A novel label-free electrochemical aptasensor was fabricated based on a three-dimensional porous electrode (3D-CS/rGO/GCE) for the detection of acetamiprid residues. The sensing signal was generated by the DNA itself. The porous electrode was prepared by electrodeposition in situ and characterized by scanning electron microscope (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). These results indicated that the porous electrode has a uniform nanoporous structure, high active area, and excellent conductivity, leading to improve the transmission efficiency of current signals. The 3D-CS/rGO/GCE was used to increase a load of acetamiprid aptamer on the electrode. Meanwhile, DNA self-assembly strategy was used to further increase the DNA amounts. Thus the electrochemical current was amplified significantly due to increased phosphate group amounts by the above synergistic effect. The determination of acetamiprid residues using square wave voltammetry (SWV) showed good sensitivity, with the linear range from 0.1 pM to 0.1 mu M and the detection limit was 71.2 fM. The label-free electrochemical aptasensor was also used to detect acetamiprid residues in tea samples with satisfactory results.