The aim was to carry out a population-based sero-prevalence survey of Epstein-Barr virus (EBV) across a wide age range in England and Wales and to identify any associations between EBV and herpes simplex virus types one and two (HSV-1 and 2). Sera from an age-stratified sample of 2,893 individuals, submitted for diagnostic purposes to 15 public health laboratories in England and Wales in 1994, were tested for immunoglobulin G (IgG) antibody to EBV. The samples had been tested previously for IgG antibody to HSV-1 and HSV-2. The serological profile of EBV was consistent with an endemic infection with peaks in transmission in those less than 5 years old and in young adults. An age adjusted analysis found a significant association between EBV and HSV-1 seropositivity that is most likely explained by similarities in their mode of transmission. The very low seroprevalence of HSV-2 in this sample complicated the comparisons of EBV and HSV-1 with HSV-2. Any associations were most likely explained by chance. Given the association between EBV and HSV-1, it is likely that recently documented epidemiological changes in HSV-1 also apply to EBV. Continuing surveillance of these herpes viruses is necessary as the predicted changes could have a significant public health impact, especially in the young adult population.