HDV infectivity particularly related to sexual activity has been difficult to establish. We investigated the prevalence of HDV in a high risk urban male population currently evaluated for HIV infection. Fourth-eight homosexual or bisexual men (96% positive for HIV) being routinely followed in the outpatient clinic, 40 sera obtained randomly from male homosexuals and 24 HBsAg carriers were examined by ELISA and Western Blot. HDV RNA was assessed by slot-blot after hybridization with cDNA probe from a recombinant plasmid (pS-1). [None of the 48 male subjects or from a recombinant plasmid (pS-1).] None of the 48 male subjects or from the randomly selected homosexuals tested positive for anti-HDV. HDV RNA searched in a selected group of sera from either high risk population or from HBsAg carriers proved also to be negative. We suggest that factors other than HBV chronic bearing and/or sexual promiscuity should be associated with HDV spread.