Aim To evaluate the role of fixation in causing pterygium and determining its laterality and location. Methods This is a prospective, observational, case-control study. Cases were defined as patients with primary pterygium who had unilateral amblyopia with eccentric fixation. Controls were age-matched patients with primary pterygium, but without amblyopia and eccentric fixation. All patients underwent complete ocular, orthoptic, and systemic examination and a detailed risk-factor assessment (latitude of residence, exposure to sunlight, sand, and a high-reflectance environment). The role of fixation in the causation, laterality and location of pterygium was evaluated. Fisher's exact test, the unpaired t-test, and odds ratio (OR) were carried out to determine the significance of the observations. Results The mean age of subjects was 47.1 ± 5.25 years in cases (n ¼ 107) and 48.2 ± 4.75 years in controls (n ¼ 310; P ¼ 0.78). As far as known risk factors were concerned, both groups were evenly matched. Among the cases, 88 (82.2%) patients demonstrated suppression of the amblyopic eye and 19 (17.8%) patients had abnormal retinal correspondence (ARC). Patients with suppression had a unilateral pterygium in the better (fixating) eye, whereas those with ARC had bilateral pterygia. Among the controls, 192 (61.9%) eyes had bilateral pterygia and 118 (38.1%) eyes had unilateral pterygium. In controls, the dominant eye had a higher prevalence of pterygium. All patients in both groups had a nasal pterygium. Pterygium and fixation were strongly associated (P ¼ 0.007; Fisher's exact test; OR À15.98; P ¼ 0.008). Conclusion Fixation appears to have an important role in causing pterygium and determining its location and laterality.