In Vitro Detection of Dissimilar Amounts of Hepatitis C Virus (HCV) Subtype-Specific RNA Genomes in Mixes Prepared from Sera of Persons Infected with a Single HCV Genotype

Abstract

ABSTRACT The level of in vitro detection of viral genomes in mixes with two different hepatitis C virus (HCV) subtypes was investigated by artificially mixing previously measured subtype-specific HCV RNA genomes. The RNAs in these mixtures were reverse transcribed and then PCR amplified by using two sets of primers corresponding to the 5′ untranslated region and digested with endonucleases to analyze the restriction fragment length polymorphism patterns. This approach facilitated detection of a wider range of type-specific HCV genomes than originally described, beyond equimolar concentrations of contributing HCV subtypes. Moreover, by using computerized image analysis, this study also demonstrated that the true contribution of each virus type—and consequently of mixed infections—may be underestimated when only visual observation is carried out. These results may be useful for comparing data obtained from this and other currently used methodologies.