Comparison of Culture with PCR for Detection of Mycoplasma hominis and Ureaplasma urealyticum in Semen Samples of Infertile Men Referring to the Royan Institute in 2009

Infection due to genital Mycoplasmas such as Mycoplasma hominis (M.hominis) and Ureaplasma urealyticum (U.urealyticum) may have harmful effects on the reproductive health of men and may lead to male infertility. This study was performed to compare culture with PCR method for detection of these bacteria in infertile men who referred to the Royan Institute. In this descriptive-analytic, cross-sectional study semen samples were collected from 220 infertile men and divided to 3 sections: first section was used for semen analysis, second section was inoculated into PPLO broth transport media and passed through 0.45µm pore-size filters, then filtrates were inoculated into specific PPLO broth and agar media and incubated at 37˚C under elevated CO2 atmosphere and the third section was used for PCR in which U4 and U5 primers were used for amplification of Urease gene of U. urealyticum and RNAH1 and RNAH2 for amplification of 16S rRNA gene of M. hominis. The results were analyzed with SPSS V.16 software. For data analysis ANOVA, Post Hoc, Crosstabs, Chi square and Mc Nemar tests were used. From a total of 220 semen samples tested, 3.2% were positive with culture and 4.1% with PCR method for only M. hominis meanwhile 27.7% were positive with culture and 29.1% with PCR for only U. urealyticum. Also 5% of the samples in culture and 11.4% in PCR method were positive for both of the bacteria evaluation of semen parameters showed that in both culture and PCR methods, pH was lower in the two groups of “only U. urealyticum positive” and “positive for both bacteria” as compared to the group “negative for both bacteria” (p=0.006 and p=0.000 respectively for culture and PCR) and the mean of sperm motility was lower in group “positive for both bacteria” than the group “only U. urealyticum positive” in two p=0.032(culture), p=0.009(PCR). Results of this study showed that higher percentage of infertile men are infected with these bacteria which may lead to infertility thus, isolation of these bacteria in infertile men with no clinical symptoms is necessary. PCR as compared to culture is a more sensitive, quick, specific and fast method./