Disease biomarkers serve as crucial indicators for objectively reflecting physiological states and pathological progression, playing a vital role in clinical diagnostics. Electrochemical sensors, with their advantages of simple construction, low cost, and rapid response, have emerged as promising tools with broad applications in clinical diagnosis, biochips, food and drug analysis, and environmental protection. This review systematically examines the application of electrochemical sensors based on diverse nanomaterials—such as graphene, noble metal nanoparticles, carbon nanotubes, quantum dots, and nano-oxides—for the detection of disease biomarkers. Finally, it outlines the research directions and future trends in this field.