A detection method for organophosphorus pesticides using electrochemical sensor based on enzyme inhibition has been developed. Acetylcholine esterase (AChE) was selected to quantify the pesticides concentration in water samples due to its activity was non-competitively inhibited in the presence of pesticides. To detect parathion and dichlorvos, optimized analytical conditions were that the substrate concentration was 3mmol/L, enzyme solution concentration was 18g/L and reaction time was 20min. Under the optimized detection conditons, a detection limit of 1.2ng/mL and 0.004ng/mL for parathion and dichlorvos was obtained, respectively. The approach was rapid, simple, accurate and of high sensitivity when compared with immunoassays or chromatographic methods.