We evaluated a rapid point-of-care test for the detection of Chlamydia trachomatis in specimens from 278 sex workers attending sexually transmitted infection clinics in Guatemala. The sensitivity and the specificity of the test compared to the results of PCR were 62.96% and 99.60%, respectively. The test performed moderately well as a screening tool in a context in which clinical follow-up visits are infrequent. Copyright © 2009, American Society for Microbiology. All Rights Reserved.