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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 determine the homology of 78 strains of Vibrio parahaemolyticus isolated by 15-tube MPN by pulsed-field gel electrophoresis (PFGE) and automated ribosome typing (RP). Methods DNA was digested with EcoRⅠ and sequenced. DNA fragments were digested with restriction endonucleases SfiⅠ and NotⅠ at 50 ℃ and 37 ℃, and analyzed by pulsed-field gel electrophoresis. The results of both were analyzed by BioNumerics software Cluster analysis. Results 78 strains of Vibrio parahaemolyticus were digested with EcoRⅠ, SfiⅠ and NotⅠ, respectively, and 64, 68 and 71 bands were produced respectively. Some isolates of the same sample belonged to the same type, but most of them belonged to different bands. To different groups. Conclusion PFGE is a rapid, powerful and reproducible typing technique, and its resolving power is higher than that of ribosome typing. In the future traceability studies, it is necessary to consider the use of MPN quantification to select multiple colonies for suspicious foods Analysis of kinship to prevent missed inspection.