The prediction model of beef’s storage time was established based on multi indexes of fresh beef, such as TVB-N, colony total, pH value, and L* parameter. Visible and near-infrared spectroscopy (Vis/NIR) combined with interval PLS(iPLS)and genetic algorithm(GA) was investigated for establishing PLS calibration model of above 4 indexes, respectively, and rapid and nondestructive prediction of the storage time of fresh beef stored at 4 ℃ was realized. PLS models of 4 indexes were built with full spectrum and effective variables selected by iPLS and iPLS-GA method, respectively. The performance of each model was evaluated according to two correlations coefficients(R) and standard error (SE) of calibration and prediction sets. Experimental results showed that the performance of all models built with effective variable selected by iPLS-GA was better than full spectrum and iPLS. The storage time of calibration and prediction sets of beef samples was predicted by storage time model with predicted values of above 4 indexes, and was achieved as follows: Rc=0.903, Rp=0.897, SEC=1.88 and SEP=2.24. The study demonstrated that the beef’s storage time can be synthetically predicted with multi-index by using visible and near-infrared spectroscopy combined with the prediction model of beef’s storage time. This provides a new method for rapid and non-destructive detection of beef’s storage time or shelf life.