An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on an anti-aflatoxin B1 monoclonal antibody was standardised and validated for aflatoxin screening in poultry feed samples and its performance was compared to high-performance liquid chromatography (HPLC). The ic-ELISA showed good linearity (r2=0.994) and detection limits of 1.25ngg−1 for broiler feed and 1.41ngg−1 for laying hen feed. Mean aflatoxin recovery rates by ic-ELISA were 102% (laying hen feed) and 98% (broiler feed). Aflatoxins were detected in 88.2% of the 34 broiler feed samples by ic-ELISA and HPLC at means of 10.48ngg−1 and 8.41ngg−1, respectively, while 92% of laying hen feed samples (n=36) showed aflatoxin contamination at means of 20.83 and 19.75ngg−1. The standardised ic-ELISA showed reliability and a high correlation with HPLC of 0.97 (broiler feed) and 0.98 (laying hen feed) indicating its potential for aflatoxin screening in poultry feed samples.