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CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples.

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资源类型：
Pubmed体系：
作者：
Wang Yuxi[1,2] ; Xue Ting[1] ; Wang Minjin[3] ; Ledesma-Amaro Rodrigo[4] ; Lu Ying[5,6] ; Hu Xinyue[2,6] ; Zhang Ting[5] ; Yang Ming[1] ; Li Yalun[1] ; Xiang Jin[2] ; Deng Ruijie[5,*1] ; Ying Binwu[3,*2] ; Li Weimin[1,2,*3] ;
机构： [1]Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China [2]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 [3]Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu 610041, China [4]Department of Bioengineering, Imperial College Centre for Synthetic Biology, Imperial College London, London, UK [5]College of Biomass Science and Engineering, Sichuan University, Chengdu 610065, China [6]State Key Laboratory of Biotherapy and Cancer Center/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China
出处：
DOI： 10.1016/j.snb.2022.131765
ISSN：

关键词： COVID-19 SARS-CoV-2 CRISPR-Cas13 RNA aptamer Nucleic acid tests

摘要：
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.© 2022 Elsevier B.V. All rights reserved.

基金：
This work was supported by Sichuan Science and Technology Program [grant numbers 2021YFS0403, 2019YFS0003, 2020YFS0572, and 2020YFS0573], Major Science and Technology Innovation Project of Chengdu City [grant numbers 2020-YF08-00080-GX], National Major Scientific and Technological Special Project for ‘Significant New Drugs Development’ [grant numbers 2018ZX09201018-021, 2018ZX09201018-020], Special Funds for COVID-19 Prevention of West China Hospital [grant numbers HX-2019-nCoV-003].
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2022]版：
大类 | 1 区 化学
小类 | 1 区 分析化学 1 区 仪器仪表 1 区 电化学
最新[2023]版：
大类 | 1 区 化学
小类 | 1 区 分析化学 1 区 仪器仪表 2 区 电化学
第一作者：
第一作者机构： [1]Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China [2]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
共同第一作者：
通讯作者：
通讯机构： [1]Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China [2]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 [3]Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu 610041, China [5]College of Biomass Science and Engineering, Sichuan University, Chengdu 610065, China [*1]College of Biomass Science and Engineering, Sichuan University, No.24, South Section, First Ring Road, Wuhou District, Chengdu 610065, Sichuan Province, China. [*2]Department of Laboratory Medicine, West China Hospital, Sichuan University, No.37, Guoxue Lane, Wuhou district, Chengdu 610041, Sichuan Province, China. [*3]Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, No.37, Guoxue Lane, Wuhou district, Chengdu 610041, Sichuan Province, China.
推荐引用方式(GB/T 7714)：
Wang Yuxi,Xue Ting,Wang Minjin,et al.CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples.[J].Sensors and actuators. B, Chemical.2022,362:131765.doi:10.1016/j.snb.2022.131765.
APA：
Wang Yuxi,Xue Ting,Wang Minjin,Ledesma-Amaro Rodrigo,Lu Ying...&Li Weimin.(2022).CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples..Sensors and actuators. B, Chemical,362,
MLA：
Wang Yuxi,et al."CRISPR-Cas13a cascade-based viral RNA assay for detecting SARS-CoV-2 and its mutations in clinical samples.".Sensors and actuators. B, Chemical 362.(2022):131765

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