Mobile genetic elements (MGEs), especially multidrug-resistance plasmids, are major vehicles for the dissemination of antimicrobial resistance determinants. Herein, we analyse the MGEs in three extensively drug-resistant (XDR) Klebsiella pneumoniae isolates from Germany. Whole genome sequencing (WGS) is performed using Illumina and MinION platforms followed by core-genome multi-locus sequence typing (MLST). The plasmid content is analysed by conjugation, S1-pulsed-field gel electrophoresis (S1-PFGE) and Southern blot experiments. The K. pneumoniae isolates belong to the international high-risk clone ST147 and form a cluster of closely related isolates. They harbour the bla_OXA-181 carbapenemase on a ColKP3 plasmid, and 12 antibiotic resistance determinants on an multidrug-resistant (MDR) IncR plasmid with a recombinogenic nature and encoding a large number of insertion elements. The IncR plasmids within the three isolates share a high degree of homology, but present also genetic variations, such as inversion or deletion of genetic regions in close proximity to MGEs. In addition, six plasmids not harbouring any antibiotic resistance determinants are present in each isolate. Our study indicates that genetic variations can be observed within a cluster of closely related isolates, due to the dynamic nature of MGEs. The mobilome of the K. pneumoniae isolates combined with the emergence of the XDR ST147 high-risk clone have the potential to become a major challenge for global healthcare.

中文翻译：

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从德国回收的广泛耐药的肺炎克雷伯氏菌序列类型147中的抗生素耐药性和移动遗传元素

流动遗传元件（MGEs），尤其是耐多药质粒，是传播抗药性决定因素的主要载体。在这里，我们分析了来自德国的三种广泛耐药（XDR）肺炎克雷伯菌的MGEs 。使用Illumina和MinION平台进行全基因组测序（WGS），然后进行核心基因组多基因座序列分型（MLST）。通过缀合，S1脉冲场凝胶电泳（S1-PFGE）和Southern印迹实验分析质粒的含量。的肺炎克雷伯菌菌株属于国际高风险克隆ST147和形成密切相关菌株的集群。他们怀有bla _OXA-181ColKP3质粒上的碳青霉烯酶和多药抗性（MDR）IncR质粒上的12种抗生素抗性决定簇，具有重组原性并编码大量插入元件。这三个分离物中的IncR质粒具有高度的同源性，但也存在遗传变异，例如紧邻MGE的遗传区域的倒置或缺失。另外，每个分离物中存在六个没有任何抗生素抗性决定簇的质粒。我们的研究表明，由于MGE的动态性质，可以在一组密切相关的分离株中观察到遗传变异。在的mobilome肺炎克雷伯菌菌株与XDR ST147高风险克隆的出现，结合具有成为全球医疗保健的一大挑战的潜力。