环介导等温基因扩增技术快速检测不耐热型肠毒素大肠杆菌反应条件的优化  

RAPID DETECTION OF HEAT-LABILE ENTEROTOXIGENIC ESCHERICHIA COLI BY LOOP-MEDIATED ISOTHERMAL AMPLIFICATION

作　　者：万红梅[1] 李玉锋[1] 李帅[1] 黄韬睿[1] 郑春亮[1] 

机构地区：[1]四川西华大学生物工程学院,成都610039

出　　处：《现代预防医学》2011年第2期321-323,329,共4页Modern Preventive Medicine

基　　金：四川省教育厅重点项目(08ZA027);西华大学食品生物技术省级重点实验室

摘　　要：[目的]建立环介导等温基因扩增技术快速检测食源性致病菌不耐热型肠毒素大肠杆菌的方法。[方法]基于产肠毒素大肠杆菌的LT基因序列设计1套(共4条)能识别6个区域的特异性LAMP引物,优化引物加入量、反应温度以及反应时间等反应条件,最终评估该方法的特异性和灵敏性。[结果]引物加入量对扩增影响较大;在扩增温度为63℃时扩增最明显;扩增反应进行到45min后即有扩增产物产生。[结论]该方法检测不耐热型肠毒素大肠杆菌具有耗时短、操作简捷、特异性较好、灵敏性高等优点。[Objective]To developed the method of rapid detection for food-borne enterotoxigenic Escherichia coli by loop-mediated isothermal amplification（LAMP）technology.[Methods]In this study,the primers were designed cording to six conservative regions from the heat-labile enterotoxin（LT）gene.The primer concentration,the reaction temperature and the reaction time of LAMP were optimized.[Results]A greater impact on the amplification was the concentration of primers;63℃ was deemed the optimal reaction temperature because of its better specificity;and the amplification produced weak bands at 45min,the amplification produced better results at 60min.[Conclusion]LAMP assay is a rapid,sample,an extremely specific and a better sensitivity method for rapid detection of food-borne enterotoxigenic Escherichia coli.

关 键 词：环介导等温基因扩增 产肠毒素大肠杆菌 肠毒素 LT基因 检测 

分 类 号：R378.21[医药卫生—病原生物学]