In a prospective controlled study, we aimed to determine the prevalence of Chlamydia trachomatis infection in Jordanian women attending an infertility clinic and whether screening is useful as part of routine investigations for infertility. Two groups of patients (152 infertile patients and 146 control patients) had endocervical swab testing by polymerase chain reaction for the presence of C. trachomatis infection. A total of 6/152 patients in the infertility group tested positive for C. trachomatis (3.9%), compared with 1/146 patients in the control group (0.7%), a difference that was not statistically significant. In view of the tendency toward increased prevalence of C. trachomatis infection, screening women for chlamydial infection as part of routine investigations for infertility is recommended.