CRISPR-Cas12-based detection of SARS-CoV-2

被引:1886
作者
Broughton, James P. [1 ]
Deng, Xianding [2 ,3 ]
Yu, Guixia [2 ,3 ]
Fasching, Clare L. [1 ]
Servellita, Venice [2 ,3 ]
Singh, Jasmeet [1 ]
Miao, Xin [1 ]
Streithorst, Jessica A. [2 ]
Granados, Andrea [2 ,3 ]
Sotomayor-Gonzalez, Alicia [2 ,3 ]
Zorn, Kelsey [4 ]
Gopez, Allan [2 ]
Hsu, Elaine [2 ]
Gu, Wei [2 ]
Miller, Steve [2 ]
Pan, Chao-Yang [5 ]
Guevara, Hugo [5 ]
Wadford, Debra A. [5 ]
Chen, Janice S. [1 ]
Chiu, Charles Y. [2 ,3 ,6 ]
机构
[1] Mammoth Biosci Inc, San Francisco, CA 94080 USA
[2] Univ Calif San Francisco, Dept Lab Med, San Francisco, CA 94143 USA
[3] UCSF Abbott Viral Diagnost & Discovery Ctr, San Francisco, CA 94143 USA
[4] Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA USA
[5] Calif Dept Publ Hlth, Viral & Rickettsial Dis Lab, Richmond, CA USA
[6] Univ Calif San Francisco, Dept Med, Div Infect Dis, San Francisco, CA 94143 USA
基金
美国国家卫生研究院;
关键词
CORONAVIRUS; DIAGNOSTICS; DISEASE;
D O I
10.1038/s41587-020-0513-4
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
SARS-CoV-2 in patient samples is detected in under an hour using a CRISPR-based lateral flow assay. An outbreak of betacoronavirus severe acute respiratory syndrome (SARS)-CoV-2 began in Wuhan, China in December 2019. COVID-19, the disease associated with SARS-CoV-2 infection, rapidly spread to produce a global pandemic. We report development of a rapid (<40 min), easy-to-implement and accurate CRISPR-Cas12-based lateral flow assay for detection of SARS-CoV-2 from respiratory swab RNA extracts. We validated our method using contrived reference samples and clinical samples from patients in the United States, including 36 patients with COVID-19 infection and 42 patients with other viral respiratory infections. Our CRISPR-based DETECTR assay provides a visual and faster alternative to the US Centers for Disease Control and Prevention SARS-CoV-2 real-time RT-PCR assay, with 95% positive predictive agreement and 100% negative predictive agreement.
引用
收藏
页码:870 / +
页数:8
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