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Fumonisin FB is produced by <i>Fusarium moniliforme</i> Sheld, of which FB₁ is the most common and the most toxic. The establishment of a rapid detection method is an important means to prevent and control FB₁ pollution. A highly sensitive fluorescent sensor based on an aptamer for the rapid detection of fumonisin B₁ (FB₁) in corn was established. In this study, 5-carboxyfluorescein (FAM) was labeled on the aptamer of FB₁ (F10). F10 was adsorbed on the surface of graphene oxide (GO) by π-π stacking. The FAM fluorescence signal could be quenched by fluorescence resonance energy transfer between fluorescent molecules and graphene oxide (GO). In the presence of FB₁, the binding efficiency of the aptamer to GO was reduced. Therefore, the content of FB₁ in corn samples was determined by fluorescence measurements of mixed FAM-labeled F10, GO and corn samples. This method had a good linear relationship in an FB₁ concentration range of 0–3000 ng/mL. The equation was y = 0.2576x + 10.98, R² = 0.9936. The limit of detection was 14.42 ng/mL, and the limit of quantification was 43.70 ng/mL. The recovery of a spiked standard in the corn sample was 89.13–102.08%, and the time of detection was 30 min.