Survey on the genital Mycoplasmosis by multiplex PCR

Document Type: Research Article

Authors

1 Ph.D. Student of Microbiology, Islamic Azad University, Science and Research branch, School of Basic Sciences, Tehran, Iran.

2 Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

3 Department of Microbiology, Islamic Azad University, North Tehran branch, Tehran, Iran.

4 Department of Microbiology, Islamic Azad University, Science and Research branch, School of Basic Sciences, Tehran, Iran.

Abstract

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 104 samples was collected from 104 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 100 μl of specimen (American, Roche Company). In addition to the kit, boiling method was used to extraction of DNA from samples. UUA2 and UUS2 primers were used for urease gene amplification of U.urealyticum, MH1 and MH2 used for 16S rRNA gene amplification of M.hominis, and Adhesionproteingene (MgPa) used for 16S rRNA gene amplification of M.genitaliumin all samples. The number of 28 samples (27%) was positive for Mycoplasmas. M.hominis, M.genitalium,and U.urealyticum were detected in 8.7, 3.9, and 14.5 percent of samples, respectively. The accumulated frequencies for M.hominis, M.genitalium,and U.urealyticum were 9(8.7%), 4(3.9%), and 15(14.5 %), 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.

Keywords

Main Subjects