铜绿假单胞菌耐药基因分析及多位点序列遗传分型

Antibiotic-resistant genes and multilocus sequencing typing of Pseudomonas aeruginosa

目的了解2011-2012年深圳市南山区医院病人,各医院、诊所内环境台面涂抹样、医护人员手涂抹样分离的铜绿假单胞菌,抗生素耐药基因分布及基因的遗传多样性。方法采用聚合酶链反应技术检测铜绿假单胞菌的20种耐药基因:TEM、VEB、CARB、OXA、SHV、PER、GES、GTX、SPM、GIM、IMP、VIM、DHA、oprD、Aac(6′)-Ⅰ、Aac(6′)-Ⅱ、Aac(3′)-Ⅰ、Aac(2″)-Ⅰ、qacE1-sull及Ⅰ类整合子基因。采用多位点序列分子分型方法进行聚类和克隆分析。结果检出11种耐药基因:TEM、SHV、IMP、DHA、Aac(6’)-Ⅰ、Aac(6′)-Ⅱ、Aac(3′)-Ⅰ、Aac(2″)-Ⅰ、qacE1-sull、Ⅰ类整合子及oprD基因,检出率分别为8.1%、6.4%、4.8%、9.7%、4.8%、14.5%、4.8%、56.5%,8.1%,8.1%,oprD基因缺失率为61.2%。52株铜绿假单胞菌检出耐药基因,形成19种耐药基因谱。多位点序列分型方法将62株铜绿假单胞菌,分为39个ST型,5个克隆群,1个优势克隆群CC244,1个优势独特型ST856。结论不同类型样本分离菌株携带耐药基因存在差异,部分病人分离株携带多种耐药基因。本研究铜绿假单胞菌具有遗传多样性,存在优势克隆群。

We investigated the antibiotic-resistant genes and genetic diversity of Pseudomonas aeruginosa from patients in hospital, the smear samples from hospital and clinic environment, and from medical staff＇ hands respectively in 2011-2012 in Nanshan District of Shenzhen, Polymerase chain reaction were used to detect the 20 kinds of antibiotic-resistant genes （TEM, VEB, CARB, OXA, SHV, PER, GES, GTX, SPM, GIM, IMP, VIM, DHA, oprD, Aac（6＇）- I , Aac（6＇）-1I , Aac （3＇）- I , Aac（2I＇） I , qacEl-sull and int- I ）. Multilocus sequencing typing was used to analyze the clonal complexes. The 11 kinds resistant genes TEM, SHV, IMP, DHA, Aac（6＇）- I , Aac（6＇） lI ,Aac（3＇）- I , Aac（2＂）- I , qacEl-sull,int- I and oprD were detected, for the positive rates respectively, and which were 8.1%, 6.4%, 4.8%, 9.7 %, 4.8%, 14.5 %, 9.7%, 56.5%, 8.1%, and 8. 1~ the loss rate of oprD gene was 61.2~. The 19 antibiotic resistance gene profiles existed in 52 Pseudomonas aeruginosa strains. Multiloeus sequencing typing found 39 sequence types and 5 clonal complexes in 62 Pseudo- monas aeruginosa strains, CC244 and ST856 were dominant. There were some differences of antibiotic resistance gene profiles between different samples, the Pseudomonas aeruginosa strains from patients carried multiple resistant genes. In our research, the Pseudornonas aeruginosa had the genetic diversity and the dominant clonal complexes existed.