Rapid, disposable, nucleic acid diagnostic device for multiplexed point of care f

描述(申请人提供)：快速准确地识别高传染性和致病性疾病，对于及早预测暴发风险和有效部署医疗和行政对策至关重要。不幸的是，目前在护理点(POC)使用的诊断方法主要是免疫学分析，通常缺乏足够的敏感性、特异性和多重能力来进行准确的早期诊断。已开发或正在开发的基于核酸的诊断产品需要一种相对昂贵的专用仪器，这种仪器往往使小型诊所和资源有限的环境望而却步，不适合大流行的高容量需求。我们建议开发一种无仪器、自给自足、低成本(20美元/台)、一次性、快速(40分钟，样本到答案)的核酸POC诊断设备(MTIDx)，能够检测甲型流感、乙型流感、呼吸道合胞病毒和其他相关呼吸道疾病。最终的MTIDx产品被设计为FDA批准的、CLIA豁免的(1988年临床实验室改进修正案)基于核酸(NA)的诊断设备，用于分析人类鼻拭子样本。MTIDx(多目标识别诊断)将是一款手机大小的设备，操作简单，用户干预最少，还可以选择由9V电池供电，用于资源有限/现场设置。它将适用于大批量生产，允许快速生产数百万台，以满足大流行爆发期间的激增产能。该诊断系统将梅萨科技的核酸提取、快速放大、基于横向流动的(基于LF的)比色检测组件与简单的流体控制和微加热器解决方案集成到一个易于使用、低成本的一次性设备中。在初步数据中，我们已经通过一个实验室原型证明了集成装置方法的可行性。第一阶段SBIR提案的工作将侧重于系统集成的优化和MTIDx实验室原型初始设计的改进，以促进实验室原型向可制造设计的过渡。现有的原型设计将过渡到注塑原型，以便能够进行短期生产，以进行严格的表征和实验室测试。我们还将开发用于放大和检测的冻干剂以实现环境存储兼容性，开发稳定的缓冲试剂和胶囊方法，设计制造优化以促进未来的大批量生产，并进行初步测试以确定实现性能目标所需的设计改进。第一阶段的主要成果是演示集成的MTIDx DFM原型(面向制造的设计)，提供样本到答案的4路诊断测试(流感A、B、RSV和RNaseP作为对照)，在40分钟内完成。在后续的第二阶段项目中，我们将纳入更多疾病靶点，进一步优化可生产的设计，生产注塑设备，并在一大批具有良好特征的临床样本上验证性能。

DESCRIPTION (provided by applicant): Rapid and accurate identification of highly infectious and pathogenic disease is extremely important for the early forecast of outbreak risk and the effective deployment of medical and administrative countermeasures. Unfortunately, current diagnostics for use at the point-of-care (POC) are largely immunological assays that often lack adequate sensitivity, specificity, and multiplex capacity for an accurate early diagnosis. Nucleic acid- based diagnostic products developed or under development requires a relatively expensive dedicated instrument that is often prohibitive to small clinics and resource-limited settings and unsuitable for pandemic high capacity demand. We propose to develop an instrumentation-free, self-contained, low cost (<$20/unit), disposable, rapid (<40 minute, sample-to-answer), nucleic acid POC diagnostic device (MTIDx) capable of detecting influenza A, influenza B, RSV and other relative respiratory diseases. The final MTIDx product is designed to be a FDA approved, CLIA-waived (Clinical Laboratory Improvement Amendments of 1988) nucleic acid (NA)-based diagnostic device for analyzing human nasal swab samples. MTIDx (Multiple Target Identification Diagnostic) will be a cell phone-sized, device that is easy to operate with minimal user- intervention and also has an option to be powered by a 9V battery for use in resource-limited/field settings. It will be suitable for high volume manufacturing allowing rapid production of millions of units to meet surge capacity during a pandemic outbreak. The diagnostic system is based on the integration of Mesa Tech's proprietary technologies of nucleic acid extraction, rapid amplification, lateral flow-based (LF-based) colorimetric detection components with simple fluid control and microheater solutions into one easily used, low cost, disposable device. In preliminary data, we have demonstrated the feasibility of integrated device approach through a laboratory prototype. Work in this Phase I SBIR proposal will focus on the optimization of system integration and the refinement of initial MTIDx laboratory prototype designs to facilitate the transition of laboratory prototypes to manufacture-ready designs. Existing prototype designs will be transitioned to injection- molded prototypes to enable short-run production for rigorous characterization and laboratory testing. We will also incorporate the development of lyophilized reagents for amplification and detection to confer ambient storage compatibility, development of stable buffer reagents and capsulation methods, design for manufacturing optimizations to facilitate future high volume production and initial testing to identify design refinements required to attain performance objectives. The primary Phase I deliverable is the demonstration of an integrated MTIDx DFM-prototype (Design For Manufacturing) that offers a sample-to-answer 4-plex diagnostic test (Flu A, B, RSV and RNaseP as a control) complete in <40 minutes. In a follow-on Phase 2 project, we will incorporate more disease targets, further optimize the manufacture-ready designs, produce injection-molded devices and validate performance on a large panel of well-characterized clinical samples.