The Netherlands has experienced large community outbreaks of Q fever since 2007. Sera and questionnaires containing epidemiological data from 5654 individuals were obtained in a nationwide seroprevalence survey used to evaluate the National Immunization Programme in 2006–2007. We tested these sera for IgG phase-2 antibodies against Coxiella burnetii with an ELISA to estimate the seroprevalence and to identify determinants for seropositivity before the Q fever outbreaks occurred. Overall seroprevalence was 1·5% [95% confidence interval (CI) 1·3–1·7]. Corrected for confirmation with immunofluorescence results in a subset, the estimated seroprevalence was 2·4%. Seropositivity ranged from 0·48% (95% CI 0·00–0·96) in the 0–4 years age group to 2·30% (95% CI 1·46–3·15) in the 60–79 years age group. Keeping ruminants, increasing age and being born in Turkey were independent risk factors for seropositivity. The low seroprevalence before the start of the outbreaks supports the hypothesis that The Netherlands has been confronted with a newly emerging Q fever problem since spring 2007.