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Development of sensitive nanosenor for organophosphorus and carbarmate pesticides detection is of great importance.We presented an assay with dual readouts (colorimetric and fluorometric) for detecting organophosphorus and carbamate pesticides in complex solutions.This assay was based on a system used to detect acetylcholinesterase (AChE).The organophosphorus and carbamate pesticides can inhibit the activity of acetylcholinesterase (AChE), thus preventing the generation of thiocholine (which turned the RB-AuNP solutions blue and unquenched the fluorescence of RB simultaneously).