Infections caused by Chlamydia trachomatis (CT) are one of the most prevalent of all sexually transmitted diseases (STD). This cross sectional study was carried out to diagnose genital CT infection on 108 (59 pregnant and 49 non-pregnant) women attending at Department of Gynaecology and Obstetrics, Mymensingh Medical College Hospital (MMCH) during the period from January 2009 to December 2009. This non- culture technique was based on detection of CT major outer membrane protein (MOMP) by Direct Fluorescence Antibody Test (DFAT) from endocervical swab. Chlamydial inclusion bodies (IB) were looked for using Iodine stain. CT antigens were detected in 45.3% (49/108) cases by DFAT; IBs were detected in 5.5% cases (06/108) by Iodine staining technique. Majority of CT positive cases (65%) were found in the younger age group (15 to 25 years). The CT infection was found 47.2 % (35/74) in symptomatic cases, 41.1% (14/34) in asymptomatic cases and 47.4% in pregnant group, 42.8% in non-pregnant group. Although high incidence of genital chlamydia infection is common both in pregnant, non-pregnant, symptomatic and asymptomatic women in Bangladesh an early and reliable diagnostic method for genital chlamydia infection in Bangladesh should be further explored.