单核细胞增生李斯特菌PFGE分型与血清学分型的联合分析  

Study on molecular typing by pulsed-filed gel electrophoresis of Listeria monocytogenes isolates and their correlation with serovar

作　　者：王冰[1] 扈庆华[1] 兰全学[1] 石晓路[1] 林一曼[1] 程锦泉[1] 马汉武[1] 叶长芸[2] 阚飚[2] 

机构地区：[1]广东省深圳市疾病预防控制中心,广东深圳518020 [2]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《中国卫生检验杂志》2010年第3期459-461,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金资助项目(30300281);广东省卫生厅资助项目(A2003709);深圳市科技局资助项目(200404139)

摘　　要：目的:采用国际标准化的脉冲场凝胶电泳(PFGE)分析方法对单核细胞增生李斯特菌(Lm)进行基因分型,同时进行血清学分型,比较分析两种分型方法的优劣和之间的关系。方法:按照标准化的Lm-PFGE方法对17株Lm进行PFGE分型,分别用限制性内切酶ApaI和AscI酶对菌株进行酶切,通过脉冲场凝胶电泳获得电泳图谱,利用B ioNum erics软件对图谱进行聚类分析。同时对上述菌株进行血清学分型。结果:17株Lm被内切酶ApaI分为13种带型,被内切酶AscI分为14种PFGE型。17株Lm分为5种血清型。结论:Lm-PFGE分型方法是一种高分辨力、稳定可靠的技术。PFGE的分辨力高于血清学分型,两种分型方法密切相关。聚类分析结果显示ApaI酶切分型与H抗原密切相关。Objective：To study the molecular typing by pulsed-filed gel electrophoresis of Listeria monocytogenes isolates and their Correlation with serovar.Methods：PFGE protocol was optimized for genomic analyses of Lm.Chromosomal DNA from 17 isolates were digested in seakem gold agrose with restriction enzyme ApaI or AscI and plugs were then analyzed by pulsed-field gel electrophoresis.Pulsed-field gel electrophoresis（PFGE） patterns of L.monocytogenes isolates were clustered using BioNumerics software.Results：Of 17 Lm strains analyzed,the enzymes AscI and ApaI established 14 and 13 unique restriction endonuclease digestion profiles（REDP）,respectively.All the Lm strains were divided into 5 serovar.Conclusion：PFGE analysis is a reproducible and highly discriminating method for characterizing Lm.Data indicates that genotypic data generated by PFGE can be directly related to phenotypic data generated by serotyping for establishing the overall relatedness of isolates.And PFGE is better than serotyping.Distinct genomic divisions by ApaI of Lm are correlated with the flagellar（H） antigen type.

关 键 词：单核细胞增生李斯特菌 脉冲场凝胶电泳 分子分析 血清学分型 分型方法 血清型 密切相关 电泳图谱 菌株 酶切 

分 类 号：R378.994[医药卫生—病原生物学]