Background: Premature rupture of membrane (PROM) is considered an important cause of perinatal morbidity and mortality with increased maternal and neonatal risks. The etiology of PROM is known to be multifactorial, however, genital infections such as Chlamydia trachomatis is a possible contributing factor to its occurrence. The aim of this study was to assess the incidence rate of Chlamydia trachomatis infection in pregnancy and its role in the etiology of PROM. Methods: Two groups of patients were included; study and control groups. The study group included twenty full term pregnant women with the diagnosis of recent premature rupture of membranes with a duration of less than twelve hours while the control group included ten normal cases having the same criteria but with intact membranes. All patients were admitted to El Shatby Maternity University Hospital and an informed written consent was taken. Serum samples were collected in red top vacutainer for all women to detect Chlamydia trachomatis Immunoglobulin M (IgM) and Immunoglobulin G (IgG). In addition, endocervical swabbing was performed to detect Chlamydia trachomatis antigen using ELISA technique. Results: The study showed no significant association between PROM and Chlamydia trachomatis infection. However, the prevalence of Chlamydia trachomatis cervicitis in women with PROM was higher as compared to the control group; 45% compared to 20% by chlamydial antigen. Conclusion: The fact that this infection statistically increases the risk of PROM could not be confirmed.