A double polymerase chain reaction, involoving nested primers deduced from NS3-5 regions of the hepatitis G virus (GBV-c/HGV) genome, was developed for a sensitive and specific detection of GBV-C (HGV) RNA. With this method, 10 anti-HGV positive patients with non A-E hepatitis and 10 healthy subjects were tested. Among 10 patients, GBV-C RNA was detected in 9, 8, 4, 9 patients by the NS-3 (1) primers, the NS3(2) primers, the NS4 primers, the NS5(1) primers and the NS5(2) primers respectively. Control sera from 10 healthy subjects did not reveal GBV-C (HGV) RNA by any set of the primers. These data suggest that the detection rate of GBV-C (HGV) RNA is different in various set of primers, the NS3(1) and the NS5(2) primers are superior to other primers and are more suitable for RT-PCR of GBV-C (HGV) RNA.