HCV RNA Quantification from Plasma

Hepatitis C virus (HCV) RNA testing was done on 200 µL of stored plasma, processed the same day as the blood used for LTL measurement. Briefly, HCV viral RNA was extracted from plasma using MagMAX™-96 Viral RNA Isolation Kit (Life Technologies, Carlsbad, CA, USA) following the manufacturer’s protocol. HCV RNA was detected by qPCR using probes and primers described elsewhere [34 (link)]. The cycling parameters for the ABI 7900HT Fast Real-Time PCR system were as follows: 1 cycle of 50 °C for 30 min, 1 cycle of 95 °C for 20 s, and 40 cycles of 95 °C for 1 s followed by 60 °C for 20 s. The limit of detection was approximately 100–200 IU/mL for genotypes 1, 2, 3, and 4.