An indirect competitive ELISA was developed using rabbit antiserum against the exoantigens (ExAgs) of , and cross-reactivities were determined with 15 ExAgs from other species and genera of fungi and with the water-soluble extracts from four grains. The assay had good sensitivity to ExAgs (1 μg per ml serum), with generally low or no cross-reactivity with grain extracts and the other fungi, including five species of , four species of , three species of , two species of and one species of Immunoblotting analysis of the ExAgs from yielded at least 20 distinct antigenic bands, with about 3 or 4 being dark, 10-12 light and 5-8 faint in colour. Most of the bands had molecular masses of about 70-90 kDa. In contrast to , the ExAgs from the other fungi either did not react in the immunoblotting assay with the antiserum, or reacted only weakly with it. One antigenic band from , however, reacted moderately strongly with the antiserum. The ELISA and immunoblotting analysis were used to detect in naturally contaminated wheat samples, and in wheat samples spiked with a second common storage fungus, The results demonstrated that the antiserum was able to detect in a background of other fungal species, and that these did not produce false positives. There was also a positive association between the concentration of ExAgs, as measured by ELISA, and the amount of mould as determined by the number of colony-forming units. Immunoblotting qualitatively confirmed the ELISA results obtained with fungi when cultured on liquid or on solid (wheat) media. The data suggest that the immunoassays developed for are useful for the detection and identification of on the basis of its ExAgs, with the advantages of being more efficient, simple and reliable than conventional techniques.


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