Author: Patchsung, Maturada; Jantarug, Krittapas; Pattama, Archiraya; Aphicho, Kanokpol; Suraritdechachai, Surased; Meesawat, Piyachat; Sappakhaw, Khomkrit; Leelahakorn, Nattawat; Ruenkam, Theerawat; Wongsatit, Thanakrit; Athipanyasilp, Niracha; Eiamthong, Bhumrapee; Lakkanasirorat, Benya; Phoodokmai, Thitima; Niljianskul, Nootaree; Pakotiprapha, Danaya; Chanarat, Sittinan; Homchan, Aimorn; Tinikul, Ruchanok; Kamutira, Philaiwarong; Phiwkaow, Kochakorn; Soithongcharoen, Sahachat; Kantiwiriyawanitch, Chadaporn; Pongsupasa, Vinutsada; Trisrivirat, Duangthip; Jaroensuk, Juthamas; Wongnate, Thanyaporn; Maenpuen, Somchart; Chaiyen, Pimchai; Kamnerdnakta, Sirichai; Swangsri, Jirawat; Chuthapisith, Suebwong; Sirivatanauksorn, Yongyut; Chaimayo, Chutikarn; Sutthent, Ruengpung; Kantakamalakul, Wannee; Joung, Julia; Ladha, Alim; Jin, Xin; Gootenberg, Jonathan S; Abudayyeh, Omar O; Zhang, Feng; Horthongkham, Navin; Uttamapinant, Chayasith
Title: Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA. Cord-id: jufff8i9 Document date: 2020_8_26
ID: jufff8i9
Snippet: 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 nasophar
Document: 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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