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Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA.
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.
  • Patchsung M; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Jantarug K; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Pattama A; Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Aphicho K; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Suraritdechachai S; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Meesawat P; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Sappakhaw K; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Leelahakorn N; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Ruenkam T; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Wongsatit T; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Athipanyasilp N; Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Eiamthong B; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Lakkanasirorat B; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Phoodokmai T; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Niljianskul N; PTT Public Company Limited, Bangkok, Thailand.
  • Pakotiprapha D; Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand.
  • Chanarat S; Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand.
  • Homchan A; Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand.
  • Tinikul R; Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand.
  • Kamutira P; Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand.
  • Phiwkaow K; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Soithongcharoen S; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Kantiwiriyawanitch C; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Pongsupasa V; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Trisrivirat D; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Jaroensuk J; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Wongnate T; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Maenpuen S; Department of Biochemistry, Faculty of Science, Burapha University, Chonburi, Thailand.
  • Chaiyen P; School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand.
  • Kamnerdnakta S; Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Swangsri J; Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Chuthapisith S; Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Sirivatanauksorn Y; Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Chaimayo C; Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Sutthent R; Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Kantakamalakul W; Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
  • Joung J; Howard Hughes Medical Institute, Cambridge, MA, USA.
  • Ladha A; Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • Jin X; McGovern Institute for Brain Research at MIT, Cambridge, MA, USA.
  • Gootenberg JS; Department of Biological Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
  • Abudayyeh OO; Howard Hughes Medical Institute, Cambridge, MA, USA.
  • Zhang F; Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • Horthongkham N; McGovern Institute for Brain Research at MIT, Cambridge, MA, USA.
  • Uttamapinant C; Department of Biological Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
Nat Biomed Eng ; 4(12): 1140-1149, 2020 12.
Article in English | MEDLINE | ID: covidwho-733522
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / CRISPR-Associated Proteins / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Nat Biomed Eng Year: 2020 Document Type: Article Affiliation country: S41551-020-00603-x

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / CRISPR-Associated Proteins / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Nat Biomed Eng Year: 2020 Document Type: Article Affiliation country: S41551-020-00603-x