Abstract
The treatment schedule (combination of compounds, doses, and duration) and the virological follow-up for management of antiviral treatment in patients chronically infected by HCV is now well standardized, but to ensure good monitoring of the treated patients, physicians need rapid, reproducible, and sensitive molecular virological tools with a wide range of detection and quantification of HCV RNA in blood samples. Several assays for detection and/or quantification of HCV RNA are currently commercially available. Here, all these assays are detailed, and a brief description of each step of the assay is provided. They are divided into two categories by method: those based on signal amplification and those based on target amplification. These two categories are then divided into qualitative, quantitative, and quantitative detection assays. The real-time reverse-transcription polymerase chain reaction (RT-PCR)–based assays are the most promising strategy in the HCV virological area.
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References
Steinhauer, D. A., Domingo, E., and Holland, J. J. (1992) Lack of evidence for proofreading mechanisms associated with an RNA virus polymerase. Gene 122, 281–218.
Simmonds, P., Bukh, J., Combet, C., Deleage, G., Enomoto, N., Feinstone, S., Halfon, P., Inchauspe, G., Kuiken, C., Maertens, G., Mizokami, M., Murphy, D. G., Okamoto, H., Pawlotsky, J. M., Penin, F., Sablon, E., Shin, I. T., Stuyver, L. J., Thiel, H. J., Viazov, S., Weiner, A. J., and Widell, A. (2005) Consensus proposals for a unified system of nomenclature of hepatitis C virus genotypes. Hepatology 42, 962–973.
Hadziyannis, S. J., Sette, H., Jr., Morgan, T. R., Balan, V., Diago, M., Marcellin, P., Ramadori, G., Bodenheimer, H., Jr., Bernstein, D., Rizzetto, M., Zeuzem, S., Pockros, P. J., Lin, A., and Ackrill, A. M. (2004) Peginterferon-α2a and ribavirin combination therapy in chronic hepatitis C: a randomized study of treatment duration and ribavirin dose. Ann. Intern. Med. 140, 346–355.
Fried, M. W., Shiffman, M. L., Reddy, K. R., Smith, C., Marinos, G., Goncales, F. L., Jr., Haussinger, D., Diago, M., Carosi, G., Dhumeaux, D., Craxi, A., Lin, A., Hoffman, J., Yu, J. (2002) Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection. N. Engl. J. Med. 347, 975–982.
Manns, M. P., McHutchison, J. G., Gordon, S. C., Rustgi, V. K., Shiffman, M., Reindollar, R., Goodman, Z. D., Koury, K., Ling, M., and Albrecht, J. K. (2001) Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet 358, 958–965.
Berg, T., Sarrazin, C., Herrmann, E., Hinrichsen, H., Gerlach, T., Zachoval, R., Wiedenmann, B., Hopf, U., and Zeuzem, S. (2003) Prediction of treatment outcome in patients with chronic hepatitis C: significance of baseline parameters and viral dynamics during therapy. Hepatology 37, 600–609.
Payan, C., Pivert, A., Morand, P., Fafi-Kremer, S., Carrat, F., Pol, S., Cacoub, P., Perronne, C., and Lunel, F. (2007) Rapid and early virological response to chronic hepatitis C treatment with IFN α-2b or PEG- IFN α-2b plus ribavirin in HIV/HCV co-infected patients. Gut 56, 1111–1116.
Poordad, F., Reddy, K.R., Martin, P. (2008) Rapid virologic response: a new milestone in the management of chronic hepatitis C. Clin. Infect. Dis. 46, 78–84.
Jorgensen, P. A., and Neuwald, P. D. (2001) Standardized hepatitis C virus RNA panels for nucleic acid testing assays. J. Clin. Virol. 20, 35–40.
Trimoulet, P., Halfon, P., Pohier, E., Khiri, H., Chene, G., and Fleury, H. (2002) Evaluation of the VERSANT HCV RNA 3.0 assay for quantification of hepatitis C virus RNA in serum. J. Clin. Microbiol. 40, 2031–2036.
Lunel, F., Cresta, P., Vitour, D., Payan, C., Dumont, B., Frangeul, L., Reboul, D., Brault, C., Piette, J. C., and Huraux, J. M. (1999) Comparative evaluation of hepatitis C virus RNA quantitation by branched DNA, NASBA, and monitor assays. Hepatology 29, 528–535.
Veillon, P., Payan, C., Picchio, G., Maniez-Montreuil, M., Guntz, P., and Lunel, F. (2003) Comparative evaluation of the total hepatitis C virus core antigen, branched-DNA, and amplicor monitor assays in determining viremia for patients with chronic hepatitis C during interferon plus ribavirin combination therapy. J. Clin. Microbiol. 41, 3212–3120.
Krajden, M., Ziermann, R., Khan, A., Mak, A., Leung, K., Hendricks, D., and Comanor, L. (2002) Qualitative detection of hepatitis C virus RNA: comparison of analytical sensitivity, clinical performance, and workflow of the Cobas Amplicor HCV test version 2.0 and the HCV RNA transcription-mediated amplification qualitative assay. J. Clin. Microbiol. 40, 2903–2907.
Morishima, C., Morgan, T. R., Everhart, J. E., Wright, E. C., Shiffman, M. L., Everson, G. T., Lindsay, K. L., Lok, A. S., Bonkovsky, H. L., Di Bisceglie, A. M., Lee, W. M., Dienstag, J. L., Ghany, M. G., and Gretch, D. R. (2006) HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial. Hepatology 44, 360–367.
Morishima, C., Chung, M., Ng, K. W., Brambilla, D. J., and Gretch, D. R. (2004) Strengths and limitations of commercial tests for hepatitis C virus RNA quantification. J. Clin. Microbiol. 42, 421–425.
Lunel, F., Mariotti, M., Cresta, P., De la Croix, I., Huraux, J. M., and Lefrere, J. J. (1995) Comparative study of conventional and novel strategies for the detection of hepatitis C virus RNA in serum: amplicor, branched-DNA, NASBA and in-house PCR. J. Virol. Methods 54, 159–171.
Leckie, G., Schneider, G., Abravaya, K., Hoenle, R., Johanson, J., Lampinen, J., Ofsaiof, R., Rundle, L., Shah, S., Frank, A., Toolsie, D., Vijesurier, R., Wang, H., and Robinson, J. (2004) Performance attributes of the LCx HCV RNA quantitative assay. J. Virol. Methods 115, 207–215.
Bertuzis, R., Hardie, A., Hottentraeger, B., Izopet, J., Jilg, W., Kaesdorf, B., Leckie, G., Leete, J., Perrin, L., Qiu, C., Ran, I., Schneider, G., Simmonds, P., and Robinson, J. (2005) Clinical performance of the LCx HCV RNA quantitative assay. J. Virol. Methods 123, 171–178.
Sarrazin, C., Gartner, B. C., Sizmann, D., Babiel, R., Mihm, U., Hofmann, W. P., von Wagner, M., and Zeuzem, S. (2006) Comparison of conventional PCR with real-time PCR and branched DNA-based assays for hepatitis C virus RNA quantification and clinical significance for genotypes 1 to 5. J. Clin. Microbiol. 44, 729–737.
Halfon, P., Bourliere, M., Penaranda, G., Khiri, H., and Ouzan, D. (2006) Real-time PCR assays for hepatitis C virus (HCV) RNA quantitation are adequate for clinical management of patients with chronic HCV infection. J. Clin. Microbiol. 44, 2507–2511.
Chevaliez, S., Bouvier-Alias, M., Brillet, R., Pawlotsky, J. M. (2007) Overestimation and underestimation of hepatitis C virus RNA levels in a widely used real-time polymerase chain reaction-based method. Hepatology 46, 22–31.
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Le Guillou-Guillemette, H., Lunel-Fabiani, F. (2009). Detection and Quantification of Serum or Plasma HCV RNA: Mini Review of Commercially Available Assays. In: Tang, H. (eds) Hepatitis C. Methods in Molecular Biology™, vol 510. Humana Press. https://doi.org/10.1007/978-1-59745-394-3_1
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DOI: https://doi.org/10.1007/978-1-59745-394-3_1
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