UNIQUE IMMUNOGENIC PROTEINS IN HETERODERA-GLYCINES EGGSHELLS

Polyclonal antibodies were raised against Heterodera glycines eggshell s to determine the feasibility of developing an immunoassay for H. gly cines eggs. An indirect enzyme-linked immunosorbent assay (ELISA) was developed from antisera collected 10 weeks after the initial injection . From serial dilutions of sonicated eggshells or whole eggs, a sensit ivity of detection to 5 ng/ml sonicated eggshells or 1 egg of H. glyci nes was determined. The method of eggshell preparation had no effect o n the antibodies produced: however, the antibodies cross-reacted with sonicated J2 of ii. glycines and eggs of Meloidogyne incognita and H. schachtii. Most of the proteins in both life stages of N. glycines and eggs of M. incognita and H. schachtii had similar migration propertie s when separated on SDS-PAGE gels and stained with Coomassie blue. Wes tern blot analysis, with antisera adsorbed with homogenized J2 of H. g lycines, showed proteins that were specifically localized to eggshells of H. glycines. Monoclonal antibodies might provide a useful immunoas say where polyclonal antibodies lack sufficient specificity.