Background: Listeria monocytogenes is a gram positive and facultative anaerobic bacteria and Listeria species have a high spread in environment. Listeria monocytogenes is agent of meningitides, encephalitis and septicemia and could accrue during pregnancy. Detection of Listeria via culture method is difficult. The aim of this study was accessing of Listeria monocytogenes in pregnant women who refer to women hospital of Tehran via culture and Real Time PCR methods. Methods: Two hundred cervix swabs samples were collected from pregnant women in seventh and eights mounts of pregnancy and transfer to laboratory in 2ml transport medium. 1ml was used for DNA extracting directly and other keep in refrigerator for cold enrichment among 4 weeks and then cultured. Primers were designed for amplifying hly gene which coded listeriolysin O. Results: The result of culture and real time PCR methods were showed of 200 samples, 28 (14%) and 41(20.5%) were positive, respectively. Conclusion: All of 13 false negative via culture, were positive in PCR technique. So, molecular method was more sensitive and recommended for applying. Moreover, the results showed high prevalence of carrier women in this time. Therefore, public screening was recommended for pregnant women