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The emergence of hypervirulent carbapenem-resistant  Klebsiella pneumoniae  (hv-CRKP) was regarded as an emerging threat in clinical settings. Here, we investigated the prevalence of CRKP strains among inpatients in a new hospital over 1 year since its inception with various techniques, and carried out a WGS-based phylogenetic study to dissect the genomic background of these isolates. The genomes of three representative  bla  NDM–1 -positive strains and the plasmids of four  bla  KPC–2 -positive strains were selected for Nanopore long-read sequencing to resolve the complicated MDR structures. Thirty-five CRKP strains were identified from 193  K. pneumoniae  isolates, among which 30 strains (85.7%) harbored  bla  KPC–2 , whereas the remaining five strains (14.3%) were positive for  bla  NDM–1 . The antimicrobial resistance profiles of  bla  NDM–1 -positive isolates were narrower than that of  bla  KPC–2 -positive isolates. Five isolates including two  bla  NDM–1 -positive isolates and three  bla  KPC–2 -positive strains could successfully transfer the carbapenem resistance phenotype by conjugation. All CRKP strains were categorized into six known multilocus sequence types, with ST11 being the most prevalent type. Phylogenetic analysis demonstrated that the clonal spread of ST11  bla  KPC–2 -positive isolates and local polyclonal spread of  bla  NDM–1 -positive isolates have existed in the hospital. The  bla  NDM–1  gene was located on IncX3, IncFIB/IncHI1B, and IncHI5-like plasmids, of which IncFIB/IncHI1B plasmid has a novel structure. By contrast, all ST11 isolates shared the similar  bla  KPC–2 -bearing plasmid backbone, and 11 of them possessed pLVPK-like plasmids. In addition,  in silico  virulome analysis,  Galleria mellonella  larvae infection assay, and siderophore secretion revealed the hypervirulence potential of most  bla  KPC–2 -positive strains. Given that these isolates also had remarkable environmental adaptability, targeted measures should be implemented to prevent the grave consequences caused by hv-CRKP strains in nosocomial settings.

frontiers in microbiology

Deciphering the Epidemiological Characteristics and Molecular Features of blaKPC–2- or blaNDM–1-Positive Klebsiella pneumoniae Isolates in a Newly Established Hospital