[Quantitative analysis of hepatitis C virus RNA by competitive nested PCR].
A method that allows the quantitation of hepatitis C virus (HCV) RNA is described. The RNA was extracted from serum sample and reverse transcribed. Target cDNA was then co-amplified by nested polymerase chain reaction with a known amount of competitive template at various concentrations. Since this internal control DNA uses the same primers as those of the target and is distinguishable from the target cDNA after amplification by size, the initial concentration of the target could be estimated by comparing the intensity of the two bands of amplified DNA fragment. That is, if the starting amount of the cDNA and the competitive template are equal, the intensity of the two bands should also be equal.
Kobayashi
M
M
Department of Liver research laboratory, Toranomon hospital.
D005838:Genotype; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000175:diagnosis; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis*
D012367:RNA, Viral