In an attempt to further define the natural history of Mycoplasma genitalium and Mycoplasma fermentans infections in humans, we used cultures and the polymerase chain reaction (PCR) to determine whether these organisms were present in the urethra and cervix of sexually active adults and in the amniotic fluid of women whose membranes were intact and collected at the time of cesarean delivery (to preclude cervical contamination). M. genitalium was detected by PCR but not by culture in 11% of patients with urethritis or cervicitis. It was not detected by either PCR or culture in the 232 amniotic fluid samples analyzed. In contrast, M. fermentans was not detected by either method in patients with urethritis or cervicitis but was detected by PCR in 4 of 232 amniotic fluid samples tested. These results indicate that in these four cases M. fermentans was transferred transplacentally. Histological evidence of chorioamnionitis was present in two of the four patients, a finding suggesting that M. fermentans may be a cause of chorioamnionitis. These results must be confirmed by other investigators, and further studies should be undertaken to determine the potential clinical significance of M. fermentans infection.