Abstract Study question How frequent is Human Papilloma Virus (HPV) infection in both partners of infertile couples referring to our ART center and whats its impact? Summary answer Only 6% of infertile couples are positive for HPV and infection has no impact on embryologic parameters. What is known already There is limited and conflicting evidence about the relationship between HPV infection in male partners and infertility. Notably, most of the previous studies did not assess the presence of the infection in both partners in the context of ART. Furthermore, even if considered, the influence of the infection of the female partner may be underestimated because only data on adverse pregnancy outcomes were evaluated. Elucidating the HPV-positive status of the two individuals belonging to the couple could open up new scenarios. Study design, size, duration This prospective cohort study was performed on 81 couples. The analysis was conducted on semen samples of male patients and on cervical samples of female partners who underwent assisted reproduction techniques (ART) treatment at the infertility unit of Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico of Milan from February 2022 to November 2022. Participants/materials, setting, methods The female partecipants were recruited before the beginning of the controlled ovarian stimulation (COS) while male participants were recruited on the day of the female ovum pick-up (OPU). The technique used to assess the presence of the HPV virus was the Real Time PCR (RT-PCR). Personal characteristics, lifestyle habits and sexual habits were investigated as possible favoring elements for the insurgence of HPV infection. Main results and the role of chance Among 81 couples recruited, 12 men and 22 women tested positive for high- and low risk HPV viruses. This streins corrisponds to a prevalence of about 15% (95% CI: 9-24%) for males and 27% (95% CI: 19-38%) for females. Among these positive couples, only 5 cases (6%, 95% CI: 3-14%) simultaneously tested positive for both partners: in two pairs they were positive for the same viral strains, in two other pairs there was no concordance between the viral strains found, and in one pair there was only a partial concordance. McNemar's test (a dichotomous evaluation test for data paired aimed at assessing possible associations) was not significant. No differences in embryo developmental failure, “top-quality” embryo formation and “top-quality” blastocyst formation were found between the two groups (positive vs negative couples). Limitations, reasons for caution A possible limitation of the study is the small sample size. Additionally we mainly focused on the impact of infection by HPV on embryological variables, without considering developmental pregnancies and live birth rate. Wider implications of the findings The very low concordance observed suggests that infection follows intricate pathways, far beyond the trivial view of a ping-pong effect between partners of the couple. These results suggest that HPV infection probably develops independently in the two individuals of the couple, as if there is not a mutual influence. Trial registration number NOT APPLICABLE