CAJ | 학술논문

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Xba Ⅰ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-Bln Ⅰ双酶切分型,获得分型共174种,D值为0.989l,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.
목적 료해광동성서상한사문균감측현황급균주분자분형특점,병초보건립수거고,위식원성질병폭발추종오염원급전파도경제공기선수거.방법 삼고국제PulseNet공포적사문균맥충장응효전영(PFGE)분형방안급7개위점적서상한사문균다위점가변수목천련중복서렬분석(MLVA)방법,분석광동성2009-2011년10개시275주서상한사문균분형특점,이용BioNumerics연건건립분자분형수거고.결과 2009-2011년광동성사문균년균검출솔위4.03％;서상한사문균년균검출솔화구성비분별위1.38％화34.29％,균정쌍봉형분포추세,류행고봉위5화9월.275주서상한사문균경Xba Ⅰ매절화PFGE분석,획득124충대형,기분형D치위0.928 6,이MLVA분형획득143충형별,D치위0.966 5,분변능력고우단매절적PFGE분형.대기중＞3주적상동대형진일보채용PFGE-XbaⅠ-Bln Ⅰ쌍매절분형,획득분형공174충,D치위0.989l,여채용MLVA방법상동,즉대동일균주적분변결과구유교고일치성.결론 광동성서상한사문균구유유전다양성.초보건립적균주분자분형수거고현시채용MLVA분형방법가완성균주취집성분석,이학인서상한사문균인기적폭발.
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 foodbome 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 Xba Ⅰ restricted 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 Bln Ⅰ 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-Xba Ⅰ but equal to PFGE-Xba Ⅰ-Bln Ⅰ.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.