BackgroundThe colonization of Ureaplasma species in genital tract is related with male infertility. However, it has been postulated based upon limited study that virulence is related to serotype specificity. The aim of this study was to determine the distribution of Ureaplasma serovars in genital tract of infertile males and analyze their role in male infertility.MethodsA total of 358 urethral swabs samples were obtained from infertile males. The culture of Ureaplasma species were performed using a commercially available Mycoplasma IST 2 kit. Serovars were determined by real‐time polymerase chain reaction (real‐time PCR).ResultsA total of 92 (25.7%) infertile males were positive for Ureaplasma spp; among them, Ureaplasma parvum (UPA) was detected in 73 (79.3%) isolates, and Ureaplasma urealyticum (UUR) was detected in 19 (20.7%) isolates. Serovars 1, 6, or in combination accounted for 63.0% (46/73) of UPA isolates. Serovar 9 (alone and in combination of other serovars) was the most common serovar in UUR (47.4%, 9/19). Multiple serovars were detected in 21 (22.8%) isolates, and serovars 4, 5, 7, and 12 were not detected in any sample.ConclusionThe distribution of 14 Ureaplasma serovars in genital tract of infertile males was identified for the first time by real‐time PCR assay. UPA serovars 1 and 6, and UUR serovar 9 are the most common serovars colonization in urogenital tract of infertile males.