Competitive Elisa for quantifying small Amounts of aflatoxin B₁

Abstract

An indirect ELISA was developed to quantify aflatoxin B1 (AFB1). The detection limit was 0.025 ng ml‐1. The test used polyvinyl chloride (PVC) plates, activated with AFB1 bound to bovine serum albumin (BSA). Polyclonal antibodies were raised in rabbits against AFB1‐BSA. The specific anti‐AFB1antibodies were recovered from the crude antiserum by affinity chromatography from a column containing immobilized BSA on Nylon 6–6. Goat anti‐rabbit IgG antibodies bound to peroxidase were used to detect the rabbit IgG anti‐AFB1 antibodies bound to PVC plates. The colour developed by the subsequent enzyme conversion of the substrate was detected by spectrophotometry. The developed colour gave clear absorbance differences at varying doses of AFB1. Cross‐reactivity with aflatoxin B2, aflatoxin G1 and aflatoxin G2 was measured, showing percentages of 5.43, 64.5 and 5.07 respectively.