Purified total RNA was reverse transcribed to cDNA using TaqMan reverse transcription reagents (random hexamers; Applied Biosystems, Foster City, CA, USA). Real-time quantitative PCR was carried out using Brilliant II Fast QPCR reagents (Stratagene, La Jolla, CA, USA) according to the manufacturer's instructions on an Mx3005P QPCR system (Stratagene). Online ProbeFinder software (Roche Applied Science) was used to find primers that would allow amplification of the HCV RNA 59 end in combination with the Human Universal Probe Library from Roche (Roche Applied Science). For amplification of the HCV RNA 59 region, 400 nM of both forward primer (59-CAT-GGCGTTAGTATGAGTGTCG-39) and reverse primer (59-GG-TTCCGCAGACCACTAT-39) were used in combination with 200 nM of probe #75 from the Human Probe Library (Roche). HCV RNA levels were relatively quantified across samples and normalized to beta-actin RNA levels using 500 nM primers (forward: 59-GCC CTG AGG CAC TCT TCC and reverse: 59 GGA TGT CCA CGT CAC ACT TC-39) and 250 nM probe (59AC TCC ATG CCC AGG AAG GAA GGC-39 with a 59 Cy5 fluorophore and 39 black hole quencher).