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【目的】建立空肠弯曲菌脉冲场电泳(pulsed-field gel electrophoresis,PFGE)图谱分型方法。【方法】在PFGE基本程序基础上,通过调整菌液浓度、Seakem Gold琼脂糖凝胶浓度、蛋白酶K浓度、洗涤方式和限制性内切酶SmaⅠ浓度,进行程序的比较与优化。应用PFGE技术对不同来源分离株进行分析。【结果】37株空肠弯曲菌脉冲场凝胶电泳图谱显示分离株均产生了6～24条电泳带,条带数量适中,清晰易读;系统进化树显示,可分为4个遗传谱系,分离株主要分布于PFGE遗传谱系Ⅳ,不同源分离株在各组群中呈交叉分布。【结论】PFGE对空肠弯曲菌具有很强的分型能力和良好的溯源性,初步数据显示鸡、牛等动物源空肠弯曲菌与腹泻病人空肠弯曲菌病密切相关。
【Objective】 To establish a method for the mapping of Campylobacter jejuni pulsed-field gel electrophoresis (PFGE). 【Method】 Based on the PFGE protocol, the programs were compared and optimized by adjusting the bacterial concentration, Seakem Gold agarose gel concentration, proteinase K concentration, washing method and restriction endonuclease Sma Ⅰ concentration. PFGE technology was used to analyze isolates from different sources. 【Result】 The results of gel electrophoresis of 37 strains of Campylobacter jejuni showed that all of the isolates had 6-24 electrophoretic bands with moderate number of bands and legibility. The phylogenetic tree showed that there were 4 genetic lines Strain mainly distributed in PFGE genetic line Ⅳ, different source isolates were cross-distributed in each group. 【Conclusion】 PFGE has strong typing ability and good traceability to Campylobacter jejuni. Preliminary data show that Campylobacter jejuni is closely related to Campylobacter jejuni in diarrhea patients.