This retrospective cohort study aimed to assess the characteristics of patients enrolled in the national diabetic retinopathy (DR) screening program and evaluate the selected program's quality elements in the first five years after implementation. Patients who underwent DR screening between February 2018 and June 2022 at the University Medical Centre Ljubljana were included. General patient characteristics (age, sex, type and therapy of diabetes mellitus (DM), duration of DM, blood HbA1c levels), best corrected visual acuity, stage of DR, presence of sight-threatening diabetic retinopathy (STDR), and selected quality indicators (quality of photographs, re-screening interval, time to treatment) were recorded and evaluated. A total of 34 654 screening examinations were performed on 13 513 patients with diabetes. The majority (77.3%) had Type 2 diabetes, with a mean DM duration of 9.74 ± 9.87 years. In 55.9% of patients, DM duration was less than ten years, and the mean HbA1c was 7.33% ± 1.49. DR signs were noted in 29.1% of patients and 0.89% of patients exhibited STDR. After establishing the diagnosis of STDR, treatment was initiated in 3.43 ± 1.98 months. Implementing the DR screening program provides valuable insights into managing diabetic retinopathy nationwide. It allows for the early detection of sight-threatening DR and facilitates timely and effective treatment, which could help prevent severe visual impairment.