【作者】 高玉梅； 何宝祥；

【机构】 广西大学临床兽医学实验室；

【摘要】 奶牛乳房炎是成年奶牛最常见的一种传染病,是引起奶牛业经济损失的主要原因。本实验根据金黄色葡萄球菌、大肠杆菌、无乳链球菌和停乳链球菌16S-23S rRNA之间的区域以及白色念珠菌18S rRNA的序列设计了5对特异性引物,建立了检测金黄色葡萄球菌、大肠杆菌、无乳链球菌、停乳链球菌和白色念珠菌5种乳房炎主要病原茵的多重PCR方法。结果表明:本实验建立的多重PCR方法具有较好的特异性,多重PCR方法检测五种病原菌基因组DNA的灵敏性分别为:金黄色葡萄球菌2.6405ng/mL、大肠杆菌0.0264ng/mL、无乳链球菌2.5455ng/mL、停乳链球菌25.8750ng/mL、白色念珠菌43.850ng/mL。对采自临床型乳房炎(22份)和隐性乳房炎(76份)的98份乳样分别用传统细菌学培养法和多重PCR方法进行检测,二者的符合率为86.4%。更多还原

【Abstract】 Bovine mastitis(BM) is the most common infectious disease of adult cows,which is the main reason of economic loss in the world.According to the 16S to 23S rRNA space region of Staphylococcus aureus,Escherichia coli,Streptococcus agalactiae and Streptococcus dysgalaciae and 18S rRNA sequence of Candida albicans,five pairs of special primers were designed and the method of multiplex PCR to detect these five main pathogens cause bovine mastitis was established.The results showed that this method had a good specificity,and the sensitivity of multiplex PCR to detect the genomic DNA of five pathogens S.aureus,E.coli,S.agalactiae,S.dysgalaciae and C.albicans were 2.6405 ng/mL,0.0264 ng/mL, 2.5455 ng/mL,25.8750 ng/mL and 43.850 ng/mL,respectively.Ninety-eight milk samples which include clinical mastitis(22 samples) and subclinical mastitis(76 samples) have been detected by the method of multiplex PCR and compared with the traditional bacteriological culture,the results suggested that the coincidence rate of the two methods is 86.4%.更多还原