SummarySwabbing the urethrae of men has been the traditional approach for collecting specimens for detection ofChlamydia trachomatis. Recently, however, urine testing using enzyme immunoassay has yielded promising results. A total of 105 patients attending the Andrology Clinic at Ga Rankuwa Hospital, Medunsa were included in the study. These patients were asymptomatic and had no urethral discharge. Three endo‐urethral swabs and first‐catch urine were collected fiom each patient. The urethral swabs were used for enzyme immunoassay (EIA) (IDEIA 111), tissue culture and direct immonufluorescent antibody (DFA) test (IMAGEN) to detectC. trachomatis. In addition about 15–30 ml of first‐catch urine, or urine collected at least 2h after the previous micturition, was collected for each patient for EIA testing. Fifteen (14.3%) of 105 patients were positive on urethral swab EIA, in comparison with the DFA test in which 14 (13.3%) were positive. Eight (7.8%) were positive in tissue culture. Urine EIA was positive in 17 (16.2%) patients, of whom five (4.8%) were positive in urine EIA only. All EIA positive urines were confirmed by DFA. We recommend that first‐catch urine or urine collected at least 2h after the previous micturition in infertile males may be considered a suitable alternative to urethral swab for chlamydial diagnosis because it is noninvasive and non