牛源金黄色葡萄球菌16S rRNA分子鉴定与随机多态性扩增分型

Identification with 16S rRNA and RAPD typing of Staphylococcus aureus isolated from bovine mastitis

利用16S rRNA保守序列对引起内蒙古地区奶牛乳房炎的金黄色葡萄球菌进行分离鉴定,并且建立引物随机多态性扩增(RAPD)体系对金黄色葡萄球菌分离株进行基因分型研究。结果表明,共分离出金黄色葡萄球菌35株,RAPD结果显示这35株菌均可得到清晰的RAPD指纹图谱,扩增产物在1～8条带之间,产物大小在350～4 500 bp之间。菌株分为6个基因型,Ⅳ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,这与牛场的环境和病原菌在牛场间流行传播情况有关。研究结果为地区性奶牛乳房炎的防治及新疫苗的研制提供了可靠的理论依据。

The 16S rRNA conserved sequence was used for the identification of Staphylococcus aureus（S.aureus） strains causing bovine mastitis in Inner Mongolia area,and system of randomly amplified polymorphic DNA（RAPD） with a oligonucleotide primers OLP13 was established for research on genetic polymorphism of this S.aureus.The result show that 35 S.aureus strains were isolated and identified.All of the 35 S.aureus strains produced fingerprint distinctly and amplified out 1～8 electrophoresis strip with sizes ranging from 350～4 500 bp after RAPD.The S.aureus strains were separated to 6 genotype according to the fingerprint.The preponderant S.aureus strains in Inner Mongolia was TypeⅣ.Difference of environment and transmission of pathogenic bacteria was the main reason for the distinct discrepancy on proportion of different genotype strains in different cattle farms.These results provide a reliable theory evidence for preventive treatment and new bacteria research of regional bovine mastitis