甘肃地区牛源金黄色葡萄球菌分子鉴定及RAPD分型

Identification and RAPD Typing of Staphylococcus aureus Isolated from Bovine Mastitis in Gansu Area

本研究目的是分离鉴定引起甘肃地区奶牛乳房炎的金黄色葡萄球菌,掌握其基因型情况。利用16S、23SrRNA保守序列PCR扩增对乳房炎奶样中的金黄色葡萄球菌进行鉴定,并进行RAPD基因分型。结果表明,310份奶样中共分离出金黄色葡萄球菌100株,RAPD结果显示这100株金黄色葡萄球菌均可得到清晰的RAPD指纹图谱,扩增产物在2～7条带之间,具有多种带型组成。通过聚类分析100株菌产生11个基因型,其中Ⅰ型4株,Ⅱ型4株,Ⅲ型10株,Ⅳ型13株,Ⅴ型7株,Ⅵ型24株,Ⅶ型16株,Ⅷ型6株,Ⅸ型4株,Ⅹ型10株,Ⅺ型2株。Ⅵ型为该地区的流行优势菌群,不同牛场各基因型菌株分布有明显差异。本研究说明牛场的环境与养殖条件对病原菌流行传播有明显的影响,这一结果对地区性奶牛乳房炎的防治提供了可靠的理论依据。

The aim of this study was to isolate and identify Staphylococcus aureus（S.aureus） strains causing bovine mastitis in Gansu province,and investigate genotype of this strains.16S rRNA and 23S rRNA sequence was used for PCR identification on S.aureus isolate from milk samples of bovine mastitis.Randomly amplified polymorphic DNA（RAPD） was utilized to research genotype of these strains.The results showed that 100 S.aureus strains were isolated and identified from 310 mastitis milk samples.All of the 100 S.aureus strains produced vivid fingerprint and number of electrophoresis strip was 2-7 with different banding pattern after RAPD.The S.aureus strains were separated to 11 genotype according to the cluster analysis and 4 strains were typeⅠ,4 strains were typeⅡ,10 strains were typeⅢ,13 strains were typeⅣ,7 strains were typeⅤ,24 strains were typeⅥ,16 strains were typeⅦ,6 strains were typeⅧ,4 strains were typeⅨ,10 strains were typeⅩ and 2 strains were typeⅪ.The preponderant genotype of S.aureus strains in this area was TypeⅥ.Distribution of different genotype S.aureus strains were obvious difference in different cattle farm.Environment and factor of cultivation of the cattle farm had a distinct effect on transmission of pathogenic bacteria.These results provide a reliable theory evidence for preventive treatment of regional bovine mastitis.