Objective To measure the changes in the peripapillary and perimacular retinal nerve fiber layei (RNFL) thickness.of patients with Leber's hereditary optic neuropathy (LHON) with spectral-domain optical coherence tomography (OCT).Methods In this retrospective case-control study,the peripapillary and perimacular RNFL thickness of clinical patients with LHON (33 people,66 eyes) and normal volunteers (67 people,67 eyes) was measured by Heidelberg OCT,and blood samples of clinical patients with LHON were collected from three primary mtDNA mutation points (G11778A,G3460A and T14484C).The patients were divided into two groups,the LHON group and those suspected of having LHON,based on their genetic diagnosis.The differences in the thickness of the peripapillary and perimacular RNFL were then compared.The comparisons included the temporal side,superior temporal quadrant,inferior temporal quadrant,nasal side,nasal superior quadrant,nasal inferior quadrant,and the average thickness of 360° RNFL.Results There were 18 LHON patients and 15 patients suspected of having LHON based on the mutation detection of the 33 clinical patients.Peripapillary RNFL had significant differences in the temporal quadrant,temporal superior quadrant,temporal inferior quadrant and nasal superior quadrant among the three groups (F=145.14,11.25,57.10,4.48,P＜0.05).Perimacular RNFL had significant differences in all quadrants (F=24.07,67.01,85.99,130.21,121.90,128.66,P＜0.05).The peripapillary RNFL of the LHON patients was thinner than that of the normal volunteers except for the nasal side and nasal inferior quadrant (P＜0.05).All quadrants of the perimacular RNFL of the LHON patients were thinner than that of the normal volunteers (P＜0.05).The peripapillary RNFL of the patients suspected of having LHON was thinner than that of the normal volunteers in the temporal quadrant,temporal superior quadrant,and temporal inferior quadrant (P＜0.05).All quadrants of the perimacular RNFL of patients suspected of having LHON were thinner than those of the normal volunteers (P＜0.05).Conclusion In later stages,the LHON patients presented with an optic atrophy,not only in the papillomacular bundle,but also in the superior and inferior temporal quadrants.The nerve fibers of the nasal quadrant are relatively unchanged.