National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Miaoli County, Taiwan, ROC.
College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC.
J Med Microbiol. 2013 Jan;62(Pt 1):101-107. doi: 10.1099/jmm.0.050492-0. Epub 2012 Sep 20.
We investigated the molecular mechanisms and clonality of imipenem-non-susceptible Klebsiella pneumoniae isolates collected during a Taiwan national surveillance programme, between 2002 and 2009. Genes for carbapenemases, plasmid-borne ampC-type genes and extended-spectrum β-lactamase (ESBL) genes were analysed by PCR. The major porin channels OmpK35 and OmpK36 were studied by SDS-PAGE. Molecular typing was performed with pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Our study revealed that all 29 of the isolates tested were ESBL producers. Of the K. pneumoniae isolates collected in Taiwan from 2002 to 2009, most (84.6 %, 11/13) imipenem-resistant (MIC >2 mg l(-1)) isolates carried the bla(IMP-8) gene. Isolates with an imipenem MIC of 2 mg l(-1) produced ESBLs with or without DHA-1 in combination with OmpK35/36 loss. PFGE analysis revealed that six small clusters of isolates were clonally related. The MLST grouping results were in concordance with the PFGE results. The predominant sequence types (ST) were ST11, ST48 and ST101. Two novel STs, ST1033 and ST1034, were found. The dominant clone in Taiwan, ST11, has been reported worldwide to be associated with various resistance mechanisms.
我们研究了 2002 年至 2009 年间台湾国家监测计划期间收集的对亚胺培南不敏感的肺炎克雷伯菌分离株的分子机制和克隆性。通过 PCR 分析了碳青霉烯酶、质粒携带的 AmpC 型基因和超广谱β-内酰胺酶(ESBL)基因。通过 SDS-PAGE 研究了主要的孔蛋白通道 OmpK35 和 OmpK36。采用脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)进行分子分型。我们的研究表明,所有 29 株受试分离株均为 ESBL 产生菌。在 2002 年至 2009 年期间从台湾收集的肺炎克雷伯菌分离株中,大多数(84.6%,11/13)对亚胺培南耐药(MIC>2mg/L)的分离株携带 bla(IMP-8)基因。亚胺培南 MIC 为 2mg/L 的分离株产生的 ESBL 可与或不与 DHA-1 结合 OmpK35/36 缺失。PFGE 分析显示 6 个小的分离株簇具有克隆相关性。MLST 分组结果与 PFGE 结果一致。主要的序列类型(ST)是 ST11、ST48 和 ST101。发现了两种新的 ST,ST1033 和 ST1034。在台湾占主导地位的克隆,ST11,已在全球范围内与各种耐药机制有关。