A new quantitative reverse transcription-polymerase chain reaction (RT-PCR) method is described for analyzing the amount of GB virus-C (GBV-C)/hepatitis G virus (HGV) RNA in serum. This multicyclic RT-PCR (MRT-PCR) method used oligonucleotide primers deduced from the 3' noncoding region (3'NCR) that is highly conserved among GBV-C/HGV isolates. Quantitation of GBV-C/HGV RNA using MRT-PCR ranged between 10(2) and 10(10) copies/ml when PCR cycle number was regulated at exponential amplification of the products. Competitive RT-PCR (CRT-PCR) was carried out with mutant RNA and sample that had been measured by MRT-PCR. Quantitation of GBV-C/HGV RNA using both methods agreed. MRT-PCR detected viral RNA in a single step PCR, and demonstrated a high degree of sensitivity that was equal to that of the RT-PCR procedure, which used nested primers deduced from the non-structural (NS) 3 region. The MRT-PCR method for quantitation of GBV-C/HGV RNA in serum may prove useful for diagnosis.
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