重庆某三甲医院2014-2016年铜绿假单胞菌耐药表型及金属酶、外膜孔蛋白耐药基因型分析  

Drug resistance profiles and metallo-β-lactamase and outer membrane protein OprD2 genotypes of Pseudomonas aeruginosa: a 2014-2016 survey in a Grade 3 Class A general hospital in Chongqing

作　　者：李进[1] 胡韦维 张峰领[1] 陈鸣[1] 邓少丽[1] 鲁卫平[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所检验科,重庆400042 [2]重庆医科大学附属儿童医院检验科,重庆400014

出　　处：《第三军医大学学报》2017年第22期2200-2205,共6页Journal of Third Military Medical University

基　　金：全军医学科研"十二五"计划面上项目(CWS13J039);国家自然科学基金重点项目(81430053)~~

摘　　要：目的检测亚胺培南耐药铜绿假单胞菌金属酶基因分布和外膜孔蛋白OprD2基因缺失情况,了解铜绿假单胞菌的耐药状况。方法收集第三军医大学大坪医院野战外科研究所2014-2016年临床分离的非重复铜绿假单胞菌1 887株,VITEK-MS质谱仪进行菌株鉴定,最小抑菌浓度(minimal inhibitory concentration,MIC)法测定其对10种常用抗菌药物的敏感性,利用WHONET5.6软件对药敏资料进行统计分析,同时采用亚胺培南-EDTA纸片法筛选金属酶表型阳性的菌株,用聚合酶链反应(polymerase chain reaction,PCR)测铜绿假单胞菌金属β-内酰酶(metallo-β-lactamase,MBL)相关基因(IMP、VIM、SPM、GIM、SIM、NDM)和外膜孔蛋白OprD2基因的情况。结果 3年中铜绿假单胞菌每年的分离数量分别为756株(9.4%)、579株(9.5%)和552株(10.7%),主要来自ICU病房和呼吸内科病房,标本类型以痰液为主;2014-2016年分离出的铜绿假单胞菌对妥布霉素、庆大霉素的耐药率逐年下降,其耐药率分别从2014年的11.3%、13.2%下降至2016年的7.8%、9.0%,对左旋氧氟沙星、哌拉西林/他唑巴坦的耐药率呈上升趋势,其耐药率从2014年的10.7%、9.2%上升至2016年的23.5%、11.6%。从基因型分布情况来看,269株亚胺培南耐药铜绿假单胞菌中MBL基因阳性51株(18.9%),外膜孔蛋白OprD2缺失158株(58.7%);51株产酶菌株中VIM型阳性29株,SPM型阳性22株,未检测出其他基因型。结论铜绿假单胞菌对亚胺培南的耐药率较高;产MBL和外膜孔蛋白OprD2基因缺失是引起铜绿假单胞菌对亚胺培南耐药的主要原因。Objective To investigate the drug resistance profiles of Pseudomonas aeruginosa isolates and survey the distribution of metallo-β-lactamase（ MBL） phenotypes and outer membrane protein OprD2 gene deletion among the strains. Methods From January 2014 to December 2016,a total of 1 887 nonduplicate clinical isolates of Pseudomonas aeruginosa were collected from Daping Hospital. All the strains were identified by VITEK-MS,and their susceptibilities to 10 antimicrobial agents were detected using the minimal inhibitory concentration method. WHONET5. 6 software was used to analyze the results of drug sensitivity tests. Imipenem-EDTA disc test was used to detect the MBL phenotypes of the strains,and the MBL genes（ IMP,VIM,SPM,GIM,SIM,and NDM） and OprD2 gene were detected by PCR amplification. Results Among the total of 1 887 Pseudomonas aeruginosa strains isolated,756（ 9. 4%） were isolated in 2014,579（ 9. 5%） in 2015,and 552（ 10. 7%） in 2016; most of the strains were isolated from the sputum samples in the Department of Respiratory Medicine and ICU. From January 2014 to December 2016,the drug resistance rates of the isolated strains to tobramycin and gentamicin decreased progressively（ from 11. 3% and 13. 2% in2014 to 7. 8% and 9. 0% in 2016,respectively）,while their resistance rates to levofloxacin and piperacillin/tazobactam tended to increase（ from 10. 7% and 9. 2% in 2014 to 23. 5% and 11. 6% in 2016,respectively）. Of the 269 imipenem-resistant isolates,51（ 18. 9%） were MBL-producing strains（ including29 VIM type MBL-positive and 22 SPM type MBL-positive isolates）,and 158（ 58. 7%） showed deletion of OprD2 gene; no other genotypes were detected among the isolated strains. Conclusion We found a high incidence of drug resistance（ particularly imipenem） in the clinical isolates of Pseudomonas aeruginosa. MBL production and the loss of OprD2 gene are the main contributors to imipenem resistance in Pseudomonas aeruginosa.

关 键 词：铜绿假单胞菌 耐药性 金属酶 外膜孔蛋白OprD2基因 

分 类 号：R181.34[医药卫生—流行病学] R378.991[医药卫生—公共卫生与预防医学]