Purpose： An indirect competitive enzyme-linked immunosorbent assay （ic-ELISA） for detection of aflatoxin B1（AFB1） in peanut was developed. Methods： The AFB1 antigen was prepared， and coated in 96 well plates with the content of 0.1 μg/well； antibody was diluted 64 000 times； the 0.1% skimmed milk powder solution was selected as the blocking buffer， 0.01 mol/L phosphate buffer solution （PBS， pH 7.4） was used as dilution buffer to establish the proposed ic- ELISA detection method. Results： The limit of detection limit （LOD， IC15） of this ic-ELISA assay was 0.0066 μg/L； the sensitivity （IC50） was 0.027 μg/L； and the linear range was 0.0097-0.088 μg/L （R2 = 0.999）. The intra-assay variation coefficient （n=6） range was 0.29%-16.9%， and the inter-assay variation coefficient （n=6） range was 0.22%-21.19%. The sensitivity（IC50） in peanut sample was 1.04 μg/kg， and the recovery range was 96.67%-106.51%. Conclusion： The ic-ELISA assay established in this study was simple， sensitive and accurate and can be used for quantitative determination of AFB1 in peanut samples.