With a reverse transcription-polymerase chain reaction method that utilized highly efficient primers, the NS5 region of hepatitis C virus (HCV) genome could be amplified from all of the 24 serum samples randomly obtained from Japanese patients with liver diseases. Subtype analysis on the basis of nucleotide sequence similarities of the amplified fragments revealed that 2 out of 24 Japanese patients (8.3%) were infected with the recently identified subtype HCV-3b, which had been thought to be rare in Japan.