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CRISPR-Cas技术在病原体核酸快速检测中的应用进展

Advances in the application of CRISPR-Cas technology in rapid detection of pathogen nucleic acid.

作者信息

Li Xiaoping, Zhong Jiaye, Li Haoyu, Qiao Yinbiao, Mao Xiaolei, Fan Huayan, Zhong Yiwu, Imani Saber, Zheng Shusen, Li Jianhui

机构信息

Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province, Shulan International Medical College, Zhejiang Shuren University, Hangzhou, China.

Faculty of Medicine, Macau University of Science and Technology, Avenida Wai Long Taipa, Macau, China.

出版信息

Front Mol Biosci. 2023 Sep 21;10:1260883. doi: 10.3389/fmolb.2023.1260883. eCollection 2023.

Abstract

(CRISPR) and (Cas) are widely used as gene editing tools in biology, microbiology, and other fields. CRISPR is composed of highly conserved repetitive sequences and spacer sequences in tandem. The spacer sequence has homology with foreign nucleic acids such as viruses and plasmids; Cas effector proteins have endonucleases, and become a hotspot in the field of molecular diagnosis because they recognize and cut specific DNA or RNA sequences. Researchers have developed many diagnostic platforms with high sensitivity, high specificity, and low cost by using Cas proteins (Cas9, Cas12, Cas13, Cas14, etc.) in combination with signal amplification and transformation technologies (fluorescence method, lateral flow technology, etc.), providing a new way for rapid detection of pathogen nucleic acid. This paper introduces the biological mechanism and classification of CRISPR-Cas technology, summarizes the existing rapid detection technology for pathogen nucleic acid based on the trans cleavage activity of Cas, describes its characteristics, functions, and application scenarios, and prospects the future application of this technology.

摘要

成簇规律间隔短回文重复序列(CRISPR)和CRISPR相关蛋白(Cas)在生物学、微生物学及其他领域被广泛用作基因编辑工具。CRISPR由高度保守的重复序列和串联间隔序列组成。间隔序列与病毒和质粒等外源核酸具有同源性;Cas效应蛋白具有核酸内切酶活性,因其能识别并切割特定的DNA或RNA序列,成为分子诊断领域的研究热点。研究人员通过将Cas蛋白(Cas9、Cas12、Cas13、Cas14等)与信号放大及转换技术(荧光法、侧向流动技术等)相结合,开发出了许多高灵敏度、高特异性且低成本的诊断平台,为病原体核酸的快速检测提供了新途径。本文介绍了CRISPR-Cas技术的生物学机制和分类,总结了基于Cas反式切割活性的现有病原体核酸快速检测技术,阐述了其特点、功能及应用场景,并对该技术的未来应用进行了展望。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c0dd/10552857/b458e6ef020a/fmolb-10-1260883-g001.jpg

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