猪链球菌2型三重PCR检测方法的建立  

Development and application of triplex-PCR assay for detection of Streptococcus suis type 2

作　　者：陈健舜[1] 李肖梁[1] 田国明[1] 马有智[1] 方维焕[1] 

机构地区：[1]浙江大学动物预防医学研究所,浙江杭州310029

出　　处：《中国兽医学报》2008年第7期791-794,共4页Chinese Journal of Veterinary Science

基　　金：浙江省科技厅重点资助项目(00112226)

摘　　要：针对猪链球菌2型gdh、mrp及cps2J基因设计3对引物,建立了该菌的三重PCR方法。30μL反应体系中3组引物gdh1、gdh2,mrp1、mrp2及cps2J1c、ps2J2的最适浓度比为1∶1∶1.67。最佳反应条件为94℃3 min;94℃1 min,55℃40 s,72℃50 s,30个循环;72℃5 min。可扩增到目的基因gdh、mrp及cps2J的片段大小分别为689、532和457 bp;猪链球菌2型BHI培养物的检测下限为450 CFU;猪肉人工污染样品经4 h增菌,其检测下限仅为4 500 CFU。以上结果显示,该三重PCR方法可用于组织样品和纯培养物中猪链球菌2型的检测,具有快速、特异和简便的特点。Three pairs of polymerase chain reaction(PCR) primers were designed for the gdh、mrp and cps2J genes,and the triplex-PCR assay was developed to identify Streptococcus suis serotype 2(SS-2) both in purified cultures and pork samples.688 bp,532 bp and 459 bp DNA products were amplified from Streptococcus suis type 2 under following conditions in 30 μL reaction systems:the concentration ratio of primer pair for gdh,mrp and cps2J is 1∶1∶1.67;and the cycle detailed circumstance are 3 min of pre-incubation at 94℃,...

关 键 词：猪链球菌2型 谷氨酸脱氢酶 溶菌酶释放蛋白 荚膜多糖 PCR 

分 类 号：S854.43[农业科学—临床兽医学]