鸡白痢沙门菌等位基因特异性PCR检测方法的建立与应用  

Development and Application of Allele-specific PCR for Detection of Salmonella pullorum

作　　者：姚远 张帆[2] 郗正林 黄忠阳 周涛 匡伟 王冰心 何宗亮 王润之 

机构地区：[1]南京市畜牧家禽科学研究所,江苏南京210036 [2]南京出入境检验检疫局,江苏南京210006

出　　处：《中国家禽》2013年第5期13-17,共5页China Poultry

基　　金：南京市科技发展计划项目(2012S02005)

摘　　要：参照GenBank公布的鸡白痢沙门菌与鸡伤寒沙门菌fimH基因序列,根据两者在第37和第544位碱基的不同,设计了一对等位基因特异性引物,建立了一种鸡白痢沙门菌等位基因特异性PCR(AS-PCR)检测方法,并应用该方法对鸡白痢沙门菌临床样品进行了检测。结果表明,该AS-PCR的扩增产物大小为543bp,核酸检出限为12.6pg/μL,菌液检出限为2.3×104cfu/mL,适用于鸡泄殖腔拭子、鲜蛋、饲料和饮水中鸡白痢沙门菌的检测。该AS-PCR检测方法具有简便、快速、特异性强、敏感度高等特点,可应用于鸡白痢沙门菌的快速检测。Based on nuclei polymorphism of fimH gene sequence of Salmonella pullorum at site 37 and 544 compared with Salmonella gallinarum from GenBank,a pair of allele-specific primers was designed,and an allele-specific polymerase chain reaction （AS-PCR） method for detecting S.pullorum was developed. Clinical isolated samples of S.pullorum were detected by PCR. The results showed that the method could specifically determine S.pullorum,and the PCR product was 543 bp. Sensitivity of PCR assay for DNA and pure cell culture of S.pullorum were 12.6 pg/μL and 2.3×104 cfu/mL, respectively. This PCR method was applicable for detecting S.pullorum in cloacal swabs, eggs,feed and drinking water. Therefore,it＇s a easy,quick,specific and sensitive method,and could be applied for rapid detection of S. pullorum.

关 键 词：鸡白痢沙门菌 等位基因特异性 PCR fimH基因 

分 类 号：S858.23[农业科学—临床兽医学]