应用单克隆抗体检测牙鲆血清中抗淋巴囊肿病毒的特异性抗体

Detection of Specific Antibody Against Lymphocystis Disease Virus in Japanese Flounder(Paralichthys olivaceus) by Using Monoclonal Antibody Technology

应用杂交瘤单克隆抗体技术制备10株分泌抗牙鲆免疫球蛋白单克隆抗体的杂交瘤细胞。利用单抗2H4,通过间接酶联免疫法(ELISA)对牙鲆血清中抗淋巴囊肿病毒(LCDV)特异性抗体进行了检测。结果表明:无外观症状的牙鲆血清中特异性抗体水平很低(平均OD值为0.092),个别鱼体偏高,证明已感染LCDV,处于潜伏期;患淋巴囊肿病且症状显现的牙鲆血清中特异性抗体水平升高(平均OD值为0.165),患淋巴囊肿病后处于恢复期的牙鲆抗体水平最高(平均OD值为0.231);健康牙鲆接种LCDV灭活疫苗后,特异性抗体水平显著升高。

By using hybridoma-monoclonal antibodytechnology, ten hybridoma cell lines which could secrete monoclonal antibodies （Mabs） against the Ig of Japanese flounder （Paralichthys olivaceus ） were developed, An enzyme linked immunosorbent assay （ELISA） was used to detect the antibody in the serum of Japaneseflounder against lymphocystis disease virus （LCDV）by using the Mab 21-14. The results indicated that most of the apparently healthy flounder showed a low level of ELISA absorbance（OD 0. 092） but some of the floundershowed high absorbances as the LCD-diseased fish being in the latent period. The flounder with remarkable symptoms of LCDV showed higher absorbance（OD 0. 165） and the absorbance of flounder recovering from the disease was the highest（OD 0. 231）. The level of the specific antibody of healthy flounder remarkably increased after immunized with LCDV inactivated vaccine.