Esmaeil Ghorbanalinezhad - Ph.D. Student of Microbiology, Islamic Azad University, Science and Research branch, School of Basic Sciences, Tehran, Iran.
Nour Amirmozafari - Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Abbas Akhavan Sepahi - Department of Microbiology, Islamic Azad University, North Tehran branch, Tehran, Iran.
Ramezanali Khavari-nejad - Department of Microbiology, Islamic Azad University, Science and Research branch, School of Basic Sciences, Tehran, Iran.

Mycoplasmas hominis, Mycoplasmas genitalium and Ureaplasma urealyticum are associated with infections of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. A multiplex PCR was developed for simultaneously detection of these Mycoplasmas species in a single amplification reaction. The total number of ۱۰۴ samples was collected from ۱۰۴ women’s genital specimens with urogenital infections for identification of M.hominis, M.genitalium and U.realyticum by multiplex PCR. The High Pure PCR Template Preparation Kit purified nucleic acids from ۱۰۰ μl of specimen (American, Roche Company). In addition to the kit, boiling method was used to extraction of DNA from samples. UUA۲ and UUS۲ primers were used for urease gene amplification of U.urealyticum, MH۱ and MH۲ used for ۱۶S rRNA gene amplification of M.hominis, and Adhesionproteingene (MgPa) used for ۱۶S rRNA gene amplification of M.genitaliumin all samples. The number of ۲۸ samples (۲۷%) was positive for Mycoplasmas. M.hominis, M.genitalium,and U.urealyticum were detected in ۸.۷, ۳.۹, and ۱۴.۵ percent of samples, respectively. The accumulated frequencies for M.hominis, M.genitalium,and U.urealyticum were ۹(۸.۷%), ۴(۳.۹%), and ۱۵(۱۴.۵ %), respectively. The results of this study revealed that Multiplex PCR is a highly sensitive, specific and cost-effective test for screening of genitourinary tract infections.

Mycoplasmosis, Multiplex PCR, Urogenital diseases