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The extended-spectrum-β-lactamase (ESBL)- and Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacteriaceae represent serious and urgent threats to public health. In a retrospective study of multidrug-resistant K. pneumoniae , we identified three clinical isolates, CN1, CR14, and NY9, carrying both bla  CTX-M and bla  KPC genes. The complete genomes of these three K. pneumoniae isolates were de novo assembled by using both short- and long-read whole-genome sequencing. In CR14 and NY9, bla  CTX-M and bla  KPC were carried on two different plasmids. In contrast, CN1 had one copy of bla  KPC-2 and three copies of bla  CTX-M-15 integrated in the chromosome, for which the bla  CTX-M-15 genes were linked to an insertion sequence, IS Ecp1 , whereas the bla  KPC-2 gene was in the context of a Tn 4401a transposition unit conjugated with a PsP3-like prophage. Intriguingly, downstream of the Tn 4401a-bla  KPC-2 -prophage genomic island, CN1 also carried a clustered regularly interspaced short palindromic repeat (CRISPR)- cas array with four spacers targeting a variety of K. pneumoniae plasmids harboring antimicrobial resistance genes. Comparative genomic analysis revealed that there were two subtypes of type I-E CRISPR- cas in K. pneumoniae strains and suggested that the evolving CRISPR- cas , with its acquired novel spacer, induced the mobilization of antimicrobial resistance genes from plasmids into the chromosome. The integration and dissemination of multiple copies of bla  CTX-M and bla  KPC from plasmids to chromosome depicts the complex pandemic scenario of multidrug-resistant K. pneumoniae Additionally, the implications from this study also raise concerns for the application of a CRISPR- cas strategy against antimicrobial resistance.

antimicrobial agents and chemotherapy

Emergence and Evolution of Multidrug-Resistant Klebsiella pneumoniae with both bla KPC and bla CTX-M Integrated in the Chromosome