2016-2018年东莞地区耐碳青霉烯类肠杆菌目细菌的耐药表型、耐药机制和分子流行病学分析  

Analysis of drug resistance phenotype,drug resistance mechanism and molecular epidemiological of carbapenem-resistant Enterobacteriaceae in Dongguan from 2016 to 2018

作　　者：郭主声[1] 吕飞 谢树金 黄亚 林偲思 徐宝华[1] 冯剑波 冯森 何芬 周谋清 Guo Zhusheng;LüFei;Xie Shujin;Huang Ya;Lin Sisi;Xue Baohua;Feng Jianbo;Feng Sen;He Fen;Zhou Mouqing(Dongguan Tungwah Hospital,Dongguan 523110)

机构地区：[1]东莞东华医院,东莞523110

出　　处：《中国抗生素杂志》2023年第7期798-805,共8页Chinese Journal of Antibiotics

基　　金：2019东莞市社会科技发展(重点)项目(No.201950715046197)。

摘　　要：目的通过研究东莞地区的耐碳青霉烯类肠杆菌目细菌(carbapenem-resistant Enterobacteriaceae,CRE)的耐药情况和基因分型,研究耐碳青霉烯类肺炎克雷伯菌(carbapenem-resistant Klebsiella pneumoniae,CRKP)的亲缘性、耐药机制和bla_(KPC-2)基因环境,为临床寻找控制和治疗该致病菌的更优方案提供参考。方法回顾性收集2016年1月—2018年12月东莞市人民医院、东莞东华医院、康华医院、东莞市中医院和东莞市妇幼保健院等十三所医院的住院患者的CRE菌株进行常规微生物培养、鉴定和药敏试验。以美罗培南或亚胺培南纸片法(K-B法)或最小抑菌浓度(MIC)测定法对肠杆菌目细菌进行初筛;采用改良Hodge试验、亚胺培南-EDTA双纸片协同试验和CIM试验检测CRE产酶情况;聚合酶链式反应(polymerase chain reaction,PCR)检测并鉴定CRE的碳青霉烯类耐药基因(bla_(KPC)、bla_(NDM)、bla_(IMP)、bla_(DHA)、bla_(CTX-M)和bla_(CMY)),以及CRKP的多位点序列分型(multilocus sequence typing,MLST)和孔蛋白(OmpK35、OmpK36和OmpK37);通过Junction PCR、Mapping PCR和Crossing PCR检测bla_(KPC-2)基因环境;使用WHONET 5.6软件统计分析CRE临床分离株在标本和科室中的分布与耐药情况,以及碳青霉烯酶基因在科室中的分布情况。结果从37217株肠杆菌目细菌中共检出131株CRE(占0.35%),其中肺炎克雷伯菌79株(占60.31%)、大肠埃希菌21株(占16.03%)和阴沟肠杆菌12株(占9.16%);检出CRE的临床科室主要为ICU(66株,占50.38%);在检出CRE的标本中,位于前3位分别是痰液标本(56株,占42.75%)、尿液标本(21株,占16.03%)、伤口分泌物标本(12株,占9.16%);CRE对目前临床中常使用的抗菌药物表现出高耐药性,仅对阿米卡星耐药率较低,为21.38%。检测104株CRE(包括肺炎克雷伯菌、大肠埃希菌和阴沟肠杆菌)发现的碳青霉烯酶有KPC-2型(57,54.81%)、NDM-1型(7株,6.73%)和NDM-5型(2株,1.92%);其中,CRKP主要为ST11型(49株,占62.03%),其Objective To provides a reference for better clinical control and treatment of such pathogenic bacteria in Dongguan by studying the drug resistance and genotyping of carbapenem-resistant Enterobacteriaceae(CRE),and the affinity,the drug resistance mechanism,and the bla_(KPC-2)gene environment of carbapenem-resistant Klebsiella pneumoniae(CRKP).Methods CRE from 13 hospitals including Dongguan People's Hospital,Dongguan Tungwah Hospital,Kanghua Hospital,Dongguan Traditional Chinese Medicine Hospital,and Dongguan Maternal and Child Health Hospital et al,were collected retrospectively for routine microbial culture,and identification and drug sensitivity tests from January 2016 to December 2018.Gram negative Enterobacteriaceae were screened by the meropenem or imipenem disk method(the K-B method)or the minimum inhibitory concentration(MIC)method.Enzyme production of carbapenem resistant Enterobacteriaceae was detected by the modified Hodge test,the imipenem EDTA double disk synergy test,and the CIM test.The detection and identification of carbapenem resistance genes(bla_(KPC),bla_(NDM),bla_(IMP),bla_(DHA),bla_(CTX-M),and blaCMY)of CRE,multi-locus sequence typing(MLST),and the detection of porins(OmpK35,OmpK36 and OmpK37)of CRKP were performed to analyze the affinities and the drug resistance mechanism by polymerase chain reaction(PCR);Simultaneously,the bla_(KPC-2)gene environment was detected by junction PCR,mapping PCR,and crossing PCR.The WHONET 5.6 software was used to analyze the distribution and drug resistance of CRE clinical isolates in specimens and departments,as well as the distribution of carbapenemase genes in departments.Results A total of 131 strains of CRE(0.35%)were detected from 37,217 strains of Enterobacteriaceae,including 79 Klebsiella pneumoniae(60.31%),21 Escherichia coli(16.03%),and 12 Enterobacter cloacae(9.16%).The main clinical departments detected CRE were ICU(66 strains,accounting for 50.38%).The top three samples of CRE were sputum(56 strains,42.75%),urine(21 strains,16.03%),and wound sec

关 键 词：耐碳青霉烯类肠杆菌目细菌 耐碳青霉烯类肺炎克雷伯菌 多位点序列分型 孔蛋白 bla_(KPC-2) 基因环境 

分 类 号：R978.1[医药卫生—药品]