犬细小病毒单克隆抗体的制备及其在胶体金免疫层析试纸研发中的应用  

Development of Monoclonal Antibody and Colloidal Gold Immunochromatographic Assay for Detecting CPV

作　　者：马辉[1] 权国辉[2] 乔宏兴[1] 郑鸣[1] 赵绪永[1] 王永芬[1] 边传周[1] 

机构地区：[1]河南牧业经济学院生物工程学院,河南郑州450011 [2]郑州职业技术学院材料工程系,河南郑州450121

出　　处：《动物医学进展》2016年第9期26-30,共5页Progress In Veterinary Medicine

基　　金：郑州市科技攻关计划项目(121PPTGG463);河南省科技攻关项目(142102310034);河南省高等学校科技创新团队项目(HUAHE2015001)

摘　　要：为制备犬细小病毒胶体金免疫层析试纸,将F81细胞中分离的CPV免疫Balb/c小鼠,取其脾细胞与骨髓瘤细胞融合,建立间接ELISA方法筛选分泌CPV单克隆抗体的杂交瘤细胞,获得1株能稳定分泌CPV单克隆抗体的杂交瘤细胞株,命名为3E2。3E2的亚类为IgG1型,腹水抗体效价为1.8×105,交叉试验表明,3E2可与CPV特异性结合,与其他犬常见病毒无交叉反应。用3E2单克隆抗体制备的检测CPV的胶体金免疫层析试纸条特异性良好,为诊断和研究CPV奠定了基础。The canine arvovirus （CPV） isolated from dog feces was cultured and purified. SP2/0 myeloma cells SP2/0 were fused with spleen cells from CPV immunized Balb/c mice. Positive cells producing antibodies were screened by indirect ELISA,and a monoclonal antibody against CPV was generated and designated as 3E2. The monoelonal antibody belongs to IgG1. The titer of aseites was 1.8 × 10^5 as detected by ELISA. No cross-reactions were observed between the McAb prepared and other canine viruses and the McAb could recognize CPV. Based on the McAb, the GICA was established by purified and gold labeled McAb 3E2. It could detect the CPV specifically. Therefore, the results indicated that the method had high specificity. It could serve as effective detection measure for clinic and further research of CPV.

关 键 词：犬细小病毒 病毒分离 单克隆抗体 胶体金免疫层析试纸 

分 类 号：S854.43[农业科学—临床兽医学] S852.659.2[农业科学—兽医学]