犬瘟热病毒单克隆抗体的制备及鉴定  

Preparation and Identification of Monoclonal Antibodies Against Canine Distemper Virus

作　　者：孙玮[1] 王铁成 杨松涛 高玉伟 王承宇 张涛 杨玉姣[1] 刘玉秀[1] 阮洋 靳晓霞 夏咸柱 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]解放军军事兽医研究所,吉林长春130062 [3]吉林省人兽共患病预防与控制重点实验室,吉林长春130062

出　　处：《动物医学进展》2011年第3期73-77,共5页Progress In Veterinary Medicine

摘　　要：将纯化的犬瘟热病毒(CDV)分别与弗氏完全佐剂(CFA)和弗氏不完全佐剂(IFA)乳化制备的乳化抗原作为免疫原,以有限稀释法和间接ELISA法进行单克隆抗体的筛选,从而获得了3株稳定分泌单克隆抗体的杂交瘤细胞株2F7、3G2、5E3。亚型鉴定结果显示,2F7为IgG2a型,3G2为IgG1,5E3为IgGM。用2F7制备的荧光抗体对感染CDV 3 d的Vero细胞进行染色观察,结果表明,荧光抗体具有很好的特异性,染色效价为1∶200,获得的单克隆抗体可用作犬瘟热病毒抗原检测的诊断试剂。With the depurated CDV and emulsified antigens of CFA and IFA as immunogen,use the limiting dilution assay and indirect ELISA for selecting,three hybridoma cell strains stably producing monoclonal antibodies against CDV which named 2F7,3G2,5E3 were obtained.Immunoglobulin subtype experiments showed that 2F7,3G2 and 5E3 were pertaining to IgG2a,IgG1and IgM,respectively.2F7,which conjugated with FITC,was confirmed to have good specificity by direct immunofluorescence assay based on the detection for the Vero cells infected with CDV.The working concentration of the FA is 1∶200.The monoclonal antibodies obtained could be used for detecting canine distemper virus antigen.

关 键 词：犬瘟热病毒 杂交瘤 单克隆抗体 

分 类 号：S852.659.5[农业科学—基础兽医学]