3种奶牛乳房炎主要病原菌PCR检测方法的建立

Development of PCR Methods for Detection of Three Main Pathogens Isolated from Bovine Mastitis

奶牛乳房炎是影响奶业健康发展的一种重要细菌性疾病,迫切需要建立一种敏感、特异的检测方法,以实现奶牛乳房炎的快速、准确诊断。现根据奶牛乳房炎主要致病菌16～23SrRNA之间的序列设计引物,建立检测金黄色葡萄球菌、无乳链球菌、停乳链球菌等3种奶牛乳房炎主要致病菌的PCR检测方法。用建立好的PCR检测方法对临床患病奶牛获取的31份样品进行特异性和敏感性检测。结果显示:PCR检测方法能快速、特异地检测出3种奶牛乳房炎主要致病菌,从而确定致病菌型。对金黄色葡萄球菌、无乳链球菌、停乳链球菌的检测灵敏度分别为1×106cfu/mL、1×106cfu/mL、1×103cfu/mL。因此,该方法可作为一种常规检测的替代方法。

Bovine mastitis was an important bacterial disease that influences the development of milk industry. To improve rapid and accurate diagnosis of mastitis in diary, establishment of a sensitive and special method was necessary. The primers were designed according to the special sequence of the 16S to 23S rRNA spacer regions, and a polymerase chain reaction （PCR） assay was developed for the detection of Staphylococcus aureus, Staphylococcus agalactiae and Staphylococcus dysgalactiae which were three major bacterial causes of bovine mastitis. Then, the pathogenic bacteria strain was isolated from the 31 milk samples and detected by PCR. The results indicated that PCR assay could detected three main pathogenic bacteria isolated from bovine mastitis and confirm the pathogen. The sensitivity of the method for detecting the Staphylococcus aureus, tiae and Staphylococcus dysgalactiae was 1×106 cfu/mL, 1×105 cfu/mL and 1×103 cfu/mL, respectively, These data suggested the method could be used as an alternative in routine diagnosis