【作者】 潘文； 薛红； 赵明秋； 琚春梅； 乔进平； 陈金顶；

【Author】 PAN Wen~1,XUE Hong~2,ZHAO Ming-qiu~1,JU Chun-mei~1,QIAO Jin-ping~1,CHEN Jin-ding~1 (1 College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China 2 Guangzhou Animal Health Inspection,Guangzhou 510440,China)

【机构】 华南农业大学兽医学院； 广州市动物卫生监督所；

【摘要】 根据GenBank中已发表的流产布鲁氏菌种特异的omp25基因序列及牛分枝杆菌特异保守的16S rRNA加工蛋白rimM基因序列设计了2对特异性引物,通过对PCR条件的优化,建立了快速鉴别检测牛布鲁氏菌和牛分枝杆菌的双重PCR方法。对建立的方法进行特异性和敏感性试验,结果显示,在牛布鲁氏菌和牛分枝杆菌中分别扩增出448bp、269bp的特异性目的条带,作为对照的大肠杆菌、沙门氏菌、八叠球菌、金黄色葡萄球菌、巴氏杆菌、军团菌、枯草杆菌及溶血性链球菌均未扩增出任何条带。牛布鲁氏菌和牛分枝杆菌的DNA最低检出量均为10pg;牛奶样品中人工污染的牛布鲁氏菌、牛分枝杆菌的检测敏感性分别为7.9×10₃CFU、3ng/mL。更多还原

【Abstract】 A multiplex PCR assay for detection of Brucella abortus and Mycobacterium bovis infection simultaneously in bovine milk was established using two pairs of primers according to the species -specific omp25 gene of B.abortus and conserved 16S rRNA rimM gene of M.bovis.The multiplex PCR assay amplified unique 448bp and 269bp amplicons from B.abortus and M.bovis respectively,while none of 8 other common bacterial species strains revealed any amplification products.Sensitivity of genomic DNA detection of B.abortus and M.bovis were both 10pg,meanwhile the limit of detection in artificially contaminated milk with B.abortus and M.bovi were as low as 7.9×10₃CFU and 3ng/mL,respectively.更多还原