猪产肠毒素大肠杆菌多重PCR快速检测试剂盒的研制及应用  

Development and Application of Rapid Multiplex-PCR Detection kit for Enterotoxigenic E.coli

作　　者：程福亮[1] 陈甜甜[1] 姜艳萍[1] 聂兆晶[1] 刘洋庆[1] 夏晓飞[1] 谷巍[1] 

机构地区：[1]山东宝来利来生物工程股份有限公司,山东泰安271000

出　　处：《畜禽业》2013年第11期12-15,共4页Livestock and Poultry Industry

摘　　要：为建立一种简单、快速、灵敏、准确的产肠毒素大肠杆菌检测方法,根据产肠毒素大肠杆菌LT、ST1、ST2保守基因序列分别设计合成了1对引物,建立了快速检测产肠毒素大肠杆菌的多重PCR方法,并对反应条件进行优化,组装成快速检测试剂盒。特异性检验表明参考菌株均可扩增出LT(282 bp)或ST1(183 bp)或ST2(360bp)的特异性条带,而非大肠杆菌均未扩增出任何条带。检测灵敏度达66 CFU,而且稳定性良好,保质期可达12个月以上。通过对临床样品的检测,证实该试剂盒具有操作简便、快速、灵敏度高、特异性强、重复性好、稳定性高等优点。To establish a simple, sensitive, accurate and rapid multiplex-PCR method for detection of Enterotoxigenic E.coli in pigs, three pairs of specific primers were designed and synthesized according to the conserved genes LT, ST1 and ST2 of Enterotoxigenic E.coli available in GenBank, and reaction parameters were optimized to develop a muhiplex-PCR detection kit. The LT gene of 282 bp, the ST1 gene of 183 bp and the ST2 gene of 360 bp were amplified in all detected Enterotoxigenic E.coli by the multiplex-PCR and no any fragments in 11 non ETEC strains. The sensitivity of PCR was 66 CFU. It could be stored over 12 months at -20 %. The results showed that the developed kit is rapid, simple, sensitive, specific, accurate and easy for detection of clinical samples with Enterotoxigenic E.coli .

关 键 词：产肠毒素大肠杆菌 多重聚合酶链式反应 试剂盒 

分 类 号：S858.28[农业科学—临床兽医学]