Fast Fourier transform (FFT) is commonly adopted as the signal processing method of laser Doppler velocimeter (LDV) system to get Doppler frequency which is the key factor of the calculation of object velocity. However, precision of FFT is restricted by phenomena caused by asynchronous sampling, such as spectral interference, fence effect and spectrum leakage. Aiming at enhancing the precision of LDV signal processing, an algorithm based on the association of spectral refinement and interpolation is put forward. Zoom-FFT was utilized to refine the local range of Doppler frequency estimated by FFT. The refined spectrum was further corrected by Rife-Vincent (Ⅲ) window interpolation and got the accurate estimate of Doppler frequency. Cubic spline fitting was applied to calculate the interpolation coefficient of Rife-Vincent (Ⅲ) window without solving a quintic equation in the computing procedure of interpolation. For experimental signals, the minimum relative error of proposed method is 0.02%. And for simulated signals, the minimum relative error reaches only 0.0013%. The proposed method has capability to significantly improve the precision of signal processing and is applicable for a LDV system.