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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 the method of pulsed-field gel electrophoresis (PFGE) of Listeria monocytogenes (LM) and analyze the correlation between the strains. Methods According to the standardized Lm-PFGE method, a series of experiments were carried out based on the results. The strains were digested with restriction endonucleases ApaI and AscI, respectively. Electrophoresis patterns were obtained by pulsed-field gel electrophoresis. BioNumerics software was used for cluster analysis. Results After optimization, the quality of electropherogram was significantly improved. Twenty-two LM strains were divided into 16 bands by endonuclease ApaI and divided into 17 PFGE types by endonuclease AscI. Two enzymes were divided into nine strains of 10 LM in Shenzhen. Conclusion A high resolution, stable and reliable Lm-PFGE typing method was established. LM in Shenzhen belongs to the distribution of multiple clonal groups.