We determined the prevalence of Chlamydia trachomatis (CT) infection and risk factors for its acquisition by women aged less than 35 years who were attending sexual health centres in the province of Barcelona in 2007. A convenience sample of 397 adolescents and young adult women aged between 16 and 35 years, stratified by age and recruitment site, were tested using realtime polymerase chain reaction by CT DNA. A standardized questionnaire was used to collect epidemiological and behavioural data on the participants, which were analysed in a multivariate logistic regression model. The overall prevalence of CT was 4%, significantly higher in those under 25 years of age (5.8% versus 1.6% over 25). The independent risk factors for CT infection were: foreign origin (odds ratio [OR] 4.7; confidence interval [CI] 1.02-21.8), having had a sexual partner in the last three months (OR 4.59; CI 1.16-18.08) and tobacco use the last 12 months (OR 6.38; CI 1.16-34.93). In conclusion, this is the first study performed in Catalonia showing a high prevalence of CT in young women, consistent with trends in the rest of Europe. Systematic monitoring of CT infection in sentinel populations such as this will inform future targeted screening programmes in our setting.