铜绿假单胞菌的GM-PFGE分型的研究  

Pseudomonas aeruginosa genotyping using genome macrorestriction analysis with pulsed field gel electrophoresis

作　　者：韩黎[1,2] 吴桂芝[1,2] 陈世平[1,2] 霍云燕[1,2] 

机构地区：[1]解放军总医院 [2]解放军第309医院

出　　处：《中华微生物学和免疫学杂志》1999年第4期338-341,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的研究全基因组ＤＮＡ稀有位点限制性内切酶酶切脉冲电场电泳图谱（ＧＭ－ＰＦＧＥ）在铜绿假单胞菌基因分型中的应用，并与表型分型比较。方法对１个月内来自两个医院的病人及环境的２０株铜绿假单胞菌进行了ＧＭ－ＰＦＧＥ图谱分析，同时进行３０种生化反应的统计分型、２３种药物的敏感性分型、胞外脂多糖的血清抗原分型；并利用数值分类软件包进行相关性比较研究。结果临床致病铜绿假单胞菌的生化表型基本稳定，但其药物抗性的获得与丢失较为明显。血清型、生化性状及药物抗性之间无明显对应关系。当２菌株ＧＭ－ＰＦＧＥ图谱条带相似系数大于８０％时，为同一菌株的不同克隆亚型，当相似系数在２５％～７０％之间时，则为不同菌株。结论ＧＭ－ＰＦＧＥ分析可显示染色体结构的区域多型性，其重复性好，分辨率明显高于表型分型，结果可靠，必将成为铜绿假单胞菌或其它病原微生物分子流行病学研究的有力工具。Objective To apply genome macrorestriction pulsed field gel electrophoresis (GM PFGE) for genotyping the Pseudomonas aeruginosa isolates, and with which patterns of phenotypes of the individual isolates are compared. Methods Twenty P.aeruginosa strains collected from patients in two hospitals and the environment of one hospital within 1 month period were analysed with GM PFGE. The individual genotypes were compared with patterns of resistance to 23 antimicrobial agents, patterns of 30 biochemical properties and lipopolysaccharide (LPS) serotypes in MINTS software. Results The biochemical properties of 20 P.aeruginosa strains were little different, whereas, the acquisition or loss of resistance to antimicrobial agents was distinct and there were no corresponding relations between biochemical properties, resistance to antimicrobial agents and serotypes. Quantitative comparison of similarities of macro restriction fingerprinting patterns disclosed two discrete ranges. Those that the similarity of patterns was equal to or over 80% could be subclones of the same strain, and those with 20% 70% similarities could be different strains. Conclusions By indexing regional polymorphisms throughout the chromosome structure, GM PFGE can monitor subclonal evolution of P. aeruginosa and identify isogenic resistance mutants. With its higher discrimination power than phenotyping and its good reproducibility, it has been a reliable tool of molecular epidemiology and nosocomial research for P.aeruginosa and other pathogens.

关 键 词：分型 铜绿假单胞菌 基因脉冲电泳 相似性系数 

分 类 号：R378.991[医药卫生—病原生物学]