Bacterial carbapenem resistance, especially when mediated by transferable carbapenemases, is of important public health concern. An increased number of metallo-β-lactamase (MBL)-producing Klebsiella pneumoniae strains isolated in a tertiary hospital in Thessaloniki, Greece, called for further genetic investigation.The study included 29 non-repetitive carbapenem resistant K. pneumoniae isolates phenotypically characterized as MBL-producers collected in a tertiary hospital in Greece. The isolates were screened for the detection of carbapenemase genes (K. pneumoniae carbapenemase (blaKPC), Verona-integron-encoded MBL-1 (blaVIM-1), imipenemase (blaIMP), oxacillinase-48 (blaOXA-48) and New Delhi MBL (blaNDM)). The genetic relationship of the isolates was determined by Random Amplified Polymorphic DNA (RAPD) analysis. The whole genome sequences (WGS) from two NDM-positive K. pneumoniae isolates were further characterized.The presence of New Delhi MBL (blaNDM) gene was confirmed in all K. pneumoniae isolates, while blaKPC and blaVIM-1 genes were co-detected in one and two isolates, respectively. The RAPD analysis showed that the isolates were clustered into two groups. The whole genome sequence analysis of two K. pneumoniae isolates revealed that they belonged to the sequence type 11, they carried the blaNDM-1 gene, and exhibited differences in the number and type of the plasmids and the resistant genes.All MBL-producing K. pneumoniae isolates of the study harbored a blaNDM gene, while WGS analysis revealed genetic diversity in resistance genes. Continuous surveillance is needed to detect the emergence of new clones in a hospital setting, while application of antimicrobial stewardship is the only way to reduce the spread of multi-resistant bacteria.

中文翻译：

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产生NDM的肺炎克雷伯氏菌在希腊一家三级医院中传播。

细菌碳青霉烯耐药性，特别是在可转移的碳青霉烯酶介导的情况下，是重要的公共卫生问题。在希腊塞萨洛尼基一家三级医院分离的产金属β-内酰胺酶（MBL）的菌株增加了数量，需要进一步的基因研究，该研究包括29个表型特征为MBL的非重复性碳青霉烯抗性肺炎克雷伯菌菌株。 -生产者收集在希腊的一家三级医院。筛选出分离物以检测碳青霉烯酶基因（肺炎克雷伯菌碳青霉烯酶（blaKPC），维罗纳-整合子编码的MBL-1（blaVIM-1），亚胺培南酶（blaIMP），草酸酶-48（blaOXA-48）和新德里MBL （blaNDM））。分离株的遗传关系通过随机扩增多态性DNA（RAPD）分析确定。进一步鉴定了两个NDM阳性肺炎克雷伯菌分离株的全基因组序列（WGS）。在所有肺炎克雷伯菌分离株中均确认了New Delhi MBL（blaNDM）基因的存在，同时共检测了blaKPC和blaVIM-1基因分别分离为一株和两株。RAPD分析表明，分离株分为两组。对两个肺炎克雷伯菌的全基因组序列分析表明，它们属于11型序列，它们携带blaNDM-1基因，并且在质粒和抗性基因的数量和类型上存在差异。该研究的肺炎支原体分离株带有blaNDM基因，而WGS分析显示其抗性基因具有遗传多样性。需要持续监视以检测医院环境中新克隆的出现，