Reduction in seroprevalence of Hepatitis A virus (HAV) is known to be associated with improvements in socioeconomic conditions of the community. National Institute of Virology, Pune has been studying seroprevalence of hepatitis viruses in Pune region over the past four decades. In total, 1438 samples were collected from urban general (UGEN), urban lower socioeconomic stratum (ULSES) and rural (RURAL) populations of the Pune district. Based on estimates in previous studies, subjects were enrolled from age groups ‘6–10’, ‘15–25’ and ‘40 + ’ years. HAV seroprevalence in younger population showed a significant decline. A significant decline in HAV seroprevalence in ‘15–25’ years age group in UGEN (from 85.9% to 73.9%; OR = 0.46, 95% CI: 0.25–0.86) and RURAL (from 98.6% to 91.4%; OR = 0.15, 95% CI: 0.05–0.45) populations suggested that the trend probably started more than a decade ago. Seroprevalence of HAV among ULSES ‘6–10’ children was found to be significantly higher (70.4%) than that among the RURAL children (44.2%; OR = 3.0, 95%CI: 1.7–5.2) and UGEN children (40.4%; OR = 3.5, 95%CI: 1.8–6.7). In view of increasing rates of urbanisation in India, ULSES population needs special consideration while designing future studies and viral hepatitis vaccination/elimination strategies. Our findings call for robust population-based studies that consider heterogeneity within populations and dynamics of socio-economic parameters in various regions of a country.