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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
Published Web Location
https://doi.org/10.1016/j.cell.2020.12.001Abstract
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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