肺炎克雷伯菌碳青霉烯酶的检测及基因型分析  

Detection of carbapenemases in Klebsiella pneumoniae and analysis of the drug resistant genes

作　　者：董大光 王健[1] 潘亚萍[1] 沈继录[1] 徐元宏[1] Dong Daguang;Wang Jian;Pan Yaping(Dept of Clinical Laboratory,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院检验科,合肥230022

出　　处：《安徽医科大学学报》2018年第11期1721-1725,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81171618)

摘　　要：目的分析耐碳青霉烯类肺炎克雷伯菌(CRKP)的耐药情况,检测常见的碳青霉烯酶的发生情况及基因型别,为临床治疗该类菌提供依据。方法采用VITEK-2型全自动微生物检测系统进行细菌鉴定和药敏试验;改良Hodge试验(MHT)、Carba NP试验和改良碳青霉烯灭活试验(m CIM)对碳青霉烯酶进行表型检测,EDTA协同试验检测金属酶;并采用聚合酶链反应(PCR)方法检测KPC碳青霉烯酶耐药基因,以及ERIC-PCR对耐药株的同源性分析。结果药敏试验显示25株CRKP仅对复方新诺明和米诺环素敏感率较高; MHT试验、Carba NP试验和m CIM试验结果一致,24株(96%) CRKP产碳青酶烯酶,1株CRKP未检出碳青霉烯酶,协同试验未检测出B类碳青霉烯酶(金属酶); 25株CRKP中有23株检测出bla KPC-2基因; ERIC-PCR分型可分为8型,其中Ⅱ型为主要的流行株,共14株,Ⅰ型3株,Ⅲ型和Ⅶ型各2株,Ⅳ、Ⅴ、Ⅵ、Ⅷ型各1株。结论 CRKP多药耐药情况严重,其碳青霉烯酶以产KPC-2型为主;改良Hodge试验、Carba NP试验和m CIM试验结果与PCR法有高度的一致性,且快速、简单,为临床提供了可靠的筛选方法。Objective To analyze the resistance of carbapenem-resistant Klebsiella pneumoniae(CRKP)to clinical common antibiotics,and to detect the commom methods for detection of carbapenemases,in order to provide the basis for the clinical treatment of such bacteria.Methods These clinical strains identificition and antibiotic suscep-tibility test were carried out with the automatic VITEK2 system.Modified Hodge test(MHT),Carbapenemase Nordmann-Poirel(Carba NP)test and modified Carbapenem inactivation method(mCIM)were usded to detect carbapenemases phenotype of carbapenems.The carbapenem class B enzymes(metallo)was tested by EDTA/IMP for joint experiments.KPC carbapenemase resistance gene was detected by PCR and the homology of resistant strains was analyzed by ERIC-PCR.Results Drug sensitivity test showed that 25 strains of CRKP had a higher susceptibility to sulfamethoxazole and minocycline.It showed that the test results were the same through comparing MHT,Carba NP test and mCIM.24 strains(96%)of CRKP contained carbapenemases.One strain of CRKP did not detect carbapenemases.Synergy experiment to test 25 strains of CRKP were not found metal enzyme-positive strains.23 strains of CRKP carrying blaKPC-2 gene were identified by PCR.ERIC-PCR results showed that 25 strains of CRKP were divided into 8 types,the main epidemic strain was typeⅡ,14 strains belonged to typeⅡ.GenotypeⅠhad 3 isolate,genotypeⅢand genotypeⅦeach had 2 strains,genotypeⅣ,Ⅴ,ⅥandⅧeach had 1 isolate.Conclusion The multi-drug resistance of CRKP is serious and carbapenemase is mainly produced KPC-2 type in CRKP.MHT,Carba NP test and mCIM can rapidly and simplely screening the carbapenemase-producing Klebsiella pneumoniae,and the result is highly consistent with PCR method,so as to provide reliable methods for clinical screening.

关 键 词：肺炎克雷伯菌 碳青霉烯酶 改良HODGE试验 CARBA NP试验 改良碳青霉烯灭活试验 耐药性 

分 类 号：R446.5[医药卫生—诊断学]