PURPOSE: This study compared performance of the Atlas io polymerase chain reaction-based, point-of-care (POC) assay for Chlamydia trachomatis (CT), to Aptima Combo 2, a standard of care nucleic acid amplification assay, and evaluated patient attitudes toward POC testing. METHODS: Women 14 years or older undergoing CT screening/testing were recruited from Teen Health Center and a sexually transmitted disease clinic. Participants provided self-obtained vaginal swabs for testing with the Atlas io and Aptima Combo 2, and completed questionnaires assessing attitudes toward POC testing. RESULTS: Of 296 women recruited, 284 (192 from sexually transmitted disease clinic, 92 from Teen Health Center) had Aptima Combo 2 and Atlas io results available; 273 completed the questionnaire. Average age was 27.4 years (SD, 10.8 years). Sensitivity and specificity of the Atlas io test were 83.9% (26/31 specimens; 95% confidence interval [CI], 70.9-96.8%) and 98.8% (250/253 specimens; 95% CI, 97.5-100%), respectively. When specimens with discrepant results were included in the analyses, adjudicated sensitivity and specificity were 92.9% (26/28 specimens; 95% CI, 83.3 to 100%) and 98.8% (253/256 specimens; 95% CI, 97.5 to 100%), respectively.A majority (70%) of women preferred to collect vaginal self-swab if a POC test were available. Most (61%) were willing to wait up to 20 minutes, and 26% were willing to wait up to 40 minutes for results, if they could be treated before leaving clinic. CONCLUSIONS: A POC polymerase chain reaction test detecting CT had high sensitivity and specificity when testing prospective, vaginal swab samples. Availability of CT results during patients' visits may decrease time to treatment.