OBJECTIVE: This study evaluates trends in antibody seroprevalences of seven high-risk human papillomavirus (hr-HPV) serotypes (HPV16, 18, 31, 33, 45, 52, and 58) between the 1995-96 and 2006-07 sero-surveys among the Dutch general population in the pre-vaccination era. METHODS: Serum samples of men and women (0-79 years of age) from two cross-sectional population-based serosurveillance studies performed in 1995-96 (n = 3303) and 2006-07 (n = 6384) were tested for HPV-specific antibodies in a VLP-based multiplex immunoassay. RESULTS: HPV16-specific antibody seroprevalence increased during adolescence and shifted to younger ages in the 2006-07 survey compared to the 1995-96 survey. This step-up in HPV16 seroprevalence was most pronounced in women, while a more gradual increase was observed in men. Also in cohorts older than 49 years, HPV16 seroprevalence was higher in 2006-07 as compared to 1995-96 survey. A higher overall seroprevalence in individuals older than 15 years of age was found for HPV16, 18, 31 and 45 in 2006-07 as compared to 1995-96. For HPV33, 52 and 58 seroprevalences were comparable over this 11-year time period. Seropositivity for one or more HPV types was significantly higher in 2006-07 (23.1%) than in 1995-96 (20.0%) (p = 0.013). Multi-seropositivity increased from 7.1% in 1995-96 up to 10.2% in 2006-07 (p<0.0001). Differences in HPV seropositivity for at least one of the seven HPV types between both surveys could be explained in addition to demographic characteristics (age, sex, urbanization degree and ethnicity), also by changes in sexual behaviour (marital status, age of sexual debut and ever reported an STI). CONCLUSION: The observed increase in particular HPV16 seroprevalence could be due to changes in sexual behaviour over the years, and especially in age of sexual debut. Seroprevalence studies provide insight into the distribution of HPV types and infection dynamics in the general population over time, which is important to assess the impact of HPV-vaccination.