使用环介导等温扩增技术快速检测食品中肠出血性大肠杆菌O157:H7的初步研究  

Rapid Detection of Enterohemorrhagic Escherichia coli O157:H7 in Food by Loop-Mediated Isothermal Amplification Assay

作　　者：易海华 赵金伟 徐波 吴萍兰 宋阳威 房超 徐政 徐继承[2] 

机构地区：[1]徐州出入境检验检疫局,江苏徐州221006 [2]徐州医学院全科医学系,江苏徐州221001

出　　处：《中国食品卫生杂志》2010年第3期206-213,共8页Chinese Journal of Food Hygiene

基　　金：江苏出入境检验检疫局科研专项基金项目(2007KJ29)

摘　　要：目的探索建立一种快速、简单的食品中肠出血性大肠杆菌O157∶H7的检测方法。方法针对编码O157∶H7脂多糖的rfbE基因(GenBank S83460)和编码H7鞭毛抗原的fliC基因(GenBank L07388)特征性保守序列,利用环介导等温扩增(loop-mediated isothermal amplification,LAMP)核酸扩增基因技术,对21株O157∶H7和非O157∶H7菌株进行特异性检测,并与聚合酶链式反应方法进行比较。同时使用部分食品样品对LAMP法检测肠出血性大肠杆菌O157∶H7的实际运用价值进行评估。结果LAMP法可以在1h内完成检测工作。LAMP法检测肠出血性大肠杆菌O157∶H7的灵敏度为96.72%,特异度为85.71%,准确性为93.26%。结论LAMP检测技术的灵敏度较聚合酶链式反应技术高。LAMP是一种简单、快速的检测技术,适用于筛选可疑大肠杆菌O157∶H7样本。Objective To develop a rapid and simple method of loop-mediated isothermal amplification（LAMP） for detecting Escherichia coli（E.coli） O157：H7 in foods.Method Based on the characteristic conserved sequence of lipopolysaccharide gene（rfbE,GenBank S83460） of O157 antigen and flagellin gene（fliC,GenBank L07388） of H7 antigen,the loop-mediated isothermal amplification method was developed to detect the specificity of twenty-one E.coli O157：H7 strains and non E.coli O157：H7 strains.The results of LAMP were compared with polymerase chain reaction（PCR）.The feasibility of LAMP was evaluated by detecting E.coli O157：H7 in food samples.Results The detection could be finished by LAMP within 1 h;The sensitivity was 96.72%,the specificity was 85.71% and the accuracy was 93.26%.Conclusion The sensitivity of LAMP was higher than that of PCR.LAMP is a simple,rapid method suitable for screening E.coli O157：H7 in suspicious samples.

关 键 词：肠出血性大肠杆菌O157∶H7 环介导等温扩增 rfbE基因 fliC基因 

分 类 号：R155.5[医药卫生—营养与食品卫生学]