多药耐药纹带棒杆菌的重复序列PCR分型与耐药机制的研究  

Research on repetitive sequence by PCR genotyping of multidrug-resistant Corynebacterium striatum strains and drug resistance mechanisms

作　　者：李科[1] 张德纯[1] 张名均[1] 杨金梅[1] 杨小容[1] 刘胜男[1] 李世超[1] 

机构地区：[1]重庆医科大学基础医学院病原生物学教研室分子医学与肿瘤研究中心,重庆400016

出　　处：《中华医院感染学杂志》2015年第5期965-967,977,共4页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金资助项目(31000075)

摘　　要：目的对引起医院感染暴发的多药耐药纹带棒杆菌(MDRCs)进行重复序列PCR研究,为MDRCs的亲缘性分析提供可靠的方法,并对MDRCs的耐药机制进行初步研究。方法用基因外重复回文序列-PCR(REPPCR)和肠细菌基因间共有重复序列-PCR(ERIC-PCR)技术对临床分离的32株MDRCs进行分型研究,根据细菌的耐药表型选择对应的耐药基因进行检测。结果 REP-PCR和改良ERIC-PCR分型技术分别将32株MDRCs分为6型和8型,两种方法的分辨指数分别为0.585和0.696;所有菌株中与大环内酯类抗菌药物相关的核糖体甲基化酶基因(ermX)检出率为90.6%,与四环素耐药相关的核糖体保护蛋白基因(tetW)的检出率为100.0%,菌株耐药基因的检出与耐药表型完全相符。结论重复序列PCR(rep-PCR)分型方法可用于纹带棒杆菌的分型研究及医院感染MDRCs的亲缘性分析,ERIC-PCR的分辨能力优于REP-PCR;中国重庆地区的MDRCs分别介导ermX和tet(W)基因对大环内酯类和四环素类抗菌药物高水平耐药。OBJECTIVE To study the repetitive sequence by PCR of multidrug‐resistant Corynebacterium striatum （MDRCs）strains causing prevalent nosocomial infections so as to provide reliable basis for analysis of affinity of the MDRCs and conduct the preliminary study of the drug resistance mechanisms of the MDRCs .METHODS The genotypes of 32 clinical MDRCs isolates were studied by using repetitive extragenic palindromic PCR （REP‐PCR） and enterobacterial repetitive intergenic consensus PCR （ERIC‐PCR） ,then the corresponding genes that were chosen based on the bacterial drug resistance phenotypes were detected .RESULTS Totally 6 genotypes and 8 geno‐types of the 32 MDRCs were respectively identified by using REP‐PCR and modified ERIC‐PCR genotyping tech‐niques ,of which the discrimination indexes were 0 .585 and 0 .696 ,respectively .Among all the strains ,the detec‐tion rate of the ribosomal methylase gene（ermX） associated with macrolides was 90 .6% ,the detection rate of the ribosomal protection protein gene（tetW） associated with tetracycline was 100 .0% ;the detected drug resistance genes were completely consistent with the drug resistance phenotypes .CONCLUSION The repetitive PCR （rep‐PCR） genotyping can be applied in the study of genotypes of the C .striatum strains as well as the affinity of the MDRCs causing nosocomial infection ;the ERIC‐PCR is superior to the REP‐PCR in the ability to distinguish .The ermX‐and tet（W ）‐mediated MDRCs are highly resistant to macrolides and tetracycline .

关 键 词：多药耐药 纹带棒杆菌 重复序列聚合酶链反应 

分 类 号：R378[医药卫生—病原生物学]