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<b>Objective:</b> To investigate the incidence and clinical features of retinopathy of prematurity (ROP) among preterm infants in a neonatal intensive care unit (NICU) of Lhasa, and to explore the screening criteria for ROP in high-altitude areas. <b>Methods:</b> It was a single-center retrospective case study. A total of 166 preterm infants of Tibetan ethnicity admitted to the NICU of Lhasa People's Hospital from May 1, 2020 to June 1, 2022 with a gestational age<36 weeks were enrolled. ROP was screened using the third-generation wide-angle digital pediatric retinal imaging system (RetCam Ⅲ). The ROP detection rate, staging, and zonal distribution were analyzed. Multivariate logistic regression was performed and risk factors for ROP in preterm infants were identified by calculating the odds ratio (<i>OR</i>) and its 95% confidence interval (95%<i>CI</i>). <b>Results:</b> There were 88 males and 78 females, among whom 33 patients (66 eyes) were diagnosed with ROP. The birth gestational age of ROP patients was 30 (29, 31) weeks, and the average birth weight was 1 188.5±219.2 g. The disease was at stage 1 in 48 eyes (72.7%), stage 2 in 16 eyes (24.2%), and stage 3 in 2 eyes (3.0%). Two eyes (3.0%) had zone Ⅰ disease, 26 eyes (39.4%) had zone Ⅱ disease, and 38 eyes (57.6%) had zone Ⅲ disease. Thirty ROP patients (90.9%) had spontaneous regression and vascularization. Three patients (9.1%) developed type Ⅰ ROP, which was treated by anti-VEGF therapy (1 patient) or spontaneously resolved during the follow-up (2 patients). Multiple logistic regression analysis showed that gestational age (<i>OR</i>=0.53, 95%<i>CI</i>: 0.33-0.84, <i>P</i>=0.007), birth weight (<i>OR</i>=0.95, 95%<i>CI</i>: 0.92-0.98, <i>P</i><0.001), and duration of invasive mechanical ventilation (<i>OR</i>=1.42, 95%<i>CI</i>: 1.06-1.90, <i>P</i>=0.018) were influencing factors of ROP. <b>Conclusions:</b> The detection rate of ROP among Tibetan preterm infants in the single-center neonatal intensive care unit in Lhasa was 19.9%, while the detection rate of type Ⅰ ROP was 1.8%. Preterm infants with shorter gestational age, lower birth weight, and prolonged invasive mechanical ventilation were liable to develop ROP.