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目的了解无锡市2009～2011年副溶血性弧菌分离株携带的主要毒力因子的流行状况,并对同血清型菌株进行脉冲场凝胶电泳(PFGE)分析。方法应用PCR方法检测分离的35株副溶血性弧菌耐热性溶血毒素基因(tdh)、耐热性溶血毒素相关的溶血毒素基因(trh)和不耐热溶血毒素基因(tlh)。根据美国CDC PulseNet实验方法,用限制性内切酶SfiⅠ对O3﹕K6血清型菌株的染色体进行酶切,通过PFGE获得电泳图谱,利用BioNumerics软件对图谱进行聚类分析。结果 35株副溶血性弧菌tdh、trh及tlh基因的携带率分别为85.7%、8.6%和100%,77.1%的副溶血性弧菌携带的毒力基因为tdh+、trh-、tlh+。PFGE图谱显示,19株O3﹕K6血清型的副溶血性弧菌共有9种PFGE带型,带型100%相同的菌株几乎都出现在同一年代相近的时间点,但也出现了跨年代菌株。结论无锡市副溶血性弧菌致病性较强,具有潜在的O3﹕K6型副溶血性弧菌暴发流行可能,需进一步加强监测管理。
Objective To understand the prevalence of major virulence factors carried by Vibrio parahaemolyticus isolates from 2009 to 2011 in Wuxi and analyze the PFGE of the same serogroups. Methods 35 strains of Vibrio parahaemolyticus heat-tolerant hemolysin gene (tdh), heat-resistant hemolysin-related hemolysin gene (trh) and heat-labile hemolysin gene (tlh) were detected by PCR. According to the CDC PulseNet experimental method in the United States, the chromosome of the O3: K6 serotype was digested with restriction endonuclease SfiI, the electrophoresis pattern was obtained by PFGE, and the pattern was analyzed by BioNumerics software. Results The carriage rates of tdh, trh and tlh genes of 35 Vibrio parahaemolyticus were 85.7%, 8.6% and 100%, respectively. The virulence genes of 77.1% Vibrio parahaemolyticus were tdh +, trh- and tlh +. PFGE patterns showed that 19 strains of Vibrio parahaemolyticus of O3: K6 serotypes had 9 PFGE patterns. Strains with 100% identical bands almost all appeared at similar time points in the same age, but cross-generation strains also appeared. Conclusion The pathogenicity of Vibrio parahaemolyticus in Wuxi City is strong. Potential O3: K6 Vibrio parahaemolyticus outbreaks are likely to occur, and the monitoring and management should be further strengthened.