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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
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)
