重症监护病房耐亚胺培南铜绿假单胞菌耐药机制和分子分型的研究

doi:10.3969/j.issn.1006-3110.2021.04.010

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摘要目的分析重症监护病房(intensive care unit, ICU)耐亚胺培南铜绿假单胞菌的耐药特征及其耐药机制。方法收集 2018年1月—2019年9月广州市某医院ICU分离的37株耐亚胺培南铜绿假单胞菌(imipenem resistant Pseudomonas aeruginosa, IRPA),使用微生物鉴定药敏分析系统分析菌株的抗菌药敏感性,脉冲场凝胶电泳(pulsed-field gel electrophoresis, PFGE)进行聚类分析,PCR检测金属β-内酰胺酶基因以及oprD2基因,并对阳性株进行测序。结果 37株IRPA的抗菌药敏感性结果显示对β-内酰胺类抗生素的耐药率均很高,而氨基糖苷类和喹诺酮类抗菌药的耐药率很低。37株IRPA有11株检出携带金属酶基因,其中7株blaVIM-2,4株blaIMP-9。oprD2缺失和有意义突变合计33株,占89.2%。PFGE 分析结果显示菌株高度克隆多样性,但携带金属酶基因的11株菌被分成了两个带型簇。结论本实验证实oprD2基因缺失,发生有义突变,是铜绿假单胞菌对亚胺培南耐药的主要机制;携带blaVIM-2和blaIMP-9的IRPA菌株有在ICU引起暴发流行的风险。

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	周勇
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	胡玉山
	吴新伟

关键词 ：铜绿假单胞菌, 亚胺培南, 耐药, 金属β-内酰胺酶, oprD2基因, 脉冲场凝胶电泳

Abstract：Objective To analyze the antimicrobial resistance characteristics and mechanisms of imipenem-resistant Pseudomonas aeruginosa isolates in intensive care units (ICUs). Methods Thirty-seven imipenem-resistant Pseudomonas aeruginosa isolates were obtained from ICUs of a hospital in Guangzhou from January 2018 to September 2019. The minimum inhibitory concentrations (MICs) of the isolates were determined by microbial identification and drug sensitivity analysis system. Pulsed-field gel electrophoresis (PFGE) was used to perform clustering analysis. Polymerase chain reaction (PCR) was applied to determining metallo-β-lactamase (MBL) production and oprD2 gene, and the positive isolates were sequenced. Results All 37 IRPA isolates were resistant to the most beta-lactam antibiotics, but susceptible to aminoglycosides and quinolones. Eleven of the 37 IRPA isolates were MBL producers, including 7 isolates with blaVIM-2 and 4 isolates with blaIMP-9. The sequencing results of oprD2 showed that the disrupted oprD2 mutations rate was as high as 89.2%(33/37). PFGE revealed a high level of clonal diversity, but there were two clones represented in 11 MBL producers. Conclusions This study confirmed that imipenem resistance was driven by oprD2 inactivation. The IRPA isolates carrying blaVIM-2 or blaIMP-9 may be prevalent in ICUs.

Key words： Pseudomonas aeruginosa imipenem drug resistance metallo-β-lactamase oprD2 gene pulsed-field gel electrophoresis

收稿日期: 2020-03-12

R378.99+1

基金资助:广州市医学重点学科建设项目(No.2017-2019-07)

通讯作者: 吴新伟,E-mail:Tomwu@126.com。

作者简介: 周勇(1978-),男,硕士研究生,副主任技师,研究方向:细菌耐药。

引用本文:

周勇, 和鹏, 侯水平, 陶霞, 张晶, 胡玉山, 吴新伟. 重症监护病房耐亚胺培南铜绿假单胞菌耐药机制和分子分型的研究[J]. 实用预防医学, 2021, 28(4): 446-449. ZHOU Yong, HE Peng, HOU Shui-ping, TAO Xia, ZHANG Jing, HU Yu-shan, WU Xin-wei. Antimicrobial resistance mechanism and molecular subtyping of imipenem-resistant Pseudomonas aeruginosa isolates in intensive care units. , 2021, 28(4): 446-449.

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