Laboratory confirmation of chlamydial antigen in clinically suspected cases of chlamydial eye infections is important, as similar clinical picture can be presented by different infective or noninfective causes. We retrospectively analyzed the presence of Chlamydia trachomatis antigen in 690 clinically suspected patients over the last 8 years (2009–2016). The chlamydial antigen was detected using direct immunofluorescence assay. Overall, Chlamydia-specific antigen positivity was 45.5%. The highest positivity was seen in 2014 (68.6%) and the least in 2016 (9.4%). The antigen positivity in years 2015 (13.4%) and 2016 (9.4%) was significantly less than in all the previous study years (P < 0.0001). Antigen positivity in patients having clinical diagnosis of trachoma was significantly higher than those having other eye manifestations suggestive of chlamydial infections (P = 0.0274). Stringent surveillance both at community level and in hospital attendees is required to know the actual load of this pathogen.