OUSD (R&E) CRITICAL TECHNOLOGY AREA(S): Combat Casualty Care OBJECTIVE: To develop an in vitro diagnostic (IVD) platform with fast and wide multiplexing capability. This tool should be able to detect at least 25 multi-omics targets in a rapid and automated fashion from single input of minimally invasive biomatrix with an insignificant risk of an erroneous result. DESCRIPTION: Recent advancements in the field of hardware miniaturization and nanotech based manufacturing industry essentially made a significant change in the conventional molecular testing landscape. The prospect of detecting multi-target profile in the clinical set up with a high precision has becoming more feasible, which is also reflected in the latest list of FDA approved detection kits [1]. Current trend in the commercial pipeline could be highlighted by the following IVD platform, namely FoundationOne CDx (F1CDx), which received FDA approval in 2020; its latest version, namely PGDx is enabled to detect the genetic aberrations linked to multiple types of cancers. These tests deploy Next Gen Sequencing (NGS) platform to detect substitutions, insertions and deletions, and copy-number alterations in 300-500 genes [2]. Such capability, namely one platform that can diagnose multiple diseases is highly beneficial in battlespace since Role 1/ 2 facilities have limited real estate. The advantages of this platform will be magnified should the technology meet the following criteria- simple laboratory examinations and procedures that have an insignificant risk of an erroneous result. This is essentially the criteria to get a COW from CLIA testing. None of the available IVD platform that can detect multiple targets can meet the criteria to get CLIA waiver. Present solicitation seeks to bridge this knowledge gap. Note: It's not necessary for a solicitor to get COW from CLIA within this project timeframe; however, these guidelines should be used as a benchmark to monitor the success criteria. Present market is dominated by polymerized chain reaction (PCR)-based detection kits that detect a handful of most eligible molecular targets. Diverging from the current position of IVD market, our proposed platform seeks a more holistic approach to detect all possible actionable targets from minimum inputs and within short turnaround time. Our approach is bolstered by a 2019 market analysis that forecasted a significant gain in diagnostic market share in near future by the wide multiplexing tools with improved sensitivity and specificity compared with traditional sequencing technologies, as well as faster identification. [3] Our objective is to develop a prototype of wide multiplexing capability. It should be able to detect at least 25 targets with high sensitivity and specificity as compared to traditional sequencing technology. Moreover, the prototype should have a small footprint and easy to executable protocol. The protocol should have an automated hands-free process that would allow a single input of minimally invasive biomatrix for a nearly error-free detection. This prototype should be developed with the aim to eventually meet the criteria to get a COW from CLIA. Till date, there are only 40 tests that have been approved for COW status at CLIA website http://www.fda.gov/cdrh/clia and none has wide multiplexing capability. Our 40-gene panel of sepsis biomarker could be used in the prototype; nevertheless, the best candidate prototype should have maximum flexibility, so that the prototype could be easily repurposed or co-diagnose additional diseases including, but not limited to traumatic brain injury (TBI), infection, and other psychological markers. A web search of SBIR and STTR solicitations (dated January 23, 2024) found no existing solicitations to develop wide multiplexed tool that will be portable, simple, automated, and can detect a wide spectrum of molecules to facilitate diagnose/monitor/predict multiple diseases. PHASE I: Phase I will demonstrate the feasibility of a fast and wide multiplexing detection platform; the proof-of-concept should explain methodologies to detect fast and wide multiplexing capability from single input of biomatrix of choice. The expectation is that the biomatrix should be minimally invasive, such as blood, saliva, urine etc. Target biomarkers could be curated from the public domain. Use of human or animal subjects is not intended, or expected, to establish/achieve the necessary proof-of-concept in Phase I. In summary, our expectations from Phase I are the following: A plan to develop a prototype that can detect at least 25 targets. The final product should be flexible to diagnose multiple diseases, such as sepsis, TBI and pathogenic infection. The expected device should be an automated and portable platform that should be readily used in far forward lab or at bedside with an insignificant risk of an erroneous result. The concept is expected to support an end-to-end methodology e.g., an integrated sample collection-to-assay-to-detection protocol. The concept should show a feasible route to develop wide multiplexing capability with high precision as compared to traditional NGS platform is essential. This metrics should be used/ tested from the beginning of protocol design. PHASE II: The proof-of-concept generated in Phase I should be transformed into a working prototype during Phase II. Phase II should start with a plan to assay the biomatrix of choice to detect a panel of multi-omics biomarkers. A comprehensive testing is expected to determine the feasibility of the platform to be operated with minimum hands-on time and least supervision. Suitable biomatrix should be finalized. The detection mode (e.g., visual inspection vs. digital record etc.) of endpoint reading should be finalized, and this process should be easily interpretable. We encourage to have a data driven analysis of the proposed capability tested using biomatrix that can inform us about the feasibility of next steps. In summary, our expectation from Phase II is the following: The input and output modus operandi should be finalized. Assay sensitivity and specificity should be characterized. Screening of limit of detection (LOD) profile in presence of potential confounders and contaminates is expected. A turn-around time should be tested. Herein the assay time includes the sample collection, assay and detection. Potential risk factors and mitigation plan should be discussed. Probable assay cost should be estimated. Plan for a path forward to secure FDA approval. PHASE III DUAL USE APPLICATIONS: The goal of this phase is to secure an FDA approved product that is intended to be suitable for use and potential procurement for primary use in the field/prehospital environment, including bedside, austere/ far forward, Role 1/2 facilities and prolonged care scenarios. At this phase, target diseases and pertinent biomarkers should be determined. Accuracy, reliability, and usability should be assessed. This testing should be controlled and rigorous. Statistical power should be adequate to document final efficacy and feasibility of the assay. Funding could be solicited from CDMRP and BARDA, who usually support such efforts focused to military health. As mentioned previously, the target disease might include those health issues that are nonexclusive to active-duty members. Realization of a dual-use technology applicable to both the military and civilian use could be achieved via making commercial partners with IVD marker leaders like Roche, Inc, Illumina, Inc., Bio-Rad, Inc. etc.The goal of this phase is to secure an FDA approved product that is intended to be suitable for use and potential procurement for primary use in the field/prehospital environment, including bedside, austere/ far forward, Role 1/2 facilities and prolonged care scenarios. At this phase, target diseases and pertinent biomarkers should be determined. Accuracy, reliability, and usability should be assessed. This testing should be controlled and rigorous. Statistical power should be adequate to document final efficacy and feasibility of the assay. Funding could be solicited from CDMRP and BARDA, who usually support such efforts focused to military health. As mentioned previously, the target disease might include those health issues that are nonexclusive to active-duty members. Realization of a dual-use technology applicable to both the military and civilian use could be achieved via making commercial partners with IVD marker leaders like Roche, Inc, Illumina, Inc., Bio-Rad, Inc. etc. REFERENCES: US Food and Drug Administration Website: Nucleic acid-based tests. Page last updated: 11 Sep 2018. Accessed 12 Dec 2023. https://www.fda.gov/medical-devices/in-vitro-diagnostics/nucleic-acid-based-tests Sternberg, A. HITTING THE TARGET: Multigene Tests Gain Foothold in More Clinical Settings. Targeted Therapies in Oncology. June 1, 2020; 9 (8), pp 65. https://www.targetedonc.com/view/hitting-the-target-multigene-tests-gain-foothold-in-more-clinical-settings Global Companion Diagnostics Market Report 2019-2024. News release. BUSINESS WIRE; Page last updated: September 24, 2019. Accessed 12 Dec 2023. https://www.businesswire.com/news/home/20190924005792/en/Global-Companion-Diagnostics-Market-Report-2019-2024---Market-to-Reach-7.3-Billion-by-2024-Registering-Massive-Growth---ResearchAndMarkets.com KEYWORDS: Fast and wide multiplexing, targeted biomarker quantification, CLIA waiver certificate, Far forward lab, minimally invasive biomatrix, simple protocol, automated hands-free protocol
