Production of Monoclonal Antibodies AgainstNosema bombycisand their Utility for Detection of Pebrine Infection inBombyx moriL.

摘要

Latex agglutination assay based on monoclonal antibodies (MCAs) described in this communication may be useful for detection of Pebrine infection in silkworm. Four murine MCAs were produced againstNosema bombycisspore. In ELISA all 4 MCAs (IgM isotype) reacted with alkali treated Nosema spores and to variable extent with acetone precipitated surface protein. However, MA-310 and MA-542 showed a low degree of cross reactivity with BmNPV. In contrast, MA-503 and MA-515 were devoid of reactivity with BmNPV,B. thuringiensis, S. marcescens, Azotobactor, Rhizobium and normal hemolymph protein in ELISA. Latex beads sensitized with a combination of MA-503 and MA-515 (50 #x3BC;g each per ml of 0.4 latex beads) could detect 1#xD7;105Nosema spores per test. Sensitization of the latex beads with the cocktail of these two MCAs through protein-A bridge further led to a 10-fold increase in the sensitivity (1#xD7;104spores/test) of the assay. No agglutination was observed in presence of BmNPV, Rhizobium, Azotobactor,E. coli, B. thuringiensis, S. marcescensand normal hemolymph protein indicating the specificity of the test. The results obtained by latex agglutination assay on hemolymph samples of infected as well as normal larvae collected from field, II instar larvae infected in the laboratory and from infected mother moth revealed 100 correlation with results by microscopic examination.