Expression of HCV RNA positive and negative (replicative) strands in PBMC samples obtained at two collections approximately 5 years apart from individuals with self-resolved hepatitis C.

<p>(A) HCV RNA positive strand identification using 2 μg of total RNA extracted from PBMC treated <i>ex vivo</i> with PHA or C5 (*). (B) Detection of HCV RNA negative strand in HCV RNA positive strand reactive PBMC shown in A. Samples were obtained during the first (1) or the second (2) collection at follow-up time (years) indicated under panel A. As positive controls for HCV RNA positive strand detection, serial 10-fold dilutions of rHCV UTR-E2 carrying indicated copy numbers/reaction were used (panel A). For HCV RNA negative strand detection, synthetic HCV RNA positive strand (sHCV RNA pos) and HCV synthetic RNA negative strand (sHCV RNA neg) at 10⁴ copies/reaction were used as positive and specificity controls, respectively (panel B). Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals demonstrated the expected 244-bp oligonucleotide fragments. Numbers under panels marked as relative DU represent relative densitometric units (DU) given by hybridization signals.