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

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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.