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To determine the feasibility and efficacy of an online-mediated syphilis screening among men who have sex with men.We developed a Web site that offered information about syphilis and motivated users to download a referral letter with which they could test for syphilis in a nonclinical setting. A week after the blood test, participants could retrieve their results online. To assess the feasibility and efficacy of the Web site we followed the users through the online procedure and compared the percentage of syphilis infected men detected online with those diagnosed at the local sexually transmitted infection (STI) clinic, during the same time frame. The trial was divided into an initial period of online advertising (4 months) and no advertising (11 months) to examine how advertisements affect usage.During 15 months, 898 visitors downloaded a referral letter. Of these, 93 (10%) men tested and 96% (90 of 93) obtained their test results online. Through the Web site we found a significantly higher percentage of men who needed treatment for syphilis compared with the STI clinic (50% online vs. 24% STI clinic, P <0.01). Of the Online users who tested positive 33% (3 of 10) had never visited the STI clinic before. In the bannered period there was a monthly average of 15 testers compared with 3 per month in the nonbannered period.Online-mediated testing for syphilis is feasible and was more successful in detecting men who have sex with men with an early or late syphilis infection than standard procedures. However, longer promotion periods are needed to generate more usage of the online service.