This retrospective study investigates prognostic factors affecting the visual acuity after a single intravitreal injection of rAAV2-ND4 gene therapy in 53 patients with Leber’s hereditary optic neuropathy (LHON) with mutation at site 11 778. Patients were reviewed one and three months after treatment and divided into improvement and non-improvement groups, with best-corrected visual acuity (BCVA) improvement ≥ 0.3 logMAR defined as improvement. Sex, onset age, duration of disease, BCVA, visual field index (VFI) and mean deviation (MD) were recorded at baseline. Correlation between BCVA prognosis and baseline characteristics were analysed by univariate analysis. Logistic regression analysis was performed on independent factors affecting BCVA. Only greater baseline VFI and better baseline BCVA predicted better visual acuity prognosis post gene therapy.