Publication: Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA
Issued Date
2020-01-01
Resource Type
ISSN
2157846X
Other identifier(s)
2-s2.0-85089869548
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Mahidol University
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SCOPUS
Bibliographic Citation
Nature Biomedical Engineering. (2020)
Suggested Citation
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, Danaya Pakotiprapha, Sittinan Chanarat, Aimorn Homchan, Ruchanok Tinikul, Philaiwarong Kamutira, Kochakorn Phiwkaow, Sahachat Soithongcharoen, Chadaporn Kantiwiriyawanitch, Vinutsada Pongsupasa, Duangthip Trisrivirat, Juthamas Jaroensuk, Thanyaporn Wongnate, Somchart Maenpuen, Pimchai Chaiyen, Sirichai Kamnerdnakta, Jirawat Swangsri, Suebwong Chuthapisith, Yongyut Sirivatanauksorn, Chutikarn Chaimayo, Ruengpung Sutthent, Wannee Kantakamalakul, Julia Joung, Alim Ladha, Xin Jin, Jonathan S. Gootenberg, Omar O. Abudayyeh, Feng Zhang, Navin Horthongkham, Chayasith Uttamapinant Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA. Nature Biomedical Engineering. (2020). doi:10.1038/s41551-020-00603-x Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/59008
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Title
Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA
Author(s)
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
Danaya Pakotiprapha
Sittinan Chanarat
Aimorn Homchan
Ruchanok Tinikul
Philaiwarong Kamutira
Kochakorn Phiwkaow
Sahachat Soithongcharoen
Chadaporn Kantiwiriyawanitch
Vinutsada Pongsupasa
Duangthip Trisrivirat
Juthamas Jaroensuk
Thanyaporn Wongnate
Somchart Maenpuen
Pimchai Chaiyen
Sirichai Kamnerdnakta
Jirawat Swangsri
Suebwong Chuthapisith
Yongyut Sirivatanauksorn
Chutikarn Chaimayo
Ruengpung Sutthent
Wannee Kantakamalakul
Julia Joung
Alim Ladha
Xin Jin
Jonathan S. Gootenberg
Omar O. Abudayyeh
Feng Zhang
Navin Horthongkham
Chayasith Uttamapinant
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
Danaya Pakotiprapha
Sittinan Chanarat
Aimorn Homchan
Ruchanok Tinikul
Philaiwarong Kamutira
Kochakorn Phiwkaow
Sahachat Soithongcharoen
Chadaporn Kantiwiriyawanitch
Vinutsada Pongsupasa
Duangthip Trisrivirat
Juthamas Jaroensuk
Thanyaporn Wongnate
Somchart Maenpuen
Pimchai Chaiyen
Sirichai Kamnerdnakta
Jirawat Swangsri
Suebwong Chuthapisith
Yongyut Sirivatanauksorn
Chutikarn Chaimayo
Ruengpung Sutthent
Wannee Kantakamalakul
Julia Joung
Alim Ladha
Xin Jin
Jonathan S. Gootenberg
Omar O. Abudayyeh
Feng Zhang
Navin Horthongkham
Chayasith Uttamapinant
Other Contributor(s)
PTT Public Company Limited
Vidyasirimedhi Institute of Science and Technology
McGovern Institute
Society of Fellows, Harvard University
Massachusetts Institute of Technology
Mahidol University
Faculty of Medicine, Siriraj Hospital, Mahidol University
Harvard University
Howard Hughes Medical Institute
Burapha University
Broad Institute
Massachusetts Consortium on Pathogen Readiness
Vidyasirimedhi Institute of Science and Technology
McGovern Institute
Society of Fellows, Harvard University
Massachusetts Institute of Technology
Mahidol University
Faculty of Medicine, Siriraj Hospital, Mahidol University
Harvard University
Howard Hughes Medical Institute
Burapha University
Broad Institute
Massachusetts Consortium on Pathogen Readiness
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.