With the demonstration of an effect of GBV-C infection on the outcome of HIV infection, it has become important to understand the epidemiology of GBV-C. The purpose of this study was to determine the prevalence in high- and low-risk populations. The following populations were tested: school children, 9 and 15 years of age (n = 901), blood donors (n = 5,203), hospital employees (n = 1,432), and prisoners and injecting drug users (n = 447). In-house RT-PCR for GBV-CRNA was used together with a commercial ELISA for anti-E2 (Boehringer, Germany). In addition, questionnaires for risk factors for transmission and serological tests for HIV and hepatitis were applied. The overall prevalence of GBV-CRNA was 1.4% among children, 2.2% among blood donors, 2.2% among hospital employees, 12.5% among non-injecting prisoners, and 34.9% among drug injectors. Correspondingly anti-E2 was found in 0.3%, 12.3%, 25.0%, and 42.7%. Among hospital employees, independent risk factors for GBV-C were professions with blood exposure and sexual risk partners. Among prisoners and drug users, injecting and a sexual risk index were associated independently with GBV-C. Based on these results, the following hypothesis is suggested: GBV-C is transmitted frequently at birth or early childhood and this leads to chronic infection in most cases. Sexual transmission is the most important route of transmission in the adult population but this infection is usually transient. Blood borne transmission plays a role among health care workers and injecting drug users and GBV-C should be further evaluated as a surrogate marker for professional blood exposure.