Rapid detection of SARS-CoV-2 with Cas13

Agrawal, Shreeya; Fanton, Alison; Chandrasekaran, Sita S.; Prywes, Noam; Lukarska, Maria; Biering, Scott B.; Smock, Dylan C. J.; Mok, Amanda; Knott, Gavin J.; Van Dis, Erik; Dugan, Eli; Kim, Shin; Liu, Tina Y.; Harris, Eva; Stanley, Sarah A.; Lareau, Liana F.; Doudna, Jennifer A.; Savage, David F.; Hsu, Patrick D.

To combat disease outbreaks such as the COVID-19 pandemic, flexible diagnostics for rapid viral detection are greatly needed. We report a nucleic acid test that integrates distinct mechanisms of DNA and RNA amplification optimized for high sensitivity and rapid kinetics, linked to Cas13 detection for specificity. We paired this workflow, termed Diagnostics with Coronavirus Enzymatic Reporting (DISCoVER), with extraction-free sample lysis using shelf-stable reagents that are widely available at low cost. DISCoVER has been validated on saliva samples to incentivize frequent testing for more widespread community surveillance and robustly detected attomolar levels of SARS-CoV-2 within 30 minutes, while avoiding false positives in virus-negative saliva. Furthermore, DISCoVER is compatible with multiplexed CRISPR probes to enable simultaneous detection of a human gene control or alternative pathogens.