HCV persistent infection in VeroE6 cells.

VeroE6 cells were transfected with pVAls6 (encoding VA RNAI) or pcDNA3, and then infected with genotype 1b-infected human plasma [28], [29], at week 0. For each of 20 weeks the cells were diluted 1∶6, split and re-transfected with pVAls6. Cells were harvested and analyzed weekly for detection of HCV RNA by RT-PCR (PCR, first 40 cycles) followed by nested PCR [62]. Positive, (+); weakly positive, (±); negative (−); negative control pcDNA3, (NC); not determined, (ND). The limit of detection for this assay was evaluated [62] and calculated at 103 RNA copies/ml.