Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA
| Autor: | Danaya Pakotiprapha, Khomkrit Sappakhaw, Chutikarn Chaimayo, Bhumrapee Eiamthong, Yongyut Sirivatanauksorn, Kochakorn Phiwkaow, Benya Lakkanasirorat, Thanyaporn Wongnate, Vinutsada Pongsupasa, Alim Ladha, Ruchanok Tinikul, Xin Jin, Thitima Phoodokmai, Thanakrit Wongsatit, Omar O. Abudayyeh, Jonathan S. Gootenberg, Suebwong Chuthapisith, Aimorn Homchan, Nootaree Niljianskul, Chayasith Uttamapinant, Feng Zhang, Sahachat Soithongcharoen, Wannee Kantakamalakul, Duangthip Trisrivirat, Surased Suraritdechachai, Sittinan Chanarat, Chadaporn Kantiwiriyawanitch, Piyachat Meesawat, Pimchai Chaiyen, Niracha Athipanyasilp, Sirichai Kamnerdnakta, Krittapas Jantarug, Navin Horthongkham, Jirawat Swangsri, Archiraya Pattama, Nattawat Leelahakorn, Ruengpung Sutthent, Philaiwarong Kamutira, Somchart Maenpuen, Maturada Patchsung, Julia Joung, Theerawat Ruenkam, Kanokpol Aphicho, Juthamas Jaroensuk |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Detection limit business.industry Loop-mediated isothermal amplification Biomedical Engineering RNA Medicine (miscellaneous) Bioengineering Nucleic acid amplification technique Virology Reverse transcriptase Virus Computer Science Applications 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Real-time polymerase chain reaction Medicine business Viral load 030217 neurology & neurosurgery Biotechnology |
| Zdroj: | Nature Biomedical Engineering |
| ISSN: | 2157-846X |
| DOI: | 10.1038/s41551-020-00603-x |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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