Integrated System for Rapid Detection of Respiratory Pathogens

描述(由申请人提供)：mBio诊断公司与加州大学圣地亚哥分校和PATH(西雅图，华盛顿州)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个主要的公共卫生需求。首先，它将建立一个易于配置的诊断工具，可以在发生生物恐怖事件或新的大流行紧急情况时迅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，与商业上可获得的基于抗原的快速诊断测试相比，该测试将提供显著的敏感性和特异性改进。虽然聚合酶链式反应和质谱学方法对于新的病原体识别和疾病诊断具有足够的敏感性和特异性，但与这些技术相关的成本和周转时间使它们在大规模筛查应用中的使用有限。下一代诊断工具需要具有实验室质量的性能，但快速、坚固、非常低的成本和易于重新配置。在这里，我们建议建立一个研究伙伴关系，用于开发一种简单的、无扩增的核酸检测系统，用于NIAID优先病原体名单中的呼吸道病原体。研究将集中在单个设备中生物样品制备和目标检测的新集成上。核心研究重点是一种创新的分析方法，它结合了直接磁粉捕获目标核酸序列和同时标记在简单阅读器上进行即时、高灵敏度荧光检测的方法。该奖项下的模式病原体将是流感病毒，开发的重点将是在临床样本中进行多重病毒RNA检测，样本到答案的周转时间不到40分钟。研究目标是(1)展示多重、多粒子核酸提取和荧光检测在日益复杂的样本中的敏感性，以30,000个目标拷贝/毫升的灵敏度；(2)建立用于分析和系统开发的具有良好特性的人类临床样本收集；(3)开发由一次性使用的一次性墨盒和简单的阅读器设备组成的强大、低成本的快速检测系统；(4)在实验室和现场条件下展示快速检测系统的可行性，并建立相对于经批准的流感快速检测的临床敏感性和特异性表现；以及(5)将该技术推广到其他NIAID优先考虑的病原体。由于拟议的技术将具有FDA批准的持续用途，关键基础设施和训练有素的用户将在发生生物防御紧急情况时到位。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颗粒的分析科学，并将建立一个具有良好特征的流感临床标本储存库，供NIAID计划继续使用。

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.