摘要
奶牛乳房炎是引起奶牛业经济损失的一种重要疫病,目前还没有快速、特异检测奶牛乳房炎主要致病原的方法。本试验根据金黄色葡萄球菌、无乳链球菌、大肠杆菌各自保守的16S或23S rRNA基因序列,合成了3对特异性引物,建立了三重PCR检测方法。特异性试验表明,该方法对所有参与测试的金黄色葡萄球菌、无乳链球菌和大肠杆菌都能扩增出各自的阳性条带,而对所有参与测试的对照菌株则不能扩增出任何条带。敏感性试验表明该方法能检测到4个菌的金黄色葡萄球菌、无乳链球菌和2个菌的大肠杆菌。对送检的乳房炎奶样36份直接进行PCR检测,金黄色葡萄球菌阳性7份,无乳链球菌阳性2份,大肠杆菌阳性6份。
Bovine mastitis could result in great loss in dairy industry, and there is no rapid and specific detection method for pathogens of bovine mastitis. Three pair primers were designed and synthesized from 16S or 23S rRNA sequence for each of pathogens. Multiplex PCR was set to detect Staphylococcus aureus （S. aureus ） , Streptococcus agalactiae （ S. agalactiae ） , and Escherichia coli（ E. coli ） with three pair primers. 1 319 bp in length of S. aureus, 663 bp in length of E. coli, 406 bp in length of S. agalactiae were amplified. Specific test results showed that all test strains of S. aureus, S. agalactiae and E. coli were positive. Other bacterial strains as a contrast were not amplified. The multiplex PCR using three pair primers was able to detect at least 4 bacteria of S. aureus, S. agalactiae and 2 strains of E. coli for detection of clinical samples, 36 milk samples from different herds were directly detected. 7 samples were positive for S. aureus, 2 samples were positive for S. agalactiae and 6 samples were positive for E. coli by this multiplex PCR.
出处
《畜牧与兽医》
北大核心
2006年第7期4-7,共4页
Animal Husbandry & Veterinary Medicine
基金
广西水产畜牧局科研计划资助(2004-233-5)
