Abstract
The use of molecular-based methodologies for the identification of viral nucleic acid has evolved into a clinically useful adjunct to the biochemical, serological, and histological assays used to evaluate patient health. The clinical application of viral-load assays for monitoring viral infections has been the impetus for the development of standardized tests for the hepatitis C virus (HCV). Standardization and precision of quantification have been instrumental in assessing the natural history of HCV, in which viral RNA levels have a large dynamic range. Assessment of the HCV viral load is now a routine part of the algorithm for the diagnosis of chronic hepatitis C (1–5). Patient’s response to anti-viral therapy can be assessed, in part, by quantification of serum viral loads (6,7).
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References
Alter M.J., Kruszon-Moran D., Nainan O.V., McQuillan G.M., Gao F., Moyer L.A., et al. (1999) The prevalence of hepatitis C virus infection in the United States 1988 through 1994. N. Engl. J. Med. 341, 556–562.
Fanning L., Kenny E., Sheehan M., Cannon B., Whelton M., O’Connell J., et al. (1999) Viral load and clinicopathological features of chronic hepatitis C (1b) in a homogeneous patient population. Hepatol. 29, 904–907.
Kenny-Walsh E. (1999) Clinical outcomes after hepatitis C infection from contaminated anti-D immune globulin. Irish Hepatology Research Group. N. Engl. J. Med. 340, 1228–1233.
Lu R.H., Hwang S.J., Chan C.Y., Chang F.Y., and Lee S.D. (1998) Quantitative measurement of serum HCV RNA in patients with chronic hepatitis C: comparison between Amplicor HCV monitor system and branched DNA signal amplification assay. J. Clin. Lab. Anal. 12, 121–125.
Nomura H., Kimura Y., Rikimaru N., Tada H., Okamoto O., Shiraishi G., et al. (1997) Usefulness of HCV-RNA assays in efficacy evaluation of interferon treatment for chronic hepatitis C: Amplicor HCV assay and branched DNA probe assay. J. Infect. 34, 249–255.
McHutchison J.G., Gordon S.C., Schiff E.R., Shiffman M.L., Lee W.M., Rustgi V.K., et al. (1998) Interferon alpha-2b alone or in combination with ribavirin as initial treatment for chronic hepatitis C. Hepatitis Interventional Therapy Group. N. Engl. J. Med. 339, 1485–1492.
Poynard T., Marcellin P., Lee S.S., Niederau C., Minuk G.S., Ideo G., et al. (1998) Randomised trial of interferon alpha 2b plus ribavirin for 48 weeks or for 24 weeks versus interferon alpha 2b plus placebo for 48 weeks for treatment of chronic infection with hepatitis C virus. International Hepatitis Interventional Therapy Group (IHIT). Lancet 352, 1426–1432.
Gordon S.C., Dailey P.J., Silverman A.L., Khan B.A., Kodali V.P., and Wilber J.C. (1998) Sequential serum hepatitis C viral RNA levels longitudinally assessed by branched DNA signal amplification. Hepatol. 28, 1702–1706.
Halfon P., Khiri H., Gerolami V., Bourliere M., Feryn J.M., Reynier P., et al. (1996) Impact of various handling and storage conditions on quantitative detection of hepatitis C virus RNA. J. Hepatol. 25, 307–311.
Fanning L., Kenny-Walsh E., Levis J., Choudhury K.R., Cannon B., Sheehan M., et al. (2000) Natural fluctuations of hepatitis C viral load in a homogeneous patient population: a prospective study. Hepatol. 31, 225–229.
Martinot-Peignoux M., Boyer N., Le Breton V., Le Guludec G., Castelnau C., Akremi R., et al. (2000) A new step toward standardization of serum hepatitis C virus-RNA quantification in patients with chronic hepatitis C. Hepatology 31, 726–729.
Saldanha J., Lelie N., and Heath A. (1999) Establishment of the first international standard for nucleic acid amplification technology (NAT) assays for HCV RNA. WHO Collaborative Study Group. Vox Sang 76, 149–158.
Davis G.L., Esteban-Mur R., Rustgi V., Hoefs J., Gordon S.C., Trepo C., et al. (1998) Interferon alpha-2b alone or in combination with ribavirin for the treatment of relapse of chronic hepatitis C. International Hepatitis Interventional Therapy Group. N. Engl. J. Med. 339, 1493–1499.
Bresters D., Cuypers H.T., Reesink H.W., Mauser-Bunschoten E.P., van den Berg H.M., Schaasberg W.P., et al. (1994) Comparison of quantitative cDNA-PCR with the branched DNA hybridization assay for monitoring plasma hepatitis C virus RNA levels in haemophilia patients participating in a controlled interferon trial. J. Med. Virol. 43, 262–268.
Hawkins A., Davidson F., and Simmonds P. (1997) Comparison of plasma virus loads among individuals infected with hepatitis C virus (HCV) genotypes 1, 2, and 3 by quantiplex HCV RNA assay versions 1 and 2, Roche Monitor assay, and an in-house limiting dilution method. J. Clin. Microbiol. 35, 187–192.
Sarrazin C., Teuber G., Kokka R., Rabenau H., and Zeuzem S. (2000) Detection of residual hepatitis C virus RNA by transcription-mediated amplification in patients with complete virologic response according to polymerase chain reaction-based assays. Hepatloogy 32, 818–823.
McHutchison J.G., Blatt L.M., Ponnudurai R., Goodarzi K., Russell J., and Conrad A. (1999) Ultracentrifugation and concentration of a large volume of serum for HCV RNA during treatment may predict sustained and relapse response in chronic HCV infection. J. Med. Virol 57, 351–355.
Roth W.K., Weber M., and Seifried E. (1999) Feasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting. Lancet 353, 359–363.
Sun R., Schilling W., Jayakar H., Ku J., Wang J., Rosenstraus M., et al. (1999) Simultaneous extraction of hepatitis C virus (HCV), hepatitis B virus, and HIV-1 from plasma and detection of HCV RNA by a reverse transcriptase-polymerase chain reaction assay designed for screening pooled units of donated blood. Transfusion 39, 1111–1119.
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Fanning, L.J. (2002). Detection and Quantification of the Hepatitis C Viral Genome. In: O’Connell, J. (eds) RT-PCR Protocols. Methods in Molecular Biology, vol 193. Humana Press. https://doi.org/10.1385/1-59259-283-X:151
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DOI: https://doi.org/10.1385/1-59259-283-X:151
Publisher Name: Humana Press
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