Abstract: To obtain porcine pseudorabies virus gB specific antibodies for PRV clinical testing and laboratory research,this study immunized mice with the mutated virus strain PRV HN1201,the spleen cells of the immunized mice were fused with SP/20 cells to prepare monoclonal antibodies against PRV gB protein.An immunoperoxidase monolayer cell assay(IPMA) and indirect immunofluorescence assay(IFA) were used to select a monoclonal cell line stably secreting PRV gB antibody.The antibody secreted by this cell line was identified as IgG1 subtype.The IFA titer of ascites purified antibody was 2-11 .The optimal dilution ratios of cell culture supernatant and ascites purified antibody for Western blot were 1∶50 and 1 ∶5 000,respectively,and the neutralization titer of hybridoma cells to mouse ascites was 1∶25.IPMA,IFA and Western blot test showed that the prepared monoclonal antibodies can specifically recognize PRV gB protein.

Key words: Pseudorabies virus variant HN1201, gB protein, Monoclonal antibody, Western blot, Immunoperoxidase monolayer cell assay, Indirect immunofluorescence assay

摘要： 为获得用于伪狂犬病毒(PRV)临床检测和实验室研究所需的PRV gB特异性抗体，以PRV变异病毒株PRV HN1201免疫小鼠，取免疫后小鼠脾脏细胞与SP/20细胞融合，制备抗PRV gB蛋白的单克隆抗体。经免疫过氧化物酶单层细胞试验（IPMA）和间接免疫荧光试验（IFA）筛选出1株稳定分泌PRV gB蛋白抗体的单克隆细胞系，经鉴定该细胞系所分泌抗体为IgG1亚型，腹水纯化抗体的IFA效价为2-11；细胞培养上清和腹水纯化抗体用于Western blot的最佳稀释比分别为1∶50和1∶5 000；杂交瘤细胞诱导小鼠腹水的中和效价为1∶25。IPMA、IFA及Western blot试验结果显示，制备的单克隆抗体能特异性识别PRV gB蛋白。

关键词: 伪狂犬病毒变异株HN1201, gB蛋白, 单克隆抗体, Western blot, 免疫过氧化物酶单层细胞试验, 间接免疫荧光试验