Résumé : Detection of HIV-1 RNA in semen is used commonly to determine the safety of semen processing procedures before assisted reproductive technology (ART). Using two panels of prepared semen samples containing HIV-1 the performances of protocols from 14 centers have been compared. No false-positive results were detected but false-negative results were frequent when the concentration was below 500 HIV-1 RNA copies/ml of seminal plasma. Frequency of HIV-1 RNA detection was higher on seminal cells than on seminal plasma. Assays (or protocols) for quantifying HIV-1 RNA in semen performed less well than standardized blood plasma assays. The HIV load in seminal plasma could be a useful marker of the risk of sexual transmission of the virus. Its use as a marker of global HAART efficiency in the HIV reservoir needs further study. Standardized assays are required for detection and measurement of HIV-1 RNA in semen samples. © 2011 Wiley Periodicals, Inc.