EAGER: Rapid Planar PCR for COVID-19 Testing

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The goal of this project is to develop the first point-of-contact molecular diagnostic as a tool for COVID-19 testing. The investigator envisions a point-of-contact test being performed before someone passes through a door, checkpoint, airport gate, or border. Many of the critical strategies for this work have already been established, including the feasibility of performing a one-step assay on various viruses, including SARS-CoV-2, the virus responsible for COVID-19. This project provides a basis for expanding the capability of the method/diagnostic tool, with the goal of establishing a reliable method for detecting COVID-19 positivity from saliva in less than 2 minutes at a cost of approximately $2/test. When functional, this tool could be used routinely to test for hidden spreaders of disease at airports, entrances to hospitals or long-term care facilities. Similarly, the method could be used to screen employees when they arrive to work at health care facilities or other large facilities to protect essential workers and patients. Finally, the method could be used for routine screening at large facilities such as factories, food processing or distribution facilities, and large government buildings, allowing our economy to return to a more normal state. The research will also provide training for a postdoctoral fellow and an undergraduateThis project focuses on establishing a new method for PCR (Polymerase Chain Reaction, a method used to rapidly make millions of copies of a DNA sample) that will realize the goal of a point-of-contact molecular diagnostic test for envelope viruses such as COVID-19. Already established for this work are critical strategies: (1) for rapid, uniform cycling using optical heating; (2) for large scale partitioning to accelerate sample preparation without micro-patterning or microfluidics; and (3) for performing a one-step lysis/rp-PCR (rapid planar-PCR) assay on envelope viruses. The Research Plan is organized under three objectives: (1) to extend the limited area heating due to use of individual LED/lens pairs to large areas through the design and fabrication of a printed circuit that will allow higher density of LEDs per zone, allow many more zones, simplify connections to the rest of the control circuitry, and allow use of surface mount LED drivers; (2) to show, once uniform, large area heating is established, that the diffusion limitation works to provide a very large level of partitioning, with a goal of achieving 1,000,000 partitioning sites in a 5 x 5 cm sample; and (3) to demonstrate the expected assay speed and detection limit (~2 minutes and ~100 copies/milliliter) by modifying reagents, temperatures, and time periods to achieve a good compromise between efficiency of the three steps (lysis, reverse transcription, PCR) overall assay speed, and limit-of detection. If successful, project results will provide the basis for rolling out a point-of-contact test that can be rapidly translated for clinical applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.