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To investigate the possible involvement of human trichomonads (Pentatrichomonas hominis and Trichomonas tenax) other than Trichomonas vaginalis in the aetiology of vaginal trichomoniasis.Vaginal swabs taken from women attending antenatal clinics were tested for Trichomonas vaginalis by traditional assays (wet-mount microscopy and InPouch culture) and nucleic acid amplification (polymerase chain reaction). These swabs were also tested for the presence of P hominis and T tenax by nucleic acid amplification. Oral and rectal swabs from these women were tested for T tenax and P hominis respectively. Data on sociodemographic characteristics, sexual and anogenital hygiene practices likely to seed P hominis and T tenax into the vagina were collected by a questionnaire.93% (161) of the 173 samples in which T vaginalis was detected by wet preparation or culture was evaluable by PCR. Of this, T vaginalis was detected in 94% (152) by T vaginalis- specific PCR. Neither P hominis nor T tenax was detected in any of the vaginal swab samples. These included nine samples for which T vaginalis had been detected by wet preparation or culture, but were negative by T vaginalis nucleic acid amplification. P hominis and T tenax were not detected in any of the rectal and oral swabs, respectively.In this group of women, there was no evidence for the involvement of trichomonads other than T vaginalis in the aetiology of vaginal trichomoniasis.