Poul F, Dunez J
Station de Pathologie Végétale, Institut National de la Recherche Agronomique, Centre de Bordeaux, Pont de la Maye, France.
J Virol Methods. 1989 Aug;25(2):153-65. doi: 10.1016/0166-0934(89)90029-3.
Stable hybridoma cell lines secreting antibodies specific for the apple chlorotic leaf spot virus (CLSV) were produced by fusing spleen cells of a Biozzi mouse immunized with CLSV P863 strain, with the non-secretory P3 X63 Ag8.653 myeloma cell line. Two hybridoma clones producing monoclonal antibodies of the IgG1 subclass were obtained. These monoclonal antibodies were used for virus detection by enzyme-linked immunosorbent assay (ELISA). In contrast to polyclonal antisera to CLSV, which always contain some antibodies to host components, monoclonal antibodies are highly specific for the virus. It was thus possible to develop a detection assay which is more sensitive and specific than the assays using polyclonal antibodies. Using monoclonal antibodies, it was possible to detect less than 0.1 ng/ml of purified virus. In addition, these two monoclonal antibodies recognize 17 strains or isolates maintained in our laboratory and representing most of the known CLSV strains.
通过将用苹果褪绿叶斑病毒（CLSV）P863株免疫的Biozzi小鼠的脾细胞与非分泌型P3 X63 Ag8.653骨髓瘤细胞系融合，产生了分泌针对CLSV特异性抗体的稳定杂交瘤细胞系。获得了两个产生IgG1亚类单克隆抗体的杂交瘤克隆。这些单克隆抗体用于通过酶联免疫吸附测定（ELISA）进行病毒检测。与总是含有一些针对宿主成分抗体的CLSV多克隆抗血清不同，单克隆抗体对该病毒具有高度特异性。因此，有可能开发出一种比使用多克隆抗体的检测方法更灵敏、更特异的检测方法。使用单克隆抗体，可以检测到低于0.1 ng/ml的纯化病毒。此外，这两种单克隆抗体识别保存在我们实验室中的17个毒株或分离株，它们代表了大多数已知的CLSV毒株。
