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Ten stable hybridoma cell lines secreting monoclonal antibodies to grapevine fleck virus (GFkV) were selected after fusing spleen cells of immunized Balb/C mice with mouse myeloma cells (SP2/0-Ag 14). All MAbs reacted positively in ELISA with leaf extracts from fifty GFkV-infected grapevines from various geographical origins. MAb 2B5 was used for routine detection of GFkV and appeared to be more sensitive than polyclonal antibodies. The first attempt to purify GFkV by immunoaffinity chromatography using MAb 2B5 led to highly purified coat protein. This procedure encompassed fewer steps and allowed the use of tissues other than rootlets for satisfactory purification.  相似文献   
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Five stable hybridoma cell lines secreting monoclonal antibodies (Mabs) specific for GLRaV-1, one of the agents involved in the aetiology of grapevine leafroll disease, were produced by fusing a nonsecreting myeloma cell line with spleen cells from BALB/c mice immunized with purified GLRaV-1. The Mabs were characterized for their recognition of virus coat protein by DAS-ELISA and Western blotting. Mab (1G10) reacted specifically in ELISA, immuno-electron microscopy and immunoblotting with both GLRaV-1 and GLRaV-3 coat proteins. Mab 1C4 detected 25 of the 33 GLRaV-1 isolates, while Mab 1B7 reacted with 32 isolates including the eight isolates not recognized by Mab 1C4. Two of these hybridoma lines (2F11 and 2F3) are now used routinely for the immunodiagnosis of GLRaV-1.  相似文献   
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Eleven stable hybridoma cell lines secreting monoclonal antibodies specific for FD-phytoplasma, the pathogenic agent of grapevine Flavescence dorée, were produced by fusing a non-secreting myeloma cell line with spleen cells from Balb/c mice immunized with Flavescence dorée phytoplasma purified by immunoaffinity. These monoclonal antibodies were characterized for their recognition of phytoplasma proteins by western blot. Six of eleven reacted specifically in ELISA and immunoblotting with Elm-yellows phytoplasma. These antibodies did not react either in ELISA or in western blot with preparations from periwinkles infected with phytoplasmas that cause GYU (Grapevine Yellows from Udine), AP (Apple Proliferation), EAY (European Aster Yellows) and StolC (Stolbur from France). Two of these hybridoma lines were used routinely for the immunodiagnosis of Flavescence dorée phytoplasma in diseased grapevines.Abbreviations ELISA Enzyme linked immunosorbent assay - FD Flavescence dorée - MLO Mycoplasmalike organism - EY Elm yellows  相似文献   
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