penile cancer is a potentially mutilating disease. although its occurrence is relatively rare worldwide, penile cancer rates can be high in developing countries. a few studies have been conducted on the involvement of human papillomavirus (hpv) in penile carcinoma, which have found hpv present in 30-70% of penile malignant lesions, with a higher prevalence of hpv 16 and 18. it has been assumed that cofactors, such as epstein-barr virus (ebv) infections, may play a role in the progression of penile neoplasia. the aim of this study was to determine hpv and ebv prevalence in 135 penile malignant lesions from brazilian men through the use of my09/11 polymerase chain reaction (pcr), type-specific pcr and restriction fragment length polymorphism analysis. hpv prevalence among the men tested was 60.7%. of the men who tested positive, 27 presented with hpv 16 (29.7%), five with hpv 18 (5.5%), 21 with hpv 45 (23.1%) and nine with hpv 6 (9.9%). seven mixed infections were detected (9.2%), while 11 cases remained untyped (13.4%). regarding ebv positivity, 46.7% of the samples contained ebv dna with ebv-1 as the most prevalent type (74.6%). more than 23% of the men were co-infected with both hpv and ebv, while 35% presented exclusively with hpv dna and 20% presented only with ebv dna. penile carcinoma aetiology has not been fully elucidated and the role of hpv and ebv infections individually or synergistically is still controversial. hence, more studies are needed to determine their possible role in carcinogenesis.