为建立快速检测引起奶牛乳房炎5种病原菌的方法，参照GenBank发表的产气荚膜梭菌、乳酸乳球菌、变形杆菌、绿脓杆菌、肠球菌序列，分别根据其16S rDN A 保守序列区间设计5对引物，进行PCR扩增。结果显示：这5对引物均能针对自身菌株扩增出目的条带，且具有较强的特异性；产气荚膜梭菌、乳酸乳球菌、变形杆菌、绿脓杆菌、肠球菌菌液D N A的最低检测量分别为3.40、0.89、0.91、3.30、3.60 ng/μL ；对临床采集的疑似患有乳房炎的奶牛所产的牛奶样品进行检测，结果显示，与传统的细菌分离鉴定相比，新建立的方法具有灵敏、高效的特性。%According to 16S rDNA conserved sequence ,referring to the GenBank sequence ,this test designed five pairs of primers ,established a PCR assay for detection of five kinds of bovine mastitis pathogens .Experimental results showed that :Five pairs of primers for their own strains could amplify positive bands ,and possessed high specificity .Sensitivity test showed that the prim-ers were able to detect Clostridium perfringens ,Lactococcus lactis ,Proteusbacillus vulgaris ,Pseudo-monas aeruginosa and Enterococcus bacteria DNA concentration of 3.40 ,0.89 ,0.91 ,3.30 ,3.60 ng/μL . The method was used to detect milk samples which was collected from suspected clinical mastitis cows . Compared with the traditional bacteria isolation and identification method ,the new method had the advan-tages of sensitivity and efficient .The results show that the new method is sensitive to detect cow mastitis pathogen and can be used in clinical practice .
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