3种奶牛乳房炎主要病原菌PCR检测方法的建立.pdfVIP

中 (iLtt. f ~星 J且 2012， 28(17): 105-1 08 Chinese Agricultural Science Bulletin 3 种奶牛乳房炎主要病原菌PCR检测方法的建立 陈玉霞，林峰 (河南农业大学牧医工程学院，郑州1450002) 摘要:奶牛乳房夫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l 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16~23S rRNA之间的序列设 计引物，建立检测金黄色葡萄球茵、无$L链球菌、停孔链球菌等3 种奶牛乳房炎主要致病菌的PCR检测 方法。用建立好的PCR检溃l 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。结采显 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 6 3 萄冻茵、无字L链球菌、停孔链球茵的检测灵敏度分别为 1 x 10 C也JmL、 1x 1Q6 旬JmL， 1X 10 cfu/mL。因此， c 该方法可作为一种常规检圳的替代方法。 关键词:PCR; 乳房炎;金黄色葡萄球茵;无乳链球茵;停乳链球菌 论文编号;2012-1421 中圄分类号:S852.6 文献标志码:A Development of PCR Methods for Detection of Three Main Pathogens Isolated from Bovine Mastitis Chen Yuxia , Lin Feng (College OJAnimal Hu.s bandry αnd Veterinary Medicine. Henan Agricultural Univer.lity, Zhengzhou 450002) Abstract: Bovine mastitis was an important bacterial disease that influences the development u[ milk Índustry. To improve rapid and accurate diagnosis of mastitis in diary , establishment of a sensitive and special method was necessary. The primers were designed according 10 the special sequence of the 16S to 235 rRNA spacer regions , and a po!ymerase chain reactÎon (PCR) assay was developed for the detectíon of Staphylococcus 8ureus, Staphylococcus agalactiae and Staphylococcus dysgalactiae which were three major bacterial causes of bovine mastitis. Then, the pathogenic bacteria strain was iso!ated frorn the 31 milk sarnples and detected by PCR. The results indicat