Background: Severe visual loss is seen in both multiple sclerosis-associated optic neuritis (ON) and neuromyelitis optica (NMO)-associated ON. NMO (aquaporin 4) antibodies have been reported to have diagnostic and prognostic value for visual and neurological outcomes of recurrent ON. We performed this study to investigate the frequency of NMO antibodies and its prognostic value for visual and neurological outcomes in Chinese patients with severe ON. Methods: Single-center prospective cohort study. Detection of NMO antibodies was by indirect immunofluorescence method using human aquaporin 4-transfected cells. Severe ON was defined as visual acuity of 20/200 or worse in at least 1 eye at the nadir of the patients' course. Clinical features at baseline, visual outcome, and sequential neurological events were compared between seropositive and seronegative groups. Results: NMO antibodies were detected in 11 of 34 patients (32.4%) with severe ON. Five seropositive patients with recurrent ON had significantly higher titers (range: 1:512 to 1:65,536; median: 1:512) than those of 6 seropositive patients with only 1 episode (range: 1:16 to 1:512; median: 1:32) (P = 0.021). Female to male ratio (10:1) and antinuclear autoantibody positivity in seropositive patients (3 of 9, 33.3%) were statistically higher than those of the seronegative group (12:11; 0 of 19; P = 0.026). The seropositive patients had significantly poorer visual outcomes than seronegative patients (P = 0.025). During the averaged 32-month follow-up, 2 of 11 seropositive patients (18.2%) developed clinically incomplete transverse myelitis, while no similar symptoms were reported in the seronegative group. Conclusion: NMO antibody positivity is relatively high in Chinese patients with severe ON and suggests a poorer visual outcome, probably higher risk of developing spinal cord lesions and a closer association with systemic autoimmune disorders.