Rapid and accurate detection of novel coronavirus SARS-CoV-2 using CRISPR-Cas3

Yoshimi, K.; Takeshita, K.; Yamayoshi, S.; Shibumura, S.; Yamauchi, Y.; Yamamoto, M.; Yotsuyanagi, H.; Kawaoka, Y.; Mashimo, T.

Novel coronavirus SARS-CoV-2 outbreaks have rapidly spread to multiple countries, highlighting the urgent necessity for fast, sensitive, and specific diagnostic tools for virus surveillance. Here, the previously unknown collateral single-stranded DNA cleavage we observed with type I CRISPR-Cas3 highlights its potential for development as a Cas3-mediated rapid (within 40 min), low-cost, instrument-free detection method for SARS-CoV-2. This Cas3-based assay is comparable with Cas12- and real-time reverse-transcriptase PCR-based assays in its speed and sensitivity, but offers greater specificity for single-base-pair discrimination while negating the need for highly trained operators. These findings support the use of CRISPR diagnostics for point-of-care testing in patients with suspected SARS-CoV-2 infections.