Multipled POC device for antigen/molecular detection of influenza & other viruses

描述（由申请人提供）： 在未来十年中，很有可能会出现流感大流行，一种新型呼吸道病毒（RV）的大流行[例如，SARS]，或导致呼吸道症状的生物恐怖袭击。此外，每年有多达50万人死于流感，并且没有方法在临床上将流感与许多其他常见的RV感染区分开来。迫切需要灵敏、高度特异性、廉价、便携、低复杂性的装置/测定，其允许流感和其他常见RV的多重检测和鉴定。我们在开发流感病毒和其他RV的多重商业检测方面有着悠久的历史，并在世界上开发了第一个针对流感病毒A、B、RSV A、B和副流感病毒1、2、3的商业多重分子检测（Hexaplex，1996），在美国开发了第一个针对SARS的检测（2003）。我们现在已经开发了基于独特的样品收集装置（SCD）、稳定的铕荧光微粒（EFM）和基于pRNA的结合化学的新颖的完全集成的技术，其允许对患者进行灵敏的（比FDA批准的测定高400倍）床边或现场多重POC抗原测试。这项技术依靠电池运行，耗时15分钟，可以同时检测多个病毒。更重要的是，SCD收集并稳定核酸样品（RNA和DNA），以根据需要用于分子测定（允许检测阴性样品或进一步分析阳性样品，同时保持生物防护）。在本申请中，我们提出了以下具体目标：！选择并优化化学、底物和硬件，以产生SCD-EFM-pRNA检测试剂盒，检测13种常见RV，导致儿童和成人LRI或重大疾病（甲型流感病毒、B流感病毒、RSV、[副流感病毒1、2、3、hMPV]、冠状病毒[NL 63、HKU 1、229 E、OC 43]、腺病毒和肠道病毒）。2.开发针对这13种病毒的多重RT-PCR电子微阵列（eMR）检测试剂盒，作为确证性分子检测试剂盒（也作为产品），确定并比较分析LOD、灵敏度和特异性与SA 1中开发的多重免疫检测试剂盒和其他已建立的分子检测试剂盒。3.确定FDA要求的最佳参数。(e.g.,确保所有组件的设计、易用性指南和非专业实验室人员的结果解释符合SA#1中CLIA豁免的测定标准; 4.工艺开发（确保诊断组件可制造性和以合理成本商业化的方法）。5.临床样本检测（冷冻样本）：确定SA#1和2中针对大多数产生“流感样”症状和LRI的RV开发的多重POC检测试剂盒与目前可用的标准诊断检测试剂盒相比，是否能够证明临床灵敏度、特异性和成本效益得到改善。在完成这些SA之后，临床医生和公共卫生官员将拥有两种重要的新工具来对抗流感和病毒性呼吸道感染，无论是从他们的医院，办公室还是地球上最偏远的地方。

DESCRIPTION (provided by applicant): There is a significant probability that in the next decade there will be either an influenza (flu) pandemic, a major epidemic of a novel respiratory virus (RV) [e.g., SARS], or a bioterrorism attack that results in respiratory symptoms. In addition, as many as 500,000 people die each year of flu and there is no method to clinically distinguish flu from the many other common RV infections. There is an urgent need for sensitive, highly specific, inexpensive, portable, low complexity devices/assays that allow for the multiplex detection and identification of flu and other common RV's. We have a long record of developing multiplex commercial assays for flu and other RVs and developed the first commercial multiplex molecular assay for flu A, B, RSV A, B, and parainflu 1,2,3 in the world (Hexaplex, 1996) and the first assay for SARS in the US (2003). We have now developed novel fully integrated technology based on a unique sample collection device (SCD), stable europium fluorescent microparticles (EFM), and binding chemistry based on pRNA that allows for sensitive (400x > than FDA approved assays) bedside or in-the-field multiplex POC antigen testing of patients. This technology runs on batteries, takes 15 minutes, and can detect multiple viruses simultaneously. Even more importantly the SCD collects and stabilizes a sample of nucleic acid (RNA and DNA) for use in molecular assays as needed (allowing for testing of negative samples or further analysis of positives while maintaining biocontainment). In this application we propose the following specific aims:! Select and optimize the chemistry, substrate, and hardware to produce a SCD-EFM-pRNA assay to detect 13 common RV's causing LRI or significant disease in children and adults (flu A, B, RSV, [paraflu 1,2,3, hMPV], corona- viruses [NL63, HKU1, 229E, OC43], adenovirus, and enterovirus). 2. Develop a multiplex-RT-PCR electronic microarray (eMR) assay to these same 13 viruses as a confirmatory molecular assay (also as a product), determine and compare analytical LOD, sensitivity and specificity with multiplex immunoassay developed in SA 1 and other established molecular assays. 3. Determine optimal parameters required by FDA. (e.g., ensure design of all components, ease of use guidelines, and results interpretation by non-professional laboratory personnel meets CLIA waived standards for assay in SA#1); 4. Process Development (methods to ensure manufacturability of the diagnostic components and commercialization at a reasonable cost). 5. Clinical sample testing (frozen samples); To determine if the multiplex POC assays developed in SA#1 & 2 to the majority of RV that produce "Flu-like" symptoms and LRI can demonstrate improved clinical sensitivity, specificity, and cost effectiveness compared to currently available standard diagnostic assays. After completion of these SAs, clinicians and public health officials will have two important new tools to fight flu and viral respiratory infections, whether from their hospital, office or the most remote locations on the planet.