Your browser doesn't support javascript.
loading
Rapid detection of SARS-CoV-2 with Cas13
Shreeya Agrawal; Alison Fanton; Sita S. Chandrasekaran; Bérénice Charrez; Arturo M. Escajeda; Sungmin Son; Roger Mcintosh; Abdul Bhuiya; María Díaz de León Derby; Neil A. Switz; Maxim Armstrong; Andrew R. Harris; Noam Prywes; Maria Lukarska; Scott B. Biering; Dylan C. J. Smock; Amanda Mok; Gavin J. Knott; Qi Dang; Erik Van Dis; Eli Dugan; Shin Kim; Tina Y. Liu; - IGI Testing Consortium; Eva Harris; Sarah A. Stanley; Liana F. Lareau; Ming X. Tan; Daniel A. Fletcher; Jennifer A. Doudna; David F. Savage; Patrick D. Hsu.
Afiliação
  • Shreeya Agrawal; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Alison Fanton; Department of Bioengineering, University of California, Berkeley, CA
  • Sita S. Chandrasekaran; Department of Bioengineering, University of California, Berkeley, CA
  • Bérénice Charrez; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA
  • Arturo M. Escajeda; Wainamics Inc., Pleasanton, CA, USA.
  • Sungmin Son; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Roger Mcintosh; Wainamics Inc., Pleasanton, CA, USA.
  • Abdul Bhuiya; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • María Díaz de León Derby; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Neil A. Switz; Department of Physics and Astronomy, San José State University, San José, CA, USA
  • Maxim Armstrong; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA
  • Andrew R. Harris; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Noam Prywes; Innovative Genomics Institute, University of California, Berkeley , California 94704 , United States.
  • Maria Lukarska; Innovative Genomics Institute, University of California, Berkeley, CA, USA
  • Scott B. Biering; Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, Berkeley, CA, USA
  • Dylan C. J. Smock; Innovative Genomics Institute, University of California, Berkeley, CA, USA
  • Amanda Mok; Center for Computational Biology, University of California, Berkeley, Berkeley, CA, USA
  • Gavin J. Knott; Innovative Genomics Institute, University of California, Berkeley, CA 94720, USA
  • Qi Dang; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA
  • Erik Van Dis; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA
  • Eli Dugan; Innovative Genomics Institute, University of California, Berkeley, CA, USA
  • Shin Kim; Innovative Genomics Institute, University of California, Berkeley, CA, USA
  • Tina Y. Liu; Department of Molecular and Cell Biology, University of California, Berkeley, California, USA
  • - IGI Testing Consortium;
  • Eva Harris; Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, Berkeley, CA 94720-3370 USA
  • Sarah A. Stanley; School of Public Health, University of California, Berkeley, CA 94720, USA
  • Liana F. Lareau; Department of Bioengineering, University of California, Berkeley, CA
  • Ming X. Tan; Wainamics Inc., Pleasanton, CA, USA.
  • Daniel A. Fletcher; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
  • Jennifer A. Doudna; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • David F. Savage; Department of Molecular and Cell Biology, University of California, Berkeley, California, USA
  • Patrick D. Hsu; Department of Bioengineering, University of California, Berkeley, CA
Preprint em En | PREPRINT-MEDRXIV | ID: ppmedrxiv-20247874
ABSTRACT
Rapid nucleic acid testing is a critical component of a robust infrastructure for increased disease surveillance. Here, we report a microfluidic platform for point-of-care, CRISPR-based molecular diagnostics. We first developed a nucleic acid test which pairs distinct mechanisms of DNA and RNA amplification optimized for high sensitivity and rapid kinetics, linked to Cas13 detection for specificity. We combined this workflow with an extraction-free sample lysis protocol using shelf-stable reagents that are widely available at low cost, and a multiplexed human gene control for calling negative test results. As a proof-of-concept, we demonstrate sensitivity down to 40 copies/L of SARS-CoV-2 in unextracted saliva within 35 minutes, and validated the test on total RNA extracted from patient nasal swabs with a range of qPCR Ct values from 13-35. To enable sample-to-answer testing, we integrated this diagnostic reaction with a single-use, gravity-driven microfluidic cartridge followed by real-time fluorescent detection in a compact companion instrument. We envision this approach for Diagnostics with Coronavirus Enzymatic Reporting (DISCoVER) will incentivize frequent, fast, and easy testing.
Licença
cc_by_nc_nd
Texto completo
Adicionar na Minha BVS
Imprimir
XML
Buscar no Google
Texto completo: 1 Coleções: 09-preprints Base de dados: PREPRINT-MEDRXIV Tipo de estudo: Diagnostic_studies / Prognostic_studies Idioma: En Ano de publicação: 2020 Tipo de documento: Preprint
Texto completo
Adicionar na Minha BVS
Imprimir
XML
Buscar no Google
Texto completo: 1 Coleções: 09-preprints Base de dados: PREPRINT-MEDRXIV Tipo de estudo: Diagnostic_studies / Prognostic_studies Idioma: En Ano de publicação: 2020 Tipo de documento: Preprint