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Rapid, multiplex detection of SARS-CoV-2 using isothermal amplification coupled with CRISPR-Cas12a
Abstract
In December 2019 an outbreak erupted due to the beta coronavirus severe acute respiratory syndrome (SARS)-CoV-2 in Wuhan, China. The disease caused by this virus (COVID-19) rapidly spread to all parts of the globe leading to a global pandemic. Efforts to combat the pan-demic rely on RT-qPCR diagnostic tests that have high turnaround times (~24h), are easily contaminated, need specialized equipment, facilities, and personnel that end up increasing the overall costs of this method. Loop-mediated isothermal amplification (LAMP) coupled with a reverse transcription step (RT-LAMP) is an alternative diagnostic method that can easily overcome these obstacles, when coupled with CRISPR/Cas it can eliminate false positives. Here we report a fast (~40 min), highly sensitive, point-of-care RT-LAMP and CRISPR/Cas12a assay to detect SARS-CoV-2. This fluorescence-based test prevents carry-over contamination using dUTP/UDG and achieved 100% specificity and 93% sensitivity using 25 positives and 50 negative patient samples for Ct
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Genes
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