Objective To study the incidence and risk factors of retinopathy of prematurity (ROP) in a single tertiary center during different times(Phase 1:2009 to 2011;Phase 2:2012 to 2014). Methods From 2009 to 2014, fundus examinations were performed on premature infants admitted to NICU of our hospital. The incidence and risk factors of ROP were compared between the two phases. Results During Phase 1,68 (11. 1% ) cases were diagnosed with ROP among 614 premature infants. During Phase 2, 121 (15. 2% ) cases were diagnosed with ROP among 794 premature infants. The incidences of ROP between the two phases were significantly different (P <0. 05). During Phase 1 and 2, 10 (14. 7% ) infants and 12 (9. 9% ) infants were diagnosed with Grade 3 and above or threshold ROP, respectively. The incidence of Grade 3 and above or threshold ROP between the two phases were also significantly different (P < 0. 05). Logistic analysis demonstrated that gestational age(GA), birth weight (BW), duration of oxygen therapy and mechanical ventilation(MV) were independent risk factors during hase 1, whereas GA, MV and blood transfusion were independent risk factors during Phase 2. Pulmonary surfactant (PS) was protective factor of ROP in both two groups. GA in Phase 2 was significant lower than Phase 1 (P <0. 05), while duration of oxygen therapy, incidence of MV, PS and blood transfusion were higher in Phase 2 ( P < 0. 05) . Conclusions The incidence of ROP has increased. Reducing the incidence of preterm delivery, duration of oxygen therapy, incidence of MV and blood transfusion has important role in preventing ROP. Reducing the use of MV and the oxygen concentrations, appropriate PS therapy and focusing on early ophthalmic screening are important preventing severe ROP.