Mycoplasmas and Ureaplasmas inhabit the genitourinary tract of sexually active males and females. Mycoplasma genitalium infection as well as Ureaplasma urealyticum are related to many reproductive health problems such as cervicitis, urethritis and  pelvic inflammatory disease (PID). the aim of this study was detection of Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma urealyticum cervical colonization among childbearing age females attending Gynecology and Obstetrics clinics at Ain Shams University Hospital by using polymerase chain reaction. This study was conducted on 145 women attending Gynecology and Obstetrics clinics at Ain Shams University Hospital. The patients were divided into three groups according to symptoms. Cervical samples were collected using a sterile swab and placed in a liquid-based transport medium. The detection of Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma urealyticum was performed by using polymerase chain reaction. This study was conducted during the period from January 2018 till August 2018 on145 women attending Gynecology and obstetrics  clinic at Ain Shams University Hospital. Ninety females (62%)were positively colonized with Mycoplasma spp and U. urealyticum. M. genitalium was detected in 25 patients (28%), 10 patients (11%) were positive for M. hominis and 55 patients (61%) were positive for U. urealyticum. Group (1): 60 patients attending STD clinic. Twenty patients (33.3%) were  positive for U. urealyticum, five (8.3%) were positive for M. hominis and seventeen (28.3%)were positive for M. genitalium. Group (2): 50patientswith premature rupture of membranes. Twenty eight patients(56%)were positive for U. urealyticum, two patients (4%) were positive for M. hominis and seven patients (14%) were positive for M. genitalium. Group (3): 35 patients with pelvic inflammatory disease. Seven patients(20%) were positive for U. urealyticum, three patients (8.6%) were positive for M. hominis and one patient (2.9%) was positive for M. genitalium. The highest prevalence of U. urealyticum was associated with premature rupture of membranes. The highest prevalence of M. genitalium was determined in STD clinic patients. This study shows a high prevalence of genitourinary infections due to U. urealyticum which was considerably higher when compared to Mycoplasma spp. and is significantly associated with premature rupture of membranes. M. genitalium was confirmed as an important cause of STD. The use of PCR for identification of Mycoplasmas and U. urealyticum on cervical samples should be recommended. Further studies are needed to definitely associates with Spontaneous rupture of membranes and U. urealyticum colonization.