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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 detect the gene distribution of the metalloenzyme gene of adriamycin-resistant Pseudomonas aeruginosa and the deletion of the OprD2 gene, and to understand the drug resistance of Pseudomonas aeruginosa. Methods A total of 1 887 non-repetitive Pseudomonas aeruginosa isolates from 2014 to 2016 were collected from the Daping Hospital of Institute of Field Surgery, Third Military Medical University. The strains were identified by VITEK-MS mass spectrometry. The minimal inhibitory concentration (MIC) Method to determine their sensitivity to 10 kinds of commonly used antimicrobial agents, the use of WHONET5.6 software for statistical analysis of susceptibility data, while the use of imipenem-EDTA paper screening of metal-enzyme-positive strains, with the polymerase chain The expression of metallo-β-lactamase (MBL) related genes (IMP, VIM, SPM, GIM, SIM and NDM) and OprD2 The case of genes. Results The numbers of Pseudomonas aeruginosa isolated each year were 756 strains (9.4%), 579 strains (9.5%) and 552 strains (10.7%) respectively, mainly from the ICU ward and respiratory medicine ward. The specimens were sputum . The resistance rates to tobramycin and gentamicin of Pseudomonas aeruginosa isolated in 2014-2016 decreased year by year with the rates of resistance dropping from 11.3% and 13.2% in 2014 to 2016 respectively The resistance rate to levofloxacin and piperacillin / tazobactam showed an upward trend with resistance rates of 7.8% and 9.0% from 10.7% and 9.2% in 2014 to 23.5% in 2016, respectively. , 11.6%. According to the distribution of genotypes, 51 strains (18.9%) of MBL gene were positive in 269 strains of imipenem-resistant Pseudomonas aeruginosa and 158 strains (58.7%) of OprD2 in outer membrane were missing. Among 29 VIM positive and 22 SPM positive, no other genotypes were detected. Conclusion The resistance rate of Pseudomonas aeruginosa to imipenem is high. The deletion of MBL and OprD2 gene is the main reason of resistance to imipenem in Pseudomonas aeruginosa.