变性高效液相色谱分析浙江沿海地区ESBLs基因类型研究  

Genotype of extend spectrum β-lactamases by denaturing high performance liquid chromatography in littoral area of Zhejiang

作　　者：陈弘 郑昌华 王爱萍 江金彪 吴福根 秦涛 林应荣 茹彩旺 王玲 

机构地区：[1]温岭市第一人民医院儿科,浙江温岭317500

出　　处：《中华医院感染学杂志》2011年第21期4415-4418,共4页Chinese Journal of Nosocomiology

基　　金：温岭市科技计划资助基金(07WLCA0048)

摘　　要：目的运用变性高效液相色谱(DHPLC)技术,检测超广谱β-内酰胺酶(ESBLs)基因型,了解浙江沿海地区ESBLs基因型分布情况。方法对171株多药耐药肠杆菌科细菌进行表型确证试验,采用多重PCR对标准菌株和ESBLs表型阳性的临床菌株进行CTX-M扩增,扩增产物经DHPLC分析,通过与标准菌株色谱峰比对进行临床菌株基因分型。结果 171株多药耐药肠杆菌科细菌,有142株临床菌株ESBLs表型阳性,经多重PCR扩增142株肠杆菌科109株携带CTX-M基因,52株为CTX-M-1产物,57株为CTX-M-9产物,检出率达79.80%,其中大肠埃希菌携带CTX-M基因比例最高,其次是肺炎克雷伯菌和阴沟肠杆菌;109株临床菌株PCR产物经DHPLC分析检测出4种CTX-M基因型,33株携带CTX-M-3、19株携带CTX-M-15、5株携带CTX-M-9、52株携带CTX-M-14。结论运用DHPLC技术检测出CTX-M型ESBLs菌株有4种基因型,其中CTX-M-14型是CTX-M型ESBLs主要型别;该地区细菌携带CTX-M型ESBLs以大肠埃希菌、肺炎克雷伯菌、阴沟肠杆菌最多见。OBJECTIVE To investigate the genotype distribution of CTX-M-type extend spectrum β lactamases （ESBLs） by denaturing high-performance liquid chromatography （DHPLC） in littoral of Zhejiang. METHODS A total of 171 multidrug resistant Enterobacteriaceae isolates were collected from our hospital. The CTX-M geaes of standard strains and clinical ESBLs-producing isolates were amplified by multiplex PCR followed by DHPLC and genotype determination. RESULTS Totally 142 ESBLs-producing isolates were confirmed by phenotypic confirmatory tests from 171 multidrug-resistant Enterobacteriaceae isolates. 109 isolates carried CTX-M genes among 142 ESBLs-producing isolates, and the rate was 76.8% （109/142）. The 109 clinical isolates included 52 from group CTX-M-1, 57 belonging to group CTX-M-9. The proportion of the Escherichia coli carring CTX-M gene was the highest, followed by Klebsiella pneumoniae and Enterobacter cloacae. The 109 clinical isolates were found present four different CTX-M genotypes by DHPLC, including CTX-M-3 （33 isolates）, CTX-M-15 （19 isolates）, CTX-M-9 （5 isolates） and CTX-M-14 （52 isolates）. CONCLUSION Genotyping of the CTX-M genes were conducted by DHPLC in littoral of Zhejiang for the first time. Four different subtypes of CTX-M-type ESBLs have been detected, which are CTX-M-3, CTX-M-15, CTX-M-9 and CTX-M-14. Among them,CTX-M-14 was the main subtype. Many species of Enterobaeteriaceae isolates are found present CTX-M ESBLs in littoral of Zhejiang and E. coli, K. pneuraoniae and E. cloaca are the most common CTX-M ESBLs-producing species.

关 键 词：变性高效液相色谱 基因分型 超广谱Β-内酰胺酶 肠杆菌科 

分 类 号：R378[医药卫生—病原生物学]