Background: Elevated HPV infection rates have been described in HIV-positive males, placing these subjects at high risk of anal neoplasia. Bivalent, quadrivalent, and nonavalent vaccines to prevent HPV infection have been developed, and recently proposed for gender-neutral immunization programs. Objectives: In order to estimate the benefit that could be obtained by vaccination of HIV-positive men who have sex with men (MSM), we aimed at describing the frequency of multiple and vaccine-targeted HPV infections in MSM enrolled in an anal cancer screening program. Study design: The anal cancer screening program was conducted between July 2009 and October 2012. Mucosal anal samples were tested for HPV DNA using MY09/MY11 PCR primers and, if positive, genotyped using the CLART2HPV Clinical Array (35HPV types). Results: A total of 220 MSM were screened and 88.6% were positive for HPV DNA: in 86.5% at least one highrisk (HR) type was found and in 13% only low-risk (LR) HPV were found. Multiple infections accounted for 84.5% of HPV DNA-positive cases and overall 160 different HPV genotype combinations were recognized (only three combinations were detected in more than one patient each). Based on strain coverage, at least one vaccine-targeted HPV type was found in 38.9%, 64%, and 78.4% of cases when considering bivalent, quadrivalent and nonavalent vaccines, respectively. At least one HR vaccine-targeted strain was found in 39% of MSM for bivalent and quadrivalent vaccines, and in 64% of cases for nonavalent prevention. Conclusions: Anal HPV infections in unvaccinated mostly HIV-infected MSM are highly prevalent. The majority of this population has multiple infections with an extremely heterogeneous number of genotype combinations. The nonavalent vaccine could theoretically have prevented a minimum of one HR HPV type in two thirds of subjects. (C) 2015 Elsevier B.V. All rights reserved.