Maturada Patchsung, Krittapas Jantarug, Archiraya Pattama, Kanokpol Aphicho, Surased Suraritdechachai, Piyachat Meesawat, Khomkrit Sappakhaw, Nattawat Leelahakorn, Theerawat Ruenkam, Thanakrit Wongsatit, Niracha Athipanyasilp, Bhumrapee Eiamthong, Benya Lakkanasirorat, Thitima Phoodokmai, Nootaree Niljianskul, D. Pakotiprapha, Sittinan Chanarat, Aimorn Homchan, Ruchanok Tinikul, Philaiwarong Kamutira, Kochakorn Phiwkaow, Sahachat Soithongcharoen, Chadaporn Kantiwiriyawanitch, Vinutsada Pongsupasa, Duangthip Trisrivirat, Juthamas Jaroensuk, T. Wongnate, Somchart Maenpuen, P. Chaiyen, Sirichai Kamnerdnakta, J. Swangsri, S. Chuthapisith, Y. Sirivatanauksorn, Chutikarn Chaimayo, R. Sutthent, W. Kantakamalakul, J. Joung, A. Ladha, Xin Jin, J. Gootenberg, O. Abudayyeh, Feng Zhang, N. Horthongkham, Chayasith Uttamapinant
Nature Biomedical Engineering, 2020
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. 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.
Cited by 95 publications.
Field of study: Medicine