基于CRISPR/Cas系统的纸基分析在快速检测食源性致病菌中的应用  

Application of CRISPR/Cas System-integrated Paper-based Analytical Devices for Rapid Detection of Foodborne Pathogens

作　　者：李鹏儒 沈兴 孟静南 罗林[1] 王涓 徐振林[1] LI Peng-Ru;SHEN Xing;MENG Jing-Nan;LUO Lin;WANG Juan;XU Zhen-Lin(Guangdong Provincial Key Laboratory of Food Safety and Quality,College of Food Science,South China Agricultural University,Guangzhou 510642,China;College of Food Engineering,Anhui Science and Technology University,Chuzhou 233100,China)

机构地区：[1]华南农业大学食品学院,广东省食品质量安全重点实验室,广州510642 [2]安徽科技学院食品工程学院,滁州233100

出　　处：《生物化学与生物物理进展》2024年第5期1147-1160,共14页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(32072316);广东省基础与应用基础研究重大项目(2020B0301030005)资助。

摘　　要：由食源性致病菌引起的食品安全事件严重影响人类健康,开发针对食源性致病菌的快速检测技术十分必要。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)是原核生物的适应性免疫系统,具有特异性识别并切割核酸序列的功能。纸基分析方法作为一种简便性好、成本低廉的分析检测工具,在快速检测领域展现出良好的前景。因此,将CRISPR/Cas系统的高效识别能力和纸基分析方法的简便性相结合可实现对食源性致病菌的快速灵敏检测。本文简要介绍了CRISPR/Cas系统用于核酸检测的概况,对第二类单Cas效应蛋白系统的特点及原理进行概述,重点综述基于CRISPR/Cas系统的试纸分析、侧向流动分析和纸基微流控装置在检测食源性致病菌方面的应用,并讨论了CRISPR/Cas系统结合纸基分析建立检测方法的优势、当前的挑战及未来的发展前景。Foods can be contaminated with foodborne pathogens through a variety of pathways,including water,air and soil.Food safety events caused by foodborne pathogens show a serious impact on human health.However,due to the diversity of foodborne pathogens and the complexity of food matrices,the rapid detection of foodborne pathogens was difficult.The conventional microbial culture and physiological and biochemical identification can hardly meet the need of rapid detection of foodborne pathogens in the field.It is necessary to develop rapid detection technologies for foodborne pathogens.Clustered regularly interspaced short palindromic repeats(CRISPR)and associated protein(Cas)are an adaptive immune systems of prokaryotes with specific recognition and cleavage of nucleic acid sequences,which shows good potential for development of nucleic acid detection and biosensing in the field.According to different forms of application,paper-based analytical devices can be categorized into test paper,lateral flow assay and microfluidic paper-based chips,etc.As a good simplicity and low-cost analytical testing tools,they show good prospects in the field of rapid testing.Therefore,the rapid and sensitive detection of foodborne pathogens can be realized by combining the efficient recognition ability of CRISPR/Cas system and the simplicity of paper-based analytical devices.In this paper,we briefly introduce an overview of the CRISPR/Cas system for nucleic acid detection,and this section focuses on an overview of the features and principles of the class 2 system,including types II,V and VI,which uses a single effector.The application of CRISPR/Cas system based test paper analysis,lateral flow assay and microfluidic paper-based chips for the detection of foodborne pathogens are highlighted in the paper,and finally the advantages,current challenges and future prospects of CRISPR/Cas system in combination with paper-based analytical devices to establish detection methods are discussed.

关 键 词：成簇间隔短回文重复序列及相关蛋白 食源性致病菌 纸基分析 快速检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]