Diverse studies demonstrate an association between Mycoplasma genitalium and urogenital pathologies. The aim of this study was to investigate the prevalence of M. genitalium in patients attending gynecological evaluation in private clinics (n = 172). DNA amplification assays of the genes 16S rRNA and MgPa were utilized. The prevalence of M. genitalium in the study population was 7.5%. M. genitalium was detected in 12.1% and 4.1% of the symptomatic and asymptomatic patients, respectively (p = 0.047). The infection was diagnosed in patients with cervicitis (17.2%) and mucopurulent secretion (16.6%) and the highest prevalence of infections was registered in the 31-40 years age group. No significant association between the presence of M.genitalium and individual clinical manifestations or the patients age was showed (p > 0.05). The high prevalence of M. genitalium infections, mostly in patients with clinical manifestations showed in this study, warrants the application of diagnostic strategies in the population to investigate the clinical meaning of these microorganisms and to reevaluate therapeutic schemes against non-gonococcal and non-chlamydial infections.