A serological survey was used to investigate the epidemiology of cytomegalovirus (CMV) infection in England and Wales. A total of 5237 sera representing the complete age range were used reflecting the general population. The sera were collected in 1991 and 2002, and screened for CMV-specific IgG by ELISA. Antibody prevalence increased with age from ~15% in those aged 1–4 years to ~80% in those aged ⩾65 years with no association with gender or region. Analysing by common birth cohort demonstrated that between 1991 and 2002 incidence was highest in children born 1985–1989 (1·62% per year, 95% CI 0·86–2·35), lower in older children and younger adults born 1950–1984 (0·75% per year, 95% CI 0·29–1·19) with little evidence of infection in older adults born pre-1950 (0% per year, 95% CI 0–0·64). Application to population and live-birth estimates for England and Wales suggested that between 1991 and 2002, 159 996 (95% CI 67922–278277) CMV infections occurred annually with an annual average of 2133 (95% CI 816–3435) infections affecting pregnant females.