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The objective of this study was to measure the prevalence of Chlamydia trachomatis (CT) infection among men in clinical and nonclinical settings across the United States.The goal of this study was to obtain data to inform recommendations regarding male CT screening.The authors conducted a cross-sectional study of CT prevalence among adolescent and adult men in 4 U.S. cities (Baltimore, Denver, San Francisco, and Seattle). CT was detected using urine-based testing, and prevalence was calculated for first testing event.Over 23,000 men were tested for CT over a 3½-year period. The majority (96%) were asymptomatic. Overall, prevalence was 7% and varied significantly between cities (range: Seattle, 1%; Baltimore, 12%), by age (peak prevalence at age 20–24 years, 9%), and between venues where CT testing was offered.At 7%, the prevalence of CT is moderately high among men opportunistically tested in nonclinical and clinical settings.