TY - JOUR
T1 - Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy
AU - Fozouni, Parinaz
AU - Son, Sungmin
AU - Díaz de León Derby, María
AU - Knott, Gavin J.
AU - Gray, Carley N.
AU - D'Ambrosio, Michael V.
AU - Zhao, Chunyu
AU - Switz, Neil A.
AU - Kumar, G. Renuka
AU - Stephens, Stephanie I.
AU - Boehm, Daniela
AU - Tsou, Chia Lin
AU - Shu, Jeffrey
AU - Bhuiya, Abdul
AU - Armstrong, Maxim
AU - Harris, Andrew R.
AU - Chen, Pei Yi
AU - Osterloh, Jeannette M.
AU - Meyer-Franke, Anke
AU - Joehnk, Bastian
AU - Walcott, Keith
AU - Sil, Anita
AU - Langelier, Charles
AU - Pollard, Katherine S.
AU - Crawford, Emily D.
AU - Puschnik, Andreas S.
AU - Phelps, Maira
AU - Kistler, Amy
AU - DeRisi, Joseph L.
AU - Doudna, Jennifer A.
AU - Fletcher, Daniel A.
AU - Ott, Melanie
PY - 2021/1/21
Y1 - 2021/1/21
N2 - 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.
AB - 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.
KW - COVID-19
KW - CRISPR Dx
KW - CRISPR-Cas13
KW - mobile phone microscopy
KW - point-of-care diagnostics
KW - SARS-CoV-2
UR - http://www.scopus.com/inward/record.url?scp=85097559267&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2020.12.001
DO - 10.1016/j.cell.2020.12.001
M3 - Article
C2 - 33306959
AN - SCOPUS:85097559267
VL - 184
SP - 323-333.e9
JO - Cell
JF - Cell
SN - 0092-8674
IS - 2
ER -