Nano-electrochemical cytosensors have attracted intensive attention and achieved huge progress in the biomedical field owing to their stability, rapidity, accuracy, and low-cost properties. Currently, most nano-electrochemical cytosensors are prepared using metal nanoparticles or carbon nanomaterials. In application, the nano-electrochemical cytosensors immobilize a bio-sensitive substance on the electrode, and convert the target molecule and its reaction signal into an electrical signal through specific recognition between the biomolecules, thereby achieving detection. Using nano-electrochemical cytosensors can help diagnose disease quickly and accurately, which could contribute to early diagnosis and clinical analysis. In this review, we concentrate on the latest development in metal nanoparticle and carbon nanomaterial based nano-electrochemical cytosensors in the last three years. Finally, we summarize the development of cytosensors and propose the future development prospects.