The aim of the present study was to investigate the utility of diffusion tensor imaging (DTI) parameters of the central visual pathway and the size of the lateral geniculate nucleus (LGN) in the diagnosis of primary open-angle glaucoma (POAG). To test this, 24 POAG patients and 24 age- and gender-matched controls underwent 3.0 Tesla magnetic resonance imaging examinations, including DTI and LGN structural imaging. Receiver operating characteristic (ROC) curves were generated and Spearman's correlation coefficients were also determined. The results indicated that regarding the discrimination of POAG patients from healthy controls, the fractional anisotropy (FA) values of the optic tract, at a cutoff of 0.412, with an area under the ROC curve of 0.931, exhibited the highest sensitivity (79.2%) and specificity (89.6%). The FA values of the optic tract and the optic radiation were significantly correlated with LGN size, while the mean diffusivity values were not. In conclusion, the FA value of the optic tract may be a sensitive and reliable biomarker for glaucoma evaluation.