IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae

Background: This study aims to explore the phenotypic and genotypic characteristics of a plasmid that co-harbours blaIMP-4 and blaKPC-2 in a carbapenem-resistant Klebsiella pneumoniae. Methods: Strain K194 was isolated from a 60-year-old patient. Species identification was performed with matrix-assi...

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Main Authors: Jiawei Zhou, Xiaohua Meng, Shujun Ni, Yunxing Yang, Qiong Zhang, Lingjiao Wu, Qiong Chen
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Journal of Global Antimicrobial Resistance
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Online Access:http://www.sciencedirect.com/science/article/pii/S2213716525000402
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author Jiawei Zhou
Xiaohua Meng
Shujun Ni
Yunxing Yang
Qiong Zhang
Lingjiao Wu
Qiong Chen
author_facet Jiawei Zhou
Xiaohua Meng
Shujun Ni
Yunxing Yang
Qiong Zhang
Lingjiao Wu
Qiong Chen
author_sort Jiawei Zhou
collection DOAJ
description Background: This study aims to explore the phenotypic and genotypic characteristics of a plasmid that co-harbours blaIMP-4 and blaKPC-2 in a carbapenem-resistant Klebsiella pneumoniae. Methods: Strain K194 was isolated from a 60-year-old patient. Species identification was performed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, followed by antibiotic susceptibility testing. Antimicrobial resistance genes were detected and S1-pulsed-field gel electrophoresis with Southern blot experiments were performed to identify plasmids. Whole-genome sequencing was executed with the Illumina and Oxford Nanopore platforms. Results: K. pneumoniae K194 was resistant to multiple antibiotics, including carbapenems. This strain carried both blaIMP-4 and blaKPC-2 on a single 163 kb plasmid (pK194-P2). pK194-P2 was capable of conjugation with an efficiency of 3.4 × 10–7 in vitro conjugation experiments. Whole-genome analysis confirmed that pK194-P2 was a novel plasmid and had both IncFII- and IncN-type replicons. Sequence alignment revealed direct repeats of the sequences (GCCCAAGG) flanking a 109-kb region bounded by two copies of IS26. In vitro, evolution experiments showed that blaKPC-2 in pK194-P2 could be stably maintained in the transconjugants after 10 days of passage, while blaIMP-4 could be lost during repeated laboratory passage. Whole-genome sequencing and alignment of two blaIMP-4-negative plasmids with pK194-P2 revealed that they had a deletion of 81 or 94 kb adjacent to IS26. Conclusions: Our study reports a novel plasmid co-harbouring blaIMP-4 and blaKPC-2 in K. pneumoniae, and highlights the potential role of IS26-mediated cointegration and deletion in plasmid formation and evolution.
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spelling doaj-art-e81463cd0b4f42e8aeb8822aca9798af2025-08-20T03:52:43ZengElsevierJournal of Global Antimicrobial Resistance2213-71652025-05-0142616510.1016/j.jgar.2025.02.009IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniaeJiawei Zhou0Xiaohua Meng1Shujun Ni2Yunxing Yang3Qiong Zhang4Lingjiao Wu5Qiong Chen6State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, ChinaState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, ChinaState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, ChinaDepartment of Laboratory Medicine, Hangzhou First People's Hospital, Hangzhou, ChinaState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, ChinaState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, ChinaDepartment of Laboratory Medicine, Hangzhou First People's Hospital, Hangzhou, China; Corresponding author. Mailing address. Department of Laboratory Medicine, Hangzhou First People's Hospital, No.261, Huansha Road, Hangzhou, Zhejiang 310003, China.Background: This study aims to explore the phenotypic and genotypic characteristics of a plasmid that co-harbours blaIMP-4 and blaKPC-2 in a carbapenem-resistant Klebsiella pneumoniae. Methods: Strain K194 was isolated from a 60-year-old patient. Species identification was performed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, followed by antibiotic susceptibility testing. Antimicrobial resistance genes were detected and S1-pulsed-field gel electrophoresis with Southern blot experiments were performed to identify plasmids. Whole-genome sequencing was executed with the Illumina and Oxford Nanopore platforms. Results: K. pneumoniae K194 was resistant to multiple antibiotics, including carbapenems. This strain carried both blaIMP-4 and blaKPC-2 on a single 163 kb plasmid (pK194-P2). pK194-P2 was capable of conjugation with an efficiency of 3.4 × 10–7 in vitro conjugation experiments. Whole-genome analysis confirmed that pK194-P2 was a novel plasmid and had both IncFII- and IncN-type replicons. Sequence alignment revealed direct repeats of the sequences (GCCCAAGG) flanking a 109-kb region bounded by two copies of IS26. In vitro, evolution experiments showed that blaKPC-2 in pK194-P2 could be stably maintained in the transconjugants after 10 days of passage, while blaIMP-4 could be lost during repeated laboratory passage. Whole-genome sequencing and alignment of two blaIMP-4-negative plasmids with pK194-P2 revealed that they had a deletion of 81 or 94 kb adjacent to IS26. Conclusions: Our study reports a novel plasmid co-harbouring blaIMP-4 and blaKPC-2 in K. pneumoniae, and highlights the potential role of IS26-mediated cointegration and deletion in plasmid formation and evolution.http://www.sciencedirect.com/science/article/pii/S2213716525000402Klebsiella pneumoniaeCarbapenem resistanceCarbapenemasesPlasmid cointegrationIs26
spellingShingle Jiawei Zhou
Xiaohua Meng
Shujun Ni
Yunxing Yang
Qiong Zhang
Lingjiao Wu
Qiong Chen
IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
Journal of Global Antimicrobial Resistance
Klebsiella pneumoniae
Carbapenem resistance
Carbapenemases
Plasmid cointegration
Is26
title IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
title_full IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
title_fullStr IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
title_full_unstemmed IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
title_short IS26-mediated cointegration generates a plasmid co-harbouring blaIMP-4 and blaKPC-2 in Klebsiella pneumoniae
title_sort is26 mediated cointegration generates a plasmid co harbouring blaimp 4 and blakpc 2 in klebsiella pneumoniae
topic Klebsiella pneumoniae
Carbapenem resistance
Carbapenemases
Plasmid cointegration
Is26
url http://www.sciencedirect.com/science/article/pii/S2213716525000402
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