A tilted fiber Bragg grating (TFBG) sensor based on surface plasmon resonance (SPR) with electrospinning nanofibers was proposed for the precise detection of glucose. In this design, a 50 nm thick gold film was deposited on the surface of a 10° TFBG. Polyacrylonitrile (PAN) nanofibers were wrapped around the sensor through electrospinning, subsequently serving as a scaffold for the immobilization of glucose oxidase (GOD). Within the dynamic range of 100–500 mg/dL glucose concentrations, the sensor exhibited good response and worked effectively. The experimental results show that the sensitivity and limit of detection (LOD) of the sensor are 12.42 pm/(mg/dL) and 100 mg/dL. Furthermore, the structural characterization was carried out using atomic force microscopy (AFM), scanning electron microscopy (SEM), and Fourier transform infrared spectrometer (FTIR). This sensor exhibits outstanding stability and high sensitivity, providing a new method for diabetes detection.