In order to assess the frequency of different human papillomavirus (HPV) types in Rome and the association between HPV and behavioural characteristics, we tested cervical scrapes of a population of sexually active women referring to university clinics for routine gynaecologic care. The presence of HPV DNA was revealed by polymerase chain reaction on two genome regions (L1 and E6/E7) followed by sequencing. Thirty different HPV types were identified; HPV 16 was the most prevalent (14.18%), followed by HPV 53 (9.21%), HPV 58 (7.80%), HPV 6 and 66 (both 5.67%) whereas all the other genotypes ranged below 5%. In univariate analysis the characteristics significantly associated with HPV DNA detection were the youngest age (P<0.01), the high number of lifetime partners (P<0.001) and the smoking habit (P<0.01). In multiple logistic regression analyses, the characteristics significantly associated with HPV DNA detection remained the younger age and the higher number of lifetime sexual partners. This study may be interesting in order to evaluate the circulation of HPV genotypes in Italy and to add a contribution to anti-cancer vaccine development.