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目的运用脉冲场凝胶电泳技术(pulsed-field gel electrophoresis,PFGE)和PCR技术对辽宁沈阳市2010年7—8月份临床粪便标本分离的副溶血性弧菌进行毒力基因检测和分子分型分析。方法运用PCR技术检测24株副溶血弧菌的耐热溶血素基因(tdh)、耐热溶血素相关溶血素基因(trh)和不耐热溶血素基因(tl);用PFGE技术对其进行分子分型和聚类分析。结果 24株副溶血弧菌均含有tdh和tl基因,表明均为有毒株;PFGE结果显示,24株菌株可分为10种图谱型,其中17株O3:K6被分为4种型别(仅相差1～3条带),表明17株O3:K6在流行病学上密切相关,提示沈阳地区可能存在副溶血弧菌腹泻病的局部暴发或流行。结论沈阳地区流行的食源性副溶血弧菌存在基因型的多样性,但以遗传关系密切的O3:K6型为优势型别。
Objective To detect virulence genes and analyze the molecular typing of Vibrio parahaemolyticus isolated from clinical stool samples from July to August in 2010 in Shenyang, Liaoning by pulsed-field gel electrophoresis (PFGE) and PCR . Methods 24 strains of Vibrio parahaemolyticus were detected by polymerase chain reaction (PCR), including heat-resistant hemolysin (tdh), heat-resistant hemolysin (trh) and heat-labile hemolysin (tl) Typing and cluster analysis. Results All 24 strains of V. parahaemolyticus contained tdh and tl genes, indicating that they were all virulent strains. PFGE results showed that 24 strains could be divided into 10 patterns, of which 17 were O3: K6 divided into 4 types (only A difference of 1 to 3 bands), indicating that 17 O3: K6 epidemiologically closely related, suggesting that there may be local outbreaks or epidemic of Vibrio parahaemolyticus in Shenyang. Conclusion The prevalence of foodborne Vibrio parahaemolyticus in Shenyang is genotyped, but O3: K6 with close genetic relationship is the predominant type.