| Literature DB >> 32848209 |
Maturada Patchsung1, Krittapas Jantarug1, Archiraya Pattama2, Kanokpol Aphicho1, Surased Suraritdechachai1, Piyachat Meesawat1, Khomkrit Sappakhaw1, Nattawat Leelahakorn1, Theerawat Ruenkam1, Thanakrit Wongsatit1, Niracha Athipanyasilp2, Bhumrapee Eiamthong1, Benya Lakkanasirorat1, Thitima Phoodokmai1, Nootaree Niljianskul3, Danaya Pakotiprapha4, Sittinan Chanarat4, Aimorn Homchan4, Ruchanok Tinikul4, Philaiwarong Kamutira4, Kochakorn Phiwkaow1, Sahachat Soithongcharoen1, Chadaporn Kantiwiriyawanitch1, Vinutsada Pongsupasa1, Duangthip Trisrivirat1, Juthamas Jaroensuk1, Thanyaporn Wongnate1, Somchart Maenpuen5, Pimchai Chaiyen1, Sirichai Kamnerdnakta6, Jirawat Swangsri6, Suebwong Chuthapisith6, Yongyut Sirivatanauksorn6, Chutikarn Chaimayo2, Ruengpung Sutthent2, Wannee Kantakamalakul2, Julia Joung7,8,9,10, Alim Ladha7,8,9,10, Xin Jin8,9,11,12, Jonathan S Gootenberg9,13, Omar O Abudayyeh9,13, Feng Zhang7,8,9,10,13,14, Navin Horthongkham15, Chayasith Uttamapinant16.
Abstract
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.Entities:
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Year: 2020 PMID： 32848209 DOI： 10.1038/s41551-020-00603-x
Source DB: PubMed Journal: Nat Biomed Eng ISSN： 2157-846X Impact factor: 25.671