The purpose of this study was to estimate the prevalence of amblyopia in subjects aged 3-93 years living in under-served rural areas of Iran. The study was conducted in 2015 in two under-served districts randomly selected from the southwest and north of Iran, followed by a random selection of villages from these areas. Amblyopia was defined as best corrected visual acuity (BCVA) of 20/30 or worse, or a difference of at least two lines in visual acuity between eyes in the absence of any pathology. Of 3850 invited subjects, 3314 took part with an 86.08% response rate. After exclusions, 2960 were analysed. Mean age was 36.90 ±2021 years (3-93) with 56.8% female. Prevalence of amblyopia was 2.73% (CI 2.17-3.38%; n=81). Amblyopia was bilateral in 15 and unilateral in 66. Types included anisometropic (53%), strabismic (29.5%) and mixed (17.5%). The lowest prevalence was in ages 6-20 years and highest in >70 years potentially linked to amblyopia screening programmes introduced in 1996. Prevalence reduced significantly with higher education level. They conclude there is a higher prevalence in rural than urban areas but acknowledge the limitations of the study with cohort effects and cross-sectional design. Further prospective studies are needed to examine causality.