碳青霉烯耐药铜绿假单胞菌ST1076中携带blakpc -2的巨型质粒的出现：基因组和功能见解

IF 3.2 3区医学 Q2 INFECTIOUS DISEASES

Journal of global antimicrobial resistance Pub Date : 2025-07-25 DOI:10.1016/j.jgar.2025.07.018

Jing Chen , Weidong Zhu , Chen Lin , Zhen Li , Haowei Ye , Danfeng Lou

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引用次数: 0

摘要

目的：耐碳青霉烯假单胞菌（Pseudomonas aeruginosa， CRPA）因其多药耐药和基因组可塑性对临床治疗构成了严峻挑战。在这里，我们报告了ST1076铜绿假单胞菌分离物在可转移质粒上携带blaKPC-2的基因组和功能特征。方法：采用基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF-MS）进行物种鉴定，采用聚合酶链反应（PCR）对blaKPC-2进行鉴定。采用全基因组测序（WGS）和药敏试验（AST）分析了铜绿假单胞菌L4884的特征。质粒通过s1 -核酸酶脉冲场凝胶电泳（S1-PFGE）、Southern印迹、转偶联实验和比较基因组分析进行鉴定。结果：铜绿假单胞菌L4884属ST1076序列型，对亚胺培南、美罗培南等β-内酰胺类抗生素耐药。blaKPC-2基因位于394 kb的IncP-2巨质粒上，具有保守的遗传结构（IS6100-ISKpn27-blaKPC-2-ISKpn6-klcA）。经过多次努力，转偶联实验成功，证实了质粒在27°C下的转移能力。结论：本研究为碳青霉烯类耐药性的遗传基础提供了新的见解，并强调了加强监测和遏制策略以防止CRPA在临床环境中传播的必要性。

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Emergence of blaKPC-2-carrying megaplasmid in carbapenem-resistant Pseudomonas aeruginosa ST1076: Genomic and functional insights

Objective

Carbapenem-resistant Pseudomonas aeruginosa (CRPA) poses a critical challenge to clinical treatment due to its multidrug resistance and genomic plasticity. Here, we report the genomic and functional characterization of an ST1076 P. aeruginosa isolate carrying bla_KPC-2 on a transferable plasmid.

Methods

Species identification was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and bla_KPC-2 was identified by polymerase chain reaction. The characteristics of P. aeruginosa L4884 were analysed by whole-genome sequencing and antimicrobial susceptibility testing. Plasmid characterization was carried out through S1-nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting, transconjugation experiments, and comparative genomic analysis.

Results

The isolate, P. aeruginosa L4884, belonged to sequence type ST1076 and demonstrated resistance to multiple β-lactam antibiotics, including imipenem and meropenem. The blaKPC-2 gene was located on a 394-kb IncP-2 megaplasmid, displaying a conserved genetic structure (IS6100-ISKpn27-bla_KPC-2-ISKpn6-klcA). Transconjugation experiments succeeded after repeated efforts, confirming the plasmid’s ability to transfer at 27°C.

Conclusions

This study provides new insights into the genetic basis of carbapenem resistance and underscores the need for enhanced surveillance and containment strategies to prevent the spread of CRPA in clinical settings.

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来源期刊

Journal of global antimicrobial resistance INFECTIOUS DISEASES-PHARMACOLOGY & PHARMACY

CiteScore

8.70

自引率

2.20%

发文量

285

审稿时长

34 weeks

期刊介绍： The Journal of Global Antimicrobial Resistance (JGAR) is a quarterly online journal run by an international Editorial Board that focuses on the global spread of antibiotic-resistant microbes. JGAR is a dedicated journal for all professionals working in research, health care, the environment and animal infection control, aiming to track the resistance threat worldwide and provides a single voice devoted to antimicrobial resistance (AMR). Featuring peer-reviewed and up to date research articles, reviews, short notes and hot topics JGAR covers the key topics related to antibacterial, antiviral, antifungal and antiparasitic resistance.