摘要
用纯化的猪繁殖呼吸综合征病毒沪1株（PRRSV-SI）病毒液作抗原,免疫BALB/C小鼠,获得1株稳定分泌抗PRRSV单抗的杂交瘤细胞株,命名为AG1。ELISA交叉试验和阻断试验表明,AGI具有高度特异性。AGI可与PRRSV-S1、美洲株VR-2332、欧洲株LV反应,对猪细小病毒（PPV）、猪伪狂犬病毒（PRV）、猪乙型脑炎病毒（JEV）均不反应。阳性杂交瘤细胞的上清液效价是1:256～512,腹水效价在1:10-3～10-5之间。AG1有100条染色体,琼脂双扩散试验结果表明AG1属于IgG2b。Western blotting试验表明 AG1是针对 N蛋白抗原表位的特异性单抗。以AG1为捕捉抗原抗体和酶标抗体,建立了快速检测PRRSV抗原的单抗夹心ELISA法,该方法具有快速、灵敏、准确、广谱等特点。
The BALB/C mice immunized with the purified antigen of PRRSV-S1 strain, and one hybridoma cell lines steadily secreting monoclonal antibody ( McAb ) against PRRSV-S1 was obtained, which was named AG1. ELISA block and cross tests showed that the AG1 had highly idiosyncrasy and it could react with antigens of PRRSV-S1,PRRSV-LV and PRRSV-VR-2332, and didn't react with Japanese encephalitis virus(JEV), pseudorabies virus(PRV) and porcine parvovirus(PPV). The ELISA titers of cell supernatant and ascites were 1:256~512 and 1:10-3 ~ 10-5. The chromosome number of the hybridoma cell lines was 100. AG1 belonged to IgG2b subgroup . Western blotting test indicated that AG1 reacted with N protein . On the basis of antigen AG1 for capturing antibody and enzyme-labelled AG1, the established sandwich ELISA for detecting PRRSV was of rapidity,sensitivity, accuracy and broad-spectrum.
出处
《上海农业学报》
CSCD
2002年第3期87-91,共5页
Acta Agriculturae Shanghai
基金
上海市农业科学院青年科技基金资助。
