神经重症医学科感染患者非多重耐药肺炎克雷伯菌的毒力基因分布与传播特征分析  

Analysis of Toxicity Genes Distribution and Transmission Characteristics of Non-multiple-resistant Klebsiella Pneumoniae in Patients with Infection in NICU

作　　者：元小冬[1] 张萍淑[1] 刘妍 李妍 耿贺梅[3] 王贺永[4] 张淑清 刘青 YUAN Xiaodong;ZHANG Pingshu;LIU Yan;LI Yan;GENG Hemei;WANG Heyong;ZHANG Shuqing;LIU Qing(Department of Neurology,Kailuan General Hospital,Tangshan 063000,China;Hebei Provincial Key Laboratory of Neurobiology Function,Tangshan 063000,China;Department of Infection Management,Kailuan General Hospital,Tangshan 063000,China;Department of Inspection,Kailuan General Hospital,Tangshan 063000,China)

机构地区：[1]开滦总医院神经内科,河北省唐山市063000 [2]河北省神经生物机能重点实验室,河北省唐山市063000 [3]开滦总医院医院感染管理科,河北省唐山市063000 [4]开滦总医院检验科,河北省唐山市063000

出　　处：《实用心脑肺血管病杂志》2021年第8期110-115,共6页Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease

基　　金：2020年度河北省创新能力提升计划项目——科技研发平台与新型研发机构建设专项(20567622H);2020年度唐山市科学技术研究与发展计划(20130210D)。

摘　　要：背景神经重症医学科(NICU)患者常存在意识及肢体运动障碍,多存在侵入性操作,免疫功能降低,因此其是肺炎克雷伯菌(KP)高危感染人群。探索非多重耐药KP的致病毒力、传播特点对指导临床治疗、减少多重耐药菌转化至关重要。目的通过全基因组测序分析,了解NICU感染患者非多重耐药KP毒力基因分布与传播特征。方法收集2015—2018年开滦总医院收治的93例NICU感染患者的非多重耐药KP 94株,所有KP应用K-B法进行抗菌药物敏感试验及脉冲场凝胶电泳法(PFGE)进行同源性分析,选出其中无同源性的KP 3株,其中2015年、2016年、2017年各1株,分别编号为P20、P39、P66;2018年,选出与P66为同一流行克隆系的KP 2株,分别编号为P90、P91,且P90与P91为同一克隆株。将5株非多重耐药KP进行二代、三代全基因组测序,将所得的基因组测序结果上传至NCBI的prokaryotic annotation pipeline系统进行基因组预测基因注释,并应用BLAST软件进行对比分析,获取病原菌毒力因子数据库(VFDB)比对基因注释结果。结果将NICU感染患者5株非多重耐药KP的基因组测序结果与VFDB进行比对,结果显示,5株非多重耐药KP中共筛选出6大类52种毒力基因,包括黏附类17种(32.7%)、分泌系统类15种(28.8%)、铁吸收类15种(28.8%)、调节类3种(5.8%)、内毒素类1种(1.9%)、侵入类1种(1.9%);其中P20携带毒力基因19种(36.5%)、P39携带毒力基因18种(34.6%)、P66携带毒力基因2种(3.8%)、P90携带毒力基因47种(90.4%)、P91携带毒力基因49种(94.2%)。NICU感染患者非多重耐药KP的基因注释结果显示:(1)黏附类毒力基因:P90、P91中均检出fimA、fimB、fimC、fimE、fimF、fimG、fimH、fimI、ecpB、ecpE、ecpR、mrkA、mrkB、mrkD、mrkF、yagZ/ecpA、fliY基因各1个,P20、P39中仅检出1个fliY基因,P66未检出此类毒力基因。(2)铁吸收类毒力基因:在同一克隆株P90、P91中均检出fepB、fepC、fepD、fepG、entA、entB、entS�Background Neurointensive Care Unit(NICU)patients are high-risk population of Klebsiella pneumonia(KP)infection due to disorders of consciousness,physical movement dysfunction,and many exist invasive procedure,and with lower immunity.It's urgent to discuss the virulence factors and communication features of non-multidrug-resistant KP in instructing clinical treatment and reducing transform non-multiple drug-resistant bacteria into a pathogenic multiple drug-resistant pathogenic bacteria.Objective To discuss the toxicity genes distribution and transmission characteristics of non-multiple-resistant KP in patients with infection in NICU via whole genome sequencing.Methods A total of 94 non-multidrug-resistant KP strains from 93 patients in NICU of Kailuan General Hospital from 2015 to 2018 were recruited.Antimicrobial susceptibility tests against antimicrobial agents were determined using the K-B diffusion method.The homology of these strains were analyzed by pulsed-field gel electrophoresis(PFGE).Three no homology KP strains were selected in 2015,2016 and 2017 respectivelly(P20,P39 and P66).Two KP strains belonged to the same clonal lineage as strain P66 were named P90 and P91.All KP strains were subjected to second-generation sequencing and whole genome sequencing.Sequencing data were uploaded to NCBI prokaryotic genome annotation pipeline for gene prediction and annotation.The BLAST software was used for sequence data.The virulence factors were identified by using the Virulence Factor Database(VFDB).Results A total of 6 kinds included 52 genes were selected from the sequencing results of 5 KP strains was applied to align against the VFDB database,included adhesion 17 kinds(32.7%),secretion systems 15 kinds(28.8%),iron absorption 15 kinds(28.8%),regulators 3 kinds(5.8%),endotoxin 1 kind(1.9%),and invasins 1 kind(1.9%);and with P20 carried 19 virulence genes(36.5%),P39 carried 18 virulence genes(34.6%),P66 carried 2 virulence genes(3.8%),P90 carried 47 virulence genes(90.4%),and P91 carried 49 virulence genes(94.2%

关 键 词：肺炎克雷伯菌 神经重症监护室 克隆株 全基因组测序 毒力基因 

分 类 号：R378.99[医药卫生—病原生物学]