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《Nature，4月16日，CRISPR–Cas12-based detection of SARS-CoV-2》

  • 来源专题：COVID-19科研动态监测
  • 编译者： zhangmin
  • 发布时间：2020-04-17

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

    James P. Broughton, Xianding Deng, Guixia Yu, Clare L. Fasching, Venice Servellita, Jasmeet Singh, Xin Miao, Jessica A. Streithorst, Andrea Granados, Alicia Sotomayor-Gonzalez, Kelsey Zorn, Allan Gopez, Elaine Hsu, Wei Gu, Steve Miller, Chao-Yang Pan, Hugo Guevara, Debra A. Wadford, Janice S. Chen & Charles Y. Chiu

    Nature Biotechnology (2020)

    Abstract

    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.

  • 原文来源：https://www.nature.com/articles/s41587-020-0513-4
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  • 《Nature，5月19日，SARS-CoV-2 detection with CRISPR diagnostics》
    • 来源专题：COVID-19科研动态监测
    • 编译者：zhangmin
    • 发布时间：2020-05-20
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  • 《Nature，8月26日，Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA》
    • 来源专题：COVID-19科研动态监测
    • 编译者：zhangmin
    • 发布时间：2020-09-02
    • Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA Maturada Patchsung, Krittapas Jantarug, […]Chayasith Uttamapinant Nature Biomedical Engineering (2020) Abstract Nucleic acid detection by isothermal amplification and the collateral cleavage of reporter molecules by CRISPR-associated enzymes is a promising alternative to quantitative PCR. Here, we report the clinical validation of the specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) assay using the enzyme Cas13a from Leptotrichia wadei for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)—the virus that causes coronavirus disease 2019 (COVID-19)—in 154 nasopharyngeal and throat swab samples collected at Siriraj Hospital, Thailand. Within a detection limit of 42 RNA copies per reaction, SHERLOCK was 100% specific and 100% sensitive with a fluorescence readout, and 100% specific and 97% sensitive with a lateral-flow readout. For the full range of viral load in the clinical samples, the fluorescence readout was 100% specific and 96% sensitive. For 380 SARS-CoV-2-negative pre-operative samples from patients undergoing surgery, SHERLOCK was in 100% agreement with quantitative PCR with reverse transcription. The assay, which we show is amenable to multiplexed detection in a single lateral-flow strip incorporating an internal control for ribonuclease contamination, should facilitate SARS-CoV-2 detection in settings with limited resources.
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