A high sensitivity fluorescence detection method for ochratoxin A (OTA) based on OTA aptamers and gold nanoparticle (AuNPs) was established, which employed OTA aptamers as recognition element and AuNPs to quench the fluorescence emission from fluorescein. The results indicated that the detection limit is 10 nmol/L and a linear relationship between the fluorescent intensity ratio and the logarithm of OTA concentration was plotted in the range of 25～1 000 nmol/L (R²=0.991). The detection method showed high feasibility and reliability. Compared with traditional method, this method greatly improved the detection efficiency and provided technical support for more effective identification of OTA.