优化与分析单核细胞增生李斯特菌的脉冲场凝胶电泳  

Optimization and Analysis of Molecular Typing by Pulsed-filed Gel Electrophoresis of Listeria Monocytogenes Isolates

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]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

出　　处：《实用预防医学》2009年第4期989-992,共4页Practical Preventive Medicine

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：目的优化单核细胞增生李斯特菌(LM)脉冲场凝胶电泳(PFGE)方法,分析菌株之间的相关性。方法按照标准化的Lm-PFGE方法进行预实验,根据结果对实验方法进行一系列调整。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BioNumerics软件对图谱进行聚类分析。结果优化后电泳图谱质量明显提高。22株LM被内切酶ApaI分为16种带型,被内切酶AscI分为17种PFGE型。两种酶分别将深圳的10株LM分为9种带型。结论建立了高分辨力、稳定可靠的Lm-PFGE分型方法。深圳的LM属于多个克隆群的散发流行。Objective To optimize and analyze the molecular typing by pulsed - filed gel electrophoresis of Listeria monocytogenes isolates, and to determine the genetic relationship between different LM isolates. Methods Preparing experiment was conducted according to L. monocytogenes- PFGE protocol, and the experiment method was optimized on the basis of the： results of preparing experiment. Chromosomal DNA from 22 isolates was digested in SeaKem Gold agrose with restriction enzyme Apal or Ascl 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 The quality of electrophore：tic patterns by optimization protocol was significantly better. 22 strains of L. rnonocytogenes were divided into 16 subtypes by enzyme Apal and into 17 subtypes by enzyme Ascl. 10 strains of L. monocytogenes from Shenzhen were divided into 9 subtypes by enzyme Apal or Ascl. Conclusions The stable, reliable and high- resolution Lm- PFGE protocol is established. The LM strains from Shenzhen are different clones.

关 键 词：单核细胞增生李斯特菌 脉冲场凝胶电泳 分子分型 

分 类 号：R378[医药卫生—病原生物学]