猪源粪肠球菌和屎肠球菌多重PCR快速鉴定方法的建立  

Development of a Multiplex PCR Assay for Detection of Swine-originated E.faecalis and E.faecium Isolates

作　　者：王亚宾[1] 陈丽颖[1] 胡慧[1] 段志刚[1] 崔保安[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《动物医学进展》2010年第S1期127-131,共5页Progress In Veterinary Medicine

摘　　要：粪肠球菌和屎肠球菌是引起猪感染发病的优势肠球菌种,以肠球菌的16 S rRNA基因设计属特异性引物,利用SodA基因多态性设计种特异性引物,同时优化反应条件,建立了能同时测定猪源粪肠球菌和屎肠球菌多重PCR方法。通过对来源于猪的临床菌株、粪便菌株和鲜猪肉菌株进行测定,均能成功扩增出属特异性片段和种特异性片段。经过与快速生化鉴定试剂盒(Vitek-32)和16 S rRNA测序方法比较,多重PCR与16 S rRNA测序方法对猪的临床菌株、粪便菌株和鲜猪肉菌株的鉴定符合率100%;与Vitek-32鉴定符合率为62.3%,其中,与分离于感染猪的临床菌株符合率仅有46.7%,特别是感染猪的屎肠球菌,符合率仅为22.3%。A method of multiplex PCR assay was developed for the quick detection of swine-originated Enterococcus faecalis and E.faecium isolates by using genus-primers based on the conserved regions 16S rRNA gene and species-speci?c primers on the SodA gene polymorphism.Genus-and species-fragments was successfully amplified from the isolates of clinical,fecal and raw flesh origins.By comparison,the multiplex PCR results showed a 100% coincidence rate to those identified by the 16S rRNA sequencing;whilst the coincidence rate between the PCR and Vitec-32 identification results was only 62.3%,the clinical isolate showing a coincidence rate of 46.7% and it was especially low among the E.faecium isolates(22.3%).

关 键 词：猪 肠球菌 多重PCR 16S RRNA VITEK-32 鉴定 

分 类 号：S852.6[农业科学—基础兽医学]