The use of non-invasive blood glucose detection techniques can help diabetic patients to alleviate the pain of intrusive detection, reduce the cost of detection, and achieve real-time monitoring and effective control of blood glucose. Given the existing limitations of the minimally invasive or invasive blood glucose detection methods, such as low detection accuracy, high cost and complex operation, and the laser source's wavelength and cost, this paper, based on the non-invasive blood glucose detector developed by the research group, designs a non-invasive blood glucose detection method. It is founded on dual-wavelength near-infrared light diffuse reflection by using the 1 550 nm near-infrared light as measuring light to collect blood glucose information and the 1 310 nm near-infrared light as reference light to remove the effects of water molecules in the blood. Fourteen volunteers were recruited for in vivo experiments using the instrument to verify the effectiveness of the method. The results indicated that 90.27% of the measured values of non-invasive blood glucose were distributed in the region A of Clarke error grid and 9.73% in the region B of Clarke error grid, all meeting clinical requirements. It is also confirmed that the proposed non-invasive blood glucose detection method realizes relatively ideal measurement accuracy and stability.