A unique nanostructured electrocatalyst based on copper oxide (CuO) and Co-Ni layered double hydroxides (LDHs) was synthesized using an electrodeposition method. The optimized CuO/Co-Ni-LDHs composite exhibited improved non-enzymatic electrochemical sensing performance for glucose. Display omitted The leaf-shaped copper oxide (CuO) was first fabricated by a direct precipitation method. Then, the flake-shaped Co-Ni layered double hydroxides (CoNi-LDHs) were grown on the leaf-shaped CuO by using an electrodeposition method to form a unique nanostructured electrocatalyst. Due to the unique structural and compositional advantages, CoNi-LDHs wrapped around the leaf-shaped CuO exhibited good electrocatalytic characteristics. The optimized CuO/CoNi-LDHs exhibited improved non-enzymatic electrochemical sensing performance for glucose, with a reliable linear range (0.1 μM–0.384 mM) and a low limit of detection (0.065 µM, S/N = 3). The utilization of the CuO/CoNi-LDHs sensor for glucose detection in human serum was also corroborated, implying promising potential for glucose monitoring. Results demonstrated that the developed sensor provides new horizons for facile and sensitive glucose detection.