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Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy.
Fozouni, Parinaz; Son, Sungmin; Díaz de León Derby, María; Knott, Gavin J; Gray, Carley N; D'Ambrosio, Michael V; Zhao, Chunyu; Switz, Neil A; Kumar, G Renuka; Stephens, Stephanie I; Boehm, Daniela; Tsou, Chia-Lin; Shu, Jeffrey; Bhuiya, Abdul; Armstrong, Maxim; Harris, Andrew R; Chen, Pei-Yi; Osterloh, Jeannette M; Meyer-Franke, Anke; Joehnk, Bastian; Walcott, Keith; Sil, Anita; Langelier, Charles; Pollard, Katherine S; Crawford, Emily D; Puschnik, Andreas S; Phelps, Maira; Kistler, Amy; DeRisi, Joseph L; Doudna, Jennifer A; Fletcher, Daniel A; Ott, Melanie.
Afiliación
  • Fozouni P; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94143, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Medicine,
  • Son S; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
  • Díaz de León Derby M; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA; UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
  • Knott GJ; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Monash Biomedicine Discovery Institute, Department of Biochemistry & Molecular Biology, Monash University, VIC 3800, Australia.
  • Gray CN; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • D'Ambrosio MV; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
  • Zhao C; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA.
  • Switz NA; Department of Physics and Astronomy, San José State University, San Jose, CA 95192, USA.
  • Kumar GR; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Stephens SI; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Boehm D; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Tsou CL; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Shu J; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Bhuiya A; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA; UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
  • Armstrong M; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
  • Harris AR; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
  • Chen PY; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Osterloh JM; J. David Gladstone Institutes, San Francisco, CA 94158, USA.
  • Meyer-Franke A; J. David Gladstone Institutes, San Francisco, CA 94158, USA.
  • Joehnk B; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Walcott K; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Sil A; Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94143, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Microbiology and Immunology, University of California, San Francisco,
  • Langelier C; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA; Division of Infectious Diseases, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Pollard KS; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA 94143, USA; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA; Institute for Human Genetics, University of California, San Francisco, San F
  • Crawford ED; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Puschnik AS; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA.
  • Phelps M; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA.
  • Kistler A; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA.
  • DeRisi JL; Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94143, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA 94143, USA; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA; Division of Biochemistry and B
  • Doudna JA; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Department of C
  • Fletcher DA; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA; UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA; Chan Zuckerberg Biohub
  • Ott M; J. David Gladstone Institutes, San Francisco, CA 94158, USA; Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94143, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Medicine,
Cell ; 184(2): 323-333.e9, 2021 01 21.
Article en En | MEDLINE | ID: mdl-33306959
ABSTRACT
The December 2019 outbreak of a novel respiratory virus, SARS-CoV-2, has become an ongoing global pandemic due in part to the challenge of identifying symptomatic, asymptomatic, and pre-symptomatic carriers of the virus. CRISPR diagnostics can augment gold-standard PCR-based testing if they can be made rapid, portable, and accurate. Here, we report the development of an amplification-free CRISPR-Cas13a assay for direct detection of SARS-CoV-2 from nasal swab RNA that can be read with a mobile phone microscope. The assay achieved ∼100 copies/µL sensitivity in under 30 min of measurement time and accurately detected pre-extracted RNA from a set of positive clinical samples in under 5 min. We combined crRNAs targeting SARS-CoV-2 RNA to improve sensitivity and specificity and directly quantified viral load using enzyme kinetics. Integrated with a reader device based on a mobile phone, this assay has the potential to enable rapid, low-cost, point-of-care screening for SARS-CoV-2.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: ARN Viral / Carga Viral / Teléfono Celular / Imagen Óptica / Prueba de Ácido Nucleico para COVID-19 Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: Cell Año: 2021 Tipo del documento: Article
Texto completo
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: ARN Viral / Carga Viral / Teléfono Celular / Imagen Óptica / Prueba de Ácido Nucleico para COVID-19 Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: Cell Año: 2021 Tipo del documento: Article