耐碳青霉烯类肺炎克雷伯菌全基因组测序及黏菌素耐药分析  

Whole-genome sequencing and colistin resistance of carbapenem-resistant Klebsiella pneumoniae

作　　者：储雯雯[1] 刘周[1] 李昕[1] 叶乃芳 龚真 曾晓娇 周强[1] CHU Wenwen;LIU Zhou;LI Xin;YE Naifang;GONG Zhen;ZENG Xiaojiao;ZHOU Qiang(Department of Laboratory Medicine,The Second Hospital of Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学第二附属医院检验科,安徽合肥230601

出　　处：《中国感染控制杂志》2025年第1期37-44,共8页Chinese Journal of Infection Control

基　　金：安徽医科大学第二附属医院临床研究培良计划项目(2021LCYB04);安徽省高校自然科学研究项目(2023AH053175);安徽省转化医学研究院科研基金项目(2023zhyx-C78);安徽省转化医学研究院科研基金项目(2021zhyx-C47)。

摘　　要：目的通过全基因组测序分析临床耐碳青霉烯类肺炎克雷伯菌(CRKP)的分子流行病学及黏菌素耐药基因,为临床诊疗提供参考。方法收集2021—2023年安徽省某三级甲等综合医院住院患者临床标本分离的57株CRKP,对所有菌株进行药敏试验。采用全基因测序技术分析CRKP菌株多位点序列分型、荚膜血清型、耐药基因和毒力基因,并对所有菌株序列进行单核苷酸多态性分析,采用聚合酶链式反应(PCR)方法扩增与黏菌素耐药相关基因。结果57株CRKP对除替加环素外的14种抗菌药物均为耐药。测序结果显示,93.0%(53/57)的CRKP携带bla KPC-2,ST11型CRKP菌株检出率最高(51/57,89.5%)。单核苷酸多态性聚类分析表明,57株CRKP分为11个克隆群,其中有4个克隆群均为ST11-KL64型CRKP。40株(70.2%)CRKP携带多种毒力基因。5株黏菌素耐药CRKP,其耐药机制是在mgr B基因70位点处插入一个ISKpn26元件。结论CRKP菌株以产KPC-2 ST11-KL64型为主要流行型,在重症监护病房有播散性流行。ISKpn26元件插入引起mgr B基因突变与该地区CRKP黏菌素耐药有关。Objective To analyze the molecular epidemiology and colistin-resistant genes of carbapenem-resistant Klebsiella pneumoniae(CRKP)by whole-genome sequencing,and to provide reference for clinical diagnosis and treatment.Methods 57 CRKP strains isolated from clinical specimens of hospitalized patients in a tertiary general first-class hospital in Anhui Province from 2021 to 2023 were collected and antimicrobial susceptibility testing was performed.Multilocus sequence typing,capsule serotype,resistance genes,and virulence genes of CRKP strains were analyzed by whole-genome sequencing technique,and single nucleotide polymorphism analysis was conducted on sequences of all strains.Colistin resistance-related genes were amplified by polymerase chain reaction(PCR).Results 57 CRKP strains exhibited resistance to 14 antimicrobial agents,with the exception of tigecycline.The sequencing results showed that 93.0%(53/57)of CRKP carried bla KPC-2,and the ST11 type CRKP strain had the highest detection rate(51/57,89.5%).Single nucleotide polymorphism clustering analysis showed that the 57 CRKP strains were divided into 11 clone groups,of which 4 clone groups were all ST11-KL64 type CRKP.40(70.2%)CRKP strains carried multiple virulence genes.Five strains of CRKP were colistin-resistant strains,the resistance mechanism involved the insertion of ISKpn26 element at site 70 of the mgr B gene.Conclusion The CRKP strain is primarily characterized by the production of KPC-2 ST11-KL64,with disseminated transmission in intensive care unit.The insertion of ISKpn26 element leading to mgr B gene mutation is related to resistance of CRKP to colistin in this region.

关 键 词：耐碳青霉烯类肺炎克雷伯菌 全基因组测序 单核苷酸多态性分析 黏菌素 耐药 ISKpn26 

分 类 号：R181.32[医药卫生—流行病学] R378.996[医药卫生—公共卫生与预防医学]