Performance of the Abbott RealTime and Roche Cobas TaqMan Hepatitis C Virus (HCV) Assays for Quantification of HCV Genotypes

Abstract

We evaluated the Abbott RealTime (ART) and Roche Cobas TaqMan Hepatitis C virus (HCV) viral load assays for quantification of HCV genotypes in patient specimens. The ART HCV assay was a more sensitive and precise tool for accurate HCV viral load quantification across the HCV genotypes tested, especially genotype 1b. Hepatitis C virus (HCV) infection is associated with significant liver-related morbidity and mortality around the world. The World Health Organization estimates that 3% of the global population is infected with HCV and that there are approximately 170 million people at risk of developing cirrhosis or hepatocarcinoma (23,24). Treatment of HCV infection typically consists of pegylated interferon plus ribavirin or pegylated interferon, ribavirin, and a direct-acting antiviral (DAA) protease inhibitor (triple therapy) for non-genotype 1 and genotype 1, respectively (11,12,13). Depending on the particular treatment regimen and the genotype of HCV, treatment success as measured by failure to detect viral replication 24 weeks after cessation of treatment can be achieved in 50 to 80% of patients (12).Measurement of HCV viral load (VL) for the different HCV genotypes is crucial to clinical management of HCV-infected patients, both treated and not, for disease staging, decisions regarding treatment initiation, and individualization of treatment strategy (i.e., dosage and duration based on response kinetics) (1,6,7,13). Furthermore, with the advent of DAAs, VL monitoring may help prevent protease inhibitor resistance development by allowing for switching or stopping therapy if VL does not decrease or returns while on treatment.There are currently several commercially available HCV VL assays; real-time PCR assays are generally preferred because of their wide dynamic ranges and good sensitivities (1, 3). Two commercial real-time PCR platforms are available, the Cobas AmpliPrep/Cobas TaqMan HCV assay version 1.0 (CAP-CTM; Roche Molecular Systems, Pleasanton, CA) and the Abbott RealTime HCV assay (ART; Abbott Molecular, Des Plaines, IL). We characterized the performance of the ART assay and compared results obtained with both assays using clinical specimens of diverse HCV genotypes in a university hospital central testing laboratory.HCV VL was determined with the ART and the CAP-CTM as per the manufacturers' recommendations. A serum specimen volume of 500 l was required for ART, compared to 850 l for CAP-CTM. Sample preparation for the ART assay was performed using the Abbott m2000sp instrument. HCV genotypes were identified using the Versant Inno-LIPA HCV Genotype 2.0 assay (Siemens Healthcare Diagnostics, Deerfield, IL); specimens with indeterminate results were tested with the RealTime Genotyping II RUO assay (Abbott Molecular, Des Plaines, IL) or by direct sequencing and phylogenetic analysis of NS5B (performed by Siemens Clinical Laboratories, Berkeley, CA). Statistical tests were performed using Prism 5.0 (GraphPad Software, La Jolla, CA).To evaluate the sensitivity, linearity, and intra-and interrun precisio...

European Multicenter Study on Analytical Performance of DxN Veris System HCV Assay

“…These results show that the Veris HCV Assay performance is in line with that seen for other currently marketed real-time PCR molecular assays (9)(10)(11)(12)(13)(14) and fulfills the recommendations in current clinical practice guidelines for predicting a patient achieving SVR at 12 and 24 weeks posttreatment. Results were generated across multiple sites and showed comparative overall precision and consistently low LOD despite the introduction of increased variability related to the use of multiple instruments, lots, operators, and preanalytical techniques, such as sample processing, handling, and storage prior to testing.…”

“…The SDs and % CVs were comparable with those seen with other approved HCV assays (9)(10)(11)(12)(13)(14). In this evaluation, within-run, between-lot, and between-site variability contributed most to the total imprecision seen.…”

“…There are several assays currently on the market for the quantification of HCV viral load (VL) based on real-time PCR and branched DNA quantification (9)(10)(11)(12)(13)(14). These methods are recommended based on their excellent analytical sensitivity, specificity, accuracy, and broad dynamic range of linear quantification.…”

Improving linkage to care of hepatitis C: Evaluation of GeneXpert® HCV Viral Load Point-of-Care Assay in Indonesia

“…Assays can perform differently by genotype [36,42,43], while detection of HCV RNA and measurement of VL for the different genotypes is crucial to clinical management of HCV-infected patients [12][13][14]. This study showed a high efficiency and accuracy of Xpert HCV VL assay for quantitation HCV RNA of GT1, GT2, GT2, GT4, and even indeterminate genotype.…”

Can the Abbott RealTime hepatitis C virus assay be used to predict therapeutic outcomes in hepatitis C virus-infected patients undergoing triple therapy?

“…Real-time polymerase chain reaction (PCR) assays are generally preferred because of their wide range of detection and sensitivity to low quantities of viral RNA (13,14). Two commercial real-time PCR platforms are the CO-BAS AmpliPrep/COBAS TaqMan HCV assay version 1.0 (CAP/ CTM v1.0) (Roche Molecular Systems, Inc., Pleasanton, CA, USA) and the Abbott RealTime HCV assay (ART) (15)(16)(17). The recently developed ART has an improved lower limit of detection of 1.08 Log international units per milliliter (IU/mL) of HCV RNA in human serum or plasma, allowing for a more sensitive quantitation of hepatitis C virus ribonucleic acid (HCV RNA) than that achieved by CAP/CTM v1.0.…”