临床产ESBLs大肠埃希菌整合子检测及基因型分析  

Detection of Integrons and Genotypes of Clinical ESBLs-Producing E. coli Strains

作　　者：蓝惠华 张玲 王玮玮 杨艳 程玲 骆园园 金丹丹 王厚照 Lan Huihua;Zhang Ling;Wang Weiwei;Yang Yan;Luo Yuanyuan;Jin Dandan;Wang Houzhao(PLA 174 Clin.Coll.of Anhui Med.Uni.,Xiamen, Fujian 361003,China)

机构地区：[1]厦门大学附属成功医院检验科,福建厦门361003

出　　处：《微生物学杂志》2017年第5期71-76,共6页Journal of Microbiology

基　　金：福建省自然科学基金面上项目(2013D006);全军医学科技创新项目(13QNP047);南京军区医学科技创新项目(2013ZD27);南京军区医学科技创新项目(14MS08)

摘　　要：了解临床分离的产ESBLs大肠埃希菌(ESBLs-producing Escherichia coli,ESBL-EC)中Ⅰ、Ⅱ、Ⅲ类整合子及ESBL-EC基因型的分布。收集2014年1月至12月某三甲医院住院患者分离的大肠埃希菌,经全自动细菌分析系统鉴定并检测其对临床常用抗菌药物的耐药性,双纸片协同试验确定ESBL-EC,采用聚合酶链反应(PCR)对整合子基因和ESBLs基因进行检测。K-B法比较整合子阳性菌株与阴性菌株的耐药率。结果发现,98株临床非重复ESBL-EC对青霉素类、头孢菌素类、喹诺酮类、单环酰胺类和庆大霉素耐药率均大于50%,对妥布霉素耐药率为31.62%,对呋喃妥因、阿莫西林/克拉维酸和阿米卡星较敏感,分别为11.11%、13.4%和6.12%;对碳青霉烯类抗菌素、替加环素和哌拉西林/他唑巴坦敏感率为100%。98株菌中检出47株含Ⅰ类整合子(47.96%),3株含Ⅱ类整合子(3.06%),所有菌株中有1株同时含Ⅰ类和Ⅱ类整合子,未检出Ⅲ类整合子。整合子阳性菌株对四环素和复方新诺明的耐药率高于整合子阴性菌株(P<0.05)。98株菌中β-内酰胺酶基因TEM、CTX-M-9、CTX-M-1、CTX-M-2和SHV阳性率分别为62.24%、53.06%、32.65%、4.08%和3.06%,ESBL-EC基因分型分布以TEM合并CTX-M-9型(共30株)最多见,占30.61%。结果表明,Ⅰ类整合子在产ESBLs大肠埃希菌中分布广泛,本研究尚不足以证明整合子的存在可影响ESBL-EC菌株抗生素耐药水平。同时携带TEM和CTX-M-9基因是安徽医科大学解放军174临床学院产ESBLs大肠埃希菌临床耐药菌株产生的主要原因。The distribution of class I,II and III integrons and the genotype of ESBLs-producing E.coli(ESBL-EC)strains isolated from clinical was studied.E.coli strains isolated from in-patients in a tertiary hospital from Jan.1to Dec.312014were collected.A France Bio-Merieux company VITEK-2compact automatic bacterial identification instrumentwas employed to identify the bacteria and determine antibiotic resistance of the E.coli.According to the NCCLS(2005),the double disk confirmatory tests were adopted to detect ESBLs.The polymerase chain reaction(PCR)was used to detect the integrons and ESBLs gene types.Comparison of drug resistance rates of the positive and negativestrains with the K-B method.The results showed that the resistance rates of penicillin,cephalosporin and quinolone,monocyclic amide and gentamycin were higher than50%in average,resistant rate to tobramycin is31.62%,to nitrofurantoin,amoxicillin/clavulanic acid and amikacin were11.11%,13.4and6.12%respectively.The sensitiverates to carbapenems,tigecycline and piperacillin/tazobactam were100%.98of the strains,47strains(47.96%)were detected containing class I integron,3strains(3.06%)containing class E integron,one of them both containing classes I and class E integron,however.class H integron was not detected.The resistance rates of tetracycline andcotrimoxazole to integron positive strains were significantly higher than those to integron negative strains(P<0.05).Of the98ESBL-EC strains,the positive rates of genes TEM,CTX-M-9,CTX-M-1,CTX-M-2and SHV were62.24%,53.06%,32.65%,4.08%and3.06%respectively.The distribution of genotype in ESBL-EC strains was TEM-1fused with CTX-M-9type was the commonest in the hospital,accounted for30.61%.The distribution of class I integronin ESBL-EC was wide.This study would not be enough yet to prove the existence of the integron will affect the antibiotic resistance level of ESBL-EC strains.Simultaneously carrying TEM and CTX-M-9genes may have been themain reason that caused ESBL-EC strains clinical drug resistance to produce.

关 键 词：大肠杆菌 Β内酰胺酶类 整合子类 聚合酶链反应 

分 类 号：Q939.93[生物学—微生物学] R378.2[医药卫生—病原生物学]