产ESBLs肺克雷伯菌基因型分析.docVIP

产ESBLs肺炎克雷伯菌基因型分析 摘要目的分析肺炎克雷伯菌临床分离株产超广谱β-内酰胺酶(ESBLs)的主要基因型。方法用表型确定临床分离产ESBLs的肺炎克雷伯菌34株，分别用TEM、SHVCTX-M-1、CTX-M-2和CTX-M-9引物进行聚合酶链反应(PCR)扩增并对PCR产物进行序列分析。结果PCR扩增结果显示TEM、SHV、CTX-M-1、CTX-M-2和CTX-M-9的阳性率分别为47.％、85.％、26.％、32.％和52.9％，CTX-M型阳性率88.24%，序列分析证实扩增为目的产物。结论34株ESBLs肺炎克雷伯菌的主要基因型是CTX-M型和SHV型，且同时携带2种以上基因占所测菌株的82.%， 关键词肺炎克雷伯菌；超广谱β-内酰胺酶；基因分型Genotype analysis of extended-spectrum β-lactamases(ESBLs) producing Klebsiella pneumoniae isolated from clinic WEI Jie-hong, HUANG Lie, LIN Guang-cheng, LIU Jian, YANG Lai-zhi and LU Xue-dong (Laboratory Department of The Affiliated shenzhen Futian Hospital of the Medical College of Guangdong ,Shenzhen 518033, Guangdong, China) Abstract:Objective To investigate the genotypical characteristics of extended-spectrum β-lactamases(ESBLs) producing clinical isolates of Klebsiella pneumoniae. Methods 34 strains of ESBLs producing isolates of Klebsiella pneumoniae with confirmed phenotype were used. PCR were performed to them by using primers specific for TEM,SHV,CTX-M-1,CTX-M-2 and CTX-M-9 respectively，and the PCR products were confirmed to be objective products by DNA sequencing. Results The PCR positive rate was 47.06% for TEM, 85.29% for SHV, 26.47% for CTX-M-1, 32.35% for CTX-M-2, and 52.94% for CTX-M-9 respectively. Among them, the CTX-M-type was 88.24% totally. Conclusions CTX-M and SHV were the prevalent genotype of 34 ESBLs producing isolates of Klebsiella pneumoniae. Among all strains, 82.35% carry more than 2 ESBLs genes. Key wolds:Klebsiella pneumoniae;Extended-spectrum beta-lactamases(ESBLs);Gene typing 基金项目：深圳市科技计划重大项目（编号：JH200505260279A） 作者简介：韦洁宏（1983-）,男，本科，检验技师，从事病原微生物分子生物学检验工作。 *通讯作者：陆学东，男，研究员，广东医学院硕士生导师，长期从事病原微生物分子生物学研究工作。 联系方式：0755E-mail地址：luxuedong2004@超广谱β-内酰胺酶(Extended-Spectrum Beta-Lactamases ESBLs)是由质粒介导的能赋予细菌对多种β-内酰胺类抗生素耐药的一类酶，它主要由革兰阴性杆菌产生[1]。它的产生及播散给临床抗感染治疗带来极大挑战，是新一代β-内酰胺类抗生素耐药的主要原因。其中肺炎克雷伯菌是临床最常见的产ESBLs菌。为此，我们以院分离的阳性菌株为对象，对其进行基因分型，为临床治疗、预防产ESBLs细菌引起的感染提供一定的参考依据。1 材料和方法1.1 菌株来源取自我院微生物实验室2005