Objective: To report our experience with two tests, anti-cardiolipin antibody test [venereal disease reasearch laboratory (VDRL) test] and specific treponemal test (Treponema pallidum hemagglutination assay), used for screening antenatal, high risk cases and cases from sexually transmitted infection in a tertiary care hospital from January 2006 till December 2008. Methods: A total of 14 639 samples received from various patient groups including antenatal cases, patients attending sexually transmitted disease (STD) clinic, blood donors and HIV positive patients were screened. Results: Among the 14 639 samples collected, 103 were positive by VDRL test. Of these 89 cases were confirmed by quantitative VDRL test and Treponema pallidum hemagglutination assay. The cumulative seroprevalence over two years was found to be 0.61% in this study. The syphilis seroprevalence reduced from 0.88% in 2006 to 0.40% in 2008. Among the various sub-populations studied, patients attending the sexually transmitted infection clinic showed a seroprevalence of 2.62%. The seroprevalence decreased significantly from 4.00% in 2006 to 1.39% in 2008. Conclusions: Our study showed a statistically significant declining rate of syphilis in STD clinics as well as the overall seroprevalence. These findings could be interpreted as indicators of improved programmes for prevention and management of STDs.