Strawberry variety is a main factor that can influence strawberry fruit quality. The use of near-infrared reflectance spectroscopy was explored discriminate among samples of strawberry of different varieties. And the significance of difference among different varieties was analyzed by comparison of the chemical composition of the different varieties samples. The performance of models established using back propagation-artificial neural networks (BP-ANN), least squares-support vector machine and discriminant analysis were evaluated on spectra range of 4 545～9 090 cm-1. The optimal model was obtained by BP-ANN with a topology of 12-18-3, which correctly classified 96.68% of calibration set and 97.14% of prediction set. And the 94.95%, 97% and 98.29% classifications were given respectively for “Tianbao” (n=99), “Fengxiang” (n=100) and “Mingxing” (n=117). One-way analysis of variance was made for comparison of the mean values for soluble solids content (SSC), titratable acid (TA), pH value and SSC-TA ratio, and the statistically significant differences were found. Principal component analysis was performed on the four chemical compositions, and obvious clustering tendencies for different varieties were found. These results showed that NIR combined with BP-ANN can discriminate strawberry of different varieties effectively, and the difference in chemical compositions of different varieties strawberry might be a chemical validation for NIR results.