Aim of present study was to determine prevalence of genital Chlamydia trachomatis in women of reproductive age group using PCR on urine specimen. First void 30 ml urine specimen from 160 women attending gynaecology OPD at KGMU, Lucknow were subjected to DNA extraction and PCR. 90 women had symptoms suggestive of genital tract infection (cases) and remaining 70 women were symptom free and served as control. An in- house PCR test by KL-1 and KL-2 plasmid primers was standardized on known positive urine specimen. Statistical analysis was done by chi square test. In our study, prevalence of genital Chlamydia trachomatis was 9.3%, it was more in cases (11.11%) in comparison to controls (7.14%) ( P >0.10). The difference between symptomatic and asymptomatic women is not statistically significant so we recommend universal screening of Chlamydia trachomatis in general population.