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

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 anti-bodies were screened by indirect ELISA,and a monoclonal antibody against CPV was generated and desig-nated as 3E2.The monoclonal antibody belongs to IgG1.The titer of ascites 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.