Background and objectives: To assess the presence of human papillomavirus (HPV) DNA in vulvar papillomatosis, since some women with this complaint have symptoms associated with HPV infection, such as itching, burning, and dyspareunia. Goal: To reassure the patients that they do not have a transmissible viral disease, by excluding a HPV origin of their condition. Study design: Vulvar biopsy specimens from 22 females with vulvar papillomatosis, from 10 females with prominent aceto-white vulvar lesions, and from 14 healthy controls were analyzed histologically for signs of HPV infection and by in situ hybridization and polymerase chain reaction for the presence of HPV DNA. Results: Specimens from women with vulvar papillomatosis showed some histological signs associated with HPV infection, but no koilocytotic atypia or dysplasia and thus resembled normal-looking vulvar mucosa. Aceto-white lesions frequently displayed histological features suggestive of HPV infection, including dysplasia. HPV DNA was detected in 6 of 10 patients with aceto-white lesions, but only in 1 of 22 patients with papillomatosis and in no healthy controls. Conclusions: According to our results, HPV DNA, which is generally found in cervical lesions and subclinical infections, is not present in vulvar papillomatosis even though symptoms associated with HPV infection are frequent complaints.