Chlamydia trachomatis screening is being considered in the Netherlands, but policy recommendations are hampered by the lack of population-based data. We studied the prevalence of chlamydia infection in 15-29-year-old women and men in a national representative sample of 21,000 inhabitants of rural and urban areas in the Netherlands. Of this sample, 41% responded by sending in urine and an answered questionnaire, while 11% returned a refusal card. The overall prevalence of chlamydia infection was 2.0% (CI: 1.7-2.3); 2.5% (CI: 2.0-3.0) in women and 1.5% (1.1-1.8) in men. Chlamydia prevalence was significantly greater in very highly urbanized areas (3.2%, CI: 2.4-4.0) compared to rural areas (0.6%, CI: 0.1-1.1). In very highly urbanized areas the greatest prevalence was found among 15-19-year-old women (4.3%) and among 25-29-year-old men (4.2%). A risk profile could be determined and a prediction rule was developed. These data suggest that nationwide systematic screening is not indicated in the Netherlands and that targeted approaches are a better option. Roll-out of selective screening is recommended.