山羊源K57荚膜型肺炎克雷伯菌毒力基因检测及遗传进化分析

doi:10.16431/j.cnki.1671-7236.2025.10.032

摘要/Abstract

摘要： 【目的】探究山羊源K57荚膜型肺炎克雷伯菌KPHN001株毒力因子携带情况，了解不同来源K57型肺炎克雷伯菌毒力基因分布和序列型(ST)分型信息，以探讨该型菌株的遗传多样性及可能的进化特征。【方法】将肺炎克雷伯菌KPHN001株复苏培养，采用菌落拉丝试验确定其毒力分型。使用PCR方法检测其荚膜血清型和毒力基因。根据多位点序列分型(MLST)方法和单拷贝同源基因构建系统发育树的方法，将KPHN001株与NCBI数据库中30株不同来源的K57型肺炎克雷伯菌进行比对分析。【结果】菌落拉丝试验结果显示，KPHN001株无法拉出≥5 mm的菌丝。PCR检测结果显示，KPHN001株为K57荚膜型，且含有khe、fimH、uge、kfuBC、wabG、ureA、entB、mrkD、repA和sopB多个毒力基因，高毒力型肺炎克雷伯菌(hvKP)的重要遗传标记基因如rmpA2、iucA、iroB等均未检出。结合菌落拉丝试验结果，表明KPHN001株为经典肺炎克雷伯菌(cKP)。MLST结果显示，牛源、羊源及环境菌株ST型分布呈现多样性，而人源菌株主要集中于ST412和ST218型，KPHN001株为ST528型。系统发育树与毒力基因热图联合分析结果表明，人源菌株在uge基因携带上与其他来源菌株存在显著差异，且人源菌株形成独立分支，表明这些菌株在进化过程中可能积累了独特的遗传变异。【结论】本研究揭示了山羊源K57型肺炎克雷伯菌的毒力基因及K57型肺炎克雷伯菌遗传多样性，为其毒力机制的理解及肺炎克雷伯菌病的诊断和预防提供理论基础。

关键词: 肺炎克雷伯菌; K57荚膜型; 毒力基因; 多位点序列分型; 遗传进化

Abstract: 【Objective】 The purpose of the experiment was to investigate the virulence gene profile of the goat-derived Klebsiella pneumoniae strain KPHN001 with the K57 capsule serotype,and examine the distribution of virulence genes and sequence types (STs) among K57 capsule serotype Klebsiella pneumoniae strains from various sources,in order to assess their genetic diversity and potential evolutionary features.【Method】 After resuscitation and cultivation,the virulence phenotype of KPHN001 strain was determined by string test.The serotype and virulence genes of the strain were examined by PCR method.According to multilocus sequence typing (MLST) and phylogenetic tree reconstruction of single copy homologous genes,KPHN001 strain was compared with 30 strains of K57 capsule serotype Klebsiella pneumoniae from different sources in NCBI database.【Result】 String test result showed that KPHN001 strain was unable to produce a mucoviscous string ≥5 mm in length.PCR results showed that KPHN001 strain was K57 capsule serotype,containing several virulence genes including khe,fimH,uge,kfuBC,wabG,ureA,entB,mrkD,repA and sopB,while none of the important genetic marker genes for high-virulence Klebsiella pneumoniae (hvKP),such as rmpA2,iucA and iroB,were detected.Based on the negative string test and the absence of hvKP marker genes,the KPHN001 strain was identified as a classical Klebsiella pneumoniae (cKP) strain.The MLST results showed that the ST type distribution of K57 capsule serotype Klebsiella pneumoniae strains from bovine,goat and environment exhibited diversity,while the strains from human mainly concentrated on ST412 and ST218.KPHN001 strain was classified as ST528.The combined analysis of phylogenetic tree with virulence gene heatmap indicated that strains from human exhibited significant differences in uge gene expression compared to strains from other sources,and strains from human formed independent branches,indicating that these strains might have accumulated unique genetic variation during evolution.【Conclusion】 This study elucidated the virulence gene profile of a goat-derived K57 capsule serotype Klebsiella pneumoniae strain and highlighted the genetic diversity of K57 capsule serotype strains,providing a theoretical basis for understanding their virulence mechanisms and the diagnosis and prevention of Klebsiella pneumoniae infection.

Key words: Klebsiella pneumoniae; K57 capsule serotype; virulence genes; MLST; genetic evolution

中图分类号:

S852.61

何美荣, 廖连杰, 何秋仪, 潘浩举, 杨映雪, 陈韬羽, 李昊洋, 蒋俊明, 陈巧玲, 高宏岩, 陈思, 杜丽, 王凤阳, 满初日嘎. 山羊源K57荚膜型肺炎克雷伯菌毒力基因检测及遗传进化分析[J]. 中国畜牧兽医, 2025, 52(10): 4874-4883.

HE Meirong, LIAO Lianjie, HE Qiuyi, PAN Haoju, YANG Yingxue, CHEN Taoyu, LI Haoyang, JIANG Junming, CHEN Qiaoling, GAO Hongyan, CHEN Si, DU Li, WANG Fengyang, MAN Churiga. Virulence Gene Detection and Genetic Evolution Analysis of Goat-derived K57 Capsule Serotypes Klebsiella pneumoniae[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4874-4883.

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