猪繁殖与呼吸综合征病毒Resp株单克隆抗体的研制  

Preparation of monoclonal antibody of PRRSV(Resp strain)

作　　者：吴家强[1] 姜平[1] 蔡宝祥[1] 陈溥言[1] 

机构地区：[1]南京农业大学动物医学院农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095

出　　处：《畜牧与兽医》2003年第2期1-3,共3页Animal Husbandry & Veterinary Medicine

基　　金：中华农业科教基金项目资助 (1 998 2 0 0 0 )

摘　　要：利用Marc1 4 5细胞增殖和凝胶层析纯化的猪繁殖与呼吸综合征 (PRRSV)Resp株 ,建立了酶联免疫吸附试验 (ELISA)单克隆抗体筛选技术。在猪肺泡巨噬细胞 (PAM)上增殖、浓缩、纯化PRRSV并免疫Balb/c小鼠 ,用PEG - 1 50 0进行细胞融合。结果表明 ,细胞融合率为 70 6 % ,初检PRRSV抗体阳性率为 6 3 %。经 2次亚克隆获得了 2株能稳定分泌单克隆抗体的杂交瘤细胞株 ,其单抗间接荧光抗体试验 (IFA)可用于检测细胞培养物中的PRRSV抗原 。An indirect enzyme linked immunosorbent assay was developed with purified PRRSV (Resp strain) and prepared from Marc145 cell line to detect monoclonal antibodies (McAbs) secreted by hybridoma cell lines. The BALB/C mouse were immunized for several times with the virus replicating on porcine alveolar macrophage, then the spleen cells of the hyper immunized mouse and SP 2/10 cells were fused with PEG 1500. The results showed that the cell fusion rate was 70 6%,positive rate was 6 3%, and two hybridoma cell lines secreting monoclonal antibodies were obtained after sub cloning. The McAbs could be used to detect PRRSV antigen in cells with IFA. It would be useful for studying the immunity of PRRSV isolates and improving present diagnostic methods.

关 键 词：Resp株 研制 PRRSV ELISA IFA 单克隆抗体 猪繁殖呼吸综合征病毒 间接荧光抗体试验 

分 类 号：S858.28[农业科学—临床兽医学] S852.65[农业科学—兽医学]