核酸切割酶在病原微生物检测中的研究进展  

Research Progress on RNA-cleaving DNAzyme in the Detection of Pathogens

作　　者：王昕 朱龙佼[1] 许文涛 翟晨[4] 王书雅 黄蔚霞 WANG Xin;ZHU Long-jiao;XU Wen-tao;ZHAI Chen;WANG Shu-ya;HUANG Wei-xia(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083;Beijing Advanced Innovation Center for Food Nutrition and Human Health of China Agricultural University,Beijing 100083;Key Laboratory of Safety Assessment of Genetically Modified Organism(Food Safety),Ministry of Agriculture,Beijing 100083;Nutrition&Health Research Institute,COFCO Corporation,Beijing Key Laboratory of Nutrition&Health and Food Safety,Beijing Engineering Laboratory for Geriatric Nutrition Food Research,Beijing Livestock Product Quality and Safety Source Control Engineering Technology Research Center,Beijing 102209)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]中国农业大学食品营养与人类健康高精尖创新中心,北京100083 [3]农业部农业转基因生物安全评价(食用)重点实验室,北京100083 [4]中粮营养健康研究院有限公司营养健康与食品安全北京市重点实验室老年营养食品研究北京市工程实验室北京市畜产品质量安全源头控制工程技术研究中心,102209

出　　处：《生物技术通报》2020年第1期182-192,共11页Biotechnology Bulletin

基　　金：国家重点研发计划资助(2016YFD0401204)

摘　　要：DNA是遗传信息的重要载体,其空间构象折叠性质使其具有很多的功能。利用核酸切割酶(cleaving DNAzyme)识别特定单链DNA分子并能够切割其中某条单链的性质来构建传感器,将特异性识别过程转化为凝胶电泳表征、释放荧光、比色现象的信号输出,同时能很好的和扩增反应结合来实现信号放大。核酸切割酶通过体外筛选技术获得,可以与靶物质(小分子、蛋白质,甚至整个细胞)特异性结合。由于具有制备简单,易于修饰和良好稳定性等优点,核酸切割酶被用于构建生物传感器以检测病原微生物,已应用到现场检测甚至医疗中的体内检测,结合已经成熟的检测设备血糖仪、横流层析试纸条带进行微生物检测,并广泛地应用到生物传感、食品安全、医疗在内的重要领域中。综述了近年来核酸切割酶在微生物检测中的应用,讨论了核酸切割酶在微生物检测中的切割机理和产物、靶标以及表征手段,探索核酸切割酶在微生物实际检测中的意义。对该技术的发展前景及其面临的问题进行展望,以期核酸切割酶在微生物检测领域能够更好的发展。DNA is an important carrier of genetic information,and its folding property of spatial conformation makes it have many functions.Using a RNA-cleaving DNAzyme to recognize a specific single-stranded DNA molecule and to cleave one of the single strands,a signal output sensor can be constructed,which converts the specific recognition process into the signal outputs of gel electrophoresis character,fluorescence,and colorimetric,and it can be combined with amplification reaction to achieve signal amplified.RNA-cleaving DNAzymes are obtained by in vitro screening techniques that specifically bind to target substances(such as small molecules,proteins,and even whole cells).Due to its advantages of convenient preparation,easy modification and fine stability,RNA-cleaving DNAzymes are used to construct biosensors to detect pathogenic microorganisms,for example in-situ detection and even in-vivo detection in medical treatment.In the future,it will be combined with mature detection,for instance,equipment blood glucose meter,cross-flow chromatography strips for microbiological testing.Furthermore,it can be used in important areas such as biosensing,food safety,and medical care.Here we review the application of RNA-cleaving DNAzymes in microbial detection.In addition,we discuss the mechanism,products,targets and characterization of RNA-cleaving DNAzymes in microbial detection,as well as the significance and problems of RNA-cleaving DNAzymes in the actual detection of microorganisms.Finally,we prospect the further application of this technology,aiming at the better development of nucleic acid cleavage enzymes in microbial detection.

关 键 词：核酸切割酶 微生物 生物传感 信号放大 检测 

分 类 号：R44[医药卫生—诊断学]