随机引物扩增DNA多态性技术在分析金黄色葡萄球菌和肺炎杆菌多态性中的应用

Application of randomly amplified polymorphic DNA on the DNA polymorphism of Staphylococcus aureus and Klebsiella pneumoniae

金黄色葡萄球菌和肺炎杆菌是医院内感染的主要条件致病菌 ,常常引起院内感染及暴发流行。本实验以一条含 10bp的随机引物将 5 2株肺炎杆菌和 2 8株金黄色葡萄球菌的DNA进行随机引物扩增DNA多态性 (RAPD)扩增 ,进行多态性分析 ,得到不同的带谱 ,在肺炎杆菌中存在很强的多态性 ,出现了 41种带型 ,通过肺炎杆菌间相似系数计算 ,绘制出表达菌株亲缘关系的系统树 ,图中可见各菌间的相似系数 ,5 2株菌分成 7个型 ;金黄色葡萄球菌出现了三条特征带。本实验说明RAPD技术能够追溯传染源 。

By using short single oligomer primer with arbitrarily chosen sequences in polymerase chain reaction, randomly amplified polymorphic DNA (RAPD) was applied in differentiating clinical isolates of 52 Klebsiella pneumoniae and 28 Staphylococcus aureus . RAPD produced multiple polymorphic DNA bands when K.pneumoniae DNA was used as template. Using DNA fingerprints technique, the similarity coefficients values of 52 K.pneumoniae was measured and calculated, and dendrogram was drawed. The 52 Klebsiella pneumoniae were divided into 7 Patterns. But in the 28 Staphylococcus aureus, 3 characteristic lands were obtained. Minor differences in the RAPD fingerprints were detected. The research findings demonstrated that RAPD has a potential as a widely useful tool in providing genotypic characters for taxonomic descriptions and molecular epidemiology study.