Background/Aims: Retinal nerve fiber layer (RNFL) thinning has been observed on histopathology and time-domain optical coherence tomography in many diseases of the central nervous system. In this study, with a higher resolution of spectral-domain optical coherence tomography (SDOCT), we detected RNFL changes in patients with multiple sclerosis (MS) in China, and compared RNFL thickness between eyes with and without optic neuritis (ON). Methods: In this retrospective, nonrandom case study, the patients were recruited from the Affiliated Sir Run Run Shaw Hospital of Zhejiang University. RNFL thickness was measured for each eye using SDOCT. The controls were recruited from the healthy population. Results: Peripapillary RNFL thickness of 24 eyes in 12 patients was detected by SDOCT. The average RNFL thickness of the MS patients was 81.9 ± 17.8 µm compared to the control value of 102.1 ± 8.1 µm (p = 0.00). The average RNFL of the patients with a history of ON was thinner than that of patients without ON (71.8 ± 19.2 µm vs. 92.0 ± 8.5 µm, p = 0.001). Conclusion: The RNFL thinning in Chinese patients with MS can be detected by SDOCT. The SDOCT scan represents a high-resolution, objective, noninvasive and easily quantifiable in vivo biomarker of MS.