嗜水气单胞菌气溶素单克隆抗体的制备及初步应用

Aeromonas hydrophila: Developing its Monoclonal Antibody against Aerolysin and Preliminary Application

以1%福尔马林灭活的嗜水气单胞菌纯化的气溶素免疫8周龄的BALB/C小鼠,取其脾细胞与SP2/0细胞融合,经间接ELISA筛选,获得1株稳定分泌气溶素单克隆抗体的杂交瘤细胞,命名为A2D3。腹水单抗ELISA效价为1∶16,384(1∶214),其能特异性抑制气溶素的溶血活性并且腹水单抗有中和特性,抑制效价和中和效价分别为1∶128(1∶27)和1∶128(1∶27)。以此单抗建立了检测嗜水气单胞菌气溶素的间接ELISA方法,对临床分离的50株嗜水气单胞菌培养上清检测,有48株呈阳性反应。

SP2/0 myeloma cells were fused with BALB/C mice spleen ceils immunized with purified aerolysin produced by Aeromonas hydrophila. One hydrophila cell strain secreting monoclonal antibodies （Mab） specific for aerolysin was screened with indirect-ELISA. The ELISA titre of Mab was 1： 16,384. The Mab strain could neutralize hemolytic activities of Aerolysin. Mab indirect-ELISA was developed for the detection of aerolysin, the 50 bacterial strains isolated from infected fish were detected, of which 48 strains were positive.