Very rapid, low cost multiplexed test for SARS, Influenza A and Influenza B Resubmission

项目总结 本项目的目标是开发一种接触点分子诊断技术(TEST)，用于检测多个 冠状病毒(SARS-CoV-2)和流感病毒的复合检测。而看护点诊断可能是 在临床环境中，在10-20分钟的时间内进行，我们设想执行接触点测试 在有人通过门、检查站、机场大门或边境之前，速度大约快10倍。我们有 确定了一种执行PCR的策略，该策略使用一种新的格式来加快热循环，同时实现AP- 接近1,000,000倍的样品分割，以加速样品准备，而不需要微图案或 微流体学。如下所述，我们已经为该项目制定了关键战略，包括(1) 使用光学加热的快速、均匀循环；(2)大规模样品分离；以及(3)一步裂解/RT- 对包膜病毒进行聚合酶链式反应检测，包括SARS-CoV-2(导致新冠肺炎的病毒)。 该项目将为可在AS中执行的多路复用点接触点诊断提供基础 只需2分钟，成本低至2美元/测试的物料清单(BOM)消耗品。当可用时，此 诊断平台可常规用于检测疾病的隐藏传播者，例如机场、 昏迷到医院，或在长期护理机构。诊断的可获得性将是变革性的。就是- 由于其速度快、成本低，该平台可以提供第一个接触点分子诊断 新冠肺炎等疾病，可以在无症状的传播者上船或下船时检测到 在机场。在边境或入境点快速部署该检测可防止疾病传播。 在最初的暴发之后。同样，当员工到达工作地点时，这项测试可以用来筛选他们 保护病人和基本工作人员的卫生保健设施。最后，该测试可以用于日常工作。 在工厂、食品加工或配送设施以及大型政府等大型设施进行筛查 建筑物，减少应对新冠肺炎等病毒性流行病的经济影响。

PROJECT SUMMARY The objective of this project is to develop a point-of-contact molecular diagnostic technology (test) for multi- plexed detection of a coronavirus (SARS-CoV-2) and influenza virus. While a point-of-care diagnostic might be performed in a period of 10-20 minutes in a clinical setting, we envision a point-of-contact test being performed about 10 times faster and before someone passes through a door, checkpoint, airport gate, or border. We have identified a strategy for performing PCR that uses a new format to speed thermal cycling while achieving ap- proximately 1,000,000-fold sample partitioning to accelerate sample preparation, without micro-patterning or microfluidics. As described below, we have already established critical strategies for the project including (1) rapid, uniform cycling using optical heating; (2) large scale sample partitioning; and (3) a one-step lysis/RT- PCR assay for enveloped viruses, including SARS-CoV-2 (the virus responsible for COVID-19). This project will provide the basis for a multiplexed point-of-contact diagnostic that can be performed in as little as 2 minutes and at a cost as low as $2/test for Bill of Material (BOM) consumables. When available, this diagnostic platform could be used routinely for detecting hidden spreaders of disease at, e.g., airports, en- trances to hospitals, or at long-term care facilities. Availability of the diagnostic would be transformative. Be- cause of its speed and low cost, the platform could provide the first point-of-contact molecular diagnostic for diseases such as COVID-19, which could detect asymptomatic spreaders, e.g., as they embark or disembark at airports. Rapid deployment of the test at borders or entry points could prevent disease from spreading be- yond an initial outbreak. Similarly, the test could be used to screen employees when they arrive to work at health care facilities to protect patients and essential workers. Finally, the test could be used for routine screening at large facilities such as factories, food processing or distribution facilities, and large government buildings, reducing the economic impact of a addressing a viral pandemic such as COVID-19.