This work explains a simple and easily modifed electrochemical sensor for sensitive detection of daclatasvir (DCV), a novel drug for the treatment of hepatitis C. In this study, DCV was oxidized at a glassy carbon electrode (GCE) surface modifed with nickel nanoparticles/multi-walled carbon nanotube (Ni-NPs/MWCNTs). Electrochemical studies were accomplished via cyclic voltammetry (CV) and square wave voltammetry (SWV). Elemental analysis and morphology of applied nanomaterials were characterized by energy-dispersive X-ray spectroscopy (EDX) and feld emission scanning electron microscopy (FESEM), respectively. MWCNTs and Ni-NPs illustrated satisfactory synergic electrocatalytical properties in DCV oxidation, which leads to DCV sensitive detection in real samples such as tablet and human serum. The limit of detection (LOD) and linear range were 15.82 nM and 0.024–300 µM, respectively, which, in comparison to other reported studies about the determination of DCV, were satisfying results. The stability of the sensor was studied and the result was very acceptable.