抗狂犬病病毒ERA株单克隆抗体的制备、鉴定及初步应用  

Preparation characterization and preliminary application of monoclonal antibodies to rabies virus ERA strain

作　　者：薛琳[1] 夏咸柱[2] 高明华[1] 高玉伟[2] 侯小强[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]解放军军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2007年第3期343-346,共4页Chinese Journal of Veterinary Science

基　　金：国家科技攻关计划课题资助项目(2004BA519A48)

摘　　要：将狂犬病病毒ERA株纯化后免疫雌性BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经克隆和间接ELISA筛选,获得3株稳定分泌抗狂犬病病毒ERA株单克隆抗体的杂交瘤细胞株,分别命名为C7、C4和H3。经过鉴定:其腹水效价分别为1∶1×105、1∶5×104及1∶1×104;且与犬瘟热病毒(CDV)、犬细小病毒(CPV)、犬腺病毒(CAV)不发生交叉反应;3株单抗均为IgG类型。采用G蛋白亲和层析柱对腹水进行纯化,将纯化后的单抗腹水用FITC(异硫氰酸荧光素)标记制备荧光抗体,建立了检测狂犬病病毒的荧光染色法。结果表明,该方法对狂犬病的实验室诊断具有快速和特异性高的特点。After immunization of BALB/c mice with purified rabies virus （RV）, three hybridoma cell strain againstRV named C7,C4 and H3,respectively,were developed after fusion between SP2/0 myeloma cell and the stimulated splenocytes. The indirect ELISA results showed that the McAbs ascites titer were 1×10^3 ,5×10^4 and 1×10^4; No cross reaction was found when McAbs reacted with CDV, CPV and CAV; the immunoglobulin G type of the three McAbs were purified by protein G and labelled with FITC, RV were tested by using direct immunofluorescent assay. The results show that this established method is of rapid and high specific for the diagnosis of RV.

关 键 词：狂犬病病毒ERA株 单克隆抗体 免疫荧光 

分 类 号：S852.4[农业科学—基础兽医学] S852.65[农业科学—兽医学]