Recently, sequences from a novel human flavivirus, termed GB virus C (GBV-C) or hepatitis G virus (HGV), have been identified in serum from patients with cryptogenic hepatitis and others. Sera from 116 patients with different clinical backgrounds were tested for the presence of GBV-C/HGV RNA by a reverse transcription-polymerase chain reaction with primers from the nonstructural (NS) 5 region. Ten (15%) patients on maintenance hemodialysis and 5 (19%) non A-C hepatitis patients were GBV-C/HGV RNA positive, along with one patient with chronic hepatitis B, one patient with chronic hepatitis C, and two asymptomatic individuals. Sequence comparison within 354 base pairs in the NS5 region showed homology rates varying from 87% to 97% among five Brazilian isolates, and from 86% to 93% between Brazilian strains and GBV-C/HGV isolates from other countries previously sequenced. Homology rates were higher at the amino acid level since most substitutions occurred at the third nucleotide position of codons without changing the codon meaning.