Cobas taqman pcr assay version 2

Patients were examined for pre-existing RASs to NS5A inhibitors and NS3/4 inhibitors by polymerase chain reaction (PCR) invader assay (BML, Tokyo, Japan). Amino acid substitutions resistant to NS3/4 protease inhibitors V36A, T54A/S, Q80L/R, R155K/Q/T, A156S/T/V and D168A/E/H/T/V and resistant to NS5A inhibitors L31F/M/V and Y93H were identified. When more than 20% of variants were detected, RASs were judged as strongly positive. When less than 20% of variants were detected, they were judged as weakly positive.9 (link) The amount of HCV RNA was measured using quantitative RT-PCR (COBAS TaqMan^® PCR assay version 2; Roche Diagnostics, Branchburg, NJ, USA) and were checked on the day of therapy initiation at weeks 1 and 2, as well as every 4 weeks up to 12 weeks after the end of therapy. Serum levels of hyaluronic acid and type IV collagen 7S were measured for assessment of liver fibrosis on the day of therapy initiation. Biochemical analyses including blood counts, serum alanine aminotransferase (ALT), aspartate aminotransferase, γ-glutamyl transferase (γ-GT), estimated glomerular filtration rate (eGFR) and α-fetoprotein levels were performed every 4 weeks up to 12 weeks after the end of therapy.

The amount of HCV RNA was measured using quantitative reverse transcription–polymerase chain reaction (RT‐PCR) (COBAS TaqMan PCR assay version 2; Roche Diagnostics, Branchburg, NJ, USA) and was checked on the day of therapy initiation and at every 4 weeks up to 12 weeks after the end of therapy. SVR was defined as a negative HCV RNA at the end of therapy, remaining negative for 12 weeks after the end of therapy.

HCV-RNA load was measured using quantitative reverse transcription polymerase chain reaction (COBAS TaqMan PCR assay version 2; Roche Diagnostics, Branchburg, NJ, United States). HCV genotype was determined using the antibody serotyping method (SRL, TKY, Japan). HCV serotypes 1 and 2 correspond to genotypes 1a/1b and 2a/2b, respectively. When HCV serotype could not be determined, genotype was examined using real-time polymerase chain reaction assay (BML, TKY, Japan). HCV-RNA was checked on the day of therapy initiation and every 4 wk during treatment. Biochemical analyses including blood cell counts, C-reactive protein level, blood sugar level, and liver and renal function tests were performed every 2 wk during treatment.