仔猪粪样中大肠杆菌肠毒素基因的检测  

Detection of Enterotoxin Gene in Feces of Piglets with ETEC Diarrhea

作　　者：孙磊[1] 赵立平[1] 

机构地区：[1]上海交通大学生命科学技术学院微生物分子生态学与生态基因组学实验室,上海200240

出　　处：《动物医学进展》2003年第6期89-91,共3页Progress In Veterinary Medicine

摘　　要：本实验用针对大肠杆菌肠毒素基因 L T( heat-labile)、STa( heat-stable )、STb( heat-stable )的引物直接对从猪场采集的腹泻仔猪的粪便总 DNA进行扩增。结果表明只有 STb基因被检测到。把检测到的 DNA片段克隆测序 ,得到 1 75bp的片段 ,与 Gen Bank中大肠杆菌热稳定性肠毒素基因 STb 1 0 0 %同源。通过 South-ern blot,STb基因也能够被特异、准确、灵敏地检测出来。本实验直接检测仔猪粪样 DNA中的肠毒素基因的片段 ,为致病性大肠杆菌的检测提供了一种更快捷的方法。PCR amplification was performed with total DNA from fecal samples of piglets with ETEC diarrhea on commercial farms as templates and three pairs of reported primers of enterotoxin genes which belong to LT(heat-labile),STa(heat-stableⅠ),STb(heat-stableⅡ)respectively. The result indicated that only the STb gene was detected. A fragment of 175 bp for STb gene was cloned and sequenced. BLAST analysis showed 100 percent homology of the cloned fragment with STb gene of enterotoxigenic Escherichia coli in GenBank. By using the cloned fragment as probe, STb gene was specifically detected in all fecal samples demonstrating a gradual reduction of ETEC population when the animals were recovering. This culture independent technique provides a quicker method for ETEC diagnosis in commercial settings.

关 键 词：仔猪 粪样 大肠杆菌 肠毒素 基因检测 

分 类 号：S858.28[农业科学—临床兽医学]