碳青霉烯类耐药肺炎克雷伯菌基因型检测和同源性分析  

Analysis of genetic homology and genotyping in Carbapenems-resistant Klebsiella pneumonia

作　　者：杨丽君[1] 陈坚[2] 杨燕[2] 林迪[2] 成军[2] 陈东科[3] 曾贤铭[2] 孙长贵[2] 

机构地区：[1]江苏大学基础医学与医学技术学院临床检验诊断学专业,镇江212013 [2]解放军第一一七医院检验科暨南京军区医学检验质量控制中心 [3]卫生部北京医院检验科

出　　处：《中华检验医学杂志》2013年第4期318-323,共6页Chinese Journal of Laboratory Medicine

基　　金：基金项目：南京军区医学科技创新重点课题（102037）

摘　　要：目的探讨临床分离的碳青霉烯类耐药肺炎克雷伯菌基因分型和菌株同源性。方法收集2011年1月至2012年6月临床分离的碳青霉烯类耐药肺炎克雷伯菌175株，采用MIC法检测菌株对药物的敏感性，纸片法表型确证试验检测超广谱β内酰胺酶（ESBL），改良Hodge试验检测碳青霉烯酶表型，PCR、DNA测序以及BLAST比对等方法确定菌株耐药酶基因型，重复序列聚合酶链反应（REP—PCR）分析菌株同源性。结果175株肺炎克雷伯菌中Hodge试验阳性140株，阳性率80．0％（140／175）。102株携带blaKPc-2基因，占58．3％（102／175），6株携带blalMP-4基因，占3．4％（6／175）。11．4％（20／175）菌株携带blaDHA-1基因。28．6％（50／175）菌株ESBL表型试验阳性。有89．1％（156／175）菌株携带bla…基因，63．4％（111／175）菌株携带blaTEM基因，50．3％（88／175）菌株携带blaCEx-M基因。66．9％（117／175）菌株同时具有3种以上耐药基因。REP—PCR指纹图谱扩增条带数3—8条，产物最长1600bp，最短200bp，分为12个谱型，其中以G型为主，有71株（40．6％），其次为F型有35株（20．0％）。结论碳青霉烯类耐药肺炎克雷伯菌以blaKPC-2基因型为主，其次为blaIMP-4基因型；ESBL以blaCRX-14基因型为主。在某些医院存在克隆株流行，北京A医院与杭州地区流行株不同，北京A医院以G型为主，杭州地区以F型为主。f中华检验医学杂志，2013，36：318—323）Objective To investigate genotyping and homology of Carbapenems-resistant Klebsiella pneumonia isolated from clinical specimens. Methods A total of 175 clinical isolates of Carbapenems- resistant Klebsiella pneumoniae were isolated from clinical specimens from January 2011 to June 2012. Antimicrobial susceptibility testing was performed by MIC method according to guidelines of CLSI. Extended-spectrum β-1actamases （ESBLs） were detected by phenotypie confirmatory test. The carbapenemase were confirmed by modified Hodge test （MHT）. 13-laetamases genes were amplified by polymerase chain reaction （PCR）, the PCR products were subjected direct sequencing of both directions, BLAST were preformed to confirm the genotypes of [3-1actamases. DNA fingerprinting based on repetitive- sequence-based PCR （REP-PCR） to investigate the homology of Carbapenems-resistant Klebsiella pneumonia. Results Of 175 chnical isolates of earbapenems-resistant Klebsiella pneumonia, 80.0% （ 140/ 175 ） strains appeared positive response of MHT. PCR and sequencing analyses showed that 58. 3% （ 102/ 175 ） strains harbored blaKPC-2 genes, 3.4% （ 6/175 ） harbored blaIMp-4 genes. There was 1 l. 4% （ 20/175 ） strains which harbored blaDHA-l genes, and 28.6% （50/175） strains which produced ESBLs by phenotypic test. There was 89. 1% （ 156/175 ） strains which harbored blasHv genes, and 63.4% （ 111/175 ） strains harbored blaTEM genes. There was 50.3% （ 88/175 ） strains which harbored blacTx-M genes. More than three kinds of resistant genes were detected simuhaneously in 66. 9% （117/175） strains. DNA fingerprintingshowed that 3 to 8 discernible DNA bands ranging in size from 200 bp to 1600 bp were generated by REP- PCR amplification. According to the results, the 175 strains were categorized into 12 types of gene pattern, in which 71 （40. 6% ） strains were pattern G,followed by 35（20. 0% ） strains were pattern F. Conclusions The major genotype causing Carbapenems resistance in Klebsiella pneumoniae

关 键 词：克雷伯菌 肺炎 卡巴配能类 抗药性 细菌 基因型 B内酰胺酶类 

分 类 号：R978.11[医药卫生—药品]