Undeclared Brazil nut residue in food products is of great concern because it can trigger life-threatening allergic reactions in sensitive patients. A rabbit polyclonal antibody-based competitive ELISA (IC(50) = 23.2 +/- 9 ng/mL, n = 76) with good sensitivity, detection range of 10-90 ng/mL, was developed. The ELISA could detect Brazil nut seed proteins over a pH range of 5-12. The optimal pH range for the detection assay was 7-10. Among the 66 tested foods/ingredients, only cinnamon exhibited statistically significant interference (1.36%, p = 0.05). Exposing Brazil nut seeds to processing did not adversely affect the nut seed protein detection using the assay. Brazil nut seed protein recovery from 100 mg of foods spiked with 10 and 1 microg of soluble Brazil nut proteins or 100 and 10 microg of defatted Brazil nut flour exhibited a wide recovery range, 63-315%, indicating protein-food matrix interaction.