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Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy

Cell, 2021-01, Vol.184 (2), p.323-333.e9 [Peer Reviewed Journal]

2020 Elsevier Inc. ;Copyright © 2020 Elsevier Inc. All rights reserved. ;2020. Not withstanding the ProQuest Terms and Conditions, you may use this content in accordance with the associated terms available at https://www.elsevier.com/legal/elsevier-website-terms-and-conditions ;2020 Elsevier Inc. 2020 Elsevier Inc. ;ISSN: 0092-8674 ;EISSN: 1097-4172 ;DOI: 10.1016/j.cell.2020.12.001 ;PMID: 33306959

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Title:
Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy
Author: 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
Subjects: Animals ; Cell Line ; Cell Phone - instrumentation ; Coronavirus Nucleocapsid Proteins - genetics ; COVID-19 ; COVID-19 Nucleic Acid Testing - economics ; COVID-19 Nucleic Acid Testing - instrumentation ; COVID-19 Nucleic Acid Testing - methods ; CRISPR Dx ; CRISPR-Cas Systems ; CRISPR-Cas13 ; Humans ; mobile phone microscopy ; Nasopharynx - virology ; Optical Imaging - instrumentation ; Optical Imaging - methods ; Phosphoproteins - genetics ; point-of-care diagnostics ; Point-of-Care Testing ; RNA Interference ; RNA, Viral - analysis ; RNA, Viral - genetics ; SARS-CoV-2 ; Sensitivity and Specificity ; Viral Load - economics ; Viral Load - instrumentation ; Viral Load - methods
Is Part Of: Cell, 2021-01, Vol.184 (2), p.323-333.e9
Description: 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. [Display omitted] •CRISPR-Cas13a can quantitatively detect SARS-CoV-2 RNA without pre-amplification•Combining crRNAs targeting multiple regions of the viral RNA enhances sensitivity•Cas13a can accurately and rapidly quantify SARS-CoV-2 RNA in patient samples•A mobile phone-based device allows for portable and sensitive readout of the assay Fozouni et al. devise a way to use CRISPR-Cas13a to detect and quantify SARS-CoV-2 RNA from patient samples without the need for a pre-amplification step. They then show how the assay’s signal can be efficiently detected with a portable, mobile phone-based device.
Publisher: United States: Elsevier Inc
Language: English
Identifier: ISSN: 0092-8674
EISSN: 1097-4172
DOI: 10.1016/j.cell.2020.12.001
PMID: 33306959
Source: Coronavirus Research Database

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Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy

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