Xanthopoulou Kyriaki, Carattoli Alessandra, Wille Julia, Biehl Lena M, Rohde Holger, Farowski Fedja, Krut Oleg, Villa Laura, Feudi Claudia, Seifert Harald, Higgins Paul G
Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, 50935 Cologne, Germany.
German Centre for Infection Research (DZIF), Partner site Bonn-Cologne, 50935 Cologne, Germany.
Antibiotics (Basel). 2020 Oct 5;9(10):675. doi: 10.3390/antibiotics9100675.
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) 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 isolates belong to the international high-risk clone ST147 and form a cluster of closely related isolates. They harbour the 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 isolates combined with the emergence of the XDR ST147 high-risk clone have the potential to become a major challenge for global healthcare.
移动遗传元件（MGEs），尤其是多药耐药质粒，是抗菌药物耐药性决定因素传播的主要载体。在此，我们分析了来自德国的三株广泛耐药（XDR）菌株中的MGEs。使用Illumina和MinION平台进行全基因组测序（WGS），随后进行核心基因组多位点序列分型（MLST）。通过接合、S1脉冲场凝胶电泳（S1-PFGE）和Southern印迹实验分析质粒含量。这些菌株属于国际高风险克隆ST147，并形成了一组密切相关的菌株群。它们在ColKP3质粒上携带碳青霉烯酶，并在具有重组性质且编码大量插入元件的多药耐药（MDR）IncR质粒上携带12个抗生素耐药性决定因素。这三株菌株中的IncR质粒具有高度同源性，但也存在遗传变异，如紧邻MGEs的基因区域发生倒位或缺失。此外，每株菌株中还存在六个不携带任何抗生素耐药性决定因素的质粒。我们的研究表明，由于MGEs的动态性质，在一组密切相关的菌株群中可以观察到遗传变异。这些菌株的可移动基因组与XDR ST147高风险克隆的出现有可能成为全球医疗保健的重大挑战。
