Production of Monoclonal Antibodies AgainstSpiroplasma citri

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Abstract
From 4 independent fusions, 13 stable cell lines secreting specific monoclonal antibodies against distinct epitopes on S. citri ''Maroc'' were selected from hydridomas produced by fusing NS-1/1-Ag4-1 murine myeloma cells with splenic cells from mice immunized with S. citri (Maroc). With indirect ELISA with biotinylated antimouse Ig, the antibodies secreted by 9 of these 13 clones reacted specifically with strains Maroc, C189, CB-1, Arizona, Aceratagallia, SCC, Scaph, L163, B106, and BR-3 but not with strains Algeria, ASP-1, Israel, and Iran of S. citri. The monoclonal antibodies also distinguished S. citri from corn stunt spiroplasma (strains 1-747 and Miss E), honeybee spiroplasma (AS 576), 277F spiroplasma (277F), green leaf bug spiroplasma (LB-12), Maryland flower spiroplasma (M55), Cocos spiroplasma (N525), P40 spiroplasma (P40) of the same serogroup (group I), and from the other 19 spiroplasma strains belonging in the 18 other serogroups. Isotyping of the monoclonal antibodies was carried out by using the Ouchterlony double-diffusion method with class- and subclass-specific antibodies for mouse IgG1, IgG2a, IgG2b, IgG3, and IgM. Three clones were of the IgM isotype, four of the IgGI isotype, and six of the IgG3 isotype. These specific monoclonal antibodies are useful for differentiating the strains of S. citri from different geographic areas.