Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA

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COVID-SHERLOCK ms_RR3.pdf

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sword-2021-12-13T16:36:33.original.xml (130 B)
Original SWORD entry document
Date Issued
December 1, 2020
Journal
Nature Biomedical Engineering
Publisher
Springer Science and Business Media LLC
Citation
2020. "Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA." Nature Biomedical Engineering, 4 (12).
Version
Author's final manuscript
Abstract
© 2020, The Author(s), under exclusive licence to Springer Nature Limited. Nucleic acid detection by isothermal amplification and the collateral cleavage of reporter molecules by CRISPR-associated enzymes is a promising alternative to quantitative PCR. Here, we report the clinical validation of the specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) assay using the enzyme Cas13a from Leptotrichia wadei for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)—the virus that causes coronavirus disease 2019 (COVID-19)—in 154 nasopharyngeal and throat swab samples collected at Siriraj Hospital, Thailand. Within a detection limit of 42 RNA copies per reaction, SHERLOCK was 100% specific and 100% sensitive with a fluorescence readout, and 100% specific and 97% sensitive with a lateral-flow readout. For the full range of viral load in the clinical samples, the fluorescence readout was 100% specific and 96% sensitive. For 380 SARS-CoV-2-negative pre-operative samples from patients undergoing surgery, SHERLOCK was in 100% agreement with quantitative PCR with reverse transcription. The assay, which we show is amenable to multiplexed detection in a single lateral-flow strip incorporating an internal control for ribonuclease contamination, should facilitate SARS-CoV-2 detection in settings with limited resources.
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DOI of Published Version
10.1038/s41551-020-00603-x
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