Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids

BackgroundCitrobacter portucalensis is an emerging multidrug-resistant (MDR) pathogen within the Citrobacter genus. Although individual occurrences of blaKPC–2 or blaNDM–1 have been sporadically reported, the coexistence of both carbapenemase genes in a single strain remains extremely rare.MethodsWe...

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Main Authors: Junwei Huang, Kai Shen, Keqiang Chen, Junliang Wu, Yijun Zhu, Jingchao Shi
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1633493/full
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author Junwei Huang
Kai Shen
Keqiang Chen
Junliang Wu
Yijun Zhu
Jingchao Shi
author_facet Junwei Huang
Kai Shen
Keqiang Chen
Junliang Wu
Yijun Zhu
Jingchao Shi
author_sort Junwei Huang
collection DOAJ
description BackgroundCitrobacter portucalensis is an emerging multidrug-resistant (MDR) pathogen within the Citrobacter genus. Although individual occurrences of blaKPC–2 or blaNDM–1 have been sporadically reported, the coexistence of both carbapenemase genes in a single strain remains extremely rare.MethodsWe performed whole-genome sequencing and conjugation assays on a bloodstream isolate of C. portucalensis (JH112) obtained from a critically ill patient. Plasmid structure, resistance determinants, and transferability were comprehensively analyzed using in vitro assays and bioinformatic pipelines.ResultsJH112 exhibited an extensively drug-resistant phenotype and carried two major carbapenemase genes, blaKPC–2 and blaNDM–1, located on distinct plasmids. The blaKPC–2 gene resided on an IncFII(Yp)-type plasmid (∼110 kb) with a complete conjugation module and was successfully transferred to a recipient strain. This plasmid also harbored an O-antigen biosynthesis gene cluster, potentially enhancing host adaptation. In contrast, the blaNDM–1 gene was located on a 340 kb IncHI2/HI2A-type megaplasmid with incomplete conjugation machinery and failed to transfer under standard conditions. Both plasmids showed unique structural arrangements compared to known references. The chromosome also carried blaCMY–49 and qnrB1, contributing to broad-spectrum resistance.ConclusionWe report a rare clinical C. portucalensis isolate co-harboring two carbapenemase genes on genetically distinct plasmids with divergent mobility. This highlights the species’ potential role as a resistance gene reservoir and the need for enhanced molecular surveillance in both clinical and environmental settings.
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spelling doaj-art-5170568380c34dbfbd00a50bd1f2b21c2025-08-20T03:50:53ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16334931633493Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmidsJunwei HuangKai ShenKeqiang ChenJunliang WuYijun ZhuJingchao ShiBackgroundCitrobacter portucalensis is an emerging multidrug-resistant (MDR) pathogen within the Citrobacter genus. Although individual occurrences of blaKPC–2 or blaNDM–1 have been sporadically reported, the coexistence of both carbapenemase genes in a single strain remains extremely rare.MethodsWe performed whole-genome sequencing and conjugation assays on a bloodstream isolate of C. portucalensis (JH112) obtained from a critically ill patient. Plasmid structure, resistance determinants, and transferability were comprehensively analyzed using in vitro assays and bioinformatic pipelines.ResultsJH112 exhibited an extensively drug-resistant phenotype and carried two major carbapenemase genes, blaKPC–2 and blaNDM–1, located on distinct plasmids. The blaKPC–2 gene resided on an IncFII(Yp)-type plasmid (∼110 kb) with a complete conjugation module and was successfully transferred to a recipient strain. This plasmid also harbored an O-antigen biosynthesis gene cluster, potentially enhancing host adaptation. In contrast, the blaNDM–1 gene was located on a 340 kb IncHI2/HI2A-type megaplasmid with incomplete conjugation machinery and failed to transfer under standard conditions. Both plasmids showed unique structural arrangements compared to known references. The chromosome also carried blaCMY–49 and qnrB1, contributing to broad-spectrum resistance.ConclusionWe report a rare clinical C. portucalensis isolate co-harboring two carbapenemase genes on genetically distinct plasmids with divergent mobility. This highlights the species’ potential role as a resistance gene reservoir and the need for enhanced molecular surveillance in both clinical and environmental settings.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1633493/fullCitrobacter portucalensisblaKPC–2blaNDM–1plasmid-mediated resistanceantimicrobial resistanceconjugation
spellingShingle Junwei Huang
Kai Shen
Keqiang Chen
Junliang Wu
Yijun Zhu
Jingchao Shi
Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
Frontiers in Microbiology
Citrobacter portucalensis
blaKPC–2
blaNDM–1
plasmid-mediated resistance
antimicrobial resistance
conjugation
title Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
title_full Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
title_fullStr Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
title_full_unstemmed Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
title_short Genomic characterization of a multidrug-resistant Citrobacter portucalensis isolate co-harboring blaKPC–2 and blaNDM–1 on distinct plasmids
title_sort genomic characterization of a multidrug resistant citrobacter portucalensis isolate co harboring blakpc 2 and blandm 1 on distinct plasmids
topic Citrobacter portucalensis
blaKPC–2
blaNDM–1
plasmid-mediated resistance
antimicrobial resistance
conjugation
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1633493/full
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