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Updated: Jul 14, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Comparison of performance characteristics of three real-time reverse transcription-PCR test systems for detection and
M Fernanda Sábato1, Mitchell L Shiffman, Michael R Langley
1Molecular Diagnostics Laboratory, Department of Pathology, Virginia Commonwealth University, Richmond, Virginia 23298-0248, USA.
Insights
Two real-time PCR systems, Roche RUO-MPLC and Abbott ASR-Q, effectively detect and quantify hepatitis C virus (HCV) RNA. These systems demonstrated good correlation and agreement with the established COBAS Amplicor HCV Monitor v2.0 assay.
Area of Science:
- Virology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Hepatitis C virus (HCV) infection remains a significant global health concern.
- Accurate viral load quantification is crucial for monitoring treatment efficacy and disease progression.
- Real-time reverse transcription-PCR (RT-PCR) assays are widely used for HCV RNA detection and quantification.
Purpose of the Study:
- To evaluate the performance characteristics of three real-time RT-PCR test systems for HCV detection and quantification.
- To directly compare the performance of these systems using the same clinical specimens.
- To assess the correlation and agreement of the evaluated systems with a reference method.
Main Methods:
- Performance evaluation of three real-time RT-PCR systems: Roche COBAS TaqMan HCV (RUO-MPLC), Abbott analyte-specific reagents (ASR-Q), and Roche ASR-HP.
- Assessment of linear range, sensitivity, and precision using commercial HCV panels (genotype 1b).
- Direct comparison of results with the COBAS Amplicor HCV Monitor v2.0 using 76 clinical specimens with various HCV genotypes and titers.
Main Results:
- Roche RUO-MPLC and Abbott ASR-Q demonstrated high sensitivity (1.0 log(10) IU/ml) and a broad linear dynamic range (1.0-7.0 log(10) IU/ml).
- Abbott ASR-Q exhibited the highest reproducibility among the three systems.
- Good correlation (R(2) = 0.84 for RUO-MPLC, R(2) = 0.93 for ASR-Q) and agreement were observed between Roche RUO-MPLC, Abbott ASR-Q, and the reference method. Roche ASR-HP showed poor correlation and significant underestimation of HCV RNA for certain genotypes.
Conclusions:
- The Roche RUO-MPLC and Abbott ASR-Q real-time RT-PCR systems provide acceptable performance for HCV RNA detection and quantification.
- These two systems demonstrate sufficient agreement with the COBAS Amplicor HCV Monitor v2.0, making them suitable for clinical use.
- The Roche ASR-HP system showed limitations in accuracy and genotype-dependent bias, suggesting it is less reliable for clinical application.
Abstract:
We evaluated the performance characteristics of three real-time reverse transcription-PCR test systems for detection and quantification of hepatitis C virus (HCV) and performed a direct comparison of the systems on the same clinical specimens. Commercial HCV panels (genotype 1b) were used to evaluate linear range, sensitivity, and precision. The Roche COBAS TaqMan HCV test for research use only (RUO) with samples processed on the MagNA Pure LC instrument (Roche RUO-MPLC) and Abbott analyte-specific reagents (ASR) with QIAGEN sample processing (Abbott ASR-Q) showed a sensitivity of 1.0 log(10) IU/ml with a linear dynamic range of 1.0 to 7.0 log(10) IU/ml. The Roche ASR in combination with the High Pure system (Roche ASR-HP) showed a sensitivity of 1.4 log(10) IU/ml with a linear dynamic range of 2.0 to 7.0 log(10) IU/ml. All of the systems showed acceptable reproducibility, the Abbott ASR-Q being the most reproducible of the three systems. Seventy-six clinical specimens (50 with detectable levels of HCV RNA and various titers and genotypes) were tested, and results were compared to those of the COBAS Amplicor HCV Monitor v2.0. Good correlation was obtained for the Roche RUO-MPLC and Abbott ASR-Q (R(2) = 0.84 and R(2) = 0.93, respectively), with better agreement for the Abbott ASR-Q. However, correlation (R(2) = 0.79) and agreement were poor for Roche ASR-HP, with bias relative to concentration and genotype. Roche ASR-HP underestimated HCV RNA for genotypes 3 and 4 as much as 2.19 log(10) IU/ml. Our study demonstrates that Roche RUO-MPLC and Abbott ASR-Q provided acceptable results and agreed sufficiently with the COBAS Amplicor HCV Monitor v2.0.
