奶牛乳房炎主要致病菌PCR检测分析

PCR Detection of the Main Pathogens of Mastitis in Cows

本研究应用特异性引物,扩增出大肠杆菌16SrRNA基因、表皮葡萄球菌rdr基因、绿脓杆菌eta基因、金黄色葡萄球菌16SrRNA基因、肺炎克雷伯氏菌khe基因、溶血葡萄球菌sodA基因、乳房链球菌cpn60基因、松鼠葡萄球菌gap基因、蜡样芽胞杆菌hblA基因、奇异变形杆菌ure基因、无乳链球菌16S rRNA基因、产色葡萄球菌sodA基因等基因的特异性片段,制备含有各病原菌特异性基因片段的阳性对照质粒,应用PCR检测方法对乳房炎样品进行病原菌检测。检测结果结果表明,大肠杆菌的检出率最高(76.19%),其次为凝固酶阴性葡萄球菌、无乳链球菌(26.19%)、乳房链球菌(45.24%)、铜绿假单胞菌(40.48%)、金黄色葡萄球菌(11.90%)、肺炎克雷伯氏菌(16.67%)、蜡样芽胞杆菌(4.76%)、奇异变形杆菌(4.76%),其中凝固酶阴性葡萄球菌中产色葡萄球菌的检出率最高(57.14%)。

In this study,specific primers were used.We amplified 16SrRNA gene of Escherichia coli,rdr gene of Staphylococcus epidermidis,eta gene of P.aerosa,16S rRNA gene of Staphylococcus aureus,khe gene of Klebsiella pneumoniae,sod A gene of Staphylococcus haemolyticus,cpn60 gene of Streptococcus mammary,gap gene of Staphylococcus squirrel,hbl A gene of Bacillus cereus.Specific fragments of Proteus mirabilis ure gene,Streptococcus agalactis 16S rRNA gene,Staphylococcus chromogenes sodA gene and other genes were used to prepare positive control plasmids containing specific gene fragments of each pathogen.PCR was used to detect pathogenic bacteria in mastitis samples.The results showed that the detection rate of E.coli was the highest(76.19%).It was followed by coagulase negative staphylococcus,Streptococcus agalactis(26.19%),Streptococcus mastosus(45.24%),Pseudomonas aeruginosa(40.48%),Staphylococcus aureus(11.90%),Klebsiella pneumoniae(16.67%),Bacillus cereus(4.76%)and Proteus mirabilis(4.76%).Coagulase negative staphylococcus had the highest detection rate(57.14%).