奶牛乳房炎常见致病菌多重PCR检测方法的建立

Development of a Multiplex Polymerase Chain Reaction Assay for Detection of Common Pathogens in Cow Mastitis

为建立同时快速检测奶牛奶样中无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌的方法,根据无乳链球菌sip基因、停乳链球菌isp基因、乳房链球菌pauA基因和金黄色葡萄球菌nuc基因各设计1对特异性引物,建立多重PCR检测体系。结果显示,该检测方法具有高特异性,无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌敏感性分别为105、104、105、105 CFU/mL。对临床采集的460份奶样检测结果表明,建立的多重PCR体系可以用于临床上无乳链球菌、停乳链球菌、乳房链球菌和金黄色葡萄球菌感染引起的奶牛乳房炎的检测。

To establish a rapid assay for Streptococcus agalactiae ,Streptococcus dysgalactiae, Streptococcus uberis and Staphylococcus aureus detection,in this study, four pairs of specific primers were designed according to the sequences published in GenBank,by the sip gene of Streptococcus agalactiae ,isp of Streptococcus d ysgalactiae ,pauA of Streptococcus uberis and nuc of Staphylococcus aureus.Development of the PCR assay for detection of these bacteria.The result showed that the assay had the advantages of high sensitivity and good specificity, the sensitivity of S.agalactiae, S.dysgalactiae, S.dysgalactiae, S. uberis and S.aureus were 10^5 CFU/mL,10^4 CFU/mL,10^5 CFU/mL and 10^5 CFU/mL,respectively.The performance of the multiplex PCR was evaluated by examining 460 clinical mastitis milk samples.The results showed that the multiplex PCR can be used for detection these four bacteria in milk samples.