采用脉冲场凝胶电泳和自动化核糖体分型技术分析MPN法分离出的副溶血性弧菌的同源性  

Homology analysis of vibrio parahaemolyticus isolated by MPN method using pulsed-field gel electrophoresis and automated ribotyping

作　　者：李薇薇[1] 王晓英[1] 裴晓燕[1] 朱海明[2] 郭云昌[1] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所,北京100021 [2]广东省疾病预防控制中心

出　　处：《卫生研究》2010年第3期374-380,共7页Journal of Hygiene Research

摘　　要：目的对15管MPN法分离的78株副溶血性弧菌进行脉冲场凝胶电泳(PFGE)和自动化核糖体分型(RP)研究,以确定其同源性。方法EcoRⅠ酶切DNA进行核糖体分型;限制性内切酶SfiⅠ和NotⅠ在50℃和37℃酶切包被的染色体DNA,进行脉冲场凝胶电泳分析,两者的结果均用BioNumerics软件进行聚类分析。结果78株副溶血性弧菌用EcoRⅠ、SfiⅠ、NotⅠ酶切后各产生64、68、71种带型,同一样品中分离的菌株有的带型完全相同,但大部分带型不同,且划分到不同的群中。结论PFGE是一种快速、有力、重复性好的分型技术,而且分辨能力要高于核糖体分型;在今后的溯源研究中,要考虑对可疑食品采用MPN定量法挑取多个菌落进行亲缘关系分析,防止漏检。Objective To study the molecular typing of 78 strains of Vibrio parahaemolyticus isolated from Guangdong by MPN method with pulsed-field gel electrophoresis（PFGE） and automated ribotyping（RP）.Methods DNAs from all strains were digested by EcoRⅠ for ribotyping and digested by SfiⅠ and NotⅠ for PFGE.The band patterns of PFGE and RP were clustering adn analyzed by BioNumerics software.Results There were 64,68 and 71 kinds of band patterns of each Vibrio parahaemolytics strain digested by restriction endonuclease EcoRⅠ,SfiⅠ and NotⅠ respectively.The band patterns of some strains from the same sample are completely identical,but the band patterns of most strains are different and are divided into different groups.Conclusion PFGE is a rapid,robust,and reproducible genotyping technique,more discriminative than automated ribotyping.Choosing more isolates from MPN method for suspected food samples should be considered in tracing the phylogenetic relatinship of strains.

关 键 词：弧菌 副溶血性 脉冲场凝胶电泳 核糖体分型 

分 类 号：TS207.3[轻工技术与工程—食品科学] R155.5[轻工技术与工程—食品科学与工程]