A non-enzymatic copper-doped zinc oxide glucose sensor, which is covered with silver nanowires (AgNWs) and modified with Nafion, can be employed to measure glucose with a concentration of 2–14 mM. In this study, X-ray photoelectron spectroscopy (XPS) was used to analyze the film of the working electrodes, and the AgNWs on the electrode surface were analyzed by a field-emission scanning electron microscope (FE-SEM). Also, the AgNW/CZO structure was investigated through the use of an atomic force microscope (AFM) and an ultraviolet/visible (UV/Vis) spectrophotometer. Finally, we used electrochemical impedance spectroscopy (EIS) and the voltage–time (V–T) measurement system to measure the sensing characteristics of the thin film. The potentiometric non-enzymatic glucose sensor has the average sensitivity of 10.31 mV/mM, and the linearity is 0.997. Also, the sensor has the response time with 15 s, and the interference experiment proved that it has good anti-interference and excellent repeatability (relative standard deviation (RSD) = 3%). Besides, the sensor still maintains a certain degree of average sensitivity at high temperatures, and it has the limit of detection (LOD) with 0.272 mM.