The association between human papillomavirus (HPV) infection and development of bladder cancer is variable. Furthermore, the prevalence of HPV DNA in bladder carcinoma subtypes varies from study to study. To clarify the impact of HPV infection on the development of bladder carcinoma, we performed a retrospective study on Tunisian patients to determine the status of HPV infection in urothelial carcinoma, squamous cell carcinoma, and adenocarcinoma. A total of 125 formalin-fixed, paraffin-embedded archival tissue specimens of bladder carcinoma were reviewed and classified according to the World Health Organization (WHO) classification of tumors (119 urothelial carcinomas, five squamous carcinomas, and one adenocarcinoma). Anogenital HPV DNA detection was performed using three different polymerase chain reaction (PCR) techniques: the first one used primers pU-2R/pU-1M specific to high-risk oncogenic HPV; the second one used primers PU-2R/PU-31B specific to low-risk oncogenic HPV; and the third one employed consensus primers (E1-547R/E1-350L). No evidence of HPV infection was detected by morphological examination and PCR in any case of bladder carcinoma. Our study shows that the anogenital HPVs investigated are not associated with the pathogenesis of bladder cancer in Tunisia; however, the question of whether other subtypes of HPV contribute to bladder carcinogenesis remains to be clarified.