Background: Chlamydia trachomatis is the most common sexually transmitted bacterial organism that is usually associated with asymptomatic infection and sequelae such as infertility. Molecular methods such as Polymerase Chain Reaction (PCR) are considered as gold standard and highly specific diagnostic test for its detection. The study aimed to determine the prevalence of Chlamydia trachomatis infection among women attending infertility clinic in University College Hospital, Ibadan. Methodology: A cross-sectional hospital based study in which endocervical swabs were collected from 150 infertile women asymptomatic for genital infection at University College Hospital, Ibadan. Information on socio-demographic and behavioural characteristics were obtained using interviewer – administered questionnaire. The presence of Chlamydia trachomatis infection was detected by Conventional PCR. Data analysis was done using SPSS version 20. Results: Among the study participants, 11(7.30%) had evidence of Chlamydia trachomatis infection. Among those who are PCR positive for Chlamydia trachomatis 4(36.4%) have secondary infertility while 7(63.6%) have primary infertility. There is no significant association between the sociodemographic factors and Chlamydia trachomatis. Conclusion: The prevalence of asymptomatic genital Chlamydia trachomatis infection among women presenting with infertility from this present study highlighted the asymptomatic nature of Chlamydia trachomatis infection and the need to ensure that it is screened for routinely