This study sought to determine factors associated with chlamydial infection in a low-prevalence college health setting and to determine the testing characteristics of a polymerase chain reaction (PCR) assay for chlamydial infection (AMPLICOR chlamydia test; Roche Diagnostic Systems, Indianapolis) in this population. Young women (n= 1,149) at a university student health clinic underwent testing for cervical chlamydial infection by PCR assay and culture; the characteristics of women with and without chlamydial infection were compared. Chlamydial infection was diagnosed for 26 students (2.3%). The sensitivity and specificity of PCR assay and culture were 85% and 100% and 54% and 100%, respectively. Students with chlamydial infection were more likely to be 20 years of age or younger, have symptoms, report prior chlamydial infection or gonorrhea, report exposure to a sexually transmitted disease (STD), be black, or have cervical signs during examination; however, none of these were significant predictors for asymptomatic women. PCR assay detected significantly more cervical infections than did culture in this college student population. These data are consistent with recommendations for testing college women with symptoms, STD exposure, or age of younger than 25 years.