利用一步法 RPA-CRISPR/Cas13a 检测试剂盒快速检测尼帕病毒。
Rapid detection of Nipah virus using the one-pot RPA-CRISPR/Cas13a assay.
机构信息
Viral Hemorrhagic Fevers Research Unit, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shangha 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Institute of Pathogenic Microorganisms, Guangdong Provincial Center for Disease Control and Prevention, Guangzhou 510440, China.
出版信息
Virus Res. 2023 Jul 15;332:199130. doi: 10.1016/j.virusres.2023.199130. Epub 2023 May 14.
Nipah virus (NiV) is a zoonotic pathogen with airborne transmission and high case fatality rates in humans. There is currently no treatment or vaccine against NiV infection approved for humans or animals, therefore early diagnosis is the key to control any potential outbreaks. In this study, we developed an optimized one-pot assay using recombinase polymerase amplification (RPA) coupled to CRISPR/Cas13a for the molecular detection of NiV. The one-pot RPA-CRISPR/Cas13a assay for NiV detection was specific and did not cross-react against other selected (re)-emerging pathogens. The sensitivity of the one-pot RPA-CRISPR/Cas13a assay for NiV detection can detect as little as 10 cp/μL of total synthetic NiV cDNA. The assay was then validated with simulated clinical samples. The results for the one-pot RPA-CRISPR/Cas13a assay could be visualized with either fluorescence or lateral flow strips for convenient clinical or field diagnostics, providing a useful supplement to the gold-standard qRT-PCR assay for detecting NiV detections.
鼻病毒（NiV）是一种具有空气传播能力的人畜共患病原体，其在人类中的病死率很高。目前尚无针对 NiV 感染的人类或动物治疗方法或疫苗，因此早期诊断是控制任何潜在暴发的关键。在这项研究中，我们开发了一种优化的一锅法，使用重组酶聚合酶扩增（RPA）与 CRISPR/Cas13a 偶联，用于 NiV 的分子检测。用于 NiV 检测的一锅式 RPA-CRISPR/Cas13a 检测法具有特异性，不会与其他选定的（再）出现的病原体发生交叉反应。一锅式 RPA-CRISPR/Cas13a 检测 NiV 的灵敏度可检测到低至 10 cp/μL 的总合成 NiV cDNA。然后用模拟的临床样本对该检测方法进行了验证。可以通过荧光或侧流条带对一锅式 RPA-CRISPR/Cas13a 检测进行可视化，便于临床或现场诊断，为检测 NiV 的金标准 qRT-PCR 检测提供了有用的补充。
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