CRISPR-Cas技术在动物病原微生物检测中的应用研究进展  

Advances in Application of CRISPR-Cas Technology in the Detection of Animal Pathogenic Microorganisms

作　　者：原青 孟茹 李国平 张瑞 YUAN Qing;MENG Ru;LI Guoping;ZHANG Rui(College of Animal Science and Veterinary Medicine of Southwest Minzu University,Chengdu 610041,China;Xining City Animal Disease Prevention and Control Center,Xining 810016,China;Datong County Animal Husbandry and Veterinary Station,Xining 810199,China)

机构地区：[1]西南民族大学畜牧兽医学院,成都610041 [2]西宁市动物疫病预防控制中心,西宁810016 [3]大通县畜牧兽医站,西宁810199

出　　处：《中国畜牧兽医》2025年第2期545-553,共9页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(32302870);西南民族大学引进人才科研启动金资助项目(RQD2023034);西南民族大学中央高校基本科研业务费专项基金项目(ZYN2023048);大通地区牦牛绿色防控技术集成与示范(2023-ZB-1)。

摘　　要：动物传染病是对养殖业危害最严重的一类疾病,不仅造成动物大量死亡及其产品的损失,而且对人类健康构成重大威胁。因此,快速、准确和廉价的病原体检测方法对动物传染病防控具有重要意义。规律成簇间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)及相关蛋白(CRISPR associated protein, Cas)系统是一种灵敏、特异的生物系统,可快速识别病原体特异性核酸,在原核生物免疫系统中发挥重要作用。基于CRISPR-Cas系统的病原体检测方法具有特异性强、灵敏度高和操作便捷等优点,在动物病原微生物快速检测中具有良好的应用前景。笔者系统介绍了Ⅱ型(cas9)、Ⅴ型(cas12a、cas12b、cas14a)和Ⅵ型(cas13a)CRISPR-Cas系统,简述其工作原理和基于CRISPR-Cas系统建立的NASBA-CRISPR、CAS-EXPAR、CASLFA、DETECTR、CORDS、HOLMESv2、SHERLOCK、SHERLOCKv2、DETECTR-Cas14等快速检测方法在动物病原微生物检测中的研究及应用现状,同时对CRISPR-Cas系统在动物病原检测应用中面临的问题进行了分析,包括crRNA的特异性、与扩增技术联用、样本采集和运输方式等因素对定量检测产生的影响,并提出了一些解决方案,旨在为更深入地了解CRISPR-Cas技术在动物病原微生物检测上的应用提供参考。Animal infectious diseases are one of the most serious types of diseases that jeopardize the aquaculture industry,not only causing a large number of animal deaths and the loss of their products but also posing a major threat to human health.Therefore,rapid,accurate and inexpensive pathogen detection methods are of great significance for the prevention and control of animal infectious diseases.The clustered regularly interspaced short palindromic repeats(CRISPR)and associated proteins(Cas)system is a sensitive and specific biological system that can rapidly recognize pathogen-specific nucleic acid that plays an important role in the prokaryotic immune system.Pathogen detection methods based on the CRISPR-Cas system have the advantages of high specificity,high sensitivity and convenient operation,and have good prospects for application in the rapid detection of animal pathogenic microorganisms.The authors systematically introduced the typeⅡ(cas9),typeⅤ(cas12a,cas12b and cas14a)and typeⅥ(cas13a)CRISPR-Cas systems,briefly described their working principles and application of NASBA-CRISPR,CAS-EXPAR,CASLFA,DETECTR,CORDS,HOLMESv2,SHERLOCK,SHERLOCKv2 and DETECTR-Cas14 established based on the CRISPR-Cas system in the detection of animal pathogenic microorganisms,and analyzed the problems faced by the CRISPR-Cas system in the detection of animal pathogens,including the impact of factors such as the specificity of the crRNA,the coupling with amplification technology,sample collection and transportation methods on quantitative detection,and proposed some solutions,aiming to provide a more in-depth understanding of the application of CRISPR-Cas technology in the detection of animal pathogenic microorganisms.

关 键 词：病原微生物 CRISPR-Cas 病原体检测 应用 

分 类 号：S852.6[农业科学—基础兽医学]