Loss and gain of ceftazidime-avibactam susceptibility in a non-carbapenemase-producing K1-ST23 hypervirulent Klebsiella pneumoniae.

IF 5.5 1区 农林科学 Q1 IMMUNOLOGY
Virulence Pub Date : 2024-12-01 Epub Date: 2024-05-02 DOI:10.1080/21505594.2024.2348251
Jiankang Zhao, Danni Pu, Ziyao Li, Yulin Zhang, Xinmeng Liu, Xianxia Zhuo, Binghuai Lu, Bin Cao
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引用次数: 0

Abstract

Objectives: This study aimed at revealing the underlying mechanisms of the loss and gain of ceftazidime-avibactam susceptibility in a non-carbapenemase-producing hypervirulent Klebsiella pneumoniae (hvKp).

Methods: Here we longitudinally recovered 3 non-carbapenemase-producing K1-ST23 hvKp strains at a one-month interval (KP29105, KP29499 and KP30086) from an elderly male. Antimicrobial susceptibility testing, whole genome sequencing, transcriptomic sequencing, gene cloning, plasmid conjugation, quantitative real-time PCR (qRT-PCR), and SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) were conducted.

Results: Among the 3 hvKp strains, KP29105 was resistant to the third- and fourth-generation cephalosporins, KP29499 acquired resistance to both ceftazidime-avibactam and carbapenems, while KP30086 restored its susceptibility to ceftazidime-avibactam, imipenem and meropenem but retained low-level resistance to ertapenem. KP29105 and KP29499 carried plasmid-encoded genes blaCTX-M-15 and blaCTX-M-71, respectively, but KP30086 lost both. Cloning of gene blaCTX-M-71 and conjugation experiment of blaCTX-M-71-carrying plasmid showed that the transformant and transconjugant were susceptible to ceftazidime-avibactam but had a more than 8-fold increase in MICs. Supplementation with an outer membrane permeabilizer could reduce the MIC of ceftazidime-avibactam by 32 folds, indicating that porins play a key role in ceftazidime-avibactam resistance. The OmpK35 of the 3 isolates was not expressed, and the OmpK36 of KP29499 and KP30086 had a novel amino acid substitution (L359R). SDS-PAGE and qRT-PCR showed that the expression of porin OmpK36 of KP29499 and KP30086 was significantly down-regulated compared with KP29105.

Conclusions: In summary, we reported the rare ceftazidime-avibactam resistance in a non-carbapenemase-producing hvKp strain. Resistance plasmid carrying blaCTX-M-71 and mutated OmpK36 had a synergetic effect on the resistance.

一种不产碳青霉烯酶的 K1-ST23 型高病毒性肺炎克雷伯菌对头孢他啶-阿维菌素敏感性的丧失和增益。
研究目的本研究旨在揭示非碳青霉烯酶生产型高病毒性肺炎克雷伯菌(hvKp)对头孢他啶-阿维巴坦易感性丧失和增益的内在机制。方法:我们从一名老年男性患者身上纵向采集了3株非碳青霉烯酶生产型K1-ST23 hvKp菌株(KP29105、KP29499和KP30086),间隔时间为一个月。研究人员对这些菌株进行了抗菌药敏感性检测、全基因组测序、转录组测序、基因克隆、质粒连接、实时定量 PCR(qRT-PCR)和 SDS-PAGE(十二烷基硫酸钠-聚丙烯酰胺凝胶电泳):结果:在3株hvKp菌株中,KP29105对第三代和第四代头孢菌素耐药,KP29499对头孢唑肟-阿维巴坦和碳青霉烯类耐药,而KP30086恢复了对头孢唑肟-阿维巴坦、亚胺培南和美罗培南的敏感性,但对厄他培南保持低水平耐药。KP29105 和 KP29499 分别携带质粒编码基因 blaCTX-M-15 和 blaCTX-M-71,但 KP30086 却失去了这两种基因。基因 blaCTX-M-71 的克隆和携带 blaCTX-M-71 的质粒的共轭实验表明,转化株和转接株对头孢他啶-阿维巴坦敏感,但 MICs 增加了 8 倍多。添加外膜渗透剂可使头孢他啶-阿维菌素的 MIC 降低 32 倍,表明孔蛋白在头孢他啶-阿维菌素耐药性中起着关键作用。这 3 个分离株的 OmpK35 没有表达,KP29499 和 KP30086 的 OmpK36 有一个新的氨基酸取代(L359R)。SDS-PAGE和qRT-PCR显示,与KP29105相比,KP29499和KP30086的孔蛋白OmpK36表达明显下调:综上所述,我们报道了非碳青霉烯酶产hvKp菌株对头孢他啶-阿维菌素的罕见耐药性。携带 blaCTX-M-71 和突变 OmpK36 的耐药质粒对耐药性有协同作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Virulence
Virulence IMMUNOLOGY-MICROBIOLOGY
CiteScore
9.20
自引率
1.90%
发文量
123
审稿时长
6-12 weeks
期刊介绍: Virulence is a fully open access peer-reviewed journal. All articles will (if accepted) be available for anyone to read anywhere, at any time immediately on publication. Virulence is the first international peer-reviewed journal of its kind to focus exclusively on microbial pathogenicity, the infection process and host-pathogen interactions. To address the new infectious challenges, emerging infectious agents and antimicrobial resistance, there is a clear need for interdisciplinary research.
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