RAPID:COVID-19: RT-LAMP-based electrical detection of SARS-CoV-2

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Since the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, the virus that causes the coronavirus-associated acute respiratory disease COVID-19) jumped from an animal reservoir to humans in December 2019, it has rapidly spread across the world, bringing death, illness, disruption to daily life, and economic losses to businesses and individuals. The rapid development of the COVID-19 pandemic highlights the shortcomings in the existing laboratory-based testing paradigm for viral diagnostics that features a widespread lack of test kits, extended time delays to obtain test results, and a high rate of false negative tests. The shortcomings of the existing laboratory-based infrastructure are contributing to uncertainty surrounding quarantine failure, confusion among health authorities, and also to a general public anxiety. To address this critical and timely need, in this work, development of point-of-care device is proposed for detecting the presence of SARS-CoV-2 from nasal fluid samples and in 10 minutes.The fundamental limitations of current assays for viral pathogens stem from their reliance upon polymerase chain reaction (PCR) analysis, which requires complex, labor-intensive, laboratory-based protocols for viral isolation, lysis, and removal of inhibiting materials. Importantly, PCR requires a large number of time-consuming and precise thermal cycles to enzymatically amplify specific RNA sequences. In this work development of an electrical, label-free and surface modification-free point-of-care device for detecting the presence of SARS-CoV-2 (1-3 copy/mL) from nasal fluid samples and in 10 min. The proposed work will combine Bst polymerase in an isothermal RT-LAMP (reverse transcription loop-mediated isothermal amplification) reaction, with target-specific primers and crumpled graphene field-effect transistors (gFET) to electrically detect the amplification event by sensing the consumption of primers. These reactions offer the possibility of rapidly detecting SARS-CoV-2 using a simple, inexpensive and portable potentiostat and avoiding the necessity of RNA extraction. In order to develop the proposed detection technology, this proposal will first validate twelve sets of RT-LAMP primers specific to SARS-CoV-2. Importantly, No RT-LAMP primers specific to SARS-CoV-2 are available commercially, neither in the literature. Then, the proposal will demonstrate the feasibility of the RT-LAMP-based electrical approach to detect SARS-CoV-2. Likewise, the proposal will demonstrate the approach as a global health technology to contribute to providing low-cost diagnostics around the world using portable and inexpensive heating block and potentiostat for virus detection and quantification.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.