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Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay

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收录情况： ◇ ESCI

作者：
Zhao Fei[1] * ; Hu Yamei[1] * ; Fan Zhangling[1] * ; Huang Baoying[2] * ; Wei Liang[1] ; Xie Yu[1] ; Huang Yu[1] ; Mei Shan[1] ; Wang Liming[3] ; Wang Lingwa[4] ; Ai Bin[3] ; Fang Jugao[4] ; Liang Chen[5] ; Xu Fengwen[1] ; Tan Wenjie[2] ; Guo Fei[1,7] ;
机构： [1]NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P.R. China [2]NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, P.R. China [3]Department of Medical Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing 100730, P.R. China [4]Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, P.R. China [5]Lady Davis Institute, Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada
出处：
DOI： 10.1016/j.crmeth.2023.100620
ISSN：

摘要：
Mpox is caused by a zoonotic virus belonging to the Orthopoxvirus genus and the Poxviridae family. In this study, we develop a recombinase polymerase amplification (RPA)-coupled CRISPR-Cas12a detection assay for the mpox virus. We design and test a series of CRISPR-derived RNAs(crRNAs) targeting the conserved D6R and E9L genes for orthopoxvirus and the unique N3R and N4R genes for mpox viruses. D6R crRNA-1 exhibits the most robust activity in detecting orthopoxviruses, and N4R crRNA-2 is able to distinguish the mpox virus from other orthopoxviruses. The Cas12a/crRNA assay alone presents a detection limit of 108 copies of viral DNA, whereas coupling RPA increases the detection limit to 1-10 copies. The one-tube RPA-Cas12a assay can, therefore, detect viral DNA as low as 1 copy within 30 min and holds the promise of providing point-of-care detection for mpox viral infection.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

基金：
This study was supported by funds from the National Key R&D Program of China (2022YFE0203100 and 2022YFC2303401), the National Natural Science Foundation of China (82271802, 82072288, 82241075, and 32200718), the PUMC Fundamental Research Funds for the Central Universities (3332022059), and the CAMS Innovation Fund for Medical Sciences (CIFMS 2021-I2M-1-038 and CIFMS 2022-I2M-JB-014).
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外文
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出版当年[2022]版：
无
最新[2025]版：
大类 | 3 区 生物学
小类 | 2 区 生化研究方法 3 区 细胞生物学
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出版当年[2021]版：
无
最新[2024]版：
Q1 BIOCHEMICAL RESEARCH METHODS Q2 CELL BIOLOGY

影响因子： 最新[2024版] 最新五年平均 出版当年[2021版] 出版当年五年平均 出版前一年[2020版] 出版后一年[2022版]

第一作者：
第一作者机构： [1]NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P.R. China
共同第一作者：
通讯作者：
通讯机构： [1]NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P.R. China
推荐引用方式(GB/T 7714)：
Zhao Fei,Hu Yamei,Fan Zhangling,et al.Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay[J].CELL REPORTS METHODS.2023,3(10):doi:10.1016/j.crmeth.2023.100620.
APA：
Zhao Fei,Hu Yamei,Fan Zhangling,Huang Baoying,Wei Liang...&Guo Fei.(2023).Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay.CELL REPORTS METHODS,3,(10)
MLA：
Zhao Fei,et al."Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay".CELL REPORTS METHODS 3..10(2023)

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