细菌通用引物在奶牛乳房炎病原菌检测中的作用研究

为了建立PCR直接检测奶牛乳房炎致病菌的方法,探索通用引物在奶牛乳房炎致病菌检测中的应用价值。利用16S rRNA基因的高度保守性,设计并合成细菌的通用引物,采用合成的引物扩增标准菌株及患有奶牛乳房炎的奶样。结果表明,通用引物扩增7 种标准菌株,370 bp处均可得到清晰的电泳条带;通用引物PCR 可检出250 ng/L的金黄色葡萄球菌标准菌株DNA;对患有奶牛乳房炎的奶样进行扩增,370 bp处也得到了清晰的电泳条带。建立在16S rRNA基础上的通用引物在奶牛乳房炎的检测中,初步显示具有特异、快速等优点,为进一步判断细菌的种类奠定了基础。

Research in the Role of Universal Primer in Detecting Pathogenic Bacteria of Cow Mastitis

To establish a PCR method which could be used to directly detect pathogenic bacterial components in the milk of cow with mastitis and to explore the value of this method diagnosis,apair of universal primers for eubacteria targeting conserved region in 16S rRNA gene were designed. The primers were used to detect the milk of dairy cow with mastitis and seven sorts of bacteria.The sensitivity and specifity of this method were tested. Seven sorts of bacteria gave a visible band after amplication at 370 bp.The sensitivity test showed that the PCR amplication technique could detect 250 ng/L of genomic Staphylococcus aureus. Amplifying the milk of dairy cow with mastitis,we also got a visible band. Universal primers based on 16S rRNA play an important role in detecting pathogenic bacterial components in the milk of diary cow with mastitis. The preliminary study showed its speed and sensitivity. This research is the base of further judging the sorts of bacteria.