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目的将食物中毒样本中分离到的5株副溶血性弧菌(VP)作分离鉴定、毒力基因测定及脉冲场凝胶电泳(PFGE)分型,了解其毒力基因特征,并做同源性分析。方法依据GB/T 4789.7-2008和WS 271-2007进行VP分离、生化鉴定;采用PCR技术检测耐热直接溶血素基因(tdh)和耐热直接溶血素相关溶血素基因(trh);菌株基因组DNA经限制性内切酶Not I酶切,用PFGE方法获得电泳图谱,利用BioNumerics软件对图谱进行同源性分析。结果共检出5株VP,从6件病人便和6件水产品中分别检出3株和2株;4株菌携带毒力基因tdh,2株菌携带毒力基因trh,有1株菌同时携带tdh和trh毒力基因;5株VP的DNA指纹图谱可大致分为4种PFGE图谱,1株病人分离株和1株水产品分离株图谱一致,具有高度的同源性,其它菌株的相似性较低。结论这次食物中毒为VP污染食品所致,本次分离的5株菌都携带毒力基因,PFGE具有很强的菌株同源性分析能力,适用于食物中毒病原菌鉴定和传染源溯源。
OBJECTIVE To isolate and identify five strains of Vibrio parahaemolyticus (VP) isolated from food poisoning samples for virulence gene determination and pulsed-field gel electrophoresis (PFGE) typing to understand their virulence genes and to make homologous Sexual analysis. Methods VP isolation and biochemical identification were performed according to GB / T 4789.7-2008 and WS 271-2007. PCR was used to detect tdh gene and heat-resistant hemolysin-related hemolysin gene (trh). Genomic DNA After digestion with restriction enzyme Not I, the electrophoresis pattern was obtained by PFGE method and the homology analysis was performed by BioNumerics software. Results A total of 5 strains of VP were detected, 3 strains and 2 strains were detected respectively from 6 patients and 6 aquatic products. 4 strains carried virulence gene tdh, 2 strains carried virulence gene trh, 1 strain While carrying the tdh and trh virulence genes; DNA fingerprinting of five VP can be roughly divided into four PFGE patterns, one patient isolates and one aquatic product isolates consistent, with a high degree of homology, other strains of Similarity is low. Conclusion The food poisoning was caused by VP contamination of food. All the five isolates contained virulence genes. PFGE had strong ability of homologous analysis of strains, which was suitable for identification of pathogenic bacteria and source of infection.