Our aim was to determine the prevalence of human papillomavirus (HPV) infection in virginal women and to establish if other forms of sexual activity not involving penetration may contribute to the transmission of this infection. Female patients attending medical practitioners, and female high school students were recruited. Each participant answered an anonymous self answer questionnaire and collected a tampon specimen. Tampons were analysed for the presence of human papillomavirus by the polymerase chain reaction (L1 general primer and probe). Fifty-five female participants who were of a mean age of 18 years (range 13-41) were recruited. Twenty-three reported non-penetrative sexual activity. None of the 55 women had evidence of HPV infection (0%, upper 95% confidence interval 5.4%). Twelve tampons from sexually active women with normal Papanicolaou smears and a mean age of 26 years (range 19-44) attending a hospital clinic were analysed at the same time and four were found to be positive (P = 0.0006). We conclude that HPV infection prior to sexual intercourse, as determined by tampons specimens, is rare, even in those participating in other forms of sexual activity.