高黏液性耐碳青霉烯类肺炎克雷伯菌分子特征和毒力分析

doi:10.3969/j.issn.1673-8640.2025.04.008

摘要/Abstract

摘要：

目的探讨高黏液性耐碳青霉烯类肺炎克雷伯菌（HM-CRKP）的分子特征和毒力水平，为此类菌株临床研究和感染控制提供参考。方法收集2009年1月—2020年12月分离自上海交通大学医学院附属新华医院住院患者的耐碳青霉烯类肺炎克雷伯菌（CRKP）1 825株。采用拉丝试验筛选HM-CRKP。采用荚膜wzi基因分型鉴定HM-CRKP的荚膜血清型别，采用聚合酶链反应（PCR）检测6种主要的碳青霉烯酶基因（bla_KPC、bla_NDM、bla_IMP、bla_VIM、bla_GES、bla_OXA-48）和4种毒力基因（rmpA2、iucA、rmpA、iroN），采用大蜡螟幼虫感染模型检测HM-CRKP毒力。结果 1 825株CRKP中共筛出96株HM-CRKP。HM-CRKP自2015年被检出后检出率呈显著上升趋势，总检出率为5.3%，2020年检出率为12.2%。有95株HM-CRKP检出碳青霉烯酶基因，包括bla_KPC-2、bla_KPC-3、bla_NDM-1和bla_IMP-4，以bla_KPC-2为主（89.6，86/96）；有1株HM-CRKP同时检出bla_KPC-2和bla_IMP-4。96株HM-CRKP存在9种wzi荚膜血清型，以wzi64为主（82.3%，79/96）。 69.8%的HM-CRKP携带rmpA2、iucA，59.4%的HM-CRKP携带rmpA、iroN。根据毒力基因rmpA2、iucA、rmpA、iroN的携带情况，可将HM-CRKP分成3种模式：59.4%的菌株同时检出rmpA2、iucA、rmpA、iroN（模式Ⅰ），10.4%的菌株同时检出rmpA2、iucA（模式Ⅱ），30.2%的菌株rmpA2、iucA、rmpA、iroN均未检出（模式Ⅲ）。HM-CRKP菌株间毒力差异较大，不同毒力基因模式的HM-CRKP间毒力差异无统计学意义（P>0.05）。结论 2009—2020年上海交通大学医学院附属新华医院住院患者HM-CRKP以携带bla_KPC-2的wzi64为主，且检出率逐年上升。HM-CRKP菌株间毒力差异大，不同毒力基因模式的HM-CRKP之间毒力没有显著差异。

关键词: 肺炎克雷伯菌, 碳青霉烯类耐药, 高黏液性, bla_KPC-2, wzi64, 毒力

Abstract:

Objective To study the molecular characteristics and virulence of hypermucoviscous carbapenem-resistant Klebsiella pneumoniae（HM-CRKP），and to provide a reference for clinical research and infection control.Methods The 1 825 isolates of carbapenem-resistant Klebsiella pneumoniae（CRKP） were isolated from inpatients of Xinhua Hospital，Shanghai Jiao Tong University School of Medicine from January 2009 to December 2020，and the string test was used to screen for HM-CRKP. The serotypes of HM-CRKP isolates were identified by capsular wzi genotyping. The 6 major carbapenemase genes（bla_KPC，bla_NDM，bla_IMP，bla_VIM，bla_GES and bla_OXA-48） and 4 virulence genes（rmpA2，iucA，rmpA and iroN） were identified by polymerase chain reaction（PCR）. The virulence of isolates were determined by Galleria mellonella larvae infection model.Results Totally，96 isolates of HM-CRKP were screened from 1 825 isolates of CRKP. The determination rate of HM-CRKP had been increasing since the first determination in 2015，reached 12.2% in 2020. The total determination rate of HM-CRKP was 5.3%. Carbapenemase genes were determined in 95 isolates of HM-CRKP，including bla_KPC-2，bla_KPC-3，bla_NDM-1 and bla_IMP-4，and bla_KPC-2 was found to be the most predominant（89.6%，86/96）. The bla_KPC-2 and bla_IMP-4 were found in 1 isolate，and 9 different capsular wzi serotypes were found in HM-CRKP，of which wzi64 was the dominant serotype（82.3%，79/96）. The 69.8% of HM-CRKP carried rmpA2 and iucA，59.4% of HM-CRKP carried rmpA and iroN. According to the distribution of virulence genes（rmpA2，iucA，rmpA and iroN），HM-CRKP could be classified into 3 modes. The 59.4% of the isolates were belonged to mode Ⅰ（rmpA2，iucA，rmpA and iroN were determined simultaneously）；the 10.4% of the isolates were belonged to mode Ⅱ（rmpA2 and iucA were determined simultaneously）；the 30.2% of the isolates were belonged to mode Ⅲ（rmpA2，iucA，rmpA and iroN were not determined）. The virulence of HM-CRKP isolates was different from each other in Galleria mellonella larvae model，but there was no statistical significance in virulence among HM-CRKP isolates with different virulence gene modes（P>0.05）.Conclusions The bla_KPC-2 carried wzi64 is the main HM-CRKP collected from Xinhua Hospital，Shanghai Jiao Tong University School of Medicine，and the determination rate of HM-CRKP is increasing year by year. The virulence of HM-CRKP is different from each other，while there is no significant virulence difference of HM-CRKP with different virulence gene modes.

Key words: Klebsiella pneumoniae, Carbapenem resistance, Hypermucoviscous, bla_KPC-2, wzi64, Virulence

中图分类号:

R446.1

李虎, 朱威南, 陈峰, 王佳玮, 刘瑛, 沈立松, 郑英霞. 高黏液性耐碳青霉烯类肺炎克雷伯菌分子特征和毒力分析[J]. 检验医学, 2025, 40(4): 358-364.

LI Hu, ZHU Weinan, CHEN Feng, WANG Jiawei, LIU Ying, SHEN Lisong, ZHENG Yingxia. Molecular characteristics and virulence analysis of hypermucoviscous carbapenem-resistant Klebsiella pneumonia[J]. Laboratory Medicine, 2025, 40(4): 358-364.

图/表 4

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抗菌药物	总体（96株）	携带bla_KPC-2的HM-CRKP（86株）	不携带bla_KPC-2的HM-CRKP（10株）
头孢呋辛	100.0	100.0	100.0
头孢噻肟	100.0	100.0	100.0
头孢曲松	100.0	100.0	100.0
头孢他啶	99.0	100.0	90.0
头孢吡肟	95.8	98.8	70.0^*
头孢美唑	95.8	98.8	70.0^*
氨苄西林-舒巴坦	100.0	100.0	100.0
亚胺培南	100.0	100.0	100.0
美罗培南	100.0	100.0	100.0
庆大霉素	81.3	89.5	10.0^*
阿米卡星	80.2	88.4	10.0^*
左氧氟沙星	92.7	100.0	30.0^*
环丙沙星	92.7	100.0	30.0^*
复方磺胺甲噁唑	59.4	65.1	10.0^*
替加环素	0.0	0.0	0.0

抗菌药物	总体（96株）	携带bla_KPC-2的HM-CRKP（86株）	不携带bla_KPC-2的HM-CRKP（10株）
头孢呋辛	100.0	100.0	100.0
头孢噻肟	100.0	100.0	100.0
头孢曲松	100.0	100.0	100.0
头孢他啶	99.0	100.0	90.0
头孢吡肟	95.8	98.8	70.0^*
头孢美唑	95.8	98.8	70.0^*
氨苄西林-舒巴坦	100.0	100.0	100.0
亚胺培南	100.0	100.0	100.0
美罗培南	100.0	100.0	100.0
庆大霉素	81.3	89.5	10.0^*
阿米卡星	80.2	88.4	10.0^*
左氧氟沙星	92.7	100.0	30.0^*
环丙沙星	92.7	100.0	30.0^*
复方磺胺甲噁唑	59.4	65.1	10.0^*
替加环素	0.0	0.0	0.0

碳青霉烯酶基因	2015年（142株）	2016年（271株）	2017年（264株）	2018年（361株）^①	2019年（291株）	2020年（353株）^②
bla_KPC-2	1（0.7）	2（0.7）	6（2.3）	11（3.0）	24（8.2）	42（11.9）
bla_KPC-3	0（0.0）	0（0.0）	1（0.4）	0（0.0）	0（0.0）	0（0.0）
bla_NDM-1	0（0.0）	0（0.0）	1（0.4）	2（0.6）	0（0.0）	0（0.0）
bla_IMP-4	0（0.0）	0（0.0）	0（0.0）	5（1.4）	1（0.3）	0（0.0）

碳青霉烯酶基因	2015年（142株）	2016年（271株）	2017年（264株）	2018年（361株）^①	2019年（291株）	2020年（353株）^②
bla_KPC-2	1（0.7）	2（0.7）	6（2.3）	11（3.0）	24（8.2）	42（11.9）
bla_KPC-3	0（0.0）	0（0.0）	1（0.4）	0（0.0）	0（0.0）	0（0.0）
bla_NDM-1	0（0.0）	0（0.0）	1（0.4）	2（0.6）	0（0.0）	0（0.0）
bla_IMP-4	0（0.0）	0（0.0）	0（0.0）	5（1.4）	1（0.3）	0（0.0）