Detection of mycoplasma infections, especially those caused by Mycoplasma genitalium (M. genitalium) is not a part of routine microbiological diagnostics, as commercial strip tests based on microculture in test wells do not include M. genitalium in their identification spectrum. Also classical methods of culture are especially demanding in the case of M. genitalium strains. PCR technique with application of specific starter oligonucleotides is currently used in the diagnostics of M. genitalium. In view of growing number of reports on the significance of this mycoplasma in etiopathogenesis of mainly urogenital disorders, an increasing number of laboratories uses molecular methods in diagnostics of M. genitalium infection. In this review we summarize epidemiological, clinical and microbiological research data of M. genitalium in the light of recent publications and our own observations.