Objective: This study was designed to evaluate the prevalence of Human Papilloma Virus (HPV) in semen and document the cycle outcomes in couples with previous intra-cytoplasmic sperm injection (ICSI)failures.Material and methods: One hundred and seventeen couples with at least two ICSI attempts were includedin the study. HPV infection in semen and DNA fragmentation in samples were analyzed by commerciallyavailable kits. The percentage of spermatozoa with fragmented DNA (DNA fragmentation index: DFI) wasdetermined during fluorescence microscopic examination as previously described. The cycle outcomes ofcouples with or without HPV infected male partners were recorded.Results: According to our results, the prevalence of HPV was 7.7% in asymptomatic males with at leasttwo previous ICSI failures. The increased DFI (>30%) was observed in 82.9% of the cases. In HPV-positivecases significantly lower number of good quality embryos were obtained. The implantation and pregnancyrates were similar in infected and non-infected males (p>0.05). The early miscarriage rate was slightlyhigher in HPV- positive group (33% vs. 10%, p>0.05).Conclusion: In cases with previous ICSI failures, the prevalence of HPV infection in semen is not higherthan previously reported infertile populations. The reproductive outcome might be impaired in HPV-positive semen due to lower number of good quality embryos, which needs to be clarified by further largepopulation-based studies.