副溶血性弧菌外膜蛋白的提取与免疫鉴定

[目的]为进一步确定致病性副溶弧菌共有的特异性抗原和保护性抗原奠定基础。[方法]通过小鼠毒力试验研究5株副溶血性弧菌菌株对小鼠的致病性,比较在不同培养基和培养时间下所提取的外膜蛋白的SDS-PAGE图谱并通过Western blotting分析研究副溶血性弧菌菌株的免疫原性。[结果]5株临床分离的副溶血性弧菌菌株明显对小鼠具有不同的致病性,同时能在兔血平板上形成明显的溶血圈,为TDH阳性菌株。通过菌体免疫获得的多克隆抗体,与除副溶血性弧菌外的其他试验菌株均无交叉反应。Western blotting分析显示该多克隆抗体与5株致病性副溶血弧菌菌株可发生程度不等的阳性反应,提取的外膜蛋白具有较好的免疫原性。[结论]该研究为制备有效预防副溶血性弧菌引起的疾病的菌苗提供了理论基础。

Extraction and Immuno-identification of Outer Membrane Protein from Vibrio parahaemolyticus

[Objective] The aim was to lay the foundation for the further determination on the mutual specific antigen and the protective antigen of the pathogenic V.parahaemolyticus.[Method] The pathogenicity of the 5 strains of V.parahaemolyticus to mouse were studied by the mice virulence experiments,the SDS-PAGE map of the outer membrane protein extracted under the different medium and culture time was compared and the immunogenicity of the V.parahaemolyticus strain was studied by the western blotting analysis.[Result] The 5 strains of V.parahaemolyticus had different pathogenicity to mouse obviously and could form obvious hemolytic cycle on rabbit blood plate,being TDH positive strain.The polyclonal antibody obtained by strain immunity all had no cross reaction with other experimental strains except V.parahaemolyticus strain.Western blotting analysis showed that the polyclonal antibody could produce positive reaction with the 5 strains of pathogenic V.parahaemolyticus to a different degree and the extracted outer membrane protein had better immunogenicity.[Conclusion] The research provided the theoretical basis for the preparation of the vaccine to prevent the disease caused by the V.parahaemolyticus effectively.