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Carbapenemase-producing  Klebsiella pneumoniae  emerged as a nosocomial pathogen causing morbidity and mortality in patients. For infection prevention it is important to track the spread of  K. pneumoniae  and its plasmids between patients. Therefore, the major aim was to recapitulate the contents and diversity of the plasmids of genetically related  K. pneumoniae  strains harboring the beta-lactamase gene  bla  KPC-2  or  bla  KPC-3  to determine their dissemination in the Netherlands and the former Dutch Caribbean islands from 2014 to 2019. Next-generation sequencing was combined with long-read third-generation sequencing to reconstruct 22 plasmids. wgMLST revealed five genetic clusters comprised of  K. pneumoniae bla  KPC-2  isolates and four clusters consisted of  bla  KPC-3  isolates. KpnCluster-019  bla  KPC-2  isolates were found both in the Netherlands and the Caribbean islands, while  bla  KPC-3  cluster isolates only in the Netherlands. Each  K. pneumoniae bla  KPC-2  or  bla  KPC-3  cluster was characterized by a distinct resistome and plasmidome. However, the large and medium plasmids contained a variety of antibiotic resistance genes, conjugation machinery, cation transport systems, transposons, toxin/antitoxins, insertion sequences and prophage-related elements. The small plasmids carried genes implicated in virulence. Thus, implementing long-read plasmid sequencing analysis for  K. pneumoniae  surveillance provided important insights in the transmission of a KpnCluster-019  bla  KPC-2  strain between the Netherlands and the Caribbean.

Plasmid diversity among genetically related Klebsiella pneumoniae blaKPC-2 and blaKPC-3 isolates collected in the Dutch national surveillance