Abstract:Objective: To analyze the mechanism of imipenem resistance in Pseudomonas aeruginosa clinical isolates. Methods: 110 imipenem-resistance Pseudomonas aeruginosa clinical isolates were collected from 2 hospitals of Dongguan between 2016 and 2017, and antibiotic resistance was analyzed using VITEK2 system. PCR was performed to amplify gene oprD. The amplified products were subject to sequencing analysis. Results: 110 strains of bacteria were identified as drug-resistant strains of imipenem, of which 3 were negative for PCR amplification of oprD gene. The oprD gene sequencing of the remaining 107 positive strains showed that 7 of them had no mutation, 84 had frameshift mutations(Eight of them were found to carry insertion sequences ISPpu21, IS1394, ISPpu29 and ISPst2), and 16 with a premature stop codon which is a new termination codon in advance of a position causes its peptide chain abnormalities. The rate of the mutation was 90.91%. Conclusions: oprD gene mutations result amino change and/or frame shift,hampering the binding of imipenem and oprD was the main imipenem resistance mechanism in Pseudomonas aeruginosa in Dongguan, Guangdong Province.
[1] Tomas M, Doumith M, Warner M, et al. Livermore and neil woodford. Efflux pumps, oprD porin, ampC β-lactamase and multiresistance in pseudomonas aeruginosa isolates from cystic fibrosis patients [J]. Antimicrob Agents Chemother, 2010, 54(5): 2219~2224. [2] 冯伟云,胡錫浩,许小敏,等.烧伤病房铜绿假单胞菌耐药谱检测及分子流行病学研究[J].中华医院感染学杂志,2010,20(14):2009~2011. [3] 李方去,刘洋,杨锦红,等.儿科铜绿假单胞菌的oprD基因突变及表达对碳青霉烯类抗生素耐药的作用[J].中国卫生检验杂志,2011,21(5):1063~1066. [4] Zhi-li Fang, Li-yan Zhang, Xi Huang, et al. OprD mutations and inactivation in imipenem-resistant pseudomonasaeruginosa isolates from China [J].Infection, Genetics and Evolution, 2014(21):124~128. [5] 闫玉兰,郭世辉,梁宏洁,等.铜绿假单胞菌外膜蛋白D2与亚胺培南耐药关系的研究[J].检验医学与临床,2014,11(1):13~17. [6] 多丽波,张联博,栾英,等.哈尔滨地区铜绿假单胞菌外膜蛋白D2与亚胺培南耐药关系研究[J].临床检验杂志,2010,28(2):147~148. [7] Li H,Luo YF, Williams BJ, et al. Structure and function of OprD protein in pseudomnas aeruginosa: from antibiotic resistance to novel therapies [J]. Int Med Microbiol, 2012, 302(2): 63~68. [8] 白永强,韩博,邢亮,等.2011年至2013年河北地区医院烧伤患者铜绿假单胞菌耐药株oprD基因突变分析[J].中国病原生物学杂志,2015,10(1):61~64. [9] 颜英俊,喻华,周忠华,等.亚胺培南耐药的铜绿假单胞菌oprD基因突变的研究[J].中华检验医学杂志,2009,32(4):451~454. [10] Chi Hyun Kim, Hee Young Kang, Bo Ra Kim, et al. Mutational inactivation of oprD in carbapenem-resistant pseudomonas aeruginosaisolates from koreanhospitals[J]. 2016, 54(1): 44~49. [11] Sun X, Dennis JJ, et al. A novel insertion sequence derepressed efflux pumpexpression and preadapts pseudomonas putida S12 for extreme solvent stress [J].Bacteriol, 2009, 191: 6773~6777. [12] Chew Chieng Yeo, Chit LaaPoh. IS/394 from pseudomonas alcaligenes N.C.I.B. 9867: identification andcharacterization of a member of the IS30 family of insertion elements [J]. Gene, 1996,175: 109~113. [13] Yoneyama H,Nakae T. Mechanism of efficient elimination of protein D2 in outermembrance of imipenem-resistant pseudomonas aeruginosa [J]. Anitmicrob AgentsChemother, 1999, 37(11):2385~2390. [14] 张娟,叶聪秀,曹开源,等.耐碳青霉烯类铜绿假单胞菌金属β-内酰胺酶基因检测及其与整合子的关系研究[J].中国抗生素杂志,2013,38(9):705~710.