The detection of glucose is important in clinic diagnosis, while the traditional methods for glucose sensing rely heavily on glucose oxidase (GOx). Herein, with the easy preparation, stability as well as selectivity, a polyvinyl borate (PVB) hydrogel was used for the non-GOx and portable detection of glucose. In the colorimetric channel, the hydrogel can be specifically decomposed in the presence of glucose, thus the electric resistance can be changed, causing significant color change of the electrochromic Prussian blue (PB) film in a very short time of 3 s. A linear relationship between the grayscale value of PB and glucose concentration was established in the range between 0.1 and 10.0 mM with the limit of detection (LOD) of 0.02 mM. In addition, the alkaline phosphatase (ALP) in the PVB gel was released to react with L-ascorbyl-2-phosphate, thus the product of ascorbic acid can reduce HAuCl4 to form the aggregation-induced emission enhancement (AIEE) of Au nanoclusters. Two separate ranges with glucose concentrations of 0.1–1.0 mM and 1.0–10.0 mM, and also a LOD of 0.018 mM were achieved. In particular, a mobile APP was designed to read the grayscale values of PB, making the sensor suitable for the on-site detection. Overall, this work may show a great potential in low-cost, stable and portable glucose detection.