Mycoplasma hominis and M. genitalium frequency of infertile men in the northeast of Iran by culture and Multiplex-PCR

Document Type: Original Articles

Authors

1 Department of Microbiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Mashhad, Iran

3 Jahad Daneshgahi Mashhad Laboratory, Mashhad, Iran

4 Department of Poultry Diseases, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

Abstract

Recently, infertility has become a growing social and economic world problem. Genital mycoplasmas such as Mycoplasma hominis and M. genitalium are most frequently associated with several adverse effects on men’s fertility. The study is going to determine the prevalence of M. hominis and M. genitalium in the semen samples in the northeast of Iran by cultivation, PCR, and Multiplex PCR assays as the first such research in the area.
During the cross-sectional study from February to May of 2018, one hundred semen samples were collected from 100 infertile men of Mashhad, Khorasan Razavi province, northeast of Iran. Some semen indexes like as color, volume, appearance, liquefaction, viscosity, concentration, pH, leukocyte concentration, progressive motility, morphological normality, motile sperm concentration, functional sperm concentration, sperm motility index, and functional sperm were analyzed according to WHO guideline for semen analysis.
After cold enrichment in PPLO transport medium, the samples were cultivated in PPLO broth and PPLO agar. Then, Diene’s staining and conventional biochemical tests were performed. Subsequent of all samples DNA extraction, the PCR was done by genus-specific primer of Mycoplasma, on the gene of 16SrRNA (GPO1& MGSO primers). Multiplex PCR was performed with specific primer for conserved regions in the 16SrRNA gene for M. hominis (RNAH1& RNAH2 primers) and the 140-kDa Adhesion Protein Gene for M. genitalium (MG1 & MG2 primers).
By the results, nine samples (9%) were positive with mycoplasma spp in PCR, while there are seven (7%) isolated by cultivation. All of isolated were M. hominis without any M. genitalium in biochemical tests. M. hominis was detected in eight samples (8%) by Multiplex PCR, while there’s no evidence for M. genitalium. The mean age of all infertile and infected men was 31 and 30 years respectively. There isn’t any statistical correlation between mycoplasma infection and abnormalities in semen in the results.
There are different reports about the prevalence of M. hominis and M. genitalium in Iran from 1.4 to 40%. There are many types of research with different results to show any relationship between mycoplasma infection and the abnormality in the semen parameters.

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