Direct detection of SARS-CoV-2 using CRISPR-Cas13a and a mobile phone-based device. Fozouni. et al.

Published: 9 December 2020| Version 1 | DOI: 10.17632/r3vwyr2w5x.1
Contributors:
Parinaz Fozouni,
Sungmin Son,
Maria Diaz de Leon Derby,
Gavin Knott,
Carley Gray,
Michael D'Ambrosio,
Chunyu Zhao,
Neil Switz,
Renuka Kumar,
Stephanie Stephens,
Daniela Boehm,
Chia-Lin Tsou,
Jeffrey Shu,
Abdul Bhuiya,
Max Armstrong,
Andrew Harris,
Pei-Yi Chen,
Jeannette Osterloh,
Anke Meyer-Franke,
Bastian Joehnk,
Keith Walcott,
Anita Sil,
Charles Langelier,
Katherine Pollard,
Emily Crawford,
Andreas Puschnik,
Maira Phelps,
Amy Kistler,
Joseph DeRisi,
Jennifer Doudna,
Daniel Fletcher,
Melanie Ott

Description

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/uL sensitivity in under 30 minutes of measurement time and accurately detected pre-extracted RNA from a set of positive clinical samples in under 5 minutes. 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.

Files

Steps to reproduce

See the Description of Supplementary Software.pdf

Institutions

University of California Berkeley, Gladstone Institutes

Categories

Software, Image Analysis

License