脉冲场凝胶电泳分子分型方法用于霍乱弧菌相关性分析

    于晓婕1，麻丽丹2，杨帆3，刁保卫4

    1.四川大学生命科学学院，四川 成都 610211；2.丹东出入境检验检疫局；

    3.大连医科大学公共卫生学院；4.国家疾病预防控制中心腹泻重点实验室

    摘要：目的用脉冲场凝胶电泳(PFGE)分子分型方法，分析辽宁丹东和广东珠海两地区从水体和水产品中分离的霍乱弧菌之间的相关性。方法对39株霍乱弧菌用内切酶Not I酶切DNA后进行PFGE分子分型，并在0.5×TBE电泳缓冲液中加入终浓度为3.8mg/L的硫脲溶液电泳。结果所有39株霍乱弧菌的基因组DNA显示出良好的分型结果，从朝鲜排污口、丹东鸭绿江水体中的鲫鱼、河蟹以及丹东排污口分离出的霍乱弧菌之间的相似度达到100%，说明这些产品之间应为同一个污染源；珠海的大头鱼和鳙鱼之间的相似度达到100%，说明这两者之间应为同一个污染源；珠海鲫鱼中检出的霍乱弧菌和鸭绿江水体中检出的霍乱弧菌之间的相似度达到88.4%，说明这两者之间可能为同一个污染源。结论PFGE技术适用于霍乱弧菌环境样品分离菌株的相关性分析和传染源的追踪。

    关键词：霍乱弧菌；分子分型；水体；水产品；脉冲场凝胶电泳

    中图分类号：R516.5　文献标识码：B

    Correlation analysis of Vibrio cholerae bypulsed-field gel electrophoresis

    YU Xiao-jie*, MA Li-dan, YANG Fan， DIAOBao-wei

    *School of life Sciences, Sicuan University,Sicuan， Chengdu 610207, China

    Abstract:   Objective  To study the correlation of Vibrio cholerae by pulsed-field gelelectrophoresis(PFGE) ,which were isolated from Liaoning Dandongand Guangdong Zhuhai in water body and seafood.  Methods   The chromosome DNA of 40 strains of Vibriocholerae were digested by NotⅠenzyme and the PFGE were carried out,and 3.8mg/L finally concentration of Thiourea solution was addedinto the Thiourea0.5×TBE for Gel electrophoresisbuffer.   Results   The chromosome DNA of 40strains of Vibrio cholerae were typed well. The similarity ofVibrio cholerae which were isolated from sewage outfall of North Korea, Crucian Carp and river crab in Yalu river of Dandong,and sewage outfall of Dandong was up to 100%, it showed they werepolluted by the same pollutant. The similarity of Vibrio choleraewhich were isolated from Aspiorhynchus Laticeps and AristichthysNobilis in Zhuhai was up to 100%, it showed they were polluted bythe same pollutant. The similarity of Vibrio cholerae which wereisolated from Crucian Carp in Zhuhai and water body in Yalu riverwas up to 88.4%, it showed they were polluted by the samepollutant. Conclusion   The fine typing ability andreliability of PFGE are suitable for analyzing the correlation andfinding out the infectious resource.

    Key words:   Vibrio cholerae;Molecule typing; Water body ; Seafood; Pulsed-field gelelectrophoresis（PFGE）

    《中国国境卫生检疫杂志》