Realtime hcv amplification reagent kit

Reverse transcriptase PCR was used for determining the serum HCV-RNA levels by a commercial kit (MagAttract virus mini M48 kit, RealTime™ HCV amplification reagent kit, Abbott) and anti-HCV antibodies were determined by ELISA chemiluminescence (Tarrytown, NY, USA). Genotype analysis of all the subjects was performed using the line-probe assay (Innolipa) strip method and the patients who were infected with HCV genotype 1b were enrolled in the study. The individuals who had undetectable serum HCV RNA in the 24 weeks of therapy were designated to have sustained virological response (SVR).

To reduce interassay variance, HCV and HIV RNA testing for a given subject were done at the same time on plasma centrifuged within 30 min of collection and stored at −20°C for up to 25 hours and then at −80°C until testing. HIV RNA testing was done using the Abbott RealTime HIV Assay (no. 02631-090). HCV RNA testing was done using the Abbott RealTime HCV Amplification Reagent Kit (no. 04J86-90). To provide information in “real time,” such as for screening into the study, additional HCV RNA tests were performed by the commercial laboratory of the Johns Hopkins Hospital using the Roche COBAS AmpliPrep/COBAS TaqMan HCV Test v. 1.0. Although both were reported in international units, analyses were primarily done on results from one or the other laboratory. CD4⁺ T cell count was measured by flow cytometry of whole blood that was delivered to the Johns Hopkins Hospital clinical laboratory.