基于CRISPR-Cas3的新冠病毒和流感病毒诊断方法
CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus.
作者信息
Yoshimi Kazuto, Takeshita Kohei, Yamayoshi Seiya, Shibumura Satomi, Yamauchi Yuko, Yamamoto Masaki, Yotsuyanagi Hiroshi, Kawaoka Yoshihiro, Mashimo Tomoji
机构信息
Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
Division of Genome Engineering, Center for Experimental Medicine and Systems Biology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
出版信息
iScience. 2022 Feb 18;25(2):103830. doi: 10.1016/j.isci.2022.103830. Epub 2022 Jan 30.
CRISPR-based diagnostics (CRISPR-dx), including the Cas12-based DETECTR and Cas13-based SHERLOCK Class 2 CRISPRs, have been used to detect the presence of DNA or RNA from pathogens, such as the 2009 pandemic influenza virus A (IAV) and the 2019 novel coronavirus SARS-CoV-2. Here, we describe the collateral single-stranded DNA cleavage with Class 1 type I CRISPR-Cas3 and highlight its potential for development as a Cas3-mediated rapid (within 40 min), low-cost, instrument-free detection method for SARS-CoV-2. This assay, which we call Cas3-Operated Nucleic Acid detectioN (CONAN), not only detects SARS-CoV-2 in clinical samples, but also offers specific detection of single-base-pair mutations in IAV variants. This tool allows rapid and accurate point-of-care testing for patients with suspected SARS-CoV-2 or drug-resistant IAV infections in hospitals.
基于CRISPR的诊断方法（CRISPR-dx），包括基于Cas12的DETECTR和基于Cas13的SHERLOCK 2类CRISPR，已被用于检测病原体DNA或RNA的存在，如2009年甲型大流行性流感病毒（IAV）和2019年新型冠状病毒SARS-CoV-2。在此，我们描述了1类I型CRISPR-Cas3的附带单链DNA切割，并强调了其作为一种用于SARS-CoV-2的由Cas3介导的快速（40分钟内）、低成本、无需仪器的检测方法的开发潜力。我们将此检测方法称为Cas3操作核酸检测（CONAN），它不仅能检测临床样本中的SARS-CoV-2，还能特异性检测IAV变体中的单碱基对突变。该工具可为医院中疑似SARS-CoV-2或耐药IAV感染的患者提供快速、准确的即时检测。
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