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CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples.
Wang, Yuxi; Xue, Ting; Wang, Minjin; Ledesma-Amaro, Rodrigo; Lu, Ying; Hu, Xinyue; Zhang, Ting; Yang, Ming; Li, Yalun; Xiang, Jin; Deng, Ruijie; Ying, Binwu; Li, Weimin.
  • Wang Y; Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Xue T; Targeted Tracer Research and development laboratory, Institute of Respiratory Health, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Wang M; Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Ledesma-Amaro R; Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu 610041, China.
  • Lu Y; Department of Bioengineering, Imperial College Centre for Synthetic Biology, Imperial College London, London, UK.
  • Hu X; College of Biomass Science and Engineering, Sichuan University, Chengdu 610065, China.
  • Zhang T; State Key Laboratory of Biotherapy and Cancer Center/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Yang M; Targeted Tracer Research and development laboratory, Institute of Respiratory Health, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Li Y; State Key Laboratory of Biotherapy and Cancer Center/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Xiang J; College of Biomass Science and Engineering, Sichuan University, Chengdu 610065, China.
  • Deng R; Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Ying B; Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Li W; Targeted Tracer Research and development laboratory, Institute of Respiratory Health, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu 610041, China.
Sens Actuators B Chem ; 362: 131765, 2022 Jul 01.
Article in English | MEDLINE | ID: covidwho-1757833
ABSTRACT
SARS-CoV-2 is one of the greatest threats to global human health. Point-of-care diagnostic tools for SARS-CoV-2 could facilitate rapid therapeutic intervention and mitigate transmission. In this work, we report CRISPR-Cas13a cascade-based viral RNA (Cas13C) assay for label-free and isothermal determination of SARS-CoV-2 and its mutations in clinical samples. Cas13a/crRNA was utilized to directly recognize the target of SARS-CoV-2 RNA, and the recognition events sequentially initiate the transcription amplification to produce light-up RNA aptamers for output fluorescence signal. The recognition of viral RNA via Cas13a-guide RNA ensures a high specificity to distinguish SARS-CoV-2 from MERS-CoV and SARS-CoV, as well as viral mutations. A post transcription amplification strategy was triggered after CRISPR-Cas13a recognition contributes to an amplification cascade that achieves high sensitivity for detecting SARS-CoV-2 RNA, with a limit of detection of 0.216 fM. In addition, the Cas13C assay could be able to discriminate single-nucleotide mutation, which was proven with N501Y in SARS-Cov-2 variant. This method was validated by a 100% agreement with RT-qPCR results from 12 clinical throat swab specimens. The Cas13C assay has the potential to be used as a routine nucleic acid test of SARS-CoV-2 virus in resource-limited regions.
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study Topics: Variants Language: English Journal: Sens Actuators B Chem Year: 2022 Document Type: Article Affiliation country: J.snb.2022.131765

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study Topics: Variants Language: English Journal: Sens Actuators B Chem Year: 2022 Document Type: Article Affiliation country: J.snb.2022.131765