仔猪水肿病的PCR快速诊断  

Rapid diagnosis for porcine edema disease by PCR

作　　者：成大荣[1] 朱善元[2] 高小攀[1] 王芳[1] 钱金梅[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏畜牧兽医职业技术学院,江苏泰州225300

出　　处：《中国兽医学报》2009年第11期1399-1401,1405,共4页Chinese Journal of Veterinary Science

基　　金：扬州大学科技创新培育基金(2007CXJ025);泰州市科技发展计划(TL0703)

摘　　要：根据GenBank发表的志贺毒素2型变异体（shiga-toxin 2e，Stx2e）的基因序列设计1对特异性引物，建立了一种检测STEC的PCR方法。通过对已知毒素类型参考菌株的扩增，证明所建立的PCR方法能够特异鉴定STEC。采集临床疑似仔猪水肿病未死亡病例（5例）的直肠棉拭子样品和死亡病例（15例）的十二指肠病料，接种LB肉汤于37℃增茵培养4～6h后，运用所建立的PCR方法对增菌培养物进行快速检测，结果表明有19例为STEC感染。通过与细菌分离后再鉴定的比较，证明了该诊断方法具有的速度快、栓出率高的特点，尤其适合于临床活体检测。Porcine edema disease is a common and important disease in postweaning piglets, which is caused by shiga-toxin 2e-producing Escherichia coli （STEC）. This disease are mainly diagnosed by clinical symptom and detailed body dissection. To further establish a rapid diagnosis method for porcine edema disease, one set of primers was designed according to the sequences of the shiga-toxin 2e （Stx2e） genes published in the GenBank. To evaluate its specificity in distinguishing STEC, the PCR was performed by using template DNAs from standard Stx2e^＋ and Stx2e^-E. coli as controls, and the data showed that the expected products of 454 bp were only detected in the Stx2e^＋ E. coli. A total of 20 samples were obtained from 5 cases of dying and 15 cases of died piglets which were suspected of edema disease. Following with first cultivating in LB broth at 37℃ for 4 - 6 h,all the samples were de- tected by the PCR method,and the data showed that 19 cass were infected with STEC. The PCR method is specific, easier and more rapid for diagnosing porcine edema disease with higher detection rates than that of the bacterial isolation and identification.

关 键 词：仔猪 水肿病 Stx2e PCR 快速诊断 

分 类 号：S855.12[农业科学—临床兽医学]