Integrated System for Rapid Detection of Respiratory Pathogens

描述（由申请人提供）：MBio Diagnostics, Inc. 与加州大学圣地亚哥分校和 PATH（华盛顿州西雅图）合作，提议开发一种低成本、无扩增的呼吸道病原体核酸诊断技术。拟议的系统将满足两大公共卫生需求。首先，它将建立一个易于配置的诊断工具，可以在生物恐怖事件或新的大流行紧急情况期间快速部署用于公共卫生筛查和诊断工作。其次，该计划将产生一种基于病毒 RNA 的流感快速测试，与市售的基于抗原的快速诊断测试相比，该测试的灵敏度和特异性将显着提高。虽然 PCR 和质谱方法对于新病原体鉴定和疾病诊断来说足够灵敏和特异，但与这些技术相关的成本和周转时间使得它们在大规模筛查应用中的用途有限。需要具有实验室质量性能、快速、稳健、成本极低且易于重新配置的下一代诊断工具。在这里，我们提议建立研究伙伴关系，开发一种简单的、无扩增的核酸检测系统，用于检测 NIAID 优先病原体列表中的呼吸道病原体。研究将集中在单个设备内生物样品制备和目标检测的新颖集成。核心研究重点是一种创新的检测方法，该方法结合了目标核酸序列的直接磁性颗粒捕获和同时标记，可在简单的读数仪上进行即时、高灵敏度的荧光检测。该奖项的模型病原体将是流感病毒，开发重点是临床样本中的多重病毒 RNA 检测，样本到答案的周转时间不到 40 分钟。研究目标是 (1) 展示在日益复杂的样品中以 30,000 目标拷贝/ml 灵敏度进行多重、多颗粒核酸提取和荧光检测； (2) 建立一个特征明确的人类临床样本库，用于检测和系统开发； (3) 开发一个强大的、低成本的快速测试系统，由一次性试剂盒和一个简单的读取器设备组成； (4) 论证快速检测系统在实验室和现场条件下的可行性，并建立相对于批准的流感快速检测的临床敏感性和特异性性能； (5) 将技术扩展到其他 NIAID 优先病原体。由于拟议的技术将具有 FDA 批准的持续用途，因此在发生生物防御紧急情况时，关键基础设施和训练有素的用户将到位。除了提供下一代诊断技术的概念验证结果外，拟议的研究还将总体上推进基于粒子的检测科学，并将建立一个特征明确的流感临床样本库，供 NIAID 项目持续使用。

DESCRIPTION (provided by applicant): MBio Diagnostics, Inc., in collaboration with the University of California, San Diego, and PATH (Seattle, WA), proposes to develop a low-cost, no amplification, nucleic acid diagnostic technology for respiratory pathogens. The proposed system will address two major public health needs. First, it will establish a readily configurable diagnostic tool that can be rapidly deployed for public health screening and diagnostic efforts during a bioterror event or new pandemic emergencies. Second, the program will yield a viral RNA based flu rapid test that will offer significant sensitivity and specificity improvements over commercially available, antigen-based rapid diagnostics tests. While PCR and mass spectrometry methods are sensitive and specific enough for new pathogen identification and disease diagnosis, costs and turnaround times associated with these technologies render them of only limited use in large-scale screening applications. Next generation diagnostic tools with laboratory quality performance, but that are rapid, robust, very low cost, and readily reconfigurable are needed. Here we propose a research partnership for the development of a simple, no amplification nucleic acid detection system for respiratory pathogens from the NIAID Priority Pathogen list. Research will focus on a novel integration of biological sample preparation and target detection within a single device. The core research emphasis is on an innovative assay approach that combines direct magnetic particle capture of target nucleic acid sequences and simultaneous labeling for immediate, high sensitivity fluorescence detection on a simple reader instrument. The model pathogen under this award will be influenza virus, and development will focus on multiplexed, viral RNA detection in clinical specimens, with a sample-to-answer turnaround time of less than 40 minutes. Research aims are to (1) Demonstrate multiplexed, multi-particle nucleic acid extraction and fluorescence detection in increasingly complex samples with 30,000 target copies/ml sensitivity; (2) Build a well characterized human clinical sample collection to be used for assay and system development; (3) Develop a robust, low cost rapid test system consisting of single use disposable cartridges and a simple reader device; (4) Demonstrate feasibility of the rapid test detection system in laboratory and field conditions, and establish clinical sensitivity and specificity performance relative to approved flu rapid tests; and (5) Extend the technology to other NIAID Priority pathogens. Because the proposed technology will have ongoing, FDA-approved uses, critical infrastructure and trained users will be in place in the event of a biodefense emergency. In addition to providing proof-of-concept results on this next generation diagnostic technology, the proposed research will generally advance the science of particle based assays and will establish a well characterized influenza clinical specimen repository of ongoing use to NIAID programs.