The color assessment cabinet has important applications in the color fields of textiles, pigments, printing and dyeing, packaging, and photography. In order to achieve the spectral matching of CIE standard illuminant D65 using monochromatic LEDs, in this paper, we use genetic algorithm (GA) as the spectral matching algorithm, measure the absolute spectra of monochromatic LEDs and establish a database, optimize the parameters of the algorithm through simulation experiments, and optimize the combination of monochromatic LEDs using decreasing experiments to achieve the purpose of using fewer kinds of monochromatic LEDs to obtain a better fit with the target spectrum. The results show that the correlation indices R^2 and RMSE of 30 monochromatic LED combinations fitting D65 spectrum are 0.905 and 0.946, respectively, with high matching degree of D65 spectrum; 16 LED combinations fitting D65 spectrum are proposed by decreasing experiments, which have good fitting effect and reduce the difficulty of engineering practice.