OBJECTIVE: To describe trends in chlamydia positivity from 2007 to 2011 among heterosexual people tested for chlamydia at selected clinics that provide services to people at high risk in Victoria, Australia. DESIGN: The Victorian Primary Care Network for Sentinel Surveillance is a prospective system that collates pathology results from laboratories and demographic and behavioural data from a questionnaire. SETTING: Two sexual health clinics and six other primary care clinics that target young people and women at high risk. PARTICIPANTS: All clients tested for chlamydia at sentinel clinics. Individuals aged less than 16 years, sex workers, or those reporting any same-sex sexual partners in the past 12 months were excluded from the analysis. MAIN OUTCOME MEASURES: Chlamydia positivity trends were assessed using three-level random-effects Poisson regression, with clinic and subject treated as nested random factors. Models were offset for total number of tests and adjusted for relevant covariates. RESULTS: Between 2007 and 2011, chlamydia positivity among 31 682 tests in women increased from 5.1% to 6.3%, and positivity among 23 771 tests in men increased from 7.4% to 8.2%. Adjusting for age, country of birth, number of sex partners, condom use, and presence of symptoms, chlamydia positivity increased between 2007 and 2011 significantly among women (incidence rate ratio [IRR], 1.29; 95% CI, 1.11-1.50) and non-significantly among men (IRR, 1.07; 95% CI, 0.92-1.23). Over time, a decrease in reported inconsistent condom use was also observed in both men and women; however, men became more likely to report more than one partner in the past year. CONCLUSION: We identified a concerning increase in chlamydia positivity over time, particularly among young women.