Glucose detection is very important for the diagnosis and management of diabetes, and nonenzymatic glucose sensors have attracted considerable attention due to their excellent stability, high sensitivity, and so on. Herein, a sensitive, cost-effective nonenzymatic glucose sensor using ultralong Ni nanowire (NW) modified glassy carbon electrode is presented. The current–time curve shows that the catalytic oxidation current is linearly dependent on glucose concentration in the range from 1 μM to 5 mM with a correlation coefficient 0.994, and a sensitivity of 367 μA mM⁻¹ cm⁻². The relative standard deviation is 3.7% for 8 successive measurements for 1 mM glucose. After 2-weeks storage in air at room temperature, the signal for 1 mM glucose maintains 97.2% of its initial value. What is more, the present Ni NW-based electrochemical sensing platform can successfully be used to detect glucose in human serum with satisfactory results.