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Rapid detection of SARS-CoV-2 RNA in saliva via Cas13.
Chandrasekaran, Sita S; Agrawal, Shreeya; Fanton, Alison; Jangid, Aditya R; Charrez, Bérénice; Escajeda, Arturo M; Son, Sungmin; Mcintosh, Roger; Tran, Huyen; Bhuiya, Abdul; de León Derby, María Díaz; Switz, Neil A; Armstrong, Maxim; Harris, Andrew R; Prywes, Noam; Lukarska, Maria; Biering, Scott B; Smock, Dylan C J; Mok, Amanda; Knott, Gavin J; Dang, Qi; Van Dis, Erik; Dugan, Eli; Kim, Shin; Liu, Tina Y; Moehle, Erica A; Kogut, Katherine; Eskenazi, Brenda; Harris, Eva; Stanley, Sarah A; Lareau, Liana F; Tan, Ming X; Fletcher, Daniel A; Doudna, Jennifer A; Savage, David F; Hsu, Patrick D.
  • Chandrasekaran SS; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Agrawal S; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Fanton A; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
  • Jangid AR; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Charrez B; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Escajeda AM; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Son S; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Mcintosh R; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
  • Tran H; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Bhuiya A; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • de León Derby MD; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
  • Switz NA; Wainamics Inc., Pleasanton, CA, USA.
  • Armstrong M; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Harris AR; Wainamics Inc., Pleasanton, CA, USA.
  • Prywes N; Wainamics Inc., Pleasanton, CA, USA.
  • Lukarska M; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Biering SB; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
  • Smock DCJ; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Mok A; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
  • Knott GJ; Department of Physics and Astronomy, San José State University, San José, CA, USA.
  • Dang Q; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
  • Van Dis E; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Dugan E; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Kim S; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Liu TY; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Moehle EA; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Kogut K; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Eskenazi B; Center for Computational Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Harris E; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Stanley SA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Lareau LF; Monash Biomedicine Discovery Institute, Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
  • Tan MX; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Fletcher DA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Doudna JA; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Savage DF; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • Hsu PD; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
Nat Biomed Eng ; 6(8): 944-956, 2022 08.
Article in English | MEDLINE | ID: covidwho-1991606
ABSTRACT
Rapid nucleic acid testing is central to infectious disease surveillance. Here, we report an assay for rapid COVID-19 testing and its implementation in a prototype microfluidic device. The assay, which we named DISCoVER (for diagnostics with coronavirus enzymatic reporting), involves extraction-free sample lysis via shelf-stable and low-cost reagents, multiplexed isothermal RNA amplification followed by T7 transcription, and Cas13-mediated cleavage of a quenched fluorophore. The device consists of a single-use gravity-driven microfluidic cartridge inserted into a compact instrument for automated running of the assay and readout of fluorescence within 60 min. DISCoVER can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in saliva with a sensitivity of 40 copies µl-1, and was 94% sensitive and 100% specific when validated (against quantitative PCR) using total RNA extracted from 63 nasal-swab samples (33 SARS-CoV-2-positive, with cycle-threshold values of 13-35). The device correctly identified all tested clinical saliva samples (10 SARS-CoV-2-positive out of 13, with cycle-threshold values of 23-31). Rapid point-of-care nucleic acid testing may broaden the use of molecular diagnostics.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Nat Biomed Eng Year: 2022 Document Type: Article Affiliation country: S41551-022-00917-y

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Nat Biomed Eng Year: 2022 Document Type: Article Affiliation country: S41551-022-00917-y