Background: Laboratory confirmation of viral hepatitis infection represents an important issue for working age populations, as early detection and treatment can help ameliorate clinical progression of the disease. On the other hand, prejudice may occur in the workplace against those identified by a positive hepatitis test. This study investigated attitudes towards viral hepatitis testing in Japanese people of working age, including their desire to undergo such testing, and prejudice against persons infected with hepatitis virus. Methods: A total of 3,129 working age individuals were recruited from a company that conducts Internet surveys in Japan. Results: Of the respondents, 21.3% had previously undergone viral hepatitis testing, most frequently when it was an additional option during a health checkup or health screening for local residents (36.2%) and when it was included in regular health checkups in their workplace (19.2%). Among the respondents with no history of testing, 68.7% expressed a desire to undergo testing, of whom 74.8% wanted to have the test as part of their regular health checkups in the workplace. According to the respondents, if a coworker tested positive for hepatitis, 36.0% reported that they would be anxious about it, 32.0% would try to avoid contact with the infected person as long as circumstances permitted, and 23.7% said they might harbor some kind of bias. Conclusions: Although further promotion of viral hepatitis testing is needed and this might be achieved during regular health checkups in Japanese workplaces, educational strategies will also be essential to help reduce bias against those who test positive.(J Occup Health 2013; 55: 392-397)