Dual-mode electrochemical aptasensor based on nitrogen-doped graphene (NG) doped with the conducting polymer polypyrrole (PPy) nanocomposite is proposed for the determination of acetamiprid. NG/PPy was electrodeposited onto the glassy carbon electrode (GCE) using cyclic voltammetry technique. NG/PPy/GCE showed outstanding electrocatalytic activity for the oxidation of nitrite due to "active region" induced by the charge redistribution of carbon atoms. The ultrasensitive dual-mode biosensor for acetamiprid could be easily developed by coupling acetamiprid aptamers with the NG/PPy hybrid. The specific binding between acetamiprid and the aptamers resulted in the increase of differential pulse voltammetry (DPV) signal change and the decrease of chronoamperometry (CA) signal, and the concentration of acetamiprid could be measured. The working potentials of DPV and CA were - 0.2 ~ 0.4 V and - 0.4 ~ 0.4 V (vs. SCE), respectively. The dual-mode acetamiprid biosensor showed a wide linear range from 10 to 10 g mL, with low detection limits of 1.15 × 10 g mL and 7.32 × 10 g mL through DPV and CA modes, respectively. Moreover, owing to high active area and superior conductivity, as well as good electrocatalytic ability, the dual-sensing platform based on NG/PPy nanocomposite supported the quantification of acetamiprid in complex samples. A dual-mode electrochemical aptasensor based on NG/PPy nanocomposite for acetamiprid detection was proposed through both the increase of differential pulse voltammetry (DPV) signal change and the decrease of chronoamperometry (CA) signal of the nitrite oxidation electrocatalyzed by NG/PPyn in sensors and biosensors.