Multiplex enzyme immunoassay system for the simultaneous detection of multiple allergens in foods

An enzyme immunoassay (EIA) system was developed for the simultaneous detection of multiple allergens in foods. This assay utilized a reverse dot blot EIA format in which egg yolk antibodies (IgY) specific for different allergens were immobilized in discrete spots on a strip of polyester cloth, which was then contacted with the test sample. The binding of antigens to a specific spot on the strip was revealed by sequential reactions with a pool of biotinylated allergen-specific antibodies, streptavidin-peroxidase conjugate and a chromogenic substrate. As an example of the applicability of this multiplex EIA system, we demonstrate the detection of three important food allergens: peanut, hazelnut and Brazil nut.