应用多位点VNTR分析进行铜绿假单胞菌院内感染溯源的研究

摘要

目的：应用多位点可变数目串联重复序列（VNTR）分析方法进行多重耐药铜绿假单胞菌分子分型，为由铜绿假单胞菌引发的院内感染的溯源和流行病学调研提供依据。方法从铜绿假单胞菌基因组中筛选12个VNTR 位点，设计合成引物；对98株多重耐药铜绿假单胞菌进行 PCR 扩增，产物毛细管电泳；根据产物大小计算各VNTR 位点的重复数，构建系统发育树，进行数据分析。结果12个位点的多态性指数（DI）为0．45～0．88，将98株铜绿假单胞菌分成5个群，88个基因型，同病区分离的菌株具有一定的同源性。结论多位点 VNTR 分析具有很高的分辨率，适用于铜绿假单胞菌基因分型和溯源研究，具有很好的应用前景。%Objective To conduct the molecular typing of multi‐drug‐resistance Pseudomonas aeruginosa by u‐sing the multi‐loci VNTR analysis to provide the basis for the traceability the nosocomial infection caused by Pseudo‐monas aeruginosa and epidemiological survey and research .Methods 12 VNTR loci were screened from the Pseudo‐monas aeruginosa genome and the PCR primers were designed .98 strains of multi‐drug‐resistance Pseudomonas aeruginosa were perfromed the PCR amplification by using the above primers and the products were conducted the capillary electrophoresis .The repeat numbers of various loci were calculated according to the products size and the phylogenetic tree was constructed for conducting the data analysis .Results The diversity index(DI) of 12 loci ranged from 0 .45 - 0 .88 ,and 98 strains of Pseudomonas aeruginosa were grouped into 5 clusters ,88 genotypes .The isolates from the same ward presented certain homology .Conclusion The multi‐loci VNTR analysis presents higher discrimi‐nation power and suitable for genotyping and traceability research of Pseudomonas aeruginosa ,and has good applica‐tion prospect .