重组副猪嗜血杆菌血清5型OmpP5高免血清制备及其杀菌活性和交叉反应性分析  

Hyperimmune serum preparation of recombinant outer membrane protein P5 of HPS serotype 5 and its bactericidal activity and cross reacivity evaluation

作　　者：秦翠丽[1] 李春玲[2] 高祥辉[1] 

机构地区：[1]河南科技大学食品与生物工程学院生物工程系,洛阳471023 [2]广东省农业科学院兽医研究所,广州510640

出　　处：《中国免疫学杂志》2013年第4期362-365,共4页Chinese Journal of Immunology

基　　金：广东省农业科技重大专项(2009A0201006)资助

摘　　要：目的:探讨致病性副猪嗜血杆菌(Haemophilus parasuis,HPS)血清5型外膜蛋白Omp P5高免血清的杀菌活性和交叉反应性。方法:制备HPS血清5型外膜蛋白Omp P5抗原,免疫BALB/c小鼠制备高免血清,间接ELISA法测定效价,体外杀菌试验测定杀菌率,Western blot检测与不同型HPS菌株之间的交叉反应性,并与制备的HPS血清5型灭活苗高免血清进行比较。结果:高免血清Omp P5效价远高于HPS灭活苗效价;Omp P5高免血清对5型菌株具有良好的杀菌特异性,杀菌效率为96%,而对4、12型菌株杀菌效率较低,仅为31%、39%,杀菌广谱性不如HPS灭活苗高免血清;Western blot检测Omp P5高免血清与不同型HPS菌体抗原均有较好交叉反应性。结论:Omp P5具有较强的免疫原性和良好的交叉反应性,是一个很好的用于鉴定HPS感染的血清学候选工具,可用于副猪嗜血杆菌病的鉴定和诊断以及新型疫苗的研究与开发。Objective： To research the bactericidal activity and cross-reactivity of the recombinant protein Omp P5 （ outer-mem- brane protein 5 ）. Methods： Hyperimmune serums were obtained from BALB/c inoculated with the recombinant protein Omp P5 （out- er-membrane protein 5 ） and inactivated strain of Haemophilus parasuis （HPS） strain serotype 5 insolated in the field, respectively. The antibody titers were detected by indirect ELISA. Bactericidal test in vitro was performed to determine the disinfection rate, and the cross-reactivity of Omp P5 with different serotypes of HPS was also examined by western-blot. Results ：The results showed that the recombinant protein Omp P5 showed a far higher titer than inactived vaccine of HPS and a strong bactericidal activity against HPS type 5 with a disinfection rate of 96%. The western-blot proved a good cross reactivity. Conclusion：In a word, present study provided an academic reference for further research of HPS Omp PS.

关 键 词：副猪嗜血杆菌 OmpP5 高免血清 杀菌活性 交叉反应性 

分 类 号：R392.11[医药卫生—免疫学]