Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147

ABSTRACT Klebsiella pneumoniae ST147 has emerged as a successful clone able to efficiently disseminate a number of carbapenemases, including blaKPC. This study compared a representative blaKPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor blaKPC-3 belonging to the high-risk...

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Main Authors: Esther Recacha, Mercedes Delgado-Valverde, Marina R. Pulido, Elena Pérez-Nadales, Patricia Pérez-Palacios, Inés Portillo-Calderón, Juan Manuel Sánchez-Calvo, Irene Gracia-Ahufinger, Álvaro Pascual
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Language:English
Published: American Society for Microbiology 2025-07-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.02338-24
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author Esther Recacha
Mercedes Delgado-Valverde
Marina R. Pulido
Elena Pérez-Nadales
Patricia Pérez-Palacios
Inés Portillo-Calderón
Juan Manuel Sánchez-Calvo
Irene Gracia-Ahufinger
Álvaro Pascual
author_facet Esther Recacha
Mercedes Delgado-Valverde
Marina R. Pulido
Elena Pérez-Nadales
Patricia Pérez-Palacios
Inés Portillo-Calderón
Juan Manuel Sánchez-Calvo
Irene Gracia-Ahufinger
Álvaro Pascual
author_sort Esther Recacha
collection DOAJ
description ABSTRACT Klebsiella pneumoniae ST147 has emerged as a successful clone able to efficiently disseminate a number of carbapenemases, including blaKPC. This study compared a representative blaKPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor blaKPC-3 belonging to the high-risk clone ST512. Five clinical isolates of KPC-3-producing K. pneumoniae belonging to ST147 isolated in Southern Spain were analyzed. The first ST147 isolate detected was compared with two previous isolates of K. pneumoniae 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)-XbaI, 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 blaKPC-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 blaKPC-3 was the same as pKpQIL, which is Tn4401_ISKpn7 upstream of blaKPC-3 and ISKpn6 downstream of blaKPC-3. Plasmids harboring blaKPC-3 in ST147 and ST512 are different, although differences are subtle. The genetic environment of blaKPC-3 in the different sequence types is very conservative and identical to pKpQIL.IMPORTANCEThe successful spread of blaKPC is primarily due to the dissemination of K. pneumoniae 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 blaKPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor blaKPC-3 belonging to the high-risk clone ST512. Plasmids harboring blaKPC-3 detected in ST512 and ST147 were different; however, the close genetic environments of blaKPC-3 in the different plasmids (ST147 and ST512) remained conserved.
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spelling doaj-art-e3ef658d56214d13b7de40fd166fe90f2025-08-20T02:44:39ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-07-0113710.1128/spectrum.02338-24Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147Esther Recacha0Mercedes Delgado-Valverde1Marina R. Pulido2Elena Pérez-Nadales3Patricia Pérez-Palacios4Inés Portillo-Calderón5Juan Manuel Sánchez-Calvo6Irene Gracia-Ahufinger7Álvaro Pascual8Unidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, SpainUnidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, SpainInstituto de Biomedicina de Sevilla/Hospital Virgen Macarena, Universidad de Sevilla/CSIC, Sevilla, SpainInstituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC), Hospital Universitario Reina Sofía, Córdoba, SpainUnidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, SpainUnidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, SpainUnit of Infectious Diseases and Clinical Microbiology, Jerez de la Frontera University Hospital, Jerez de la Frontera, SpainCIBER de Enfermedades Infecciosas-CIBERINFEC (CB21/13/00049), Instituto de Salud Carlos III, Madrid, SpainUnidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, SpainABSTRACT Klebsiella pneumoniae ST147 has emerged as a successful clone able to efficiently disseminate a number of carbapenemases, including blaKPC. This study compared a representative blaKPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor blaKPC-3 belonging to the high-risk clone ST512. Five clinical isolates of KPC-3-producing K. pneumoniae belonging to ST147 isolated in Southern Spain were analyzed. The first ST147 isolate detected was compared with two previous isolates of K. pneumoniae 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)-XbaI, 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 blaKPC-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 blaKPC-3 was the same as pKpQIL, which is Tn4401_ISKpn7 upstream of blaKPC-3 and ISKpn6 downstream of blaKPC-3. Plasmids harboring blaKPC-3 in ST147 and ST512 are different, although differences are subtle. The genetic environment of blaKPC-3 in the different sequence types is very conservative and identical to pKpQIL.IMPORTANCEThe successful spread of blaKPC is primarily due to the dissemination of K. pneumoniae 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 blaKPC-3-carrying plasmid of an ST147 clone with plasmids that usually harbor blaKPC-3 belonging to the high-risk clone ST512. Plasmids harboring blaKPC-3 detected in ST512 and ST147 were different; however, the close genetic environments of blaKPC-3 in the different plasmids (ST147 and ST512) remained conserved.https://journals.asm.org/doi/10.1128/spectrum.02338-24K. pneumoniaeblaKPC-3ST147
spellingShingle Esther Recacha
Mercedes Delgado-Valverde
Marina R. Pulido
Elena Pérez-Nadales
Patricia Pérez-Palacios
Inés Portillo-Calderón
Juan Manuel Sánchez-Calvo
Irene Gracia-Ahufinger
Álvaro Pascual
Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
Microbiology Spectrum
K. pneumoniae
blaKPC-3
ST147
title Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
title_full Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
title_fullStr Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
title_full_unstemmed Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
title_short Characterization of a blaKPC-3-carrying plasmid in a clinical isolate of Klebsiella pneumoniae belonging to the emerging successful clone ST147
title_sort characterization of a blakpc 3 carrying plasmid in a clinical isolate of klebsiella pneumoniae belonging to the emerging successful clone st147
topic K. pneumoniae
blaKPC-3
ST147
url https://journals.asm.org/doi/10.1128/spectrum.02338-24
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