Detection and quantitation of HCV RNA using real-time PCR after automated sample processing

There is considerable evidence that the loss of hepatitis C virus (HCV) RNA during the first 3 months of treatment with pegylated interferon plus ribavirin is a prognostic marker of response to therapy. Real-time polymerase chain reaction (PCR) assays for quantifying HCV RNA in plasma or serum are n...

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Bibliographic Details
Published in:Journal of medical virology 2007-12, Vol.79 (12), p.1821-1826
Main Authors: Sandres-Sauné, K, Abravanel, F, Nicot, F, Peron, J.M, Alric, L, Boineau, J, Pasquier, C, Izopet, J
Format: Article
Language:English
Subjects:
automated extraction
Automation
Biological and medical sciences
COBAS Ampliprep
COBAS Taqman
Fundamental and applied biological sciences. Psychology
Genotype
HCV RNA quantitation
Hepacivirus - genetics
Hepacivirus - isolation & purification
Hepatitis C virus
Human viral diseases
Humans
Infectious diseases
Linear Models
Medical sciences
Microbiology
Miscellaneous
Polymerase Chain Reaction
Reproducibility of Results
RNA, Viral - analysis
RNA, Viral - genetics
Sensitivity and Specificity
Specimen Handling - instrumentation
Specimen Handling - methods
Viral diseases
Virology
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Summary:There is considerable evidence that the loss of hepatitis C virus (HCV) RNA during the first 3 months of treatment with pegylated interferon plus ribavirin is a prognostic marker of response to therapy. Real-time polymerase chain reaction (PCR) assays for quantifying HCV RNA in plasma or serum are now commercially available. The extraction of HCV RNA can also be automated. This report analyses the performance of the COBAS Ampliprep-COBAS Taqman 48 (CAP/CTM) real-time PCR assay and compares this new test with the COBAS Amplicor HCV Monitor v 2.0 assay (CAM). CAP/CTM was 100% specific. The assay was linear across a wide range of HCV RNA concentrations without sample dilution. The intra-assay variation was 0.3-3.3% and the interassay variation was 1.5-6.7%. A total of 118 clinical samples with different HCV genotypes were assayed using both methods. The results obtained using the two methods were well correlated (r = 0.89, P < 0.001). The mean difference [CAP/CTM-CAM] was 0.17 log IU/ml and it was not influenced by the HCV genotype or by the subtype. It is concluded that the new CAP/CTM system is adequate for quantifying HCV RNA in clinical practice. J. Med. Virol. 79:1821-1826, 2007. © Wiley-Liss, Inc.
ISSN:0146-6615
1096-9071
DOI:10.1002/jmv.21003