广东省2009-2011年鼠伤寒沙门菌监测及菌株分子分型的研究

黄燕惠; 柯碧霞; 孙九峰; 何冬梅; 陈清; 柯昌文; 俞守义

Abstract

黄燕惠,柯碧霞,孙九峰,何冬梅,陈清,柯昌文,俞守义.广东省2009-2011年鼠伤寒沙门菌监测及菌株分子分型的研究[J].Chinese journal of Epidemiology,2014,35(8):917-924

广东省2009-2011年鼠伤寒沙门菌监测及菌株分子分型的研究

Molecular typing and surveillance on Salmonella typhimurium strain in Guangdong province,2009-2011

Received:March 14, 2014

DOI：10.3760/cma.j.issn.0254-6450.2014.08.010

KeyWord: 鼠伤寒沙门菌加强监测分子分型

English Key Word: Salmonella typhimurium Enhance Salmonella Surveillance Molecular typing

FundProject:广东省医学科技研究基金（2013A074）；中美新发和再发传染病合作项目（1U2GGH000018-01）

Author Name	Affiliation	E-mail
Huang Yanhui	Department of Epidemiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China Department of Skin Venereal Disease Prevention and Control, Baoan District Chronic Disease Hospital
Ke Bixia	Guangdong Provincial Center for Disease Control and Prevention, Guangzhou	kebixia@live.cn
Sun Jiufeng	Guangdong Provincial Institute of Public Health
He Dongmei	Guangdong Provincial Center for Disease Control and Prevention, Guangzhou
Chen Qing	Department of Epidemiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China
Ke Changwen	Guangdong Provincial Center for Disease Control and Prevention, Guangzhou
Yu Shouyi	Department of Epidemiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China

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Abstract:

目的了解广东省鼠伤寒沙门菌监测现况及菌株分子分型特点，并初步建立数据库，为食源性疾病暴发追踪污染源及传播途径提供基线数据。方法参考国际PulseNet公布的沙门菌脉冲场凝胶电泳（PFGE）分型方案及 7 个位点的鼠伤寒沙门菌多位点可变数目串联重复序列分析（MLVA）方法，分析广东省2009-2011年10个市275株鼠伤寒沙门菌分型特点，利用BioNumerics软件建立分子分型数据库。结果 2009-2011年广东省沙门菌年均检出率为4.03%；鼠伤寒沙门菌年均检出率和构成比分别为1.38%和34.29%，均呈双峰形分布趋势，流行高峰为5和9月。275株鼠伤寒沙门菌经XbaⅠ酶切和PFGE分析，获得124种带型，其分型D值为 0.928 6，而MLVA分型获得143种型别，D值为0.966 5，分辨能力高于单酶切的 PFGE分型。对其中>3株的相同带型进一步采用PFGE-XbaⅠ- BlnⅠ双酶切分型，获得分型共174种，D值为0.989 1，与采用MLVA方法相同，即对同一菌株的分辨结果具有较高一致性。结论广东省鼠伤寒沙门菌具有遗传多样性。初步建立的菌株分子分型数据库显示采用MLVA 分型方法可完成菌株聚集性分析，以确认鼠伤寒沙门菌引起的暴发。

English Abstract:

Objective To understand the distribution and the characteristics on molecular typing of Salmonella (S.) typhimurium isolates gathered from the surveillance program and to construct the standard S. typhimurium databank in the laboratory through surveillance network PulseNet,in Guangdong province to improve the capability of detection on laboratory-based foodborne outbreaks. Methods With the application of standard pulse-field gel electrophoresis (PFGE) and multiple loci VNTR analysis (MLVA) including seven VNTRs loci protocols on PulseNet International Network,275 isolates of S. typhimurium from ten cities in Guangdong province were typed and their patterns analyzed. The molecular typing databank was constructed by BioNumerics. Results With S. typhimurium the most common serotypes,the average annual positive rate of Salmonella strains and S. typhimurium were 4.03% and 1.38% respectively. The positive rate and proportion presented a double-peak trend,appearing in May and September. The chromosomal DNA of S. typhimurium was digested with XbaⅠrestricted endonucleotidase and 124 PFGE patterns were observed after pulse-field gel electrophoresis,with the discrimination index (D) as 0.928 6. The patterns including more than three S. typhimurium isolates and were further digested with the second enzyme BlnⅠ to achieve 174 patterns,with the D value as 0.989 1. Under MLVA method,143 variant patterns were obtained,with the D value reaching 0.966 5. Data showed that the discriminatory ability of the MLVA typing method in S. typhimurium was superior to PFGE-XbaⅠ but equal to PFGE-XbaⅠ- BlnⅠ. In addition, when S. typhimurium strains were respectively analyzed by PFGE under double enzymes digestion and MLVA,the results appeared coincident and relative. Conclusion The variant patterns showed by the two molecular typing methods indicating a genetic diversity existed among the clinical S. typhimurium isolates in Guangdong province. Databank of S. typhimurium was constructed and could be used in laboratory surveillance programs. Under the characterization of analyzing similarity and evolution among S. typhimurium isolates,MLVA was suitable for cluster analysis on early detection of outbreaks caused by S. typhimurium.

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