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为了建立一种基于免疫反应检测茶尺蠖核型多角体病毒的方法,以纯化后的茶尺蠖核型多角体病毒作为抗原,免疫BALB/c小鼠,将小鼠脾脏细胞与小鼠骨髓瘤细胞Sp2/0融合,经间接ELISA筛选及克隆得到了一株稳定分泌单克隆抗体的杂交瘤细胞株,命名为7D3。同时克隆并在大肠杆菌中表达了EoNPV多角体蛋白基因,获得重组多角体蛋白。经Western blotting鉴定,该抗体可与EoNPV的多角体蛋白特异性结合。利用制备EoNPV多角体蛋白的单克隆抗体,建立了间接ELISA测定EoNPV的方法。
In order to establish a method for detecting polyhedrosis virus (PCDV) based on immune response, BALB / c mice were immunized with the purified polycrop was used as antigen. The mouse spleen cells were incubated with mouse myeloma cells Sp2 / 0 fusion. After screening and cloning by indirect ELISA, a hybridoma cell line stably secreting monoclonal antibody was obtained and named as 7D3. At the same time, EoNPV polyhedrin gene was cloned and expressed in E. coli to obtain recombinant polyhedrin. Western blotting showed that the antibody could specifically bind polyhedrin of EoNPV. An indirect ELISA method for the determination of EoNPV was established by using monoclonal antibodies to prepare EoNPV polyhedrin.