2型猪链球菌溶血素多克隆和单克隆抗体制备及鉴定  

Preparation and identification of polyclonal and monoclonal antibody of Suilysin in highly virulent strains of Streptococcus suis serotype 2

作　　者：孙雯[1] 郑峰[2] 

机构地区：[1]扬州科技学院医学院,江苏扬州225000 [2]南京军区军事医学研究所流行病室,江苏南京210002

出　　处：《现代预防医学》2017年第13期2415-2419,共5页Modern Preventive Medicine

基　　金：国家自然科学基金青年科学基金项目(No.31300119)

摘　　要：目的制备并鉴定2型猪链球菌(Streptococcus suis serotype 2,S.suis 2)四川分离菌株05ZYH33的溶血素(Suilysin,SLY)多克隆和单克隆抗体。方法利用重组SLY蛋白制备SLY兔多抗,优化间接ELISA体系检测多抗效价,Western blot分析SLY多抗特异性,检测SLY兔多抗对人外周血杀菌作用的影响。利用杂交瘤细胞融合技术制备SLY单抗,筛选杂交瘤细胞株并制备小鼠腹水,间接ELISA和Western blot鉴定SLY单抗的效价和特异性。结果间接ELISA显示SLY多抗效价高达1∶204 800,Western blot显示SLY多抗有较高的特异性和免疫反应性,人外周血杀菌试验显示,SLY多抗能加强人外周血对05ZYH33的杀伤作用。间接ELISA检测杂交瘤细胞2B6和5F7培养上清效价分别是1∶800和1∶3 200,单抗效价都为1∶204 800。间接ELISA和Western blot鉴定显示,单抗2B6和5F7具有很强的特异性和免疫反应性。结论成功制备了效价高、特异性强的S.suis 2 05ZYH33 SLY多克隆和单克隆抗体。Objective To prepare and identify the polyclonal and monoclonal antibody of Suilysin （SLY） in Streptococcus suis serotype 2 （S. suis2 ） from Sichuan strain 05ZYH33. Methods The rabbit polyclonal antibody of SLY was prepared by recombinant SLY protein, and the titer of SLY polyclonal antibody was detected by optimized indirect ELISA. The specificity of the SLY polyclonal antibody was identified by western blot, and the effect of the SLY polyelonal antibody on bactericidal of human peripheral blood was detected. SLY monoclonal antibody was prepared by hybridoma cell fusion technology. The titer and specificity of SLY monoclonal antibody were identified by indirect ELISA and Western blot. Results Indirect ELISA showed the titer of SLY polyclonal antibody was up to 1：204 800. Western blot showed SLY polyclonal antibody had higher specificity and immunoreactivity. The bactericidal test of blood showed SLY polyclonal antibody could strengthen the killing effect of blood on 05ZYH33. Indirect ELISA showed the titers of the hybridoma cells 2B6 and 5F7 were 1：800 and 1：3 200, The titers of SLY monoclonal antibodies were both 1 ： 204 800. The indirect ELISA and Western blot identification showed SLY monoclonal antibodies 2B6 and 5F7 had strong specificity and immunoreaetivity. Conclusion With high titer and specificity, SLY polyclonal and monoclonal antibody from S. suis 2 05ZYH33 were successfully prepared.

关 键 词：2型猪链球菌 溶血素 多克隆抗体 单克隆抗体 

分 类 号：R113[医药卫生—公共卫生与预防医学]