Electrochemical aptasensors have gained significant attention due to their exceptional sensitivity, selectivity, stability, and rapid response, combining the advantages of electrochemical techniques with the specific recognition ability of aptamers. This review aims to provide a comprehensive summary of the recent advances in electrochemical aptasensors. Firstly, the construction method and the advantages of electrochemical aptasensors are introduced. Subsequently, the review highlights the application progress of electrochemical aptasensors in detecting various chemical and biological molecules, including metal ions, small biological molecules, drugs, proteins, exosomes, tumor cells, bacteria, and viruses. Lastly, the prospects and challenges associated with electrochemical aptasensors are discussed.