When an acute myocardial infarction (AMI) occurs, myoglobin (Mb) is the biomarker whose concentration firstly increases, and the high sensitive detection of Mb is critical for early diagnosis of AMI. Herein, a sandwich-type electrochemical aptasensor for the sensitive detection of Mb was constructed by using Pt@Cu1.33OCo0.83O as the signal marker. On one hand, nano-flower-like Cu1.33OCo0.83O was synthesized by hydrothermal method and Pt nanoparticles (Pt NPs) were loaded on its surface. Pt@Cu1.33OCo0.83O could immobilize aptamer 2 (Apt2) successfully by the Pt-S bond. And because of the synergistic effect between Pt and bimetallic oxide, Pt@Cu1.33OCo0.83O had an excellent catalytic effect on the signal source of hydrogen peroxide (H2O2) to amplify the current signal, which enhance the sensitivity of the aptasensor. On the other hand, the screen-printed gold electrode (SPGE) was used as the sensing base, which had good conductivity and ensured the immobilization of aptamer 1 (Apt1). The quantitative detection of Mb was achieved by specific recognition between Mb and Apt1, Apt2. As a result, the constructed electrochemical aptasensor had a good linear range (1-1500 ng/mL) with a low detection limit (LOD) of 0.128 ng/mL (S/N = 3), and a high sensitivity of 29.47 μA dec-1. The aptasensor also realized the detection of Mb in human serum samples with good accuracy, and the results were consistent with the hospital's biochemical indicators, which demonstrated the potential application of the prepared sensor in the clinical detection of Mb.