, and citrate. Blood samples intended for laboratory analysis are usually collected in glass or plastic tubes that have been partially evacuated so that their internal air pressure is lower than atmospheric pressure. These tubes have color-coded polymer stoppers that indicate their contents. Plain tubes with no anticoagulants have red stoppers and are used in the preparation of serum after clotting and centrifugation. Purple or lavender top tubes contain EDTA, and green top tubes have heparin in them. Blue top collection tubes containing citrate exist as well, but it far more common to encounter citrate containing plasma as reclaimed plasma from blood donations. When people donate blood for therapeutic purposes (e.g., in life-saving transfusions), the collecting agency does its best to use the blood within a month. Inevitably, due to a need to have enough blood on hand to meet unexpected demand, some of the blood cannot be transfused because it has been stored too long. When blood is collected, it fills a plastic bag that contains a solution of dextrose and citric acid. The dextrose is a nutrient for some of the cells in the blood, and the citric acid complexes with or chelates calcium which prevents the blood from clotting, Minutes after collection, the blood is centrifuged into packed RBCs and citrated plasma for better storage. Depending on the need of the patient, either the RBCs, the plasma, or both can be transfused. Outdated citrate plasma becomes an industrial commodity and is processed to provide a number of components, including a synthetic normal human serum that is the starting point for many of the calibrators used in commercial immunoassay products. The segments of tubing used to transfer separated plasma from the red cells can sometimes be obtained from vendors of outdated plasma. These segments contain about 500 uL of plasma and are invaluable for informal normal-range studies as they come from ostensibly healthy normal donors, who can be identified by age and gender. The entire bag of plasma associated with the tubing segments may also be available for further studies, and it holds approx. 220 mL. This can be a valuable resource in the process of developing an assay. Labs usually have time limits on how long serum or plasma can remain in touch with the red blood cells before being physically separated—usually by pouring off into another tube—while maintaining the identity of the patient from whom the blood was taken. This is necessary because RBCs can rupture over time, and the contents can interfere with a number of assays. RBCs contain high levels of potassium and the thyroid hormone thyroxine. Assays for either of these substances and others can be skewed by the contents of lysed red blood cells. Hemoglobin is the most abundant protein in blood. It can interfere in assays once released from RBCs. One technical solution to the problem of RBCs remaining in contact with the serum or plasma is to use serum separator tubes (SSTs). These are evacuated blood drawing tubes that contain a silicone gel that has a density intermediate between serum and red blood cells. In a centrifuge, this silicone gel forms an impermeable layer between the red blood cells at the bottom of the tube and the serum above. Caution is advised in using these SSTs with hydrophobic analytes such as some drugs. [1] The silicone gel can act as an extracting organic solvent, and the drug can be removed from the plasma and end up in the gel, leading to misleading low drug levels in the serum. Whole blood, serum, and various plasmas are not interchangeable sample matrices. While some analytes may give similar results, equivalence can only be ensured by testing matched samples. Whole blood contains RBCs that may occupy as much as 60% of the volume. Some analytes, such as Parathyroid Hormone (PTH), partition freely between the red blood cells and the plasma so that whole blood and plasma values are the same within experimental accuracy. Others, such as lipoproteins, are strongly influenced by red blood cell content. [2] Still others, such as some immunosuppressant drugs like cyclosporin, partition between RBCs and plasma in a temperature-dependent manner [3] that reflects the temperature history of the sample. To avoid this problem, whole blood is the preferred sample for measuring immunosuppressant drugs. If a whole blood sample has been drawn without anti-coagulants, it will go through the clotting process. The resulting serum will therefore be depleted of various clotting factors and, after centrifugation, all cellular material. Citrated plasma drawn in a blue-topped tube is the sample preferred for coagulation factor testing. Complete blood count samples are drawn in a purple- or lavender-topped EDTA tube. Plasmas are also not interchangeable. EDTA plasma has an excess of a powerful chelating agent that will sequester metal ions such as calcium and magnesium. EDTA plasma will also inactivate some enzymes that require a metal ion for their activity. This includes alkaline phosphatase used as a label in some immunoassays. To avoid this assay interference, EDTA plasma and the alkaline phosphatase containing label must never come in contact for any length of time. Heparin can also interfere with some analytes. Troponin I values measured in heparinized plasma are often lower than when measured in EDTA plasma or serum from the same patient taken at the same time. Troponin I is a positively charged molecule, and there has been speculation that the highly negatively charged polymeric heparin effectively shields Troponin I from being recognized by antibodies. If you are a user of an assay, you should consult the manufacturer’s instructions for use (IFU) to determine what sample types have been validated for use in the assays. If the sample type you wish to test is not listed in the IFU, consult the manufacture’s technical support staff as it is likely someone else has asked the same question before you. If you are the developer of an assay, you should validate the use of the sample types you believe will be used in the assay. Assuming that a certain sample type will work in a given assay without proper validation is poor science and may elicit an unfavorable response from customers and regulatory agencies. We work with a variety of sample types. Whether you’re developing a blood collection device or a blood-based LFA, save valuable time with the DCNovations Blood Collection/Separation Materials Kit. The kit has everything you need for fingerstick blood diagnostic tests and blood sample collection devices. Perfect for everyone from startups to academicians to large companies. [1] Effect of serum separator blood collection tubes on drug concentrations. Quattrocchi F, Karnes HT, Robinson JD, Hendeles L., Ther Drug Monit. 5:359-62. (1983) [2] Influence of hematocrit on the measurement of lipoproteins demonstrated by the example of lipoprotein(a) Kronenberg F, Trenkwalder M, Kronenberg M, Koenig P, Utermann G and Dieplinger H,Kidney International, 54, 1385–1389 (1998) [3] Cyclosporin Therapeutic Drug Monitoring – an Established Service Revisited, Morris, RG, Clin Biochem Rev 24 33-46 (2003) --- ## Standards, Calibrators, and Controls in Immunoassays URL: https://dcndx.com/insights/standards-calibrators-controls-immunoassays/ Type: insight Published: 2020-01-27 Standards, calibrators, and controls: if you’re reading this, you probably know that they are all essential to immunoassay development, manufacturing, and quality control. At times, some of these terms are used interchangeably to refer to the same thing (or nearly the same thing). That’s a mistake. In truth, standards, calibrators, and controls are all quite different—and knowing the difference could make or break your next immunoassay project. Knowing the Difference Can Make or Break Your Project First, a quick primer: an immunoassay involves chemical reactions between clinical samples and reagents that are performed under standardized conditions. The resulting response is related to the concentration of analyte in the sample. Regardless of the form the immunoassay takes, the relationship between response and concentration needs to be estimated (i.e., “calibrated”). What are Standards in the Context of Immunoassays? A standard is a material that contains the same analyte as the intended target analyte and is in the same matrix. Standards represent an ideal that is very difficult—if not impossible—to attain in a high-volume commercial manufacturing environment. An example of a standard is the drug digoxin dissolved in an authentic human serum matrix. What are Calibrators in the Context of Immunoassays? Often, what are called standards in an immunoassay method are actually calibrators: either they are not in the same matrix as the target analyte (as is a standard), or else an analytically pure and immunologically active analyte is not available. Digoxin standards/calibrators provide a case-in-point. The matrix in which they are prepared closely mimics normal human serum, but it is prepared from recovered Acid Citrate Dextrose (ACD) plasma by clotting the pooled plasma with the addition of calcium chloride and removing the clots by filtration. The resultant material is only about 60 mg/mL protein rather than the 70 mg/mL protein of authentic normal human serum due to the dilution of the plasma by the anticoagulant ACD solution used during the blood collection. Preservatives such as sodium azide are also added to prevent bacterial degradation during storage. This is not a substance anyone would want in their circulatory system, but it does the job of providing reference points in a digoxin immunoassay. (If you’re already thinking this is a complicated topic, you’re not wrong: the scientists at DCN Dx have spent decades becoming experts in this field. Fortunately, DCN Dx offers courses that provide startups, students, and faculty who are starting development programs with the fundamental knowledge and skills they need for successful lateral flow projects. However, if you’ve gotten to this point and you’re hungry for more, read on—we dive a lot deeper into this topic below.) Sometimes this synthetic human serum must be further processed to make the desired calibrators. Thyroxine (T4) calibrators are prepared in charcoal-stripped defibrinated plasma because the pooled plasma contains normal amounts of T4. To make calibrators in the hypothyroid range requires a starting material that contains little or no T4. Charcoal stripping removes protein, so the resultant material is usually about 50 mg/mL protein. For total T4 calibrators, this lower protein level does not matter. However, for free T4 calibrators where the analyte concentration is a cross-product of the analyte level and the endogenous binding protein level, the protein concentration must be increased by ultrafiltration to remove excess water. Charcoal stripping works well for small molecules such as drugs and some hormones, but protein markers such as human chorionic gonadotrophin (hCG) or thyroid-stimulating hormone (TSH) cannot be removed by charcoal stripping. Normal plasma contains typical amounts of TSH and some hCG. Again, the creation of calibrators in the lower ranges requires starting with a matrix that has no or very little analyte. Immunoaffinity stripping using antibodies specific for the analyte has been demonstrated on a research scale, but it is impractical for high-volume commercial use. One solution that has been used in commercial practice is to use animal serum. These matrices may contain the animal version of the hormone in question (e.g., equine TSH), but antibodies raised against the human version of the hormone do not recognize the animal hormone. Sourcing the analytes for preparation of calibrators can also be challenging. Drugs and small hormones are often commercially available in pure form and can be quantitated by weight or UV/Vis measurement. Protein hormones are much more problematic. Originally, analytes such as hCG and TSH were isolated from human tissues obtained from cadavers. This approach works, but it presents supply and biohazard issues—human tissues can (and do) contain human pathogens. The advent of genetic engineering has provided an excellent solution to this supply problem— human hormones can be made in a mammalian cell culture. Attempts to make human hormones at a lower cost in bacterial cell cultures have failed. This is because mammalian hormones are glycosylated and the presence of these sugars changes both their biological function and their recognition by antibodies raised to recognize authentic human hormones. Quantitating protein hormones so that one can make defined calibrators is a challenge. UV/Vis measurements that work well for drugs and small molecule hormones do not suffice for larger protein molecules. Pure proteins tend to aggregate, and aggregated proteins do not quantitate reproducibly in immunoassays. Adding irrelevant proteins such as albumins can reduce aggregation, but these proteins have UV/Vis absorbances as well and are often added in huge excess over the concentrations of the target analytes. The common practice for calibrating these analytes is to use international reference preparations (IRP). These materials are prepared and assigned values by organizations such as the World Health Organization (WHO). They are prepared in large batches and lyophilized for long-term storage. The user reconstitutes the material and spikes it into their own matrix and then value assigns their own calibrators based on that international reference preparation. Because the original preparations were calibrated in bioassays, and given the difficulties cited above for precise quantitation of protein hormones, the calibration given by these IRPs was frequently in international units (IU) or derivatives of IUs such as milliIU or even microIUs. These units bear no relationship to moles or exact weights of analytes, but the package insert that comes with the IRP will often provide an approximate conversion of IU to micrograms. Nonetheless, these IRPs do help achieve some degree of consistency between different methods. Despite attempts to build large lots of IRPs, the stocks of these materials inevitably run out, prompting serial IRPs that all differ slightly from one another, further complicating the analysis. What Are Controls in the Context of Immunoassays? Controls used in immunoassays are a very different material. Their sole purpose is to measure whether a given method is providing the same results day after day and month after month. They may, in fact, be of the same composition as the calibrators supplied by the manufacturer. However, they are not used to calibrate the assay—only to check the assay’s consistency over time. Commercial controls that contain a bewildering array of analytes in the same vial are also used, and they provide an important independent reference point for a commercial assay. Calibrators and controls made in the same manner can drift over time in the same way, and this drift can go undetected. An independent control is best when it is available. Controls also provide a window into the different ways various methods see the same analytes. Controls usually come with a very long list of values for a given analyte, depending on which method is used for the analysis. This does not mean that some manufacturers are correct and others are not; it simply means that different methods using different antibodies and formats report different results. To be useful in commercial practice, reagents need to have a reasonable shelf life under defined conditions. Eighteen months is a typical goal as longer shelf lives generate unsold materials that are costly to prepare and store. Liquid ready-to-use calibrators and controls are the easiest to use and provide the least opportunity for user error. Lyophilization is used when the material is not stable enough to ship in a liquid form. Reproducible lyophilization is an art form that takes careful formulation and meticulous attention to the manufacturing process. Packaging can play a major role in the performance of calibrator and controls. Some analytes must be packaged in brown glass containers because they are light sensitive. Others require antioxidants to maintain activity. Even the stopper in the vial can have a critical influence: slight changes in the polymer formulation of the stopper can change a stopper from being an inert part of the packaging to an absorber of the analyte in question. Serum and plasma are not the only matrices that are tested in immunoassay. Other clinical matrices include urine, cerebrospinal fluid, amniotic fluid and saliva. Each presents their own analytical, base material supply, production, and stability issues that must be addressed by the assay development team as they craft an assay for real world use. In summary: Standards are the ideal method for evaluating assay performance but are seldom a realistic option Calibrators are a pragmatic solution that mimic a standard and allow reference points in comparing assay results to analyte levels in patient specimens Controls provide a means of evaluating an assay’s consistency, or reproducibility, in providing accurate analytical results from a patient specimen Need to get a head start on your organization’s assay development? Looking to quickly overcome a frustrating development issue? DCN Dx’s customized hands-on lateral flow training courses are the answer. These private courses are customized to your specific needs and delivered in DCN Dx’s lab using your own assay. Perfect for startups and university research groups in the beginning stages of lateral flow development projects; manufacturing groups; and research groups in mid- to large-size organizations. --- ## Order Your Lateral Flow Supplies Online: A New Way of Doing Things at DCN Dx URL: https://dcndx.com/insights/online-ordering-lateral-flow-supplies/ Type: insight Published: 2019-10-12 Several months ago, DCN Dx quietly rolled out a powerful new online ordering system that allows our customers to order lateral flow supplies from us in a faster, more efficient way. Don’t get us wrong—we love hearing from our customers via phone or fax, just like the old days. But if you have a desktop computer, tablet, or smart phone—and we’re sure many of you do—purchasing DCNovations products online just might be the way to go. It’s easy, accessible, and it means you’ll no longer be at the mercy of our lunch breaks. Following are just a few of the benefits of purchasing DCNovations products online: DCN Dx’s new e-commerce site makes it easy to order the products you need anytime, anywhere. Ease of Use: Our entire product catalog is available online, 24/7. When time is of the essence, there’s no need to wait for someone to answer the phone: just place your order when you need it, and it will be placed in our shipping queue. Existing POs: Do you have an existing purchase order (PO) with DCN Dx? Our online store accepts them! We also accept credit cards. Special Promotions: We regularly run limited web-only promotions, saving you valuable funds on large and small orders alike. Easy Account Set-Up: New user? No problem! Our account setup process was designed to be fast and easy for everyone from corporate lab directors and purchasing agents, to busy startup scientists. Purchase History at Your Fingertips: Our system saves your favorite and frequent orders and displays them for you, so there’s no more trouble remembering the lateral flow supplies you need: simply click and re-order. More Secure Than Fax or Email: The security of your information is more important today than ever before. Our online store uses PayPal’s encrypted payment processing to ensure that your billing information stays safe, secure, and confidential. Head on over to dcndx.com/store to find out how our new online ordering system can streamline your business. --- ## Common Assay Development Issues (And How to Avoid Them!) URL: https://dcndx.com/insights/assay-development-issues/ Type: insight Published: 2019-08-19 (As published 2019-08-19; regulatory status may have changed since.) When beginning any assay development program, you must consider a number of factors before you are on the road to a finished product. Between reagent selections, regulatory requirements, controls, sample collection, cassette design, and end user requirements, details can slip through the cracks. Even the smallest missed detail can halt progress and delay your development timeline. At DCN Dx, we have helped many clients prevent problems with assay developments in order to make assay development as efficient as possible. Whether you are just getting started and want to avoid delays or you are experiencing problems in your current program, our team of scientists and engineers can help you take the right steps. Here we discuss three common pitfalls during each stage of development and the simple steps to avoid them. Common Assay Development Issue #1: Reagent Selection Selecting suboptimal reagents for your assay is a pitfall with a long road to recovery. During the Feasibility stage, you must screen and optimize reagents and materials in order to determine the basic performance of the system and decide whether the assay is feasible. This will set the groundwork for the future direction of development. To ensure you choose the right reagents, keep in mind these tips. Each assay is unique Antibodies that performed well in other platforms, such as an ELISA, may not translate to lateral flow. Incubation time, washing steps, and sample dilutions can result in significantly different performance than other platforms. Materials and buffers that worked in similar assays may not be optimal for a new assay. To achieve full performance of your system, always screen antibodies, materials, and buffers for each individual assay. Think long-term Consider any limitations that will affect your assay in the long-term, such as reagent cost and availability in large quantities for the duration of manufacturing. If cross-reactivity is an issue, test the antibodies with the potential cross reactants early in development. Exchanging even a simple reagent down the road can require extensive re-optimization, adding time and cost to your development program. It is also important to keep the end user in mind. If you use fluorescent particles for maximum sensitivity, the end user will need a fluorescent-based reader. If you need to collect a set volume of sample such as saliva, you will need an efficient and user-friendly method for sample collection. Consider the end user and design the device for usability. Re-evaluation of critical reagents and materials at a later point in development is a significant detour. Common Assay Development Issue #2: Samples, Samples, Samples A lack of real samples or waiting too long to test real samples can halt progress and cause a serious setback in development. To avoid this often-underestimated setback, follow these general tips. Source samples Before you get started, identify where you will source your samples, how much they will cost, and what quantities and volumes you will require. The difficulty of getting real samples varies significantly based on what you are detecting. There are many sources for obtaining endogenous analytes in normal state samples (i.e., non-disease samples), as well as disease state samples. Certain sample types may be more difficult to obtain than others (e.g., stool vs. plasma), while others may require extremely fresh collection for testing (e.g., saliva). It is critical to make sure you have a verification method for the desired analyte, such as an ELISA or outside lab reference. Transition to real samples early Postponing the transition to real samples is another common hindrance to efficient assay development. Tackle this hurdle earlier rather than later in the development cycle. There are times when the purified antigen spiked into buffer or the sample medium does not mimic the native antigen. Introducing real samples also introduces interfering substances, potential cross-reactants, and sample variation issues that can have a significant impact on assay results. Optimization and screening may need to be repeated to mitigate these effects, so it is important to discover this early on in the assay program. Common Assay Development Issue #3: Transitioning to Manufacturing Thinking about the transition to manufacturing early in development can avoid many delays, save time, and save money. Considerations include the supply of reagents and materials, transitioning materials to reel-to-reel manufacturing, lot-to-lot variability of materials and reagents, scale-up manufacturing of buffers and assay components, the stability of each component, and the final assay. Scaling up Many processes during the R&D stage, from conjugating small volumes to striping a single membrane, are done at a small scale. Although it may seem like a negligible issue, scaling up these processes could alter your assay results and require optimization. To avoid this issue, compare the small-scale to the large-scale procedures side-by-side and determine the appropriate specifications and tolerances for each procedure. Consider the materials that will be used in scaling up to reel-to-reel manufacturing. Make sure the tensile strength of materials, such as a conjugate pad or sample pad, are compatible with reel-to-reel and that the materials are available in roll format. Variability and stability The development stage should include thorough evaluation of lot-to-lot variability of all materials used. Before Verification, conduct interim stability studies to identify any issues that may need optimization prior to transfer. Evaluate the stability of the reagents and materials before processing to ensure steady and stable storage of all required components during manufacturing. At DCN Dx, our manufacturing capabilities help transition assays from small-scale to full-scale lot sizes. Our in-house reel-to-reel system can help identify any issues quickly and we help find the optimal solutions. If you are interested in learning more about lateral flow assay development, join us for our Advanced Lateral Flow Course this October, where we will discuss assay development issues in greater depth. --- ## Plasma Separation: Why Do You Need It? How Do You Achieve It? URL: https://dcndx.com/insights/plasma-separation-why-you-need-it/ Type: insight Published: 2019-07-17 Plasma Separation: Why Do You Need It? How Do You Achieve It? Analyzing the components of blood is a key diagnostic step in the detection of diseases and accurate separation of plasma from blood cells plays a crucial role in the precision of a lateral flow diagnostic test. Why do you need Plasma Separation? Lateral Flow Devices are a rapid, inexpensive, and simple diagnostic tool intended to be utilized at the point of care. Due to the simplicity of use and rapid results obtained, the list of assays being utilized on this platform is ever expanding. As the types of LFA’s expand, there is increasing demand for the use of whole blood as the sample matrix. Being able to separate the plasma from cells and generate sufficient quantity of plasma is one of the most basic, central and crucial steps in the field of diagnostic to maximize performance of the test devices. Utilizing a blood separation step minimizes the interference of cells in the process of analyte detection while improving sensitivity and selectivity of the assays in lateral flow tests. How do you separate the plasma? Generally, plasma can be separated from a whole blood sample by mechanical methods using sedimentation or centrifugation, which requires a laboratory setting and additional equipment for sample processing. A very attractive alternative technique that can be adapted for POCT test strips utilizes fiber based filtration membranes that can be incorporated directly into the LFA test strip and do not require any additional equipment or sample processing. Vertical or lateral filtration methods using fiber based filtration membranes have been successfully employed for decades in lateral flow devices. In general, the lateral filtration techniques of plasma separation can be classified in two different categories: Biochemical filtration: This method consists of applying a pretreatment to the sample pad (commonly made of glass fibers) to agglutinate red blood cells and allow the plasma to further flow along the conjugate pad and into the nitrocellulose membrane. Mechanical filtration: This method relies upon the fibrous components and porous matrix, as well as, the specific pores size distribution, optimized for red blood cells retention. In order to achieve the best separation results, you need to consider the fibrous components, physical properties, sample volume and potential limitations that need to be considered. How to choose the right plasma separation pad? When it comes to lateral filtration for plasma separation, the key physical characteristics of fiber-based materials that need to be considered to get good and sufficient quantities of plasma are the absorption capacity linked to the thickness of the plasma separation pad, the red blood cell retention capacity and the overall dimension of the pad for the sample volume. For a fixed volume of blood, as thickness of blood separation pads increases, amount of recovered plasma decreases. However, by using appropriate pads dimensions and a specific chemical treatment, significant improvement in plasma recovery can be achieved. If the correct fibrous components are not chosen, hemolysis or rupturing of the red blood cells can occur. Hemolysis of any type should be limited within the diagnostic tests, as the rupture of the cells leads to the release of hemoglobin molecules, causing the serum or plasma to have a pink to red color, thus making the readout difficult to judge. Leakage of the cells content along the membrane can also cause problems such as low sensitivity, selectivity and inaccurate results. One of the main reasons causing hemolysis can be the use of an inappropriate type of glass separator pads, especially the ones with small diameter glass fibers. Ahlstrom-Munksjö Plasma Separation offering: Ahlstrom-Munksjö original single layer blood separator, CytoSep®, was developed in early 1990’s to address industry demand for whole blood testing. Working closely with our customer base we continued to develop improved fiber-based materials and methodically upgrade the performance of CytoSep®, meeting the increasing demand for sensitivity, consistency and non-interference. CytoSep® is widely used for rapid separation of plasma from whole blood samples in lateral flow applications, retaining blood cells while allowing serum to flow rapidly. There are three historical grades which differ in thickness: CytoSep® 1663, CytoSep® 1662, CytoSep® 1660. These untreated media contains no chemical-interfering substances and show no significant binding of plasma components. Going into our 4 th decade of developing improved and innovative components for Lateral Flow Immuno Assays, we continue to lead the industry providing new materials capable of keeping pace with this rapidly evolving industry. The newest CytoSep® products, HV 1667 and chemically treated HV Plus 1668, are designed to separate increased volumes of whole blood within the same area of media while limiting hemolysis. Focus on: CytoSep® HV plus 1668 combines both chemical and mechanical filtration processes for higher performance. The chemicals help to deform shape of red blood cells and so limit their diffusion while the specific pore size blocks the residual mobile RBC. On a defined sample pad area, CytoSep® HV plus 1668 is able to accept 25% more in whole blood volume compare to traditional plasma separation pads. Key Features: Faster blood absorption and plasma separation Higher plasma yield for increases in line sensitivity Designed with hemolysis prevention in mind Ability to separate larger volume blood samples so that less material can be used in new designs More blood – more plasma – more analytes – higher sensitivity Discover more about Ahlstrom-Munksjö CytoSep® and other plasma separation pads: https://www.ahlstrom-munksjo.com/products/medical-life-sciences-and-laboratory/lateral-flow-test-pads/plasma-separation-pads/ --- ## Holistic Detective Work: The Interconnectedness of Materials and Performance in Lateral Flow URL: https://dcndx.com/insights/holistic-detective-work-lateral-flow/ Type: insight Published: 2019-07-17 “The term `holistic’ refers to my conviction that what we are concerned with here is the fundamental interconnectedness of all things. Let me give you an example. If you go to an acupuncturist with toothache he sticks a needle instead into your thigh. Do you know why he does that, Mrs Rawlinson? No, neither do I, but we intend to find out.” Douglas Adams The proper performance of a lateral flow assay depends on the interconnected performance of a wide variety of materials, chemistries, biological reagents and processing steps. As a result, assay development and troubleshooting in manufacturing require a holistic approach. It is necessary to fundamentally understand the system with which we are working to isolate and fix problems and a willingness to suspend disbelief at times when attempting to do so. You’d be amazed at the things that can go wrong and the things that can work to fix them. There is rarely just one cause or just one solution to an issue. That’s why there is no substitute for experience when it comes to working with lateral flow systems. That’s why, having worked with this technology for close to 30 years now, it often feels to me like it wasn’t invented – more like it was discovered and allowed to evolve slowly on its own. People have tried to generate algorithms for development of lateral flow products and recently applied advanced programming and AI to the task. Robots have been created to assist with the development process. Computer models of lateral flow systems have been generated. Design of Experiments (DOE) principles have been tried. While some benefits have accrued from these efforts, nothing has replaced the experience and intuition of a good lateral flow development scientist as yet. When designing an assay, we start with first principles. Carefully evaluate the target performance requirements, understand the user needs and the application environment. Then begin the process of selection of materials and reagents. Each material used in an assay is specifically screened and selected in the context of every other material and reagent in the system. We know what each material needs to do, we know the pros and cons of the available selections and we have a basic menu from which we select and then we begin the process of developing the treatments we need to use to make them work. Nothing is selected in isolation. Let’s take the example of sample collection pads and conjugate release pad materials, what they have to do and how they can impact your assay. Understanding sampling in the context of small volumes and rapid assays is critical For many applications traditional lateral flow formats are capable of providing sufficient sensitivity. However, there is a growing demand for sensitivity in many applications that approaches that of nucleic acid amplification and detection methods. Standard approaches to labeling and detection in lateral flow are unlikely to reach the required sensitivities for these applications. Quantification and ease of use for consumer applications are also growing in importance. Sample collection and preparation is one key to improved sensitivity and overall performance in many instances. It should be remembered that a large element of the appeal of lateral flow and other point of need assay systems is that they should provide where possible a complete “sample-to-answer” solution in a single step. It is therefore critical to consider the system as a whole, including the sample, the sampling method, the sample pre-treatment methodology and the concentration of analyte in the system. Analyte concentration can be a confounding factor either when it is too high or too low for detection, and sample treatment can and must be used to overcome related issues. Sampling refers to the generation of a representative sample of an inhomogeneous object. This inhomogeneity presents a challenge to the success of the analytical method. As applied to highly sensitive rapid diagnostics, it is not the absolute sensitivity of the system that is the most critical factor, rather it is the ability to acquire as representative a sample as possible, and that, ultimately, it is the concentration of the analyte that one can detect in the primary sample that is critical. Sampling and pre-treatment methods, primarily concentration and the removal of potential cross reactive agents and reduction of background, are critical to determining the availability of many analytes for detection in an assay. Additionally, in certain circumstances, high concentrations of analyte can be a confounding factor in an immunoassay. In any assay system to be deployed in a decentralized testing environment, the sample collection, treatment and delivery method must be simple, robust, foolproof, and ideally an integrated component of the test device. Minimal user dependent steps should be required. Let’s look at fingerstick whole blood collection and plasma separation as an example. The primary processing steps for the blood sample are metering of quantitative or semi quantitative amounts of blood from the fingerstick separation of plasma from the sample without significant hemolysis delivery of the sample, either neat, neat with a chase buffer, or pre-diluted, to the device The creation of solutions that achieve several or all of these steps in an intuitive, user friendly way, is a constant challenge, and the needs of every assay system are different. Plasma Separation and Delivery to the Test Let’s assume a quantitative volume of fingerstick blood can be collected and delivered to the test. Plasma separation can be achieved in a variety of ways in rapid assays. The most common approach is to use filtration membranes as part of the strip architecture. Many lateral flow type systems utilize an in-line blood separation membrane such as Ahlstrom-Munksjö Cytosep membranes. These systems are relatively efficient, although they are limited in the volume of sample that they can handle based on useful surface area. Often the system will require either a pre-dilution of the sample or a chase/wash buffer to follow in order to wash the plasma clear of the membrane and to provide enough volume to wet the system completely. Filtration material for red blood cell separation For many current and next-generation applications in lateral flow, quantification is important. So the ability to deliver a quantitative and highly reproducible amount of plasma to a test is one of the holy grails of this type of material. The volume of plasma that can be separated and delivered is of course confounded by hematocrit however the material should ideally be capable of accepting a defined volume of whole blood and delivering plasma to the test strip with consistent efficiency and reproducibility. There are other ways to address the hematocrit problem in the context of assays that are to be evaluated on reader systems. Getting the plasma delivery right – minimizing hemolysis, maximizing reproducibility and separation efficiency, ensuring that analyte is not bound by the separation material and that the system flows in a timely and reproducible way – are the keys to success in this type of assay. Conjugate Stabilization and Release Switching to conjugate release pads, these materials – typically glass fibers and polyesters – can make or break your assay and your manufacturing process, and give you some gray hairs along the road. The role of the conjugate pad in a typical assay system is to accept the conjugate, hold it stable over the shelf life of the product, and release it efficiently and reproducibly during that entire shelf life. In practical application, the variations in conjugate deposition, drying and release from the material demonstrably contribute the greatest sources of variation in assay performance, as measured by within and between lot coefficient of variation (CV). Assay sensitivity can also be adversely affected by poor conjugate mixing in, and release from, the conjugate pad. Depending on the system, it may be more important to achieve fast release or slow release of the conjugate, however release must always be consistent. Because of the nature of the materials used, it is often necessary to pre-treat conjugate pads to ensure the appropriate release and stability characteristics. Pad pretreatment is typically performed by immersion of the pad in an aqueous solution containing proteins, surfactants and polymers followed by drying. This process can be performed either in manual batch mode or in continuous inline mode, the latter giving the best opportunity for homogeneous processing of entire batches of materials. The addition of conjugate to the treated pad is critical to the final performance of the test. Two methods are typically used: Immersion: where the treated conjugate pad is immersed in the conjugate suspension. Dispensing using quantitative non-contact dispensers such as the BioDot AirJet Quanti. In relation to the conjugate system, the choices of label and conjugation methods are also critical. The most commonly used labels include colloidal gold, cellulose nanobeads and monodisperse latex, tagged with either a visual or a fluorescent dye. A variety of labels are now available, which can be covalently or passively coupled, and quantitatively read. Covalent coupling can be important to the ability to perform quantitative assays due to the inherently more stable bonds between the ligand and particle vs. typical passive adsorption methods. The materials most commonly used are glass fibers, polyesters or rayons. The materials ideally should be hydrophilic and flow consistently. The materials used, however, are typically very hydrophobic in nature, and as a result must be treated to make them hydrophilic. This is typically done during the manufacturing of the assay rather than by the material manufacturer, although there are exceptions to that. This treatment involves the immersion of the pads in a solution of proteins, polymers and surfactants, followed by drying at high temperatures. The goal is to use materials and treatments that enable efficient and reproducible release of the conjugate over the shelf life of the product. Typically some variation in release occurs due to binding of the particle conjugate to the fibers of the material. It is important during assay optimization to generate stabilization chemistries that minimize this effect and create the most efficient release of particles possible. Finally, it is important that the material used should not destabilize the conjugate over entire shelf life (up to 2 years). Assay optimization therefore involves the testing of multiple materials for compatibility with the protein-particle conjugate being used. The conjugate pad system is demonstrably responsible for the majority of variation in lateral flow assays when particulate labels are being used. This is due to inconsistency in the material, resulting in inconsistent uptake of the pretreatments and conjugates, destabilization of conjugates by binders, inconsistent release of the conjugate due to binding of particles to the hydrophobic fibers of the materials used. Great care must be taken in the optimization of the conjugates, the pad pre-treatment process and the conjugate deposition process to minimize these effects. The interplay of these materials with each other and with the other materials in the assay will define how your assay and your manufacturing system will perform. Selecting materials from high quality suppliers who will work with you to provide the material characteristics and quality you need and who understand the interconnectedness of all things in the performance of the assay will be critical to success. DCNovations is a line of high-quality precision rapid diagnostic test products and components distributed by DCN Dx, located in Carlsbad, California. DCN Dx is recognized globally as the go-to company for contract development and commercialization of rapid diagnostic tests. The company’s cross-functional team of scientists and engineers develop and integrate all aspects of assay systems including cassettes, sample handling devices, and reader systems. DCN Dx also provides services to researchers and labs that assist in the development of rapid diagnostic tests from concept to commercialization. --- ## NALF and Beyond: Lateral Flow Goes Molecular URL: https://dcndx.com/insights/nalf-lateral-flow-goes-molecular/ Type: insight Published: 2019-06-30 Antibodies and other immunoreagents are so tied to lateral flow assays that we frequently use the terms LFA (lateral flow assay) and LFIA (lateral flow immunoassay) interchangeably. However, lateral flow assays may have no immunoreagents whatsoever onboard. Over the past 15 years, a subset of lateral flow testing using nucleic acids as targeting molecules or analytes, including nucleic acid lateral flow or NALF, has quietly emerged and entwined principles of both immuno- and molecular recognition systems. Nucleic acid techniques, immensely sensitive because of enzymatic exponential amplification, offer unrivaled performance. However, their detection comes at a cost. Described here are some assay architectures that incorporate nucleic acids to take advantage of the many assets LFAs offer, such as low cost, simplicity, and low labor requirements. Nucleic Acid Lateral Flow (NALF) The strictest definition of nucleic acid lateral flow (NALF) incorporates no immunoreagents on the nitrocellulose membrane. Signal originates from a sandwich hybridization assay occurring at the test line, at which a DNA probe is commonly immobilized by a terminal biotin that interacts with streptavidin or NeutrAvidin on the nitrocellulose. As in lateral flow immunoassays, signaling moieties can be oligonucleotide-decorated gold or latex particles, or simply a fluorescent dye that modifies the tag sequence. One potential drawback of NALF is that recognition of the target strand by the probe and tag strands must occur quickly and is highly dependent on Watson-Crick base pairing hybridization kinetics. Any secondary structure in any of the three strands will greatly destabilize these interactions, as might common additives (e.g., surfactants) that promote flow up the membrane. These parameters must be individually optimized for each discrete sequence. Nucleic Acid Lateral Flow Immunoassay (NALFIA) The nucleic acid lateral flow immunoassay (NALFIA) hybrid format incorporates together the selectivity of Watson-Crick base pairing, the sensitivity of molecular techniques, and the well-known and well-characterized immunochromatographic assay. NALFIA is commonly used to detect amplicons in a point-of-care setting and eliminates the need for time-consuming agarose gels or the expensive optics that accompany a real-time system. In a NALFIA scenario, base modifications are incorporated into the primers, and therefore, the amplicons, using a given amplification technique. After the cycles are complete, the amplicon then acts as an antigen in a lateral flow assay, with species on the nitrocellulose membrane and the conjugate able to bind the amplicon in a sandwich via the modifications on the primers. Nucleic acid modifications for NALFIA are widely commercially available and include fluorescent dyes (FITC/FAM, Cy5, Texas Red) and small molecules (biotin, digoxigenin, DNP). Antibodies against these modifications are widely available from numerous sources. Because detection is independent of sequence and instead relies upon recognition of tagged primers by antibodies, universal lateral flow tests can be applied to the detection of multiple different amplicons. Anti-digoxigenin, anti-FAM, and anti-biotin are common capture antibodies, and these strips can be purchased from various vendors or produced in-house. This universal recognition also means that NALFIA can be applied towards the detection of amplicons produced by many different amplification scenarios. Common methods include: Symmetrical and asymmetrical polymerase chain reaction (PCR) SNP detection via ligase chain reaction (LCR) Isothermal techniques, such as loop-mediated isothermal amplification (LAMP), rolling-circle amplification (RCA), and recombinase polymerase amplification (RPA) Newly-emerging isothermal techniques are especially attractive, as they eliminate thermocycling and can potentially be executed at room temperature, greatly simplifying equipment requirements. These methods paired with the speed of LFIA move molecular diagnostics more towards a point-of-care scenario and a cheaper price tag. Lateral Flow with Aptamer Technologies Though lateral flow tests developed with aptamers do not fall strictly into the definition of molecular diagnostics, they do offer a nucleic acid-based alternative to antibodies, with some notable advantages. Aptamers represent a class of nucleic acid probes that are specifically engineered to adopt a certain conformation and bind targets of interest. The Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method, developed by the teams of Tuerk and Gold and also Ellington and Szostak in 1990, enabled the discovery of the first RNA aptamer probes against T4 DNA polymerase and organic dyes, respectively. A random library of DNA or RNA is incubated with the target for interaction to occur and then the bound sequences are separated from the unbound sequences and amplified. This process is cycled numerous times, with higher-affinity sequences emerging at the end of each cycle. Due to the varied nature of the targets available (small molecules, proteins, peptides, cells), aptamer-target affinities can vary from sub-nanomolar up to micromolar, making Kd values potentially on-par with antibody-antigen interactions. Further, aptamers offer a number of potential advantages over antibodies, including automated and reproducible synthesis, increased stability, and ease of labeling and scale-up during the fabrication process. Assay architectures with aptamer lateral flow are similar to those found in traditional antibody-based LFIA. If two aptamers are available for a given large molecular weight target, and the aptamers bind to different regions of the analyte (dubbed “aptatopes”), a sandwich assay can be designed. Competitive assays are also popular, and because aptamers can bind either their target or their nucleic acid complement, there is more than one option for a test line. Signal-on assays can also be designed by incorporating a complementary and sacrificial quencher sequence that is freed from a fluorophore-labeled aptamer once the target is bound. While lateral flow cannot currently rival the sensitivity of molecular techniques such as PCR, one could easily argue that given resource, time, and cost constraints, ultrasensitive detection is not lateral flow’s space. Lateral flow shines in its ability to supplement and support molecular techniques in a point-of-care scenario. DCN Dx’s scientists and engineers are skilled in the development of many different immunological and molecular assay architectures, including NALF. We now offer a Nucleic Acid Lateral Flow Kit from DCNovations, a generic lateral flow platform for rapid point-of-care detection of nucleic acids from amplification reactions, including those for infectious diseases like COVID-19. Learn more. --- ## Make or Buy an Anitibody? URL: https://dcndx.com/insights/make-buy-antibody/ Type: insight Published: 2019-06-13 To make or buy an antibody? Good question. To develop an immunodiagnostics test, you need an antibody. Assuming you’ve done your homework to specify the characteristics of the antibody you need, a classic business decision must be made: do you make or buy an antibody? Buying an antibody off-the-shelf is certainly the fastest route to getting that critical reagent. All the expensive and time-consuming work of generating an antibody has already been done. Someone else has assumed the risk that a particular antibody generation campaign might not work or may not have worked the first time. An online search for a mouse monoclonal antibody to human hemoglobin for a lateral flow assay produced 76 candidate antibodies in seconds. A similar search for hemoglobin monoclonals for ELISA produced 456 results. Clicking on a particular monoclonal will bring you to the vendor’s website with information on the purification method (if any), isotype, purity, storage buffer, vial sizes and cost. “ Linscott’s Directory of Immunological and Biological Reagents ” is one excellent resource for finding the necessary antibody. As attractive as buying an off-the-shelf antibody sounds, there are some drawbacks to this approach. When you buy an antibody from a catalog, continued supply is a calculated gamble. A critical question to ask the vendor is: “Do you audit the antibody supply?” If the answer is yes, then the manufacturing process for making the antibody can be examined in person or on paper to ensure long-term viability of its supply. If the answer is no, the risk of using that antibody increases considerably. Most major diagnostic companies consider not being able to audit an antibody supplier a showstopper. In the early stages assay development for a new analyte, the antibody you need may not be commercially available yet. In this case, the alternative to an off-the-shelf purchase is to scour the literature to what has been published on an antibody or assay of interest. If the authors are academics, they may be willing to make their antibody available. Sometimes this can be done informally with the scientists who developed the antibody, but more usually this will involve the technology transfer or business development office of the institution that owns the antibody. Depending on how the antibody will be used, securing a clear legal pathway for access may be advisable. There is no sense in investing time and energy developing an assay around a critical reagent if then is no legal guarantee of continued access. Academic institutions are not well equipped to ensure continuous supply of antibodies. Even if you secure access to a novel antibody for evaluation, the process should not stop there. Efforts to license the cell line should follow the successful evaluation of the antibody, and then that cell line should be transferred to a commercial entity that is experienced in antibody production. There the cell line will be tested for pathogens that could eventually affect yield, monoclonality, isotype and other parameters. A master cell bank and a working cell bank will be established to ensure reliable continued production. Offsite storage of some of the master cell bank is insurance against a disaster (fire, flood, extended power outage) at the primary storage facility. This process can be expensive but not as expensive as redeveloping and revalidating an assay if a critical raw material is no longer available. Buying catalog antibodies is also expensive. Relatively common antibodies—those with hundreds of candidate antibodies—can often be found for several hundred dollars per milligram. At a coating concentration of 3 ug/mL with a coating volume of 100 uL, a milligram of antibody translates into more than 3000 microtiter wells or several thousand lateral flow strips. Less common antibodies can command prices of thousands of dollars per milligram. You can often achieve lower per milligram antibody costs by investing in the development of your own antibodies. The first step in generating your own antibodies is securing a reliable supply of the antigen in question. For small molecules, such as drugs and hormones, this requires some organic chemistry to attach the analyte or more often an analog of the analyte to a carrier protein. Molecules with a molecular weight of less than 6000 daltons are not immunogenic unless coupled to a carrier protein. A favorite carrier protein is Keyhole Limpet Hemocyanin (KLH), which is isolated from the hemolymph fluid of a mollusk that lives off the coast of California. It is highly immunogenic, has many accessible chemical attachment moieties and has a molecular weight of approximately 390 kD. To source human protein antigens, researchers often start with either human tissues, cell cultures or body fluids such as urine, amniotic fluid, semen or blood as a raw material. Note that the protein antigen, drug or hormone is also required for calibrators and controls. Next comes the immunization campaign itself, in which multiple rabbits, mice or goats are injected with the antigen in a carefully prepared cocktail to boost the immune response. One popular mixture is Freund’s Complete Adjuvant. Multiple animals are used because some animals do not produce usable antibodies and failures of entire antibodies campaigns are not uncommon. Multiple boosts at multi-week or monthly intervals are needed to increase the quantity and affinity of the antibody. Some mouse monoclonal antibodies can be obtained in as little as three months. The same cell banking precautions as described above for purchased antibodies should be done for newly developed antibodies. Polyclonal can take 6-9 months to mature and then developing a pool from periodic bleeds can take a few months more. This work can be done in-house if the facilities and expertise are available; otherwise, multiple vendors can provide these services. Polyclonal campaigns can cost as little as $1,000 for a relatively simple small molecule antigen. Monoclonals are more complicated and, as a result, more expensive, with campaign costs averaging $6,000-15,000. How do you decide which route to take? One decision-making approach is to make an Excel model of the two different antibody procurement paths and a likely outcome in terms of sales and net revenues. Consider your development costs, the timing of when the product will be marketed, and R&D costs as factors. The production cost per test, including the antibody costs, should also be inputs. Costs are entered as negative numbers and revenues are entered as positive numbers over a number of years. Negative development cash flows in the beginning are hopefully balanced out by positive revenues in later years. Making an antibody incurs greater costs in the beginning and lower antibody costs over time. Buying an antibody trades higher antibody costs over time for lack of antibody development costs upfront and has a time-to-market advantage. All of these uneven cash flows can be combined in a net present value (NPV) function embedded in Excel. NPV takes into account that $100 today is worth more than a promise of $100 next year. The NPV calculation includes an estimate of the cost of money or discount rate. A discount rate of 5% can be used for relatively low risk assays (existing market, little competition) and a higher rate of 15% or higher can be used for a newer analyte of unproven value. The NPV function calculates the sum-total effect of all of the variables and can provide guidance for this antibody procurement decision. Note that an NPV model can also shed light on the advisability of a hybrid model, in which an early entry assay is developed with a purchased antibody, which is then followed by a cost-reducing, newly developed antibody assay. Just be sure to include the costs of redevelopment and revalidation (and, in some, cases re-registration) of the new assay. Also consider the opportunity cost of allocating your resources on this project versus what they could develop instead. DCNovations (products by DCN Dx) offers a wide selection of antibodies for your assay – shop now! --- ## Polyclonal Antibody or Monoclonal Antibody Selection for Lateral Flow Rapid Tests URL: https://dcndx.com/insights/polyclonal-monoclonal-antibody-selection-lateral-flow/ Type: insight Published: 2019-02-21 Should you select polyclonal or monoclonal antibodies for your lateral flow rapid test? Some readers are probably wondering why polyclonal antibodies are not a thing of the past, like manual transmissions on cars and telephone landlines. However, the older models still have many advantages. This blog explores these advantages in the context of lateral flow assays. Both monoclonal and polyclonal antibodies start out in much the same way: immunization of a live animal. Rabbits, goats and sheep are the most common subjects for making polyclonal antibodies and mice are the starting point for most monoclonals. Repeated immunizations are key for making good antibodies in any species. Detectable antibodies can be seen in a few months of boosting, but a really good polyclonal response can take 6 to 9 months to fully mature. Once the desired response has been achieved, production bleeds can begin. It is possible to take about 40 mLs of blood from a healthy rabbit once a month for several years with no ill effects on the animal’s health. This blood yields about 20 mLs of useable serum after processing. These sera are accumulated and tested for titer and other relevant parameters before being pooled if the individual sera are similar enough. A pool of 100 mL developed over a year is not unusual. With a coating titer of 1:10000 and a coating volume of 100 uLs, that size pool should be sufficient to produce 100,000 96-well microtiter plates or >1,000,000 lateral flow strips. A company in Massachusetts recently quoted a cost of $1,000 to develop a rabbit polyclonal antibody to a small molecule in about a month. Additional booster injections and keeping the animals longer were offered at additional yet modest costs. Production of monoclonals is more technically complex. Once the mouse has shown an adequate polyclonal response, the animal is euthanized and the spleen cells are fused with a mouse myeloma line to yield immortal cell lines. A single fusion can lead to several hundred individual cell lines, all of which must be screened and isolated so that a true monoclonal antibody with only one specificity can be assured. Good manufacturing practices demand that cell banks be created and stored in different locations to ensure a continuing supply of a given monoclonal antibody. A central European company offers a monoclonal antibody project for under $6,000 and cites a time frame of about 3 months from immunization to initial samples of antibody. Monoclonal antibodies have several advantages over polyclonals for routine production of diagnostic tests. Their specificity is the most obvious one. They only react with one epitope on the target molecule. Polyclonals contain hundreds or even thousands of specificities that can lead to cross reactions that can compromise an otherwise excellent test. An example of this would be a test for hCG, the hormone that shows rapidly increasing levels in blood and urine during the early stages of pregnancy. hCG shares common subunit building blocks with TSH, LH and FSH. Early polyclonal pregnancy tests could show false positives because the assay detected an LH surge during ovulation due to cross reactions between an antibody raised to hCG with LH. Ovulation and pregnancy are related but clearly different conditions. Monoclonals have another important advantage over polyclonals in that once the monoclonal line is established, the supply of that particular antibody is essentially infinite. It is true that more antibody must be produced either in cell culture or in ascites, but the difficult and risky task of isolating the desired cell line never has to be repeated. That is not the case with polyclonals. Pools of polyclonals eventually run out and there is no guarantee that immunizing other animals will yield a useable antibody. Estradiol is a good case study to illustrate some of these issues. Attempts to make mouse monoclonals to estradiol have been uniformly unsuccessful. Researchers have tried different immunogens with differing attachment chemistries, but the seemingly insurmountable problem is that mice appear to metabolize the estradiol immunogens to their sulfate and glucuronide derivatives and then they make antibodies to the derivatives. Since these metabolites are present in blood, these antibodies may cross react with estradiol but will give test results that are too high because they recognize both authentic estradiol and its sulfate and glucuronide derivatives. Ratios of analyte to metabolite also vary from patient to patient, further complicating that analytical problem. Rabbits immunized with estradiol immunogens also produce antibodies to the metabolites in most cases, but brute force and pure luck can sometimes yield a good antibody. Campaigns in which over 500 rabbits have been immunized to get a useable estradiol antibody have failed to produce a single, useful antibody, but a European academic was lucky: he got a rabbit that produced an antibody that met the sensitivity and cross reactivity requirements. He built a pool of more than 100 mLs and then, figuring he had several lifetimes of antibody, he then exsanguinated the rabbit. That pool lasted about 5 years when it was used to manufacture tests for a high throughput automated analyzer. Finding a replacement antibody was a difficult and expensive process. Monoclonals are also easier to label than polyclonals. The highest percentage of a specific polyclonal antibody I have ever seen was with a hyperimmunized rabbit antibody to Fluorescein Isothyocyanate (FITC). Roughly 0.4% of the total IgG was directed against FITC. The rest were the antibodies that rabbit had raised to defend itself from its environment. Affinity purification can be used to prepare a polyclonal antibody preparation that is suitable for labeling, but that step adds complexity and cost. Sometimes monoclonals can be too specific. A good example of this is the silent LH problem that surfaced when a major manufacturer introduced a sandwich assay for LH that used a monoclonal antibody for the capture antibody and another monoclonal as the detector antibody [1]. It was a very good assay that seemed to work well until they tried it in Scandinavia. Using the test, it seemed to indicate that 25% of the women there did not produce any detectable LH. Tests that used at least one polyclonal antibody to LH had no problem detecting LH. Investigation soon showed that the women with LH undetectable with monoclonals to LH had a deletion mutation in their LH that had no effect on the function of the hormone. As luck would have it, the region that was deleted is immunodominant in the mouse. That means that immunizing mice with wild type LH without the deletion mutation is going to preferentially produce antibodies to that epitope that is missing in the Scandinavian women with the mutation. Either by accident or design an antibody company made a monoclonal to LH with the deletion mutation and the problem was solved. LH is not the only analyte where monoclonals can be too specific. Modern troponin I assays use cocktails of monoclonals to overcome this problem. Sometimes two monoclonals are used as capture antibodies and two other monoclonals are used as detector antibodies [2]. Various subspecies of phosphorylated or oxidized troponin are thought to be in play. Established practice can also keep a polyclonal antibody in the market when a more specific monoclonal antibody is available. I once developed a drug assay for an automated platform that used a monoclonal. The competition was an established assay that used a polyclonal antibody that cross reacted with a metabolite. Laboratories and their clinicians had grown accustomed to the results provided by the cross reacting polyclonal and would not accept the lower results provided by the analytically superior monoclonal. Polyclonals were the first antibodies used in immunodiagnostics and their simplicity and relatively lower costs are going to keep them in play for some time to come. Monoclonals solve some of the supply and purity issues inherent in polyclonals but bring their own set of problems of narrow specificity, added costs and complexity. Landlines allow calls to specific locations and cannot be easily turned off. Manual transmissions dominate in areas where fuel costs are extremely high. As with most technological options, flexibility in choosing the right solution in a given environment is prudent and cost effective. Our team of engineers and scientists can help you determine the specification for monoclonal or polyclonal antibody selection for your test. Feel free to contact us with any questions about antibody selection. [1] Heterogeneity of plasma gonadotropins. Consequences on immunological properties of LH. Roger M, Lalhou N., Nucl Med Biol. 21(3):349-57 (1994). [2] Analytical Characteristics of High-Sensitivity Cardiac Troponin Assays, Apple FS and Collinson PO, Clin Chem 58:1 54–61 (2012) --- ## Antibody Seletion for Optimal Lateral Flow Assay Development URL: https://dcndx.com/insights/antibody-selection-lateral-flow-assays/ Type: insight Published: 2018-12-09 Antibody selection is key to creating a robust lateral flow immunoassay that consistently provides reliable analytical information. As discussed in a previous blog, proper antibody selection calls for a rigorous design process in which the assay’s intended use is defined with enough detail so that specific information is gathered and considered by all stakeholders. Some “give and take” during this process is normal and stretch goals should be included and clearly identified. No one really knows what they can do unless they try. Before venturing into the lab to begin the antibody selection process, one should be immersed in the literature of the assay in question. There is no need to reinvent wheels that already exist and that are documented in the literature. This should include the peer-reviewed literature and competitors’ package inserts, material safety data sheets, and the patent literature. This process yields parameters that are useful in antibody selection. The parameters could include molecular weight of the analyte and the desired analytical range, sample size, reaction volume, and assay time. It is important to consider the analyte’s molecular weight when deciding whether the assay is a competitive or a sandwich one. Antibody binding sites typically recognize six to eight amino acids, so a compound such as Angiotensin I (which has 10 amino acids) can only be addressed with a competitive assay simply because there is not enough space on the molecule to accommodate more than one antibody. Small molecules, such as steroids and drugs, can also only be addressed with competitive assays. A 30 amino acid peptide such as GLP-1 can give rise to a whole family of antibodies that recognize different regions of the peptide. Proteins that are bigger than GLP-1 obviously have the space to also bind to multiple antibodies. Once the likely assay architecture (competitive or sandwich) is established, it is time to consider the desired analytical range. There are often one or more medically relevant concentrations that are encompassed in the analytical range. For example, a lateral flow assay for Thyroxine or T4 should have a range such that both the hypothyroid cutoff (4.6 ug/dL) and the hyperthyroid cutoff (12 ug/dL) are in areas of the assay where adequate reproducibility is possible. Competitive assays typically span a range of concentrations of about a factor of 10 (1 log), so a good T4 assay would have a range of 2–20 ug/mL with a midpoint of about 8 ug/mL. Sandwich assays have a better range with about a factor of 30 (1.5 logs) from bottom to top, so a screening assay for prostate-specific antigen (PSA) with a medical decision point of 4 ng/mL ought to go from 0.5 ng/mL to 15 ng/mL with a midpoint around 4 ng/mL. Two parameters determine how an antibody behaves. One is the affinity constant, which is fixed. The other is the concentration of the antibody in the reaction, which can be varied. The best way of deciding which concentration of antibody to use is to do a titration of antibody concentration while holding all the other parameters (e.g., sample size, total assay volume, label concentration, timing) constant. While this can be done with the entire standard curve range of calibrators, it is often sufficient to probe the system with just a few calibrators such as the medical decision point, a zero point, and a high calibrator point. For a T4 assay, this would be 0 ug/dL, 4 ug/dL, and 16 ug/dL. For a PSA assay, these concentrations would be zero ng/mL, 4 ng/mL, and 15 ng/mL. The abbreviated three-point standard curve is quite useful for screening multiple antibodies. A few words of caution are in order about varying the antibody concentration. Closest packing of antibodies on a surface provides a saturation coating at about 1 ug/cm2. 1 On plastic surfaces, this coated concentration can be achieved with coating concentrations of about 3 ug/mL. Exceeding this concentration can leave loosely bound antibodies that can interfere in the desired reaction and give rise to paradoxical low-dose hook effects. This appears as diminished assay sensitivity in a sandwich assay or as increased signal with increased analyte concentration in the lower end of the standard curve. An antibody concentration that is too low can also cause problems. In addition to the obvious problem of low binding, trying to coat antibodies at less than 0.5 ug/mL can lead to bizarre antibody stability problems that manifest as increasing antibody activity over time. Spiking the coating antibody with an irrelevant antibody at 0.5 ug/mL can obviate this problem without affecting binding levels. Screening antibodies on a platform other than the one the assay is intended to be ultimately used on is often a waste of time. For reasons that are not well understood, antibodies can behave differently on different platforms. Antibodies that behave well in a microtiter plate platform often do not perform as well on magnetic particles. Once a reliable standard curve has been established, the real work of selecting an antibody begins. Since all other parameters depend on having a reproducible result, rigorous reproducibility studies must be conducted. A look at almost any FDA approved immunoassay’s package insert will provide examples of what data is needed. The real test of any antibody is how it behaves with real world samples, preferably ones with relevant assay values already known from a well-validated reference method. A minimum set of at least 100 samples spanning the range of interest should be assembled. Adequate volumes of these samples will ensure the ability to test multiple antibodies several times, if necessary. Smaller subsets of samples (10–20) can be used in an initial screen. There is no perfect assay, and that includes any putative reference method. In almost any method comparison, there will be discrepant results in which the reference method yields a result in one bin (e.g., above 12 ug/dL T4) and the test method yields a result in another bin (e.g., below 12 ug/dL T4). A “tie-breaker” assay from another source can be useful in this case to determine which assay is correct. If the tie breaker can be another technology (such as liquid chromatography-mass spectrometery [LC-MS]), so much the better. A good correlation between the new method and the reference method—but with a slope greater or less than unity—can indicate problems with calibration or with a binding partner in the sample. Spiking and recovery experiments can help determine if a binding partner in the sample is in play. Binding partner problems can be solved with the addition of releasing agents such as anilonaphthalene sulfonic acid salts 2, salicylate, or synthetic analogs such as danazol 3 that are not recognized by the antibody in the new method. Selection of the right vendor for the antibody is also crucial. Since a lot of time and money are going to be expended in developing an assay, it makes sense to ensure that the selected antibody is going to be available over time. A shorthand question to ask is, “Is this antibody supply or cell line auditable?” If the answer is “No,” then selecting this vendor increases the risk that the antibody supply could be compromised over time. Auditing the vendor either in person or remotely with a questionnaire can identify weak spots in the vendor’s practices that can be addressed. The issues of monoclonal antibody versus polyclonal antibody and whether to make or buy the antibody are also important and will be discussed in future blogs. Our team of engineers and scientists can help you determine the specification for antibody selection for your test. Feel free to contact us with any questions about antibody selection. 1 Binding of protein to polystyrene in solid-phase immuno assays Pesce AJ, Ford DJ, Gaizutis M, and Polak VE, Biochim. Biophys. Acta 492, 399 (1977) 2 Chopra IJ, Ho RS, Lam R. An improved radioimmunoassay of triiodothyronine in serum: its application to clinical and physiological studies. J Lab Clin Med;80:729-739 (1972) 3 Enzyme Immunoassay of Estradiol in Serum of Women Enrolled in an In Vitro Fertilization and Embryo Transfer Program; Bouve J, De Bouver J, Leyseele D, Bosmans E, Dubois P, Kohen F, and Vandekerckhove D; CLIN.CHEM.38/8, 1409-1413(1992) --- ## Antibody Labeling and Analyte Detection for Lateral Flow Assays URL: https://dcndx.com/insights/antibody-labeling-analyte-detection/ Type: insight Published: 2018-08-17 If the period at the end of this sentence were printed on paper, it would probably contain less than 10 19 molecules or atoms of ink [1]. By contrast, a lateral flow pregnancy test that shows a positive is detecting about 10 10 molecules of human chorionic gonadotropin (hCG) [2], assuming a cutoff of about 6 mIU/mL and a sample size of roughly 50 uL. In a lateral flow assay (LFA) it is typically not the analyte such as hCG that is detected but an antibody labeling the analyte. hCG has a molecular weight of about 38 kD and mouse IgG antibodies have a molecular weight of about 150 kD, so even if several antibodies are bound to each analyte molecule, it is going to be difficult to detect such a small amount of material. The task is made even more difficult by the fact that proteins such as hCG and antibodies do not absorb visible light. Uncooked egg whites offer proof of this statement. Even though egg whites contain about 85% protein, they are translucent. Only when egg white proteins are denatured by cooking or beaten with air to make a meringue do they scatter ambient light and appear to be white. Fortunately, innovative chemistry has developed several antibody labeling materials that can be used to modify antibodies to enhance their detectability [3]. These antibody labeling materials include gold nanoparticles, colored cellulose particles, colored latex particles, magnetic particles, carbon nanoparticles, quantum dots, fluorophores and various enzymes. All of these antibody labeling materials can be used to modify detector antibodies without adversely affecting their ability to bind to the relevant target analyte. Antibodies are incredibly durable reagents and can be modified with as many as 40 added label molecules without affecting their affinity or specificity. Antibody labeling with as many as 60 labels is chemically feasible but then reagent stability and shelf life may be compromised. Antibody labeling materials also can be optimized to minimize non-specific binding to other components of the LFA which could adversely affect sensitivity. Gold nanoparticles are the most commonly used label for LFA. While the exact color depends on the size of the nanoparticle, particles that appear red are most commonly used in LFAs. Colored latex beads offer a wide range of colors that can be used to implement multiplexing of LFA where more than one analyte at a time can be detected. Cellulose nanobeads offer many of the same advantages as latex beads but according to the manufacturer offer 8-10 times greater sensitivity than conventional gold nanoparticles [4]. All of these labels rely on detecting the color of the label on the surface of the LFA. What that really means is that some of the ambient light is being absorbed by the label and converted into invisible heat. What is detected is the light that is not absorbed by the label but reflected back. More sensitivity can be gained by using labels that are fluorescent. In fluorescence, a certain color or wavelength of light is absorbed by the label and after a very short time, a slightly different color of light is emitted. Filters can be used to stop the original light from being detected, and only the emitted light is thus seen against a diminished background. Direct labels such as fluorescein isothiocyanate or fluorescently labeled latex particles have been used. Magnetic particles can be used as visible labels in LFAs, but the technologies used in reading magnetic audio tapes and computer disks can also be employed to achieve even better sensitivities [5] Carbon nanoparticles derived from soot obtained by burning various materials including toluene have been used as labels in LFAs [6]. One strategy employed by a company named Vivacta was to use the thermal properties of carbon nanoparticles when irradiated by infra-red light to change the signal detected on a piezoelectric film. Enzymes such as horseradish peroxidase and alkaline phosphatase have long been used in other immunoassay formats such as microtiter plate ELISA. They have been adapted for use in LFAs as well [7], and they provide impressive sensitivity down to 0.8 ng/mL of progesterone. In contrast to the enzyme substrates used in microtiter plate ELISAs, the substrate used in the reference cited in this reference produces an insoluble product that precipitates on the device. However, this increase in sensitivity comes at the cost of another reagent (the substrate) and another step in the assay process. Unbound enzyme in the labeled reagent must be washed away either with continued sample flow or an additional wash step before the substrate can be added. This added complexity is a disadvantage in a resource limited setting and adds operator training and compliance requirements. Added complexity is the cost of all of the labels cited above that increase sensitivity over that achieved with gold nanoparticles or other labels that can be read by the naked human eye. Fluorescence reading requires instrumentation that can generate high intensity light of a certain wavelength or color, filters that can block stray light from that light source, and a light detection device to capture the emitted light. Magnetic particles need instrumentation to detect their tiny magnetic fields. Carbon nanoparticles are not as visually detectable as gold nanoparticles, dyed latex particles or cellulose nanobeads and only outperform when used with high intensity infrared light sources and specialized detection systems. The unaided human eye is a remarkably efficient device for detecting light including reflected light from LFAs. It is sensitive to light from 380 nm (violet) to 800 nm (red) [8] but is most sensitive to green light at a wavelength of 555 nm. The human eye cannot see light outside of these ranges. Men and women also differ in their ability to see colors. Woman have been found on average to have better sensitivity to color and to better differentiate between colors than men [9]. Quantitation using unaided LFAs means that the user has to be able to differentiate between different intensities of color. In a sandwich assay such as one for hCG, a darker color band means that more analyte has been detected. However, quantitation is not needed for pregnancy tests. There is no such thing as being a little pregnant. The test and the condition for which it is testing is binary. Either the test is positive or it is negative. However, other tests can benefit from quantitation as more than one condition can exist. Thyroid stimulating hormone (TSH) is one such test. Someone who has normal thyroid function will have a TSH blood level between 0.1 and 4 uIU/mL in their blood. Levels above the high cutoff are at higher risk of being hypothyroid. Levels below the low cutoff indicate a higher risk of being hyperthyroid. Printed color charts can be helpful in estimating levels, but reliable quantitation requires instrumentation, which will be the subject of a future blog. Our team of engineers and scientists can help you determine the specification for antibody labeling for your test. Feel free to contact us with any questions. [1] https://what-if.xkcd.com/106/ [2] Cole LA, Sutton-Riley JM, Khanlian SA, Bokovskaya M. Rayburn BB and Rayburn WF, Sensitivity of Over-the-Counter Pregnancy Tests: Comparison of Utility and Marketing Messages, J Am Pharm Assoc 45, 608-615 (2005) [3] Sajid M, Kawde A and Daud M, Design, Formats and Applications of Lateral Flow Assays, J Saudi Chem Soc., 19, 689-705 (2015) [4] https://www.asahi-kasei.co.jp/asahi/en/news/2014/e141216.html [5] Barnet JM, Wraith P, Kiely J, Persad R, Hurley K, Hawkins P and Luxton R, An Inexpensive, Fast and Sensitive Quantitative Magneto-Immunoassay for Total Prostate Specific Antigen, Biosensors(Basel) 4(3), 204-220, (2014) [6] Posthuma-Trumpie G, Wichers JA, Koets M, Berendsen LBJM and van Amerogen A, Amorphous carbon nanoparticles: a versatile label for rapid diagnostic (immune) assays, Anal Bioanal Chem 402(2) 593-600 (2012) [7] Samsonova JV, Safronova VA and Osipov AB, Pretreatment-Free Lateral Flow Immunoassay for Progesterone determination in whole cow’s milk, Tantala 132, 685-689 (2015) [8] https://light-measurement.com/spectral-sensitivity-of-eye/ [9] https://www.livescience.com/22894-men-and-women-see-things-differently.html --- ## The Case for User-Centric Assay Design URL: https://dcndx.com/insights/user-centric-assay-design/ Type: insight Published: 2018-06-01 Origins of User-centric Assay Design Until the advent of two-line lateral flow pregnancy tests in the late 1980s, home hCG tests were essentially miniaturized chemistry sets with complex instructions and a test tube rack. Inverness Medical’s incorporation of on-board colloidal gold immunoconjugates forever shifted the paradigm of pregnancy testing in a number of ways: Reduced complexity. No longer would the user be required to perform sample collection and treatment. Reduced time-to-result. Reaction times (not including sample preparation or manipulation time) of 30 minutes were reduced to 10-15 minutes which made the test more user-friendly. Integration of more robust materials. Eliminating liquid and glass components in favor of plastic and paper facilitated storage and extended shelf lifetime and decreased cost to the consumer. Mitigation of risk. The inclusion of a control line, reduction of steps, and simplification of instructions all compounded to give a leap forward in pregnancy-testing technology by making the results easier to interpret by an untrained user. Since these disruptive improvements were made, there have been additional incremental advances in lateral flow tests, such as the Clearblue cross and rudimentary yes/no integrated readers. However, the canonical two-line test has so disrupted the diagnostic world that it now pervades pop culture, even earning itself a country song written by Grammy-nominated artist Eric Church. Why was this such a revolution? After all, it was still testing for hCG, a straightforward assay even by laboratory standards and these improvements did not improve the sensitivity of the assay. These improvements weren’t about the sensitivity; rather, they were about how the user acquired, used and read the test. The busy mother who was preoccupied with children and did not have 30 minutes to devote to a complex test. The new mother or young adult who, with nerves and shaking hands, would no longer have to worry about adding a component to the wrong tube. Women in complex situations, who could not risk partners or relatives uncovering test remnants. As developers, we run the risk of preoccupying ourselves with achieving maximum sensitivity at the sacrifice of user-friendliness or cost point. Sometimes, maximum performance isn’t always the top deliverable, and sub-optimal sensitivity can easily be counterbalanced by fulfilling other end user requirements. Though the pregnancy test is lateral flow’s most well-known achievement to date, there are other areas where the versatility of the technology shine and who’s utility is based in large part on the user experience. It is well worth it to explore some commonalities in these markets. User-centric Assay Design for Biowarfare and Defense In preparing for chemical or biological warfare or terrorism, first responders require tools that can be used in confusing, chaotic, and uncertain environments. Testing systems should therefore incorporate the ability to identify a broad range of possible agents in a non-ideal field setting. Lateral flow tests are optimal for such a scenario, as noted by the following attributes: Multiplex test strips – Commercially-available tests commonly incorporate multiple test lines to assay multiple targets of interest. Appropriate treatment can follow accurate identification. Barcoding – Operators can track patients by unique codes and test them in quick succession. Portable and robust readers – Readers take the guesswork out of diagnostics in stressful or light-limited situations, and battery operation permits their performance in the field, without electricity. Disposability and speed – A short run time of ≤ 15 minutes enables simultaneous screening of multiple patients and a quick sample-to-answer. Results can be read and tests discarded, with no specimens to carry back to a centralized lab. User-centric Assay Design for Infectious Disease Testing in Low-Resource Environments Low-resource environments can also present challenging situations for testing. Labor- and time-intensive culturing is the diagnostic gold standard for a number of infectious diseases, but for situations where this is too cumbersome, immunoassays and molecular diagnostics have stepped in to fill the gap. Diagnostic tests in these situations must endure harsh temperatures, potential contaminants, and inexact volumes while at the same time they must produce a robust and unambiguous signal. Currently, there are a number of lateral flow immunoassays for diseases such as tuberculosis, malaria, Dengue, HIV, and Chikungunya that fulfill these criteria in the following ways: Point-of-care – Short run times ensure no need for return visits and allow physicians to administer a diagnosis and concomitant treatment. Visual readout – Colorimetric signals overcome the need for a reader, leading to simplified testing procedure. Rugged and non-demanding – Onboard or accompanying buffers and qualitative readout circumvent requirements for amenities such as electricity and running water. Cost – At under $1 USD/unit, these tests fit the price point for many developing markets. User-centric Assay Design for Environmental and Health Monitoring Lateral flow is amenable not only to isolated interrogation of a sample at a certain point in time, but also extended monitoring of a patient, workplace, environment, or other situation. The large volumes of tests that can produced per manufacturing run can translate into a high number of tests per lot, along with other advantages: Consistency – CVs of typically <10% and high reproducibility make lateral flow an ideal technology for prolonged surveillance. Shelf lifetime and storage – When stored under the proper conditions of desiccation, lateral flow tests have been shown to possess shelf lifetimes of multiple years. In most cases, refrigeration is not necessary. Little-to-no footprint – Reader technology has evolved to produce machines with increased capabilities, but lower power and space requirements. Visual qualitative tests, while less useful for quantitative monitoring, have even fewer equipment restrictions. These are just some examples of how lateral flow, which at its core is a simple technology, can be amenable to testing diverse analytes in numerous environments. The biological measurement should be robust and as sensitive and specific as possible. However, technical performance is only part of the equation needed to have strong market uptake. Usability and customer experience are equally important. If the product cannot be distributed in a timely and cost effective manner or if the instructions for use require an advanced degree to comprehend, or if the results are confusing, the test will have little value. The development of the two-line pregnancy test was the key step-function change which allowed lateral flow immunoassays to become the format of choice for point-of-care testing, and user-centric assay design improvements continue to be realized on an annual basis. If you have questions about user-centric assay design, contact us. Our team of engineers, scientists and marketers will help you identify the goals, use and consumers for your test. --- ## The Critical Components of Successful Assay Design URL: https://dcndx.com/insights/successful-assay-design-2/ Type: insight Published: 2018-04-17 A rapid diagnostic test is a tool that, like any other tool, must be designed and built for its intended purpose. Before going into the lab to develop an assay, one must start at the end and correctly identify the goals and use of the product. Only then can point of care diagnostic products be designed correctly. This is the critical first step in successful assay design. For successful assay design, the following items should be clearly articulated prior to development: A written description of the assay’s intended use What the analyte will measure What sample—that may or may not contain the analyte—will be collected How much sample is required How much experience the operator must have and what support resources, such as instrumentation for quantitation, are available The availability of critical raw materials, including antibodies and calibration materials, is another important consideration of successful assay design. Since developed rapid diagnostic tests may have useful lives measured in decades, reliable sources of these critical raw materials must be assured. In addition, development resources, such as experienced lateral flow assay developers and adequate laboratory facilities are critical. Finally, a manufacturing infrastructure with the requisite personnel, documentation and management to reproducibly produce this assay for years to come must also be available or built. All of these requirements are assembled into a multi-page specification document that defines the desired parameters of the diagnostic product. Contact us for more information about filling out a product requirements document for your diagnostic test. Diagnostic assays require resource teams with different perspectives for successful development and transfer to manufacturing. Technical resources can address questions, such assay feasibility and performance characteristics. Marketing represents the end user and provides inputs to the team as to what the end user needs the assay to do. Manufacturing suggests how best to make the assay in a consistent, reliable and cost-efficient manner. Quality defines how best to test the assay to ensure it is meeting specifications. During the development process, R&D will challenge the rapid diagnostic test with various conditions known to interfere with similar assays. One classic parameter that can affect urinalysis assays is pH. In humans, urine samples can have pH values ranging from 4.5 to 8.[1] Antibodies work best around a pH of 7.4 and can usually only tolerate deviations of plus or minus one pH unit. A sample with a pH of 5 may give erroneous or misleading results in an assay not designed to handle these extremes. Temperature also affects antibody reactions. An assay that works well at room temperature or body temperature may not give the same results in colder temperatures. Other substances in the sample can also interfere with the assays. Assays that use blood or blood fractions, such as serum or plasma, must be tested to ensure that they will perform adequately in the presence of hemoglobin released from disrupted red blood cells, or lipids from dietary sources, or bilirubin from liver abnormalities. Exogeneous substances, such as biotin and fluorescein, can also interfere with some assays.[2] Lateral flow assays can be hardened to address some, but not all of these issues. For example, the pH parameter can be dealt with by ensuring that sufficient buffering capacity is present either on the device or in the solutions intended for use with the device. Written warnings in the instructions for use can help address temperature fluctuation and naturally occurring substances in blood. Intelligent assay design can help manage biotin interference. One critical aspect of assay design is the provision of adequate controls. Controls are often assay specimens that have been engineered to mimic actual samples in composition and concentration of the analyte under investigation while maintaining stability and ease of use. In single use assays, including many lateral flow assays, this type of control is clearly impractical and other measures must be taken to ensure the assay is run appropriately. Over the counter pregnancy tests routinely include a control line that ONLY appears if there has been an adequate amount of sample added to the device and that it is not of a pH that would interfere with an immunoassay. This is often accomplished by adding a second line of reagent in the reaction area of the device. The first line is the capture antibody for the analyte of interest in this case human chorionic gonadotropin (hCG). The second line is usually an anti-species IgG antibody that can capture the labeled detector antibody. For example, most pregnancy assays use mouse monoclonal antibodies for both capture and detector antibodies. A second line that contains goat anti-mouse antibody will capture any excess detector antibody. What the user sees is that a single line on the device indicates that the test has been run properly with enough sample to carry the reagents to their intended capture lines, but the analyte is not present in detectable amounts. If the analyte were present, then the user would see two lines, one from the capture antibody line and one from the anti-mouse line. If no lines are visible the test is invalid and no conclusion can be drawn. An inadequate amount of sample is frequently the cause of a failed assay. A recently published paper provides a real-world example of how an assay was developed without consideration of how it was to be used.[3] An alert and well-trained operator noticed that a certain urine specimen failed to give either a result or control line on a Point of Care (POC) pregnancy test from one manufacturer. The urine specimen was tested on six other commercially available POC pregnancy tests and they all gave appropriate test and control results. Usually when a POC pregnancy test is run the colors of the test and control lines are the same—but not in this case. The test line was red and the control line was blue. The authors speculated that the usual anti-IgG capture line had been replaced by an immobilized streptavidin line and the capture antibody reagent had been supplemented by that addition of a biotin labeled blue dye. The suspect sample was thought to contain excess biotin ingested by the patient from an over the counter preparation that are supposed to promote good growth of hair and nails. Excess biotin in the sample overwhelmed the streptavidin in the control line, and it could not bind the biotin labeled blue dye. The other POC pregnancy tests did not use the biotin-streptavidin chemistry and were not affected by the presence of biotin. The above represents a failure in assay design, but it was a soft failure. No patients were harmed in the incident. It did, however, cause financial harm in that it triggered an investigation requiring people and resources to unravel. What did we learn from the failure? Obviously, that incorporating a biotin-avidin linkage in a test that uses human urine as a sample is inherently a risky decision. Six other tests were unaffected by the excess biotin which is good news because it points a way forward to a more robust design. The biotin-avidin linkage was probably introduced into the failed test to provide a different color for the control strip to better differentiate it from the test strip. Another way to accomplish this would be use a different species antibody for the control strip chemistry. Let’s assume the test strip is a mouse antibody as is the detector labeled antibody. If the control material were a blue dyed rabbit antibody and the control strip were a goat anti-rabbit antibody, that should provide a robust chemistry for the control strip while preserving the different colors for test and control results. This incident and the excellent scientific detective work that went into determining root causes provide a good lesson in what can happen if good assays design is not implemented and rigorously tested. If you are not sure whether your test is sufficient for its intended purpose, contact us. Our team of engineers, scientists and marketers will help you identify the goals, use and consumers for your test. [1] Moeller, KE; Kissack, JC; Atayee, RS and Lee, KC, ”Clinical Interpretation of Urine Drug Tests: What Clinicians Need to Know About Urine Drug Screens” Mayo Clin Proc., 92(5):774-796 (2017) [2] Li D; Radulescu A, Shrestha R, Root M, Karger A, Killeen A, Hodges J et al., ”Association of Biotin Ingestion With Performance of Hormone and Nonhormone Assays in Healthy Adults” JAMA 318(12):1150-1160 (2017) [3] Williams G, Cervinski M, Nerenza, R “Assessment of biotin interference with qualitative point-of-care hCG test devices” Clin Biochem https://www.ncbi.nlm.nih.gov/pubmed/29395091 (2018). --- ## DCN Dx 2018 Events Calender URL: https://dcndx.com/insights/dcn-dx-2018/ Type: insight Published: 2018-04-11 Spotlight Event: The Advanced Lateral Flow Course October 23-25, 2018 San Diego, California, USA Join us for this 3-day educational seminar co-hosted by DCN Dx and QIAGEN Lake Constance. The content of this event is aimed at developers and manufacturers of rapid diagnostic tests and their associated products. During this seminar, some of the most experienced professionals in the diagnostics industry will guide you though every aspect of producing and commercializing a point of care diagnostic device. Registration is Open! Upcoming Events: Food Safety Summit, May 7-10, 2018 in Rosemont, IL, USA BIO International Convention, June 4-7, 2018 in Boston, MA, USA American Biomanufacturing Summit, June 14-15, 2018 in San Francisco, CA, USA Biodefense World Summit, June 27-29, 2018, Bethesda, MD, USA American Veterinary Medical Association Convention, July 13- 17, 2018, Denver, CO, USA AACC Annual Scientific Meeting, July 29-August 2, 2018, Chicago, IL, USA Next Generation Diagnostics Summit, August 20-24, 2018, Washington, D.C., USA AOAC Annual Meeting, August 27-29, 2018, Toronto, Canada The Advanced Lateral Flow Course, October 23-25, 2018, San Diego, CA, USA ASTMH Annual Meeting, October 28-31, 2018, New Orleans, LA, USA --- ## DCN Dx is Exhibiting at the 2018 ASTMH Annual Meeting URL: https://dcndx.com/insights/2018-astmh/ Type: insight Published: 2018-04-11 October 28 – November 1, 2018 New Orleans, Louisiana, USA DCN Dx will be joining over 4,600 tropical medicine and global health professionals at the American Society of Tropical Medicine and Hygiene educational conference. Stop by our booth in the 2018 ASTMH exhibit hall to talk to a technical specialist about your specialty and why lateral flow is the ideal device for point of care diagnosis in low resource environments. For more information about ASTMH, visit the EVENT WEBSITE or contact us. --- ## DCN Dx is Exhibiting at the 2018 AACC Scientific Meeting URL: https://dcndx.com/insights/2018-aacc/ Type: insight Published: 2018-04-11 July 29 – August 2, 2018 Chicago, IL, USA DCN Dx will be exhibiting at the 70th AACC Annual Scientific Meeting & Clinical Lab Expo. Stop by booth #3452 to meet with one of our technical specialists and discuss DCN Dx’s portfolio of products and services geared towards developers of point of care diagnostic tests. Our private education and training courses are a great way to get your development team up to speed and get your program moving in the right direction. Contact us to set up a meeting with one of our representatives. For more information about AACC, visit the EVENT WEBSITE or contact us. --- ## DCN Dx is Sponsoring the 2018 Hands-On Lateral Flow Workshop URL: https://dcndx.com/insights/dcn-dx-is-sponsoring-the-2018-hands-on-lateral-flow-workshop/ Type: insight Published: 2018-04-11 May 15-17, 2018 Zaragoza, Spain Join DCN Dx at this 3-day, practical lateral flow workshop co-hosted by BioDot and OPERON. An agenda has been created to address a variety of topics including reagents and materials selection, conjugation methods, readers and troubleshooting. You will also get the opportunity to produce a working lateral flow test in the practical session. DCN Dx will be participating as an sponsor and will also be delivering educational talks in the main agenda. For more information, visit the EVENT WEBSITE or contact us. --- ## DCN Dx is Exhibiting at the 2018 American Biomanufacturing Summit URL: https://dcndx.com/insights/2018-aacc-2/ Type: insight Published: 2018-04-11 (As published 2018-04-11; regulatory status may have changed since.) June 14 – 15, 2018 San Francisco, CA, USA DCN Dx will be exhibiting at the 2018 American Biomanufacturing Summit. Join us at this senior level networking event to discuss strategic insights in manufacturing, outsourcing, capacity management, quality assurance, quality control, regulatory compliance, operational excellence, supply chain and logistics. Stop by our booth, or contact us to schedule a meeting with one of our representatives in the San Francisco area. --- ## Lateral Flow Assays (LFAs): How Does a Lateral Flow Device Work? URL: https://dcndx.com/insights/lateral-flow-rapid-diagnostic-test/ Type: insight Published: 2018-02-23 Lateral Flow Assays Explained Over the years, lateral flow assays (LFAs) have become an indispensable element of the rapid, point-of-care diagnostic test market. Initially gaining prominence through their early implementation in home pregnancy test kits, lateral flow immunoassays have since diversified to encompass a wide array of devices designed for point-of-care settings, removing the need to send samples to a laboratory or clinic for analysis. These tests skillfully detect an extensive range of biomarkers, pathogens, and environmental contaminants in numerous industries, making them an invaluable tool in modern diagnostics. The underlying technology of lateral flow assays dates back over three decades, yet these systems have continually evolved to meet the complex diagnostic demands of today. State-of-the-art lateral flow assays boast high sensitivity and multiplexing capabilities, effectively addressing the high error rate that often arises at the point of sampling. By incorporating user-centric designs, LFAs have become more accessible and easier to use, even for individuals with minimal technical expertise. The scope of these assays now encompasses a wide variety of diagnostic devices, ranging from simple, cost-effective, and lower complexity qualitative tests to intricate designs that facilitate the quantification of multiple biomarkers in real-world settings. These advanced devices eliminate the need for capital-intensive equipment and infrastructure, enabling rapid and accurate diagnostics in various field applications. In recent years, the development of customizable readers, smartphone-based readers, and digital platforms has further enhanced the capabilities and applications of LFAs. These advancements have not only improved the accuracy and reliability of test results but also enabled the integration of diagnostic data with electronic health records and other digital systems. This seamless data integration facilitates better decision-making, improved patient care, and more efficient resource allocation in healthcare settings. The expanding applications of LFIs have led to their adoption in diverse sectors beyond healthcare, such as food safety, agriculture, environmental monitoring, and veterinary medicine. The versatility and adaptability of lateral flow assays make them a powerful diagnostic tool, with the potential to revolutionize various industries by delivering rapid, accurate, and accessible testing solutions. What Is a Lateral Flow Assay? Lateral flow assays, a popular format for Rapid Test Devices or RDT’s, are immunosensors that rely on the use of low cost materials to achieve the flow of reactants. At their simplest, they are dipsticks with a sample application pad at one end, a reading zone where the signal is generated in the middle and an absorbent material at the other end. Tests may be incorporated into housings (cassettes) that support higher functionality, branding and user-centric design goals. Lateral flow immunoassays may be qualitative (indicating the presence or absence of an analyte), semi-quantitative, or fully quantitative. Each type of assay offers distinct advantages, depending on the specific diagnostic requirements and desired outcomes. Qualitative lateral flow immunoassays are designed to indicate the presence or absence of an analyte in a sample. These assays are often used for rapid, preliminary screening purposes, providing a simple yes or no answer regarding the existence of a target substance. Such tests are particularly useful in situations where a quick assessment is needed, such as detecting the presence of pathogens, allergens, or drugs of abuse. Semi-quantitative lateral flow immunoassays offer an intermediate level of information, providing an estimation of the analyte’s concentration within the sample. While not as precise as fully quantitative assays, semi-quantitative tests can still offer valuable insights into the severity or progression of a condition or the effectiveness of a treatment. This type of assay is frequently employed in monitoring chronic diseases, evaluating immune responses, or assessing environmental contamination levels. Fully quantitative lateral flow immunoassays deliver the most comprehensive results, accurately measuring the exact concentration of an analyte within a sample. Quantitative assays are essential in situations where precise measurements are required to make informed decisions regarding diagnosis, prognosis, or treatment. These tests are commonly used in the monitoring of hormone levels, the quantification of specific biomarkers, and the determination of viral loads, among other applications. Lateral flow assays contains several important components: Sample pad: The sample pad is the point of entry for the test sample, where it is first applied to the assay. This pad is typically made of cellulose or glass fiber materials, which are designed to facilitate the even distribution and consistent flow of the sample through the test strip. Additionally, the sample pad often contains reagents or buffers that help to prepare the sample for further processing, such as filtering out contaminants, reducing viscosity, or adjusting the pH. Conjugate pad: The conjugate pad contains immobilized detection reagents, such as antibodies or antigens conjugated to signal-generating particles, like gold nanoparticles or colored latex beads. When the sample reaches the conjugate pad, the target analyte binds to the conjugated detection reagent, forming a complex. This complex then moves through the assay via capillary action, allowing for further interactions with other assay components. Nitrocellulose membrane: The nitrocellulose membrane is the core of the lateral flow assay, where the actual detection of the target analyte occurs. This membrane is embedded with two distinct lines: the test line and the control line. The test line contains immobilized capture reagents that selectively bind to the target analyte-conjugate complex, forming a visible signal if the analyte is present in the sample. The control line, on the other hand, is used to confirm the proper functioning of the assay by capturing any remaining conjugate, regardless of the presence or absence of the target analyte. Absorbent pad: The absorbent pad is located at the end of the assay strip and serves as a reservoir for excess sample and reagents. Its primary function is to maintain the flow of the sample through the assay by drawing it past the nitrocellulose membrane via capillary action. This ensures that the sample moves at a consistent rate, optimizing the assay’s performance and increasing the accuracy of the results. These materials, each serving one or more purposes, overlap onto one another and are mounted on a backing card using a pressure sensitive adhesive. Figure 1 shows a typical configuration. Figure 1: Typical strip configuration, including a sample pad, particle conjugate on the conjugate pad, nitrocellulose membrane, wick and backing card. How Does a Lateral Flow Test Work? When a lateral flow assay is run, a sample is added to a Sample Application Pad. Here, the sample is treated to make it compatible with the rest of the test. The treated sample migrates through this region to the Conjugate Pad. Here, a particulate conjugate has been immobilized, typically a colloidal gold or a colored, fluorescent, or paramagnetic monodisperse latex particle or cellulose nanobead. This particle has been conjugated to one of the specific biological components of the assay, either antigen or antibody depending on the assay format. The sample re-mobilizes the dried conjugate, and the analyte in the sample interacts with the conjugate as they both migrate into the next section of the strip. This zone, known as the Reaction Matrix is a porous membrane onto which the other specific biological components of the assay have been immobilized. These are typically proteins, either antibody or antigen that have been laid down in bands in specific areas of the membrane where they serve to capture the target and conjugate as they migrate up the strip. Excess reagents move past the capture lines and are entrapped in the Wick or absorbent pad. Results are interpreted on the Reaction Matrix as the presence or absence of lines and can be read either by eye or using a reader. Assay formats are either sandwich (direct) or competitive (competitive inhibition) in nature, and the assay format can accommodate qualitative, semi-quantitative, or fully quantitative assays. Direct assays are typically used when testing for larger analytes with multiple antigenic sites, such as hCG, Dengue antibody or antigen or HIV. In this case, a positive result is indicated by the presence of a test line. Less than an excess of sample analyte is desired, so that some of the conjugated particles will not be captured at the capture line, and will continue to flow toward the second line of immobilized antibodies, the control line. This control line typically comprises a species-specific anti-immunoglobulin antibody specific for the conjugated antibody. Competitive formats are typically used when testing for small molecules with single antigenic determinants, which cannot bind to two antibodies simultaneously. In this format, a positive result is indicated by the absence of a test line on the reaction matrix. A control line should still form, irrespective of the result on the test line. The two formats are illustrated schematically in Figures 2a and 2b below. Figure 2a: Direct Solid Phase Immunoassay Figure 2b: Competitive solid phase immunoassay Why Consider Lateral Flow Immunoassays for Your Rapid Diagnostic Testing Needs? Lateral flow immunoassays are easy to use. The user simply needs to dip the sample pad into the liquid they want to test and then lay the test flat or apply the sample directly to the cassette. If the test is positive, a colored or fluorescent line will show up on the test along with a control line. If the test is negative, only the control line will be present. For many years, women with no medical training have successfully used this kind of test to detect pregnancy or ovulation at home. More advanced lateral flow tests provide additional benefits, including higher sensitivity, multiplexing and the ability to detect multiple biomarkers from a single finger stick. These advanced tests are useful for in-the-field testing. Lateral flow rapid tests provide results quickly. In many cases, the test and control lines show up almost immediately with definitive results available within a few minutes. This rapidity is important in medical settings where doctors have limited time available to spend with each patient, as well as in manufacturing process quality control checks in pharmaceutical companies, highly sensitive human performance testing for athletes, and bioprocessing and GMO testing in agriculture and food safety. Why DCN Dx? DCN Dx is the premier choice for designing and developing point-of-care rapid diagnostic assays with challenging specifications, such as: Exceptional sensitivity Quantitative analysis Multiplex capabilities Complex sample matrices Integrated reader systems Our cross-functional of assay development, engineering, manufacturing, and IVD clinical research professionals allows us to assist you in the development of your entire rapid diagnostic test from concept to commercialization. Our staff has unparalleled experience in developing the highest performing point-of-care assays in lateral flow markets, ranging from medical diagnostics to veterinary, food science, bio-defense, agricultural, and environmental health tests. DCN Dx operates under ISO 9001:2015 and ISO 13485:2016. All of our development programs are done under Design Controls, and all production activities for reagents, components, assays, and electronics are performed under our EN 13485 system. --- ## Beyond Pregnancy Tests: Current Applications of Point of Care Diagnostic Testing URL: https://dcndx.com/insights/current-applications-of-point-of-care-diagnostic-assays/ Type: insight Published: 2018-02-14 Although a medic with a bag of simple medical supplies is not a fully equipped emergency room, he or she is the right solution for a wounded soldier on the battlefield. Medics can supply information and treatment that can enable that soldier to survive long enough to get more advanced medical treatment. Similarly, the lateral flow assay (LFA) is the right solution for many point of care diagnostic testing applications not appropriate for a central laboratory. Because LFAs do not require electricity, running water or users with years of training, they can provide analytical information on the spot to inform decisions cost-effectively. LFAs have advanced from their earlier versions both in terms of detection limits and areas of application. Here, we trace the history of LFAs and provide four examples of their modern applications in point of care diagnostic testing. The Humble Beginnings of Immunoassay Development In 1977, the Nobel Prize for Physiology or Medicine was awarded to Rosalyn Yalow and Solomon Berson for a discovery that they had made in 1959. They were the first practitioners of a then-new technique they called radioimmunoassay. An astounding array of applications have followed, allowing the detection and quantitation of substances of interest in a wide variety of disciplines and settings. Clinical diagnostics has received the most attention, and the intervening years have seen the development of large automated instruments with impressive menus of assays. The radioisotopes in Yalow and Berson’s assay have been replaced with chemiluminescent and other non-isotopic marker molecules, and separations are now done with micron sized superparamagnetic particles. These instruments can produce hundreds of results an hour in random access using samples of serum or plasma from primary blood drawing tubes. On-board reagents for as many as 30 different assays can be loaded on the instrument. The automated systems work well in a central laboratory setting, such as a hospital or a reference laboratory, but they are not appropriate in all situations where an analytical result is needed. For example, these instruments are often the size of two washing machines side by side. While most of these instruments are acquired as reagent rental units, high sample throughput is necessary to make placement economically viable for both the user and the instrument supplier. Lateral flow technology helps fill the gap between central laboratory testing and use in point of care diagnostic testing. Modern lateral flow tests enable high sensitivity and multiplexing, and can clearly detect the presence of a wide range of biomarkers, pathogens, and environmental contaminants. Importantly, lateral flow assays can be deployed in a wide variety of industries. Lateral Flow as Rapid Point-of-Care Diagnostic Testing for EMTs Every year about 790,000 people have a heart attack and 114,000 die as a result. Rapid diagnosis is a critical first step in effective treatment of this condition. Zhu and colleagues have prototyped an LFA for point of care diagnostic testing that can detect relevant levels of Troponin I and Myoglobin in serum in an assay that takes only 15 minutes. Troponin has been called the “dying scream of cardiac tissue” and is released from heart cells when they die from lack of oxygen immediately after a heart attack. Myoglobin is also released from dying muscle tissue faster than Troponin, but it is not cardiac specific. Taken together, the two markers in an LFA can provide rapid and accurate, potentially life-saving diagnostic information even before the patient can be evacuated to a hospital. Lateral Flow and Infectious Disease Point of Care Diagnostic Testing Cruise ships allow busy people to sail away from their usual lives. However, sometimes non-human passengers embark on these ships, also. One such fellow traveler that makes the news every few months is Norovirus. This organism causes severe gastroenteritis with a wide range of unpleasant symptoms that can be easily confused with bacterial food poisoning. Effective treatment and prevention of food poisoning and Norovirus infection are quite different. The challenge in a very resource limited setting such as a cruise ship is how to tell them apart. R-Biopharm in Darmstadt, Germany has developed an LFA for point of care diagnostic testing that can detect the antigens shed by Norovirus in stool samples from sufferers. The assay can be run in 15 minutes and is claimed to have an accuracy of 96%. Lateral Flow for Next-Generation Fertility Testing Probably the most familiar application of the LFA is the home pregnancy test. Someday, there may be a home test also based on the LFA technology that will allow a woman to determine how much time she has left to become a mother. Viable ova develop from primordial follicles in the ovary. At birth, a baby girl has about one million such follicles. In adolescence, that number falls to about 400,000. At menopause, she may have only a thousand. The chronological ages at which these milestones occur varies from individual to individual. A good biomarker in blood for the number of primordial follicles is anti-Mullerian Hormone, and US Patent Application US20130224771A1 describes an LFA for this molecule. Interestingly, the sample can be either a fresh fingerstick whole blood sample or a dried blood spot (DBS). A DBS allows the collection and storage of blood samples in very resource limited settings and extend the utility of LFAs for point of care diagnostic testing. We will have more to say about DBS in later blogs. Lateral Flow for Cows? Yes! Agriculture Applications of Point-of-Care Diagnostic Testing Human pregnancy is a subject that attracts a lot of attention, but the reproductive status of other species is of considerable interest as well. Dairy cows only give milk after they have had a calf and for a 10-month period after calving. The cow must be bred again to produce milk and successful breeding can only occur when the cow is ovulating. Since modern breeding techniques rely on artificial insemination, determination of this fertile period is critical to keeping a cow in production. Biomed Diagnostics, Inc. of White City, OR has introduced a cow-side LFA that can tell the farmer within 5 minutes if the cow is ready to be bred. The assay detects the hormone, progesterone in the cow’s milk without the need for specialized equipment or operator training. These are only some of the modern applications of LFAs. If you have an analytical challenge for which central laboratory-type technology is not the answer, please contact us to learn whether lateral flow could be the solution to your point of care diagnostic testing needs. [1] Heart Disease and Stroke Statistics 2017 At-a-Glance, https://healthmetrics.heart.org/wp-content/uploads/2017/06/Heart-Disease-and-Stroke-Statistics-2017-ucm_491265.pdf [2] Simultaneous Detection of High-Sensitivity Cardiac Troponin I and Myoglobin in a Modified Sandwich Lateral Flow Immunoassay: Proof of Principal, Zhu J, Zou N, Zhu D, Wang J, Jin Q, Zhao and Mao H, Clin Chem 57:12, 1732-1736 (2011) [3] Fleming R et al, Reprod BioMed Online 2015;31:486-496 [4] La Marca et al, Hum Reprod Update 2010;16;113-130 --- ## Seven Reasons why you should Consider Lateral Flow Technology for your Next Rapid Test Development Project URL: https://dcndx.com/insights/lateral-flow-technology-rapid-diagnostic-test/ Type: insight Published: 2018-02-01 Your number one corporate objective in 2018 is to build a rapid assay for a critical application. You’re in charge. Maybe it’s a test for use in malaria eradication programs. It has to be qualitative, single-plex, fast, analytically very sensitive, scalable, easy to use by untrained users and have extremely high clinical predictive value. Or maybe it’s a test for cardiac or cancer biomarkers. It has to be multiplexed, quantitative, for use in doctor’s offices, emergency rooms or ambulances. It has to be easy to use, coupled with a digital reader, designed with the stressful application in mind with user centric design, performance and branding as key specifications. Or you might be working in veterinary, pharmaceutical, agricultural, food, bio-defense or consumer testing markets and need devices designed specifically for the unique needs of your users and markets. For example, a test for protein conformation in a bioprocessing plant. It should be easy to find the right platform, right? Then you realize that the product needs to be in market in less than two years for a reasonable cost of development with low cost of goods. It has to perform better than the competition and lab based predicates. You don’t want to operate in a restrictive IP environment. You also don’t want to risk pioneering basic development of a novel microfluidic platform or detection technology. OK, well the platform options just became more limited. So what do you do? How Do You Hit Your Rapid Test Product Development Objectives with Lateral Flow Technology? Maybe you’ve never heard of lateral flow technology. Or maybe you’ve heard of lateral flow in connection with simple pregnancy tests but don’t think it’s suitable for your complex application. After all it’s only a cheap solution to easy problems, right? And your problems are not the easy ones. It’s not surprising that you may be unaware of lateral flow technology, or that you may be unaware of its actual capabilities. You’re not alone. Lateral flow is a 30-year old technology that is best known for use in pregnancy and fertility tests for consumers. Pee on a stick and get a yes or no answer. Early products were often not designed for use and required cumbersome processing by the user. They were limited in their ability to multiplex or quantify, so they were limited in application. When you think of modern, user-centric diagnostic technology with high accuracy for tough challenges, you don’t automatically think “lateral flow technology”. That’s a pity, because you’re likely thinking of what lateral flow was 30 years ago, not what it is today. The technology has moved on. The industry, however, has not done a great job of making you aware of what lateral flow technology can do. We’re sorry for not bringing this to your attention before now. Our bad. We want you to hit your objective for 2018 so here are some things to know: Lateral flow technology is in many cases the only rapid immunoassay platform technology that can meet your product’s needs. Here are some examples of things you may not know about lateral flow technology that might provide the answer to your product development challenges: 1. Multiplexing and Quantification: It’s very possible. For example, Astute Medical’s Nephrocheck TM assay system for two biomarkers indicative of acute kidney injury (TIMP-2 and IGFBP-7) is FDA approved, commercialized and is saving lives today. 2. Robust, Field Deployable, Quantitative, Digitally Integrated Platforms Designed Specifically for Use: It’s been done. For example, the Biodosimeter platform, developed with SRI International under funding from BARDA, is a fully quantitative 4-plex biomarker assay system intended to assess radiation exposure in the event of a nuclear incident. Whole blood from a fingerstick is assayed in a single step for up and down regulation of 4 biomarkers using a lateral flow assay. The assay is interrogated by a robust digital assay reader that is connected locally and to the cloud. And the best part – all of this is in a system designed for ease of use for personnel wearing full Personal Protective Equipment (PPE) in a high stress, low resource, uncontrolled environment for testing of mass populations. A truly unique test system that – if it is ever needed (hopefully it’s not!) – will save a lot of lives. 3. High sensitivity, low cost, highly scalable assays for infectious disease diagnostics: Hundreds of millions of tests are produced annually in lateral flow format for these applications. For example, the next generation of malaria tests, which will have to be extremely low cost and highly scalable to hundreds of millions of units annually, will need to detect only a few organisms in an infected individual. This is being achieved using highly sensitive lateral flow assays. Several organizations are working with DCN and other companies to develop and transfer these highly sensitive assays to large scale production and deployment. Another lateral flow product that will save millions of lives over time. 4. Low Cost of Development and Speed to Market: Lateral flow is an established, proven technology. There is very little basic research involved in the development of a lateral flow product. The fundamentals of the technology are well understood. The entire supply and manufacturing ecosystem is in place. It’s primarily Development rather than Research once your target and your reagents are understood. Most lateral flow products can be developed and transferred to manufacturing significantly faster and cheaper than microfluidic or lab-on-a-chip-type products. This increases the attractiveness of the risk profile of the program. 5. Regulatory and Market Recognition: After decades of proven performance lateral flow is an accepted technology. 6. Uncluttered IP Space: The IP space is – relative to microfluidics and molecular diagnostics – relatively unrestrictive. 7. Enormous versatility in application and sample type: Lateral flow assays have been developed for many applications involving an enormous array of sample types. Application areas include: a. Human clinical diagnostics: Using blood, plasma, serum, urine, sweat, feces, cerebro-spinal fluid, interstitial fluid, nasal swabs, vaginal swabs, wound fluids, saliva, semen, skin cells b. Veterinary diagnostics: Using the same array of samples as clinical diagnostics c. Female Health (Fertility and Pregnancy): Using urine, saliva, blood, vaginal secretions d. Environmental testing: Using water, soil samples, air samples, powders e. Chemical industry testing: Using bulk fluids and surface swabs f. Pharmaceutical manufacturing: Using in-process manufacturing materials and reagents and finished product g. Companion diagnostics: Using the same array of samples as clinical diagnostics h. Consumer wellness testing: Biological fluids, breath, skin samples, swabs i. Agricultural testing: Plant materials, soil samples j. Food pathogen testing: Food samples of all kinds, solid and liquid, raw and prepared. Also food-chain safety testing for adulterants, hormones, pathogens etc in animals and vegetables. k. Bio-defense: All kinds of biological fluids, air samples, soil samples, bulk liquids, powders l. Chemical defense: All kinds of biological fluids, air samples, soil samples, bulk liquids, powders m. Industrial health and safety: Surface swabs, air and bulk fluids Now you know, tell us how we can help you If you need to develop a rapid test in 2018, we hope you think seriously about lateral flow, no matter what market segment you are in. At DCN we strive to stay at the forefront of the technology and to develop the most ground-breaking applications using this platform. Let us know what challenge you are trying to address and we’ll tell you if it’s possible using lateral flow technology. Often it is. If it is, we’ll work with you to educate you on the best technology options and on the process of getting your product developed. We can then fully develop your product and transfer it into manufacturing for you under our ISO 9001 and EN 13485 compliant quality system. Email or call us to find out more about lateral flow technology, DCN and why this is actually the rapid diagnostic platform you need. Check out our YouTube channel for some related videos. About DCN Diagnostics: DCN Diagnostics is a full-service development company devoted to developing, validating and transferring to manufacturing all aspects of a lateral flow platform, including reagents, assays, cassettes, sample collection and handling devices and reader systems for any application. --- ## DCN Dx at SLAS 2018 URL: https://dcndx.com/insights/slas-2018/ Type: insight Published: 2018-01-30 If you are planning on attending SLAS 2018 next week, let us know. Members of the DCN Dx team will be walking the SLAS exhibit hall throughout the week. Let’s schedule a time to catch up on existing work or discuss the possibility of collaborating on the development of a new rapid diagnostic test. We can meet at the show or at our facility in Carlsbad. Schedule a Meeting --- ## DCN Dx Charitable Fund Donations 2017 URL: https://dcndx.com/insights/dcn-charitable-fund-donations-2017/ Type: insight Published: 2018-01-03 As the holiday season comes to a close, we would like to thank you for your support over the course of the past year as we work to advance the use of diagnostic tests in point of care situations. As you may know, the core of our mission at DCN Dx is to deliver positive outcomes for our clients and for our broader community. That is why we established the DCN Charitable Fund when we founded the company. For many families, 2017 was a difficult year due to natural disasters, illness and poverty. Thanks to your support, this year we have been able to expand our list of 2017 charitable contributions to include the following organizations: BOYS AND GIRLS CLUBS OF CARLSBAD WORLD VISION NOTHING BUT NETS WATER.ORG TEMPLE STREET CHILDREN’S HOSPITAL OF DUBLIN MAKE A WISH FOUNDATION WOUNDED WARRIOR PROJECT BOYS AND GIRLS CLUB OF GARDEN GROVE CHOC–CHILDREN’S HOSPITAL OF ORANGE COUNTY ST JUDES CHILDREN’S RESEARCH HOSPITAL HOPELINK–SEATTLE FONDATION SENEGAL SANTE MOBILE PEOPLE’S KITCHEN NEWCASTLE, UK CARLSBAD UNIFIED SCHOOLS HEIFER INTERNATIONAL SAN DIEGO FOOD BANK CANCER FUND FOR CHILDREN–DAISY LODGE–BELFAST, N. IRELAND HABITAT FOR HUMANITY –WAKE COUNTY, NC CURLEY’S HOUSE OF STYLE FOOD BANK, MIAMI, FL RONALD MCDONALD HOUSE OF CT AND MA ELAZARKI CHILDREN’S HOME–ISRAEL ANGKOR HOSPITAL FOR CHILDREN IN CAMBODIA HELP MITO KIDS,ORG 28JUNE.ORG FEEDING SAN DIEGO VITAL CONNECTIONS CATERINA’S CLUB –BRUNO SERATO AMERICAN RED CROSS–SAN DIEGO CHAPTER TO SUPPORT LILAC FIRE VICTIMS GRAND RAPIDS CATHOLIC CENTRAL HIGH SCHOOL CARLSBAD CHRISTMAS BUREAU –DCN ADOPTED FAMILY INTERACT OF WAKE COUNTY, NC The DCN Dx Charitable Fund is entirely financed by DCN Dx. We do not solicit or accept outside donations. We do, however, accept suggestions from our loyal clients in regard to deserving recipients. If you would like to suggest a recipient for our 2018 list, please contact us at customercare@dcndx.com. Once again, thank you for your trust in us and for your continued support. We look forward to continuing to work with you in 2018! Sincerely, Brendan O’Farrell, on behalf of the entire DCN Dx team --- ## Multiplex Assays Part 2: The Power of Spot Arrays in Multiplex Assays URL: https://dcndx.com/insights/part-2-arent-multiplexed-lateral-flow-tests/ Type: insight Published: 2017-08-28 In part 1 of this article series, we discussed general factors impacting the development of multiplexed lateral flow assays (multiplexed LFAs) and some approaches used in their design. In this second part, we delve deeper into a spot-array approach, which moves away from standard line-based architecture, as a promising technique for developing multiplexed LFAs. At DCN Dx, we utilize Symbolics TM, a process that pixelates binding reagents using high-precision dispensers to create spot arrays in lateral flow fields. This approach offers several advantages to multiplexed LFAs. Spot Arrays Enhance Quantification and Enable Dense Multiplexing There is a growing requirement in point of need or point of care diagnostics for the generation of assays that can detect more than one analyte in a single device. In a standard configuration, this means dispensing multiple lines perpendicular to the flow direction, separated by distances of 1 or more millimeters. A typical issue seen in multiplexed LFIs of this nature is “line bleed.” This condition occurs when the signal generated on one line merges with the next line resulting in the formation of background in the device, which lowers the sensitivity of the assay and can result in false positives. Quantification in multiplexed assays is even more difficult due to the fact that the dynamics varies from assay to assay depending on their location on the strip. Lateral flow assays are extremely time sensitive assays. The reaction begins as soon as the sample and conjugate mix in the conjugate or sample pad. The mixing continues during migration through the device to the test and control lines. The reaction at the test line occurs quickly, typically in less than 30 seconds. The flow rate of the reactants through the device can be extremely important to the performance of the assay. Flow rate through an analytical membrane, typically nitrocellulose decreases in a non-linear fashion with distance from the origin. As a result, the time taken for the reaction on first capture line versus the last capture line can be significantly different. This has implications for the ability to generate quantitative assays in multiplexed formats. These issues can be overcome by using spot arrays in lateral flow fields rather than lines. One of the most powerful potential applications of the Symbolics TM lateral flow arraying process is the generation of multiplexed arrays for different analytes on a single strip with the added power of generating internal controls and replicates. This approach brings huge potential to increase the power of the lateral flow format. Spot Arrays Allow for Alpha-numerical Symbols Instead of Lines in Multiplexed LFIs Generating letters, symbols, and lines in orientations other than perpendicular to the flow direction in lateral flow assays can be challenging due to flow dynamics and conjugate binding. However, by pixelating dispensed reagents into individual small dots, spaced appropriately, these difficulties can be overcome. The patented SymbolicsTM technology that we use at DCN Dx is based on the concept of the pixilation of reagents on the analytical membrane in a fashion that allows for the even development of each pixel. This appears, at first glance, to be a rather simple concept similar to dot matrix printing. The issue in lateral flow however, is that the formation of one feature in a flow path causes flow perturbations in the system that prevents the even formation of features behind it, or at least that is the general understanding. The key to the pixilation approach is in the controlled dispensing of the reagent spots for size and pitch (the center-to-center distance between spots in any axis). These factors must be balanced for each reagent based on the binding characteristics of the reagents. When well optimized and controlled, it is possible to position individual spots of the correct size and pitch in the flow path of a lateral flow assay in such a way that each feature develops evenly and, most importantly, does not prevent the development of the other features surrounding it. With even development of individual features, it becomes possible to reproduce larger patterns in a lateral flow field. Symbolics TM, Practically Speaking, for Multiplexed LFIs The shift to a spot array based process, which effectively represents a digital approach to feature creation in lateral flow fields, will represent a major shift in thinking for lateral flow developers and manufacturers. In latter years, there has been a growing awareness of the capability of the lateral flow system in diverse markets, from commercial applications that have a requirement for simplicity in use and interpretation, to very highly specified quantitative and multiplexed biomarker panels. Although these applications require high performance from the assays, they are produced using relatively standard, although highly controlled processes. Symbolics TM technology takes the requirement for process control to the next level, melding the lessons acquired in years of protein array development with the high speed, high throughput manufacturing approaches used in standard lateral flow production. Manufacturers will have some mental hurdles to cross in adopting this technology, as will assay developers. However, the potential application advantage that Symbolics TM can bring is huge. The first challenges will come in optimization. As mentioned previously, the key to even macro feature generation is the ability to develop micro features in the flow field that do not perturb the flow of liquid and conjugate and prevent even formation of the next feature in the pathway. The ability to do this depends on the size of the micro feature and the distance between the features, but also the binding affinity of the reagents. Each reagent therefore requires some optimization for feature size, reagent concentration and feature spacing. In most lateral flow applications, when screening for reagents, the focus tends to be on isolating antibodies with high on-rates (Kon). This remains true for Symbolics TM. Features are small, so reagent on-rates must be high to ensure adequate sensitivity. Balancing that, the small size of the features means that it may be possible to reduce the amount of binding reagent significantly, resulting in savings in reagent costs. Materials must likewise be carefully specified and screened in order to best optimize performance in the Symbolics TM format, however this is not significantly different to a standard lateral flow approach. Overall, the development approach is similar between the Symbolics TM format and the standard lateral flow format. The major difference between the approach to optimization, development and manufacturing of the Symbolics TM format is in the dispensing of capture reagents. Optimization and manufacturing requires high quality, precision dispensing systems, and levels of process control that are not typical in standard lateral flow applications. Conclusion: Multiplexed LFIs Lateral flow is a continuously evolving format. Multiplexing is an obvious evolutionary step, allowing for a whole new swathe of potential applications. However, applications being considered should be carefully researched in terms of market need, regulatory complexity, reimbursement conditions as well as for the appropriate technical assay architecture and format to pursue. As one potential multiplexing format, spot arraying facilitates another step in the evolution of the lateral flow system. It has the potential to enable a variety of applications, including: More intuitive, easy to interpret consumer assays Better controlled quantitative assays Multiplexed assays in a variety of formats, including array, parallel multiplexing and “thermometer style” formats More complex pattern recognition based detection methods for high density arraying or for quantification As such, this approach provides a means to differentiate products, improve performance and to give end users an improved, intuitive and less error-prone experience. More information on the Symbolics TM approach, applications and intellectual property portfolio surrounding this pixilation approach to lateral flow multiplexing is available at www.symbolicsdx.com. Please also contact us for more information on our capabilities in regard to multiplexed LFI development and production. You may also be interested in our DCNovations lateral flow products— available online. More on multiplexed LFIs will also be presented and discussed in detail at the Advanced Lateral Flow Conference, so please join us in San Diego! --- ## Out of the Mouths of Babes URL: https://dcndx.com/insights/smartphones-lateral-flow-alfc/ Type: insight Published: 2017-08-23 This apparently intriguing string was recently published on LinkedIn from my account. When it was brought to my attention, I was going to delete it, but I noticed it was getting more and more hits. Eventually it got more views than many of the things I’ve deliberately posted through the years. Replies came in to my LinkedIn account and my personal email wondering what this was a comment on and how it should be interpreted. Was it work-related or political? Some were concerned that a stroke had taken out my language center. Some thought my love of wine had finally intersected with my late-night LinkedIn posting habit. Most were confused, but all were interested to know the hidden meaning. The less-than intriguing truth is that my 15-month-old daughter has a way with electronics. She can unlock any device, delete pretty much any application, or Facetime with her Grandad in Ireland at 3am local time. And now her first post on social media has become quite popular. I was hoping that wouldn’t happen for well over another decade. Parenting fail. And, of course, now I’m left wondering what it means that she has gotten more hits than many of my work posts have. No need to add your opinions on that to the Comments section, thank you! Beyond that, though, the fact that my daughter–despite our best efforts to minimize her exposure to screens in general at this stage of her life–has identified the phone as just about the most useful tool in her parents’ lives, making it about the most attractive thing to her, got me thinking again about the cell phone as a tool in our industry. Why there have been no commercial successes in applying cell phone readers (without added hardware!) to the toolbox of point of care, regulated, lateral flow based diagnostics. Why, despite years of development effort and marketing spin is there still no industry standardization of hardware or software specifications or a demonstrated, acceptable approach to calibration and validation that will be necessary for regulators to buy in to this concept for regulated medical diagnostic applications? Isn’t it time for that? Rather than a head-in-the-sand approach to this issue or an anecdotal acceptance that “this is just going to be difficult to sell to regulators so why bother”, isn’t there a need for an industry-wide discussion on this topic? I don’t pretend to have all the answers to this one, but I’m curious to understand the general temperature of the industry on this topic. (Feel free to contact us on this topic and let me know!) In the meantime, I find solace in knowing that my unintentional post generated concern for my well-being from so many people. As for my daughter’s intentions behind the post, I can only guess that she was urging me to put work aside and join her in playtime outdoors. Smart kid. If anyone has any thoughts on the use of smartphones in LFA interpretation in regulated applications, or suggestions as to the meaning of please or join us at the Advanced Lateral Flow Conference in San Diego to discuss. --- ## The Multiplex Assay, Part 1: Where are all the Multiplexed LFIs? URL: https://dcndx.com/insights/multiplex-assay-lateral-flow-tests/ Type: insight Published: 2017-06-06 At DCN Dx, the demand for developing multiplex lateral flow devices is on the rise. Multiplex assays, which detect or measure multiple analytes in a single device, are a growing area of application development. However, designing, developing, and commercializing these products involve specific considerations. The term “multiplexing” can have different meanings, depending on the context. In general, a “multiplexed” assay detects or measures more than one analyte in a single device. However, it can also refer to generating multiple replicates of a single test within a single device or using positive, negative, or kinetic controls within a test, along with the analyte-specific assay. Despite the interest in multiplex assays, the number of true multiplex lateral flow applications commercialized to date remains relatively low. This is due to various technical, manufacturing, and commercial considerations. In many cases, technical and manufacturing challenges are less of an obstacle than commercial ones. In this article, we will discuss these factors impacting the success of multiplexed devices. Technical Issues Multiplexing analytes is easier for certain classes of assays, such as serology tests, which target a single class of antibody directed against multiple analytes. In contrast, biomarker assays, which detect several different proteins from a single sample and may require multiple conjugates, are more complex. Let’s take a closer look at a simple duplex system designed to detect two protein antigens in a whole blood sample. The primary technical considerations include: the specificity of the available binding reagents potential interferences in the sample the availability of the analytes for detection. In other words, does the sample have to be pre-treated and does the same sample treatment work for both analytes? the required LOD and dynamic range for each analyte. Are they present in roughly the same concentrations so that both can be detected from the same sample? the stability of the binding reagents for each analyte. Is any difference in degradation rates of those binding reagents? Compatibility between the assays is crucial for multiplexing analytes from a single sample. If that compatibility exists, we can move on to the lower order technical hurdles. Not all of these issues are deal-killers. Many can be overcome. However, the more analytes we add to the mix, the greater the complexity of answering some of these questions becomes. If we add quantification to the mix, the system becomes even more complex. In that case, we are concerned with variability in the system that can limit the accuracy of the results. We have to work to generate highly reproducible results in each of our assays and understand the issues around degradation of reagent performance in the context of what that can do to the standard curves in the system. Again, more analytes, more complexity. Manufacturing Issues Manufacturing a multiplexed lateral flow assay can be complex. The more assays on a device, the greater the potential risk of manufacturing failure and post-market failure of the product. Processing equipment, materials, manufacturing, and QC processes may differ from those used in standard single-plex assays, requiring manufacturers to have deep technical capabilities to ensure ongoing quality and production yields. Commercial Issues From a technical perspective, it may be possible to multiplex many different assays onto a single rapid assay product. However, the utility and market demand for the product must be considered. Questions to ask include: What is the added regulatory complexity of the multiplex relative to single-plexed assays? What is the added manufacturing complexity and risk? What is the likely cost of the device? Can we get reimbursement for the multiplexed assays? Does the market, the clinician, or the end user want or need all the data from our multiplex system? Although there is a relatively small number of panels that make sense from all these perspectives, many assays can benefit from the power of replicates and built-in controls offered by a multiplexed assay. So, why aren’t there more multiplexed lateral flow tests on the market? In short the ability to develop, manufacture and commercialize a multiplexed point of care test requires a balance between technical capability, design and market need. But there is demand in many application areas that can help to overcome the commercial issues, so developers and manufacturers need to be ready to meet the technical challenges. In part 2 of this series, we will discuss some effective multiplexed assay and product architectures and some novel approaches to overcome those challenges. In the meantime, do not hesitate to contact us to discuss this topic further. Learn more about our lateral flow assay development services and our lateral flow education courses. We also invite you to join us at the Advanced Lateral Flow Conference in San Diego, where lateral flow professionals from all over the world are gathering to learn, collaborate, and share ideas about lateral flow technology and the future of IVD the market. Learn more here. --- ## The Power of Collaboration in Lateral Flow: Educational Programs from DCN Dx URL: https://dcndx.com/insights/dcn-article-jack-trades/ Type: insight Published: 2017-06-06 (As published 2017-06-06; regulatory status may have changed since.) “The discovery of immunoassay was part mathematics, part physics and part biology. I learned about biology from Sol (Solomon Berson) and he learned about physics from me.” – Rosalyn Yalow, co-inventor of radio-immunoassay and Nobel Laureate for Physiology or Medicine in 1977. Rosalyn Yalow played a crucial role in the development of the core technology that became the multi-billion-dollar immunoassay industry. From the very beginning, she understood that the development of immunoassays demanded a collaborative approach and proficiency in numerous disciplines. Fast forward almost 60 years since the publication of the groundbreaking Yalow and Berson paper on the immunoassay of endogenous plasma insulin 1, this principle is more relevant than ever. For instance, when it comes to developing a lateral flow assay system, one must possess knowledge in immunology, organic chemistry, materials science, fundamental biology, fluid dynamics, plastics, physics, biophysics, and optics. Additionally, a profound understanding of mechanical and design engineering, molding, manufacturing process design, and various other disciplines are required to transform a conceptual assay into a usable, reproducible, robust, commercial product. This doesn’t even take into account regulation, trials, quality systems, user experience studies, validation, and so on! This might explain why there are limited places to learn how to develop commercial lateral flow products outside of a few industrial sources like DCN Dx. With this technology, experience is invaluable when developing a product that can be robustly manufactured and produced to meet specifications in high volume. It’s challenging to find people who are willing to share the benefits of their hard-earned experience. Consequently, it’s tough to break into this industry from scratch or even enhance our knowledge base when we’re already in it! At DCN Dx, we understand that fact, which is why we make such an effort to provide education and training services to the industry alongside our development services. We’ve taken an approach that many through the years have deemed counter-intuitive—an OEM company that actually teaches others how to do what it does. How crazy is that? We believe that our clients place immense trust in our promise of delivery. Part of building and maintaining that trust is transparency, and information transfer is an essential aspect of transparency. We have always aimed to help, in good faith, build this market and make our clients successful. As the market grows, so do we. That’s why we offer various educational offerings for lateral flow technology for our clients. The Basic Lateral Flow Training is a three-day, hands-on course that combines interactive lectures from our assay scientists and engineers, with guided, practical experience in our laboratory. Limited to 12 participants and run only a handful of times a year, this training sells out fast! We also offer a custom version of this course that is tailored to an individual IVD developer’s needs. Contact us for more information. Basic Lateral Flow Training course participants during a hands-on laboratory session. April 2023. It’s also why we organize the Advanced Lateral Flow Conference (ALFC), originally with our partners at QIAGEN Lake Constance and now with more than 30 partners from the lateral flow supply chain. This 2-day seminar is designed to educate the industry on the best lateral flow development and manufacturing practices. You can ALFC 2021, Keynote address from Brendan O’Farrell, Ph.D. Networking between industry, government, and academia is a highlight of the ALFC The ALFC encourages participation by start-up companies ALFC exhibitors and attendees network throughout the event The ALFC’s scientific sessions include a variety of relevant topics Collaboration and cooperation are essential to success in this industry. It really does take a village to bring a high performance lateral flow test to the market. Come meet the neighbors. Yalow, RS; Berson, SA (1960). “Immunoassay of endogenous plasma insulin in man”. J Clin Invest 39: 1157–75. doi: 1172/JCI104130. PMC 441860. PMID 13846364. --- ## Lateral Flow Assay Cassette Design: Why Do LFI Cassettes Give Problems? URL: https://dcndx.com/insights/lateral-flow-assay-cassette-design/ Type: insight Published: 2017-06-06 The design of a lateral flow cassette is a critical factor in ensuring the performance and functionality of a lateral flow test. However, many developers treat the lateral flow assay cassette as an afterthought, which can lead to a host of issues. DCN Dx has pioneered a cassette design and development process that emphasizes the importance of lateral flow cassettes, optimizing cost, quality, and performance. The traditional approach to lateral flow cassette design often neglects the cassette’s role in the test’s overall performance. This oversight can result in problems such as sample leakage, improper flow patterns, and uneven line development. By focusing on a well-designed lateral flow assay cassette, developers can achieve better integration between the cassette and the test strip, leading to improved performance and functionality. The “Standard” Lateral Flow Assay Cassette The standard approach to lateral flow assay cassette (LFI cassette) design is for the strip developer to transfer the specifications for a strip to an industrial design house or an engineering firm that handles the design of the cassette. Often times, this happens towards the end of the assay’s development phase. The cassette design process is then handled in an iterative fashion, moving from rapid prototype parts through low cost molds with inserts, and on into a production mold. Unfortunately, this can be a slow, costly process that often yields subpar results. As test developers, DCN Dx long ago realized that this element of the development process can lead to annoying delays and cost drift when it is handled separately from the test. This comes from a variety of factors, including starting the design and integration of the cassette too late in the process, the need for multiple suppliers to develop the assay and the cassette, and the typical process that OEM cassette developers tend to follow, where each cassette tends to be designed from scratch using “industry standard” design features, low cost molding and assembly methods. We’ve has never been happy with that approach, as we know from experience that when it comes to product performance the cassette is an essential part of the product. It’s the key to functionality of the ultimate product, to usability, to user satisfaction, as well as to branding and value proposition. This component should not be an afterthought. This is why DCN Dx’s approach integrates engineering and assay development teams and utilizes a proven base cassette design, allowing for a faster, more efficient process that results in a higher quality final product. So while investing in the development of a high performance lateral flow test, why design another piece of cheap white plastic that is poorly molded and prone to failure? Why follow the “industry standard process”? At DCN Dx, we don’t. We’ve designed a process for cassette design and development that supports our clients’ needs for cost, quality and performance that is a hassle-free and effective. Let’s examine the common issues and talk about some potential solutions. LFI Cassettes Tend to be an Afterthought One of the most common – and biggest – mistakes made in lateral flow device design and development is that the LFI cassette is considered as an afterthought. The focus is placed on making the test work and it is assumed that one can drop the assay into a cassette, possibly even a generic cassette that is taken “off the shelf”, and everything will work fine. After all, the LFI cassette is just a holder for the strip….Not! The lateral flow assay cassette and the test itself are intertwined in their design and functionality. A good cassette design will rarely overcome problems in the test architecture or design (the test does need to work independently of the cassette first!). A bad cassette design will definitely exacerbate those problems. Typical problems seen when developers try to drop a strip into a cassette that has not been designed for the strip architecture include: Sample leakage and flooding of the strip Improper or uneven flow patterns Incomplete flow (tests don’t run) Uneven line development Bad background development Red blood cell leakage if it’s a whole blood-based test Most of these failures are catastrophic to the overall performance of the product, and all of them can be fixed relatively simply by designing the cassette to match the strip architecture and the needs of the sample. “Standard Practice” Does Not Yield Well-designed or Well-molded LFI Cassettes In the lateral flow OEM segment, generation of LFI cassette designs for years has depended on a relatively standard process. Assay developers engage an engineering firm to design a cassette once the test has been developed to a certain point. Initial conceptual and functional designs result in the generation of SLA’s (stereolithographic, rapid prototype parts) that are used to “dial in” the key pressure points and closure pressures in a lateral flow cassette. The next step is typically to move into a cheap mold insert (or MUD) that is used to produce parts for testing. This mold is modified, sometimes repeatedly, until the parts it produces work adequately. This is a slow, tedious, iterative process that eats up laboratory, engineering and machine shop time and it costs money. Also, this process doesn’t work very well in many cases. SLA parts don’t allow for good approximations of closure of molded parts and the materials used are different and behave differently to the final molded parts. In addition, this process is often performed by contracted engineering firms who do not have intimate knowledge of the test and how lateral flow actually works. Finally, the design of the part is often considered separately to the rest of the manufacturing process including cassette closure methods. This is a potentially disastrous mistake (see #4 below). DCN Dx’s engineering and assay development teams work hand-in-hand all the way through the process. Our industrial designers produce designs that address functionality and user needs. SLA parts are produced to assess look and feel and initial functionality. We then move very quickly into the use of Protomold parts that behave like the final molded part. Crucially, our process starts with a proven base cassette design that within two or three iterations at most will typically generate the final part. This yields a highly cost-effective and very fast process. Additionally, the design is done in conjunction with the development of the assembly process, so that closure issues are considered from the beginning. We’ll talk more about that later. The Focus Historically Has Been on Cost vs. Quality This can affect the design of the part and the design of the molding process. Cost pressure tends to force molders to skimp on measures that can produce better quality parts. Poor gating, poor mold flow, poor part ejection are all aspects that all lead to part failures in the final test. They are easily fixed for a penny or two per part, so it’s short sighted not to do it right. In this day and age, for any except the very lowest quality qualitative tests, this is no longer an acceptable approach. Proper attention needs to be paid to mold design and molding process. Spend the money to do it right! Short term pain will yield serious long term gains. Closure Design, Closure Process, Closure, Closure, Closure… Do we need to emphasize this more?! Many issues happen because of what has become an industry standard cassette closure design and closure method. Old school pin and boss closures still predominate, often with simple round pins and round holes. There’s too much detail to go into here, but give us a call and ask us why this is a problem and how it’s easily fixed. Also, the closure process is often ignored during the design of the part. This is a CRITICAL mistake. The entire functionality of the device depends on the fidelity and reproducibility of the closure and the pressure points within the cassette. The cassettes created by DCN Dx are always designed with the closure process in mind. If pins are used they are never round pins going into round holes. Pins and bosses are placed at locations that ensure that the strip can’t be over or under compressed. Guide features ensure that the pins go into the holes properly during closure and don’t shear or deform. It’s simple stuff, but it’s got to be considered. Performance is always tested in our labs using the closure method for which the part is designed and that whole process is transferred to the manufacturer. Oh, and don’t ever rely on the pins to define the closure pressure of the cassette. Ask us why and how it can be done better. Don’t Waste the Opportunity to Design the LFI Cassette Properly DCN Dx’s process for lateral flow assay cassette design and development results in high-performance cassettes that are user- and application-centric, providing added value to the product. By prioritizing the design and development of lateral flow assay cassettes, DCN Dx can help you maximize the value of your product without compromising quality. Leveraging the expertise of DCN Dx for your lateral flow cassette design and development process can lead to significant improvements in the performance and value of your product. Discover our DCNovations LFI Cassette, a solution to the “cheap white plastic” trap. Don’t settle for “industry standard”—let DCN Dx help you get more value from your lateral flow assay cassette design and development process. --- ## Advances in Lateral Flow Technology for Point or Care & Field-Based Applications URL: https://dcndx.com/insights/advances-lateral-flow-technology-applications/ Type: insight Published: 2017-06-06 Lateral flow immunoassays (LFIs) have revolutionized the way we perform rapid diagnostic testing in point of care and field-based applications. These rapid, cost-effective tests have proven invaluable in various industries, from healthcare to environmental monitoring. DCN Dx’s President and Co-founder, Brendan O’Farrell, Ph.D., authored a book chapter on recent advances in lateral flow technology in the book, Portable Biosensors and Point-of-Care Systems. This blog post will delve into some of the key advancements in LFIs and explore how they are transforming the field of point of care diagnostics. Click to Get Your Copy Advances in Lateral Flow Technology: Materials and Design One of the major advances in lateral flow technology is the development of novel materials and designs for LFIs. Traditionally, LFIs have relied on nitrocellulose membranes to facilitate the flow of samples and reagents. However, recent advancements in materials science have led to the development of new types of membranes, including cellulose nanofibers and nanoporous membranes. These materials offer improved performance, sensitivity, and reliability, making them ideal for use in next-generation LFI devices. In addition to new materials, advances in LFI design have also contributed to improved performance. For example, multiplexing capabilities now allow for simultaneous detection of multiple analytes in a single test. This feature is particularly useful in applications where multiple targets must be detected, such as infectious disease diagnostics or environmental monitoring. Enhanced Sensitivity and Quantitative Capability One of the key benefits of advances in lateral flow technology is the improvement in sensitivity and the ability to provide quantitative results. Traditional LFIs are known for their qualitative nature, providing a simple positive or negative result. However, recent advancements in detector molecules, such as gold nanoparticles, fluorescent labels, and magnetic particles, have greatly increased the sensitivity of LFI tests. Moreover, advances in readout technology, including portable readers and smartphone-based platforms, now enable the capture of quantitative results from LFIs. This capability is crucial in applications where precise measurements are necessary, such as monitoring drug levels, determining viral load, or measuring environmental contaminants. Integration with IoT and Data Management Another significant advance in lateral flow technology is the integration of LFI devices with the Internet of Things (IoT) and data management systems. This connectivity allows for real-time data collection, remote diagnostics, and centralized data storage. The ability to track and analyze diagnostic results over time is invaluable for tracking disease progression, evaluating treatment efficacy, and guiding public health interventions. Portable Biosensors and Point-of-Care Systems Dr. O’Farrell’s book chapter, found in Portable Biosensors and Point-of-Care Systems, provides a overview of the latest advances in lateral flow technology and their impact on point of care applications. As LFI technology continues to evolve, the potential applications for these rapid, sensitive tests will only continue to expand. Need Support for Your Next Assay Development Program? Don’t miss out on the benefits of cutting-edge lateral flow technology for your point of care applications! DCN Dx specializes in contract assay development, helping you leverage the latest advancements for your specific needs. Visit our Contract Assay Development page to learn more about our services and how we can help you transform your diagnostics capabilities. Get started on your path to innovation with DCN Dx today! --- # ===== PRODUCTS ===== ## Blood Collection/Separation Materials Kit URL: https://dcndx.com/store/product/blood-collection-separation-materials-kit/ Type: product Published: 2020-06-04 Blood Collection/Separation Materials Kit All-in-one kit for blood collection and RBC separation. Whether you’re developing a blood collection device or a blood-based LFA, save valuable time with the DCNovations Blood Collection/Separation Materials Kit. This all-in-one kit provides the materials you need for your blood-based project to filter out RBCs without hemolysis. Don’t waste time researching materials, contacting individual suppliers, and coordinating shipments: the Blood Collection/Separation Materials Kit has everything you need for fingerstick blood diagnostic tests and blood sample collection devices. Perfect for everyone from startups to academicians to large companies. Product Details --- ## Nucleic Acid Lateral Flow Kit URL: https://dcndx.com/store/product/nucleic-acid-lateral-flow-kit/ Type: product Published: 2020-03-31 The DCNovations NALF kit is economical, fast, and easy-to-use. It provides for the detection of amplicons without expensive laboratory equipment. Users perform the amplification reaction using two primers: one labeled with either Fluorescein or Digoxin and the other labeled with Biotin. Successful amplification will result in a line formed. Unlike our previous NALF product, NALF D is compatible with up to 5mM DTT, a reagent common to molecular reactions. DCNovations NALF kits are available in packs of 50 individually pouched cassettes. --- ## Basic Lateral Flow Training URL: https://dcndx.com/store/product/basic-lateral-flow-education/ Type: product Published: 2020-01-20 Take your lateral flow skills to the next level. If you’re ready to get started developing lateral flow assays or improve your skills, the experts at DCN Dx can help you on your way. Our Basic Training Course is a three-day lateral flow training program that combines hands-on practical lab work with lectures to provide you with the fundamental knowledge needed to kick-start your development project or improve your current skill set. This course is designed for startups, established life science companies, students, and faculty who want to learn the fundamentals of lateral flow development. With a small class size limited to 12 attendees from diverse backgrounds, you’ll have ample opportunity to interact with your instructor and classmates, ensuring that you leave with a solid foundational understanding of lateral flow assay development. Looking for a custom course designed for your specific lateral flow challenge? to inquire about our custom training program. In this comprehensive course, you’ll learn about: The development process and design control Materials and reagent selection Processing essentials and treatments Reader technologies and quantification Manufacturing considerations Stability studies Cassette design and molding Troubleshooting strategies Fee [pricing on request] (regular); [pricing on request] (early bird). Course fees include lectures, copies of lecture materials, access to the lab and scientific staff, copies of all protocols developed or used during the course, use of equipment, all standard materials and reagents, and lunch each day. Travel and accommodations are not included (though we are happy to assist with arrangements). State-Of-The-Art Facilities Our headquarters in Carlsbad, California is fully equipped to enhance your learning experience. 35,000+ sq. ft. custom-designed facility with dry room capability 6,626 sq. ft. BSL-2 development lab Reel-to-reel development, validation, and production capability Engineering lab and workshop space with SLA capability ISO 9001:2015 and EN 13485:2016 certified 100+ full-time employees and trusted partners --- ## MICA-200C LFA Cassettes URL: https://dcndx.com/store/product/cassettes/ Type: product Published: 2019-08-29 DCNovations MICA-200C LFA Cassettes are DCN Dx’s flagship off-the-shelf cassettes for 5mm x 60mm test strips. Our LFA cassettes are engineered and designed to control flow, present the correct pressure points, enable sample addition in a controlled fashion, and fit tightly together without warp. Designed by DCN Dx scientists to meet the specifications for some of the most popular materials used in lateral flow assays, our off-the-shelf cassette accommodates approximately 85% of lateral flow assay strip designs. Avoid the “cheap white plastic trap”—trust DCNovations MICA-200C LFA Cassettes. Contact us for bulk pricing. Learn more about why DCN Dx designed our DCNovations MICA-200C LFA Cassettes and lateral flow assay cassette design best practices. --- ## Lateral Flow Materials Kits URL: https://dcndx.com/store/product/lateral-flow-materials-starter-kit/ Type: product Published: 2019-01-08 Unfortunately, there is no set of “one size fits all” materials that will work for every assay. For this reason, it is necessary to evaluate several different combinations of conjugate pads, nitrocellulose and sample pads during the early stages of development. This process can be cumbersome because of the extensive catalog of options available on the market, large minimum order requirements and long lead times from manufacturers. Thus, the DCNovations Lateral Flow Materials Kits are essential for the beginning stage of any lateral flow development program. Our lateral flow materials kits come in two sizes. Each kit contain packs sourced from various vendors and offers distinct capabilities. Mini Materials Kit: This streamlined, budget-friendly kit is ideal for the first time lateral flow developer who needs to learn the basics before launching a large-scale materials evaluation. The Mini Materials Kit includes 4 packs of membranes, 2 packs of conjugate pads, 1 pack of absorbent pads, and 2 packs of backing cards. Materials Kit: Our full-sized materials kit is developed for research groups or startups with lateral flow development projects already underway. This kit contains all the necessary dry materials for assembling a lateral flow strip; containing 8 packs of membranes, 3 packs of conjugate pads, 4 packs of sample pads, 3 packs of wick pads, 4 packs of backing cards, and cassettes. Have a question? Contact our technical team here. --- ## Colloidal Gold URL: https://dcndx.com/store/product/colloidal-gold-lateral-flow/ Type: product Published: 2019-01-08 DCNovations Colloidal Gold has been used at DCN Dx for more than 15 years in the generation of stable conjugates for various sample matrices, including whole blood, plasma, urine, saliva, and alcoholic beverages. Manufacturing groups, university researchers, start-ups/spin-outs, and research groups in mid-to large-size companies alike find DCN Dx’s cost-efficient version of this respected standby an ideal addition to their LFA products. Not sure if DCNovations Colloidal Gold is right for you? Contact us. --- ## Cellulose NanoBeads URL: https://dcndx.com/store/product/cellulose-nanobeads/ Type: product Published: 2019-01-08 The unique characteristics of these particles allow for increased sensitivity, faster detection time, and improved reproducibility when compared to colloidal gold in typical lateral flow tests. These high-performance visual labels are available in a variety of bright colors ideal for multiplexing, making them ideal for multiplexing. Colors available: Red Blue Green Red ( Covalent) Blue (Covalent) All options sold at 1% solids (1mg/mL) in water and the average particle size is 300nm. Try these particles in your own assay with our CNB Conjugation Kit. This product comes equipped with all of the protocols and chemistries required for protein conjugation to cellulose nanobeads. Not sure which cellulose nanobead is right for your project? Contact us with your questions. --- ## CNB Conjugation Kit URL: https://dcndx.com/store/product/cellulose-nanobead-conjugation-kit/ Type: product Published: 2019-01-08 Each CNB conjugation kit contains the reagents necessary to prepare and optimize several protein conjugations to red and blue cellulose nanobeads using protocols recommended by DCN Dx’s expert team of scientists. There are enough reagents to prepare 6mL of 0.1% conjugates. Particles to be conjugated (0.3mL of red (RE2) NanoBeads and 0.3mL of blue (BL2) NanoBeads) Blocking buffer (long-term storage at -20°C; expiration date 3 mo from thaw date, store at 2-8°C) Conjugation buffer Wash buffer Protocols for conjugation & testing To purchase the nanobeads separately, please visit our Cellulose NanoBeads page. Unsure if the CNB conjugation kit is right for your lateral flow development program? Contact us with your questions. --- ## Colloidal Gold Conjugation Kit URL: https://dcndx.com/store/product/colloidal-gold-conjugation-kit/ Type: product Published: 2019-01-07 Each kit contains all of the reagents necessary for the optimization of most protein-colloidal gold conjugates, as well as the protocols recommended by DCN’s expert team of scientists. There are enough reagents in the kit to make up to 50mL of OD 10 conjugates. This kit contains: Conjugation, optimization & testing protocols Particles to be conjugated (0.5L of DCNovations Colloidal Gold) Blocking buffer (3 mo expiry date from date of manufacture stored at 2-8°C) Storage buffer For more information about DCNovations Colloidal Gold, visit our colloidal gold product page. ---