In Burkina Faso, as in other African countries, infertility has become a social burden for the population and a public health problem. Male infertility accounts for 30% to 40% of all infertility cases. The diagnosis of male infertility or hypofertility is often made by a simple laboratory analysis of sperm to explore sperm parameters. In most African countries, such as Burkina Faso, microbiological analysis in the context of sperm analysis is still not developed, and is carried out solely based on microscopy and traditional culture, which does not allow the growth of fragile and demanding bacteria. Our study investigated the microorganisms of sperm that may be involved in male infertility, using conventional bacteriology techniques and real-time PCR. However, it did not intend to perform a multivariate statistical association analysis to estimate the association of microorganisms with abnormal semen parameters. This prospective cross-sectional pilot study was carried out on patients who visited the bacteriology laboratory of Centre MURAZ, a research Institute in Burkina Faso, for male infertility diagnosis between 2 August and 31 August 2021. Bacteria were isolated and identified using standard bacteriology techniques. In parallel, common pathogenic microorganisms known to be associated with male infertility were targeted and detected in the sperm using a multiplex real-time PCR assay. A total of 38 sperm samples were analyzed by bacteriological culture and bacteria isolated were Staphylococcusaureus (S.aureus) 5.55%, Klebsiellapneumoniae (K.pneumoniae),Enterococcusfaecalis (E.faecalis), Streptococcusagalactiae (S.agalactiae) and Staphylococcushoemalyticus (S.hoemalyticus) respectively 2.70%. Real-time PCR targeted and detected Chlamydiatrachomatis (C.trachomatis) at 7.89%, Ureaplasmaurealyticum (U.urealyticum) at 21.05%, Ureaplasmaparvum (U.parvum) at 18.42%,Mycoplasmahominis (M.hominis) at 15.79%, Mycoplasmagenitalium (M.genitalium) at 10.53% and Trichomonasvaginalis (T.vaginalis) at 2.63%. Neisseriagonorrhoeae (N.gonorrhoeae) was targeted by the real-time PCR assay and was not detected (0%) in the tested semen samples. Our study highlights critical limitations of culture
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