A portable minimally-invasive human blood glucose detection instrument was designed by a microfluidic chip based interstitial fluid transdermal extraction system. On the basis of the minimally-invasive method, the instrument utilizes a vacuum pressure to extract the interstitial fluid from human body, adopts the Surface Plasmon Resonance (SPR) technology to measure the glucose concentration of the interstitial fluid, and then predicts the blood glucose concentration with the glucose concentration of the interstitial fluid. Moreover, by immobilizing D-galactose/D-glucose Binding Protein (D-GGBP) which has a specific adsorption function to the glucose, it preprocesses the surface of the SPR sensor and realizes the specific adsorption of glucose molecules. In the experiment, the glucose solution with different concentrations was prepared and detected. According to the experimental results, the curve illustrating the relationship between glucose concentration and refractive index was obtained. In addition, the glucose solution and interstitial fluid were measured by using minimally-invasive blood glucose detection instrument designed by our research group. The test result was compared with that measured by a glucose meter. The experimental result indicates that the glucose detection resolution could reach 0.625 mg/dL using the SPR sensor decorated with the GGBP. It shows a good linearity when the glucose concentration ranges from 0.625 mg/dL to 5 mg/dL. The experiment verifies the feasibility of the instrument, and demonstrates that the SPR technology with immobilized GGBP will obtain a wide application in the field of minimally-invasive glucose measurement.