A method for detecting hepatitis C virus (HCV) in a sample includes the steps
of
(a) extracting RNA from the sample; (b) contacting the RNA to a reverse transcriptase
and a downstream primer specific to the HCV RNA sequence to create cDNA; (c) forming
an amplification medium by mixing the cDNA with a PCR reaction mixture, a nucleic-acid-binding
fluorescent entity, and the downstream primer and an upstream primer specific to
the HCV RNA sequence; (d) thermally cycling the amplification medium between at
least a denaturation temperature and an elongation temperature; (e) illuminating
the amplification medium with a selected wavelength of light that is absorbed by
the fluorescent entity during the thermally cycling step; (f) determining the amount
of fluorescence generated by the fluorescent entity; and (g) detecting the presence
of the target nucleic acid by analyzing the amount of luminescence determined after
at least one amplification cycle.
