Characterization of a <i>bla</i> _KPC-3 -carrying plasmid in a clinical isolate of <i>Klebsiella pneumoniae</i> belonging to the emerging successful clone ST147

<i>Klebsiella pneumoniae</i> ST147 has emerged as a successful clone able to efficiently disseminate a number of carbapenemases, including <i>bla</i>_KPC. This study compared a representative <i>bla</i>_KPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor <i>bla</i>_KPC-3 belonging to the high-risk clone ST512. Five clinical isolates of KPC-3-producing <i>K. pneumoniae</i> belonging to ST147 isolated in Southern Spain were analyzed. The first ST147 isolate detected was compared with two previous isolates of <i>K. pneumoniae</i> ST512/KPC-3 and KPC-3-encoding plasmid pKpQIL as reference plasmid. Microdilution, disk diffusion, β-CARBA test, and NG-Test CARBA 5 were used for antimicrobial susceptibility analysis and phenotypic characterization of the isolates according to EUCAST guidelines. Molecular characterization was performed using pulsed-field gel electrophoresis (PFGE)-<i>Xba</i>I, sequenced (by Illumina and Oxford Nanopore) and annotated with open-source databases (CGE tools and RAST). Plasmid incompatibility groups were analyzed using PlasmidFinder; BRIG was used to compare the <i>bla</i>_KPC-3-harboring plasmids from three of the selected clinical isolates with each other and with pKpQIL. Clinical isolates of ST147/KPC-3 showed resistance to β-lactams, fluoroquinolones, and aminoglycosides. Clinical isolates ST147/KPC-3 and ST512/KPC-3 had identical PFGE patterns in each clone. Plasmid replicon analysis showed a plasmid with the formula IncFII (K2:A-:B-) in the ST512 isolates, identical to pKpQIL; the ST147 isolates harbored IncFII (K1:A-:B-), although the genetic environment of <i>bla</i>_KPC-3 was the same as pKpQIL, which is Tn<i>4401</i>_IS<i>Kpn7</i> upstream of <i>bla</i>_KPC-3 and IS<i>Kpn6</i> downstream of <i>bla</i>_KPC-3. Plasmids harboring <i>bla</i>_KPC-3 in ST147 and ST512 are different, although differences are subtle. The genetic environment of <i>bla</i>_KPC-3 in the different sequence types is very conservative and identical to pKpQIL.IMPORTANCEThe successful spread of <i>bla</i>_KPC is primarily due to the dissemination of <i>K. pneumoniae</i> isolates belonging to high-risk clonal complex 258 (ST258, ST11, and ST512); however, new clones are emerging globally as the ST147 clone. In this study, we compare the genetic environment of a representative <i>bla</i>_KPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor <i>bla</i>_KPC-3 belonging to the high-risk clone ST512. Plasmids harboring <i>bla</i>_KPC-3 detected in ST512 and ST147 were different; however, the close genetic environments of <i>bla</i>_KPC-3 in the different plasmids (ST147 and ST512) remained conserved.