河南省2011-2014年D群宋内志贺菌病原学监测与分子流行病学研究

赵嘉咏; 张白帆; 穆玉姣; 苏佳; 黄丽莉; 夏胜利; 许汴利

Abstract

赵嘉咏,张白帆,穆玉姣,苏佳,黄丽莉,夏胜利,许汴利.河南省2011-2014年D群宋内志贺菌病原学监测与分子流行病学研究[J].Chinese journal of Epidemiology,2016,37(4):558-562

河南省2011-2014年D群宋内志贺菌病原学监测与分子流行病学研究

Surveillance on the etiology and molecular epidemiology of Shigella sonnei isolated in Henan province from 2011 to 2014

Received:September 30, 2015

DOI：10.3760/cma.j.issn.0254-6450.2016.04.024

KeyWord: 志贺菌,D群毒力基因药敏测试脉冲场凝胶电泳

English Key Word: Shigella sonnei, D group Virulence gene Antimicrobial susceptibility test Pulsed field gel electrophoresis

FundProject:国家科技重大专项(2012ZX10004201,2013ZX10004203)

Author Name	Affiliation	E-mail
Zhao Jiayong	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Zhang Baifan	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Mu Yujiao	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Su Jia	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Huang Lili	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Xia Shengli	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China
Xu Bianli	Institute of Infectious Disease Control and Prevention, Henan Provincial Center for Disease Control and Prevention, Zhengzhou 450016, China	xubl@hncdc.com.cn

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Abstract:

目的分析2011-2014年河南省腹泻患者粪便中分离的216株D群宋内志贺菌毒力基因携带情况、耐药谱及代表性菌株的PFGE指纹图谱特征,为志贺菌病的病原学监测及暴发溯源提供基线数据与方法学参考。方法采集腹泻患者粪便样本,SS平板分离培养18~24 h,采用克氏双糖铁(KIA)/动力-吲哚-尿素(MIU)与API20E系统进行生化鉴定,利用志贺菌分型血清进行玻片凝集;使用热裂解法制备DNA模板,采用多重PCR鉴定毒力基因种类。根据国际PulseNet细菌性传染病分子分型监测网络公布的宋内志贺菌PFGE分型技术方案进行PFGE分子分型与聚类分析。结果 216株D群宋内志贺菌中,98株为宋内Ⅰ型,118株为宋内Ⅱ型;各菌株均携带不同的毒力因子编码基因,包括SHET-1B、SHET-2、ial、ipaH,具有4种毒力基因组合类型;216株宋内志贺菌均为耐2种以上抗生素的多重耐药菌株,其中耐2~4种为34株(15.7%),耐5~8种为147株(68.1%),耐9~10种为24株(11.1%),耐11种为7株(3.2%),耐13种为4株(1.9%)。100株宋内志贺菌经XbaⅠ酶切与PFGE后共分为31种不同带型,相似度为68.6%~100.0%,各带型包含菌株数为1~13株不等。结论 2011-2014年河南省宋内志贺菌均携带致病性毒力基因,菌株耐药现象比较严重,其PFGE指纹图谱呈现多样性的同时又具有较显著的优势带型特点,部分带型与其对应的耐药谱具有一定的关联性与相同的聚集性。

English Abstract:

Objective To detect and analyze the distribution of virulence factors, antimicrobial resistance and pulsed field gel electrophoresis(PFGE) patterns of Shigella sonnei, isolated in Henan province from 2011 to 2014. Methods Samples of diarrhea patients were collected and isolated with SS selective culture medium in 37℃ for 18-24 hours. All strains were identified under the Kligler iron agar/motility-indol-urea biochemical action and API20E biochemical system. Serological typing and prepared DNA template were carried out with thermal cracking method and multiplex PCR, to detect the virulence genes of Shigella sonnei. According to the molecular typing method and K-B drug susceptibility testing method published by the international PulseNet bacterial infectious disease monitoring network and USA clinical and Laboratory Standards Institute, antimicrobial susceptibility tests and PFGE molecular characteristics of these positive strains isolated from sentinel hospitals patients stool samples were analyzed. Results Among the 98 strains of Sonnei typeⅠand 118 strains of Sonnei typeⅡ, all the strains carried carry different virulence genes including SHET-1B, SHET-2, ial, ipaH genes, with 4 kinds of virulence gene combination types. All the 216 strains of Shigella sonnei belonged to the multi-drug resistant strains, including 34 isolates resistant to 2-4 kinds of antibiotics(15.7%), 147 isolates to 5-8 kinds of antibiotics(68.1%), 24 to 9-10 kinds of antibiotics(11.1%), 7 to 11 kinds of antibiotics(3.2%), and 4 to 13 kinds of antibiotics(1.9%). A total of 100 strains of Shigella sonnei were divided into 31 molecular patterns, digested by XbaⅠand PFGE. Each pattern contained 1-13 strains with similarities ranged from 68.6%-100.0%. Conclusions All the Shigella sonnei strains carried virulence pathogenic factors, presenting serious status on drug resistance. PFGE fingerprinting patterns showed high polymorphism and dominant characteristics. PFGE patterns of partial strains and corresponding antidrug spectrum presented certain relevance and with same aggregation relationship.

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