Journal
NATURE BIOMEDICAL ENGINEERING
Volume 4, Issue 12, Pages 1140-1149Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41551-020-00603-x
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Funding
- PTT
- PTTEP
- Kasikorn Bank
- Siam Commercial Bank
- GPSC
- VISTEC
- Siriraj Hospital
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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 fromLeptotrichia wadeifor 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. The specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) assay detected severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA with high sensitivity and specificity in hundreds of nasopharyngeal and throat swab samples collected at Siriraj Hospital in Thailand.
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