Journal
CELL
Volume 184, Issue 2, Pages 323-+Publisher
CELL PRESS
DOI: 10.1016/j.cell.2020.12.001
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Funding
- NIH: NIH/NIAID grant [5R61AI140465-03]
- NIH: NIH/NIDA grant [1R61DA048444-01]
- NIH Rapid Acceleration of Diagnostics (RADx) program
- National Heart, Lung and Blood Institute, National Institutes of Health
- National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health
- Department of Health and Human Services [3U54HL143541-02S1]
- UCSF Medical Scientist Training Program [NIH/NIGMS T32GM007618]
- NIH/NIAID [F30AI143401]
- UC MEXUS-CONACYT Doctoral Fellowship
- NHMRC Investigator Grant (EL1) [APP1175568]
- American Australian Association Fellowship
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This study developed an amplification-free CRISPR-Cas13a assay for direct detection of SARS-CoV-2, showing rapid, highly sensitive, and mobile phone-based readout capabilities.
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 car riers 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 similar to 100 copies/mu L 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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