In this research, dual-strategy biosensing of glucose was proposed based on multifunctional CuWO nanoparticles (CuWO NPs), which were prepared for the application of electrochemical and colorimetric sensing of glucose. CuWO NPs show large specific surface area and good conductivity as well as excellent peroxidase-like activity. A sensitive and selective electrochemical glucose biosensor was fabricated with the immobilization of glucose oxidase (GOx) on a CuWO NP modified electrode for enhancing the direct electron transfer behavior. A wide linear range of 0.005-1.8 mM with a low detection limit of 1.5 μM and a high sensitivity of 28.02 mA M cm were achieved by using the electrochemical biosensor. Meanwhile, a colorimetric and visual glucose biosensor was constructed based on the GOx/CuWO cascade nanozyme, which shows a linear range of 0.05-1.0 mM with excellent selectivity. CuWO NPs as a promising matrix open up a dual-strategy biosensor for sensitive and selective detection of glucose.