Prevalence and molecular characterization of carbapenemase-producing gram-negative bacteria from a university hospital in China
Prevalence and molecular characterization of carbapenemase-producing gram-negative bacteria from a university hospital in China
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中国某大学附属医院产碳青霉烯酶革兰氏阴性菌的流行情况及分子特征
DOI：
10.3109/23744235.2015.1094822
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发表时间：
2016-02
影响因子：
5.8
通讯作者：
Tieli Zhou
中科院分区：
文献类型：
--
作者：
Jianzhong Ye;Yapei Zhang;Chuanling Ma;Tieli Zhou
摘要背景：与碳青霉烯酶相关的革兰氏阴性菌中碳青霉烯类耐药性的不断出现促使了这项研究的启动。方法：2008年至2012年从温州某大学医院3380个床位中回收革兰氏阴性菌3139株，采用VITEK2紧凑型系统和琼脂稀释法测定其药敏。使用改良的 Hodge 检验、PCR 和测序证明了碳青霉烯酶的表型和基因型。进行接合实验以揭示抗性基因的可转移性。通过质粒分析和Southern印迹杂交研究了碳青霉烯酶基因的位置。通过脉冲场凝胶电泳（PFGE）和多位点序列分型（MLST）研究分离株的克隆相关性。结果：总体而言，3139 株分离株（71/2055 肠杆菌科细菌、510/620 鲍曼不动杆菌和 170/464 铜绿假单胞菌）中的 751 株表现出对碳青霉烯类抗生素的耐药性。在70.4%（50/71）的碳青霉烯类耐药肠杆菌科细菌中检测到碳青霉烯酶编码基因，其中包括blaKPC（80%）和blaIMP（20%）。所有接受基因型分析的鲍曼不动杆菌均呈 blaOXA-51 样阳性，并共存有 blaOXA-23 样（80.4%）和 blaIMP（7.8%）。 ISAba1 被发现位于 blaOXA-23-like 和 blaOXA-51-like 的上游。 170 株铜绿假单胞菌中有 8 株和 7 株分别携带 blaIMP 和 blaVIM。 PFGE 分析确定了某些物种中至少一种显性基因型。四种产KPC-2的肺炎克雷伯菌属于相同的序列类型ST11。携带blaKPC的质粒成功转移至受体菌株中。结论：本研究强调了中国温州与碳青霉烯酶基因相关的碳青霉烯耐药性日益流行以及流行性克隆传播所面临的挑战。
Abstract Background: The increasing emergence of carbapenem resistance in gram-negative bacteria associated with carbapenemase prompted the initiation of this study. Methods: A total of 3139 gram-negative bacteria were recovered from a 3380-bed university hospital in Wenzhou during 2008 and 2012. Antimicrobial susceptibility was determined using the VITEK2 Compact System and agar dilution method. The phenotype and genotype of carbapenemase were demonstrated using the modified Hodge test, PCR and sequencing. A conjugation experiment was performed to reveal the transferability of resistant genes. The location of the carbapenemase gene was studied by plasmid analysis and southern blot hybridization. Clonal relatedness of the isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results: Overall, 751 of 3139 isolates (71/2055 Enterobacteriaceae, 510/620 Acinetobacter baumannii and 170/464 Pseudomonas aeruginosa) exhibited resistance to carbapenem. Carbapenemase-encoding genes were detected in 70.4% (50/71) of carbapenem-resistant Enterobacteriaceae, including blaKPC (80%) and blaIMP (20%). All A. baumannii subjected to genotype analysis were positive for blaOXA-51-like and co-harboured blaOXA-23-like (80.4%) and blaIMP (7.8%). ISAba1 was found upstream of blaOXA-23-like and blaOXA-51-like. Eight and seven strains of 170 P. aeruginosa carried blaIMP and blaVIM, respectively. PFGE analysis identified at least one dominant genotype in certain species. Four KPC-2-producing Klebsiella pneumoniae belonged to the same sequence type ST11. The plasmids carrying blaKPC were successfully transferred into recipient strains. Conclusion: This study highlights the challenge of increasing prevalence of carbapenem resistance associated with carbapenemase genes and dissemination of epidemic clones in Wenzhou, China.
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