亚胺培南耐药PA的基因型及耐药机制研究  

Research on genotypic characterization and anti-resistant mechanism of imipenem-resistant Pseudomonas aeruginosa

作　　者：陈宏标 苏丽贤 吴紫娟 

机构地区：[1]深圳市龙华新区疾病预防控制中心,广东深圳518109 [2]深圳市福田区妇幼保健院,广东深圳518000

出　　处：《中国热带医学》2015年第8期928-931,共4页China Tropical Medicine

摘　　要：目的了解亚胺培南耐药铜绿假单胞菌分离株的基因型特征，探讨OprD2基因及金属β-内酰胺酶（MBL）在亚胺培南耐药机制中的作用。方法2011年1月至2013年8月，收集福田区妇幼保健院的医院感染铜绿假单胞菌菌株，采用纸片扩散法进行药敏试验并筛选亚胺培南耐药菌株；脉冲场凝胶电泳法对亚胺培南耐药菌株进行基因分型，电泳结果用BionumericsV4．0软件进行聚类分析；PCR法对耐亚胺培南铜绿假单胞菌的金属β-内酰胺酶（IMP、VIM及SPM）和外膜孔蛋白OprD2基因等4种主要耐药基因进行检测。结果共分离到97株铜绿假单胞菌，其中有10株为亚胺培南耐药株，当中4株为多重耐药菌。对10株耐亚胺培南菌株进行脉冲场凝胶电泳，PFGE图谱显示10株菌分为6个型，相似度在75．7％～100．0％之间。PCR法检测到10株亚胺培南耐药菌中有7株表现为外膜孔蛋白OprD2缺失，4株携带VIM-2型金属β-内酰胺酶，其中有3株在产金属酶的同时还伴有外膜孔蛋白OprD2缺失，IMP及SPM金属酶基因未检出。结论分离的亚胺培南耐药铜绿假单胞菌的基因多态性集中，外膜孔蛋白OprD2缺失及携带VIM-2型金属β-内酰胺酶是导致该院亚胺培南耐药的主要原因。Objective To understand the molecular characterization of imipenem-resistant Pseudomonas aeruginosa, then made a research of detecting OprD2 and metallo-β-lactamase （MBL）among these collected strains. Methods Strains were collected from one tertiary hospital in Shenzhen from January 2011 to August 2013. Disc diffusion method was conducted to select imipenem-resistant strains; A Pulsed-field gel electrophoresis （PFGE） method was used for molecular typing on these isolates, then analyzed by using Bionumerics 4.0 software; Detected metallo-β-1actamase（including IMP,VIM and SPM） and outer membrane porin OprD2 by PCR on these imipenem-resistant Strains. Results A total of 97 Pseudomonas aeruginosa strains were obtained, 10 ones resistant to imipenem and 4 ones were multiple antibiotic-resistant （resistant to 3 antibiotics or more）. The PFGE result of these 10 Pseudomonas aeruginosa strains showed it can be clustered into 6 patterns, with a clustering similar value between 75.7% and 100.0%. PCR results revealed that 7 strains were loss of outer membrane porin OprD2 and 4 strains were carrying VIM-2 metallo-β-1actamase, 3 strains were loss of OprD2 in company with carrying VIM-2. No IMP and SPM metallo-β-lactamase was detected out among these 10 imipenem-resistant Pseudoraonas aeruginosa . Conclusion The results indicate that a high degree genetic homology was among imipenem-resistant Pseudomonas aezuginosa in this hospital, loss of outer menabran porin OprD2and carrying VIM-2 metallo-β-lactamase were the main reason to contribute to imipenem-resistance.

关 键 词：铜绿假单胞菌 亚胺培南 脉冲场凝胶电泳 基因型 耐药机制 

分 类 号：R378.99[医药卫生—病原生物学]