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[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] The research aimed to establish a method for rapid detection of heat-labile enterotoxigenic Escherichia coli (E. coli) by food-borne pathogens by loop-mediated isothermal gene amplification. [Method] Based on the LT gene sequence of enterotoxigenic Escherichia coli, 1 set of 4 specific LAMP primers were designed and optimized. The reaction conditions such as the amount of primer, reaction temperature and reaction time were optimized. Finally, the method was evaluated The specificity and sensitivity. [Result] The amount of primer added greatly affected the amplification. The amplification was the most obvious when the amplification temperature was 63 ℃. The amplification product was produced after 45 min. [Conclusion] The method for detecting heat-labile enterotoxin-producing Escherichia coli has the advantages of short time-consuming, simple operation, good specificity and high sensitivity.