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中国宁波临床分离株中碳青霉烯类耐药机制的分子特征及表型相关性

Molecular characterization of carbapenem resistance mechanisms and phenotypic correlations in clinical isolates from Ningbo, China.

作者信息

Qiu Xuedan, Jiang Min, Xu Jianqiang, Wu Qiaoping, Lin Chenyao, Li Weiying, Li Qingcao

机构信息

Department of Clinical Laboratory, The Affiliated Li Huili Hospital of Ningbo University, Ningbo, China.

Department of Clinical Laboratory, Langxia Street Health Service Center, Ningbo, China.

出版信息

Front Microbiol. 2025 May 14;16:1546805. doi: 10.3389/fmicb.2025.1546805. eCollection 2025.

DOI:10.3389/fmicb.2025.1546805
PMID:40438220
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12116675/
Abstract

OBJECTIVE

The purpose of this study is to understand the antimicrobial susceptibility and molecular distribution characteristics of carbapenem-resistant (CRKP) in the region, and to evaluate their correlation. Additionally, the study aims to investigate the transmission status of these strains.

METHODS

A total of 150 CRKP collected from January 2019 to December 2021 in the Ningbo region were included in this study. Antimicrobial susceptibility testing was performed using broth microdilution method following CLSI guidelines (CLSI, 2023). The tested agents included: (1) basic antimicrobials (tigecycline, polymyxin B, ceftazidime-avibactam); and (2) combination therapy candidates (ertapenem, imipenem, levofloxacin, piperacillin-tazobactam, ceftriaxone, cefepime, trimethoprim-sulfamethoxazole, fosfomycin, amikacin, aztreonam, chloramphenicol, amoxicillin-clavulanate, ceftazidime). Resistance genes were detected using polymerase chain reaction (PCR). Multi-locus sequence typing (MLST) was employed to analyze the molecular characteristics and evolutionary trends of the strains to determine their clonal relationships.

RESULTS

The 150 strains of CRKP exhibit high resistance rates to various conventional drugs; The sensitivity rates to tigecycline, polymyxin B, and ceftazidime-avibactam were 98.7, 98.0, and 68%, respectively; Conversely, the sensitivity rates to fosfomycin, amikacin, and chloramphenicol were 72.0, 40.0, and 16.7%, respectively; The main proportions of carbapemen genes producing in CRKP are as follows: (61.3%), (14.7%), (8.0%), (6.0%), and (1.3%); The main proportions of β-lactamase resistance genes are as follows: (13.33%), (25.33%) (17.33%) (34.67%) (26.66%), (66.66%), (18.66%), and (10.00%); CRKP carrying class A, B, and D carbapenemases had a sensitivity rate greater than 96% for tigecycline and polymyxin B, while their sensitivities to ceftazidime-avibactam, aztreonam, and amikacin varied significantly ( < 0.01). Analysis of the MLST results for CRKP revealed that ST11 strains were predominant in the region. There was a significant difference in the resistance genes carried by ST11 strains compared to non-ST11 strains. While different healthcare institutions exhibited variations in ST types, the strains generally showed high homogeneity.

CONCLUSION

In the region, CRKP showed high sensitivity to tigecycline, polymyxin B, ceftazidime-avibactam, fosfomycin, amikacin, and chloramphenicol. The main carbapenemase genes identified were and . The inhibitory effects of ceftazidime-avibactam, aztreonam, and amikacin varied for CRKP carrying different enzyme types. ST11 strains were predominant in the region. There was a significant difference in the resistance genes carried by ST11 strains compared to non-ST11 strains. Clonal dissemination was observed both within the same healthcare institution and between different institutions.

摘要

目的

本研究旨在了解该地区耐碳青霉烯类肺炎克雷伯菌(CRKP)的抗菌药物敏感性及分子分布特征,并评估它们之间的相关性。此外,该研究还旨在调查这些菌株的传播状况。

方法

本研究纳入了2019年1月至2021年12月在宁波地区收集的150株CRKP。按照美国临床和实验室标准协会(CLSI)指南(CLSI,2023),采用肉汤微量稀释法进行抗菌药物敏感性试验。所测试的药物包括:(1)基础抗菌药物(替加环素、多粘菌素B、头孢他啶-阿维巴坦);(2)联合治疗候选药物(厄他培南、亚胺培南、左氧氟沙星、哌拉西林-他唑巴坦、头孢曲松、头孢吡肟、复方磺胺甲恶唑、磷霉素、阿米卡星、氨曲南、氯霉素、阿莫西林-克拉维酸、头孢他啶)。使用聚合酶链反应(PCR)检测耐药基因。采用多位点序列分型(MLST)分析菌株的分子特征和进化趋势,以确定它们的克隆关系。

结果

150株CRKP对各种传统药物表现出较高的耐药率;对替加环素、多粘菌素B和头孢他啶-阿维巴坦的敏感率分别为98.7%、98.0%和68%;相反,对磷霉素、阿米卡星和氯霉素的敏感率分别为72.0%、40.0%和16.7%;CRKP中产生碳青霉烯酶基因的主要比例如下: blaKPC(61.3%)、 blaNDM(14.7%)、 blaIMP(8.0%)、 blaVIM(6.0%)和 blaOXA-48(1.3%);β-内酰胺酶耐药基因的主要比例如下: blaSHV(13.33%)、 blaCTX-M(25.33%)、 blaTEM(17.33%)、 blaCMY(34.67%)、 blaDHA(26.66%)、 blaOXA-1(66.66%)、 blaOXA-2(18.66%)和 blaOXA-10(10.00%);携带A类、B类和D类碳青霉烯酶的CRKP对替加环素和多粘菌素B的敏感率大于96%,而它们对头孢他啶-阿维巴坦、氨曲南和阿米卡星的敏感性差异显著(P<0.01)。对CRKP的MLST结果分析显示,ST11菌株在该地区占主导地位。与非ST11菌株相比,ST11菌株携带的耐药基因存在显著差异。虽然不同医疗机构的ST型存在差异,但菌株总体表现出高度同源性。

结论

在该地区,CRKP对替加环素、多粘菌素B、头孢他啶-阿维巴坦、磷霉素、阿米卡星和氯霉素表现出较高的敏感性。鉴定出的主要碳青霉烯酶基因是 blaKPC和 blaNDM。头孢他啶-阿维巴坦、氨曲南和阿米卡星对携带不同酶型的CRKP的抑制作用有所不同。ST11菌株在该地区占主导地位。与非ST11菌株相比,ST11菌株携带的耐药基因存在显著差异。在同一医疗机构内和不同医疗机构之间均观察到克隆传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0367/12116675/f2abb992b3e6/fmicb-16-1546805-g006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0367/12116675/501ac19786c5/fmicb-16-1546805-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0367/12116675/e3e6d21cb805/fmicb-16-1546805-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0367/12116675/7a795ef19876/fmicb-16-1546805-g003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0367/12116675/f2abb992b3e6/fmicb-16-1546805-g006.jpg

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