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Hepatitis B and D Protocols pp 3–13Cite as

Overview of Commercial HBV Assay Systems

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Part of the book series: Methods in Molecular Medicine ((MIMM,volume 95))

Abstract

Measurement of viral nucleic acid in serum is often a valuable adjunct to the management of viral infections (1). In hepatitis B, tests for hepatitis B virus (HBV) DNA have been used widely (Table 1), but their interpretation and significance have yet to be defined. HBV DNA assays are limited by lack of standardization and variable sensitivity. Because HBV may circulate in serum at high levels (as high as 1010 virions/mL), direct molecular hybridization assays are capable of detecting HBV DNA in a high proportion of patients, particularly those with active disease and both HBsAg (hepatitis B surface antigen) and HBeAg (hepatitis B e antigen) in serum. Commercial assays comprise the liquid hybridization assay (Genostics™, Abbott Laboratories, Chicago, IL), the hybridization capture assay (Digene, HC II), and branched DNA (bDNA) signal amplification assay (Versant, Bayer Diagnostics). Furthermore, a quantitative polymerase chain reaction (PCR) assay for HBV DNA has been developed (Amplicor Monitor HBV, Roche Diagnostics); it detects HBV DNA in a higher proportion of patients with chronic hepatitis B and often yields positive results, even in HBsAg carriers without apparent disease.

Table 1 Different Principles of HBV DNA Quantification
Full size table

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References

  1. Lok, A.S., Heathcote, E.J., Hoofnagle, J.H. (2001) Management of hepatitis B: 2000—summary of a workshop. Gastroenterology 120, 1828–1853.

    Article  PubMed  CAS  Google Scholar 

  2. Kuhns, M.C., McNamara, A.L., Cabal, C.M., et al. (1988) A new assay for the quantitative detection of hepatitis B viral DNA in human serum. In: Zuckerman, A.J. (ed.) Viral Hepatitis and Liver Disease, Alan Liss, New York, 258–262.

    Google Scholar 

  3. Kuhns, M.C., McNamara, A.L., Perrillo, R.P., Cabal, C.M., and Campbel, C.R. (1989) Quantitation of hepatitis B viral DNA by solution hybridization: comparison with DNA polymerase and hepatitis B e antigen during antiviral therapy. J. Med. Virol. 27, 274–281.

    Article  PubMed  CAS  Google Scholar 

  4. Kuhns, M., Thiers, V., Courouce, A., Scotto, J., Tiollais, P., and Bréchot, C. (1984) Quantitative detection of HBV DNA in human serum. In: Vyas, G., Dienstag, J., Hoofnagle, J. (eds.) Viral Hepatitis and Liver Disease, Grune and Stratton, Orlando, FL, 665–670.

    Google Scholar 

  5. Hendricks, D.A., Stowe, B.J., Hoo, B.S., et al. (1995) Quantitation of HBV DNA in human serum using a branched DNA (bDNA) signal amplification assay. Am. J. Clin. Pathol. 104, 537–546.

    PubMed  CAS  Google Scholar 

  6. Chen, C.H., Wang, J.T., Lee, C.Z., Sheu, J.C., Wang, T.H., and Chen, D.S. (1995) Quantitative detection of hepatitis B virus DNA in human sera by branched-DNA signal amplification. J. Virol. Methods 53, 131–137.

    Article  PubMed  CAS  Google Scholar 

  7. Urdea, M.S. (1992) Theoretical aspects of nucleic acid standardization for HBV DNA detection. Report on the Sixth Eurohep Workshop, 3.1.2c.

    Google Scholar 

  8. Butterworth, L.-A., Prior, S.L., Buda, P.J., Faoagali, J.L., and Cooksley, G.E. (1996) Comparison of four methods for quantitative measurement of hepatitis B viral DNA. J. Hepatol. 24, 686–691.

    Article  PubMed  CAS  Google Scholar 

  9. Pawlotsky, J.M., Bastie, A., Lonjon, I., et al. (1997) What technique should be used for routine detection and quantification of HBV DNA in clinical samples? J. Virol. Methods 65, 245–253.

    Article  PubMed  CAS  Google Scholar 

  10. Niesters, H.G., Krajden, M., Cork, L., et al. (2000) A multicenter study evaluation of the digene hybrid capture II signal amplification technique for detection of hepatitis B virus DNA in serum samples and testing of EUROHEP standards. J. Clin. Microbiol. 38, 2150–2155.

    PubMed  CAS  Google Scholar 

  11. Kessler, H.H., Pierer, K., Dragon, E., et al. (1998) Evaluation of a new assay for HBV DNA quantitation in patients with chronic hepatitis B. Clin. Diagn. Virol. 9, 37–43.

    Article  PubMed  CAS  Google Scholar 

  12. Gerken, G., Gomes, J., Lampertico, P., et al. (1998) Clinical evaluation and applications of the Amplicor HBV Monitor test, a quantitative HBV DNA PCR assay. J. Virol. Methods 74, 155–165.

    Article  PubMed  CAS  Google Scholar 

  13. Noborg, U., Gusdal, A., Pisa, E.K., Hedrum, A., and Lindh, M. (1999) Automated quantitative analysis of hepatitis B virus DNA by using the Cobas Amplicor HBV Monitor test. J. Clin. Microbiol. 37, 2793–2797.

    PubMed  CAS  Google Scholar 

  14. Ide, T., Kumashiro, R., Hino, T., et al. (2001) Transcription-mediated amplification is more useful in the follow-up of patients with chronic hepatitis B treated with lamivudine. Hepatol. Res. 21, 76–84.

    Article  PubMed  CAS  Google Scholar 

  15. Trippler, M., Hampl, H., Goergen, B., et al. (1996) Ligase chain reaction (LCR) assay for semi-quantitative detection of HBV DNA in mononuclear leukocytes if patients with chronic hepatitis B. J. Viral Hepatitis 3, 267–272.

    Article  CAS  Google Scholar 

  16. Yates, S., Penning, M., Goudsmit, J., et al. (2001) Quantitative detection of hepatitis B virus DNA by real-time nucleic acid sequence-based amplification with molecular beacon detection. J. Clin. Microbiol. 39, 3656–3665.

    Article  PubMed  CAS  Google Scholar 

  17. Erhardt, A., Schaefer, S., Athanassiou, N., Kann, M., and Gerlich, W.H. (1996) Quantitative assay of PCR-amplified hepatitis B virus DNA using a peroxidase-labelled DNA probe and enhanced chemiluminescence. J. Clin. Microbiol. 34, 1885–1891.

    PubMed  CAS  Google Scholar 

  18. Lai, V.C., Guan, R., Wood, M.L., Lo, S.K., Yuen, M.F., and Lai, C.L. (1999) Nucleic acid-based cross-linking assay for detection and quantification of hepatitis B virus DNA. J.Clin. Microbiol. 37, 161–164.

    PubMed  CAS  Google Scholar 

  19. Cane, P.A., Cook, P., Ratcliffe, D., Mutimer, D., and Pillay, D. (1999) Use of real-time PCR and fluorimetry to detect lamivudine resistance-associated mutations in hepatitis B virus. Antimicrob. Agents Chemother. 43, 1600–1608.

    PubMed  CAS  Google Scholar 

  20. Defoort, J.P., Martin, M., Casano, B., Prato, S., Camilla, C., and Fert, V. (2000) Simultaneous detection of multiplex-amplified human immunodeficiency virus type 1 RNA, hepatitis C virus RNA, and hepatitis B virus DNA using a flow cytometer microsphere-based hybridization assay. J. Clin. Microbiol. 38, 1066–1071.

    PubMed  CAS  Google Scholar 

  21. Loeb, K.R., Jerome, K.R., Goddard, J., Huang, M., Cent, A., and Corey, L. (2000) High-throughput quantitative analysis of hepatitis B virus DNA in serum using the TaqMan fluorogenic detection system. Hepatology 32, 626–629.

    Article  PubMed  CAS  Google Scholar 

  22. Chen, R.W., Piiparinen, H., Seppanen, M., Koskela, P., Sarna, S., and Lappalainen, M. (2001) Real-time PCR for detection and quantitation of hepatitis B virus DNA. J. Med. Virol. 65, 250–256.

    Article  PubMed  CAS  Google Scholar 

  23. Pas, S.D., Fries, E., de Man, R.A., Osterhaus, A.D.M.E., and Niesters, H.G.M. (2000) Development of a quantitative real-time detection assay for hepatitis B virus DNA and comparison with two commercial assays. J. Clin. Microbiol. 38, 2897–2901.

    PubMed  CAS  Google Scholar 

  24. Kapke, G.E., Watson, G., Sheffler, S., Hunt, D., and Frederick, C. (1997) Comparison of the Chiron Quantiplex branched DNA (bDNA) assay and the Abbott Genostics solution hybridization assay for quantification of hepatitis B viral DNA. J. Viral Hepat. 4, 67–75.

    Article  PubMed  CAS  Google Scholar 

  25. Zaaijer, H.L., ter Borg, F., Cuypers, H.T., Hermus, M.C., and Lelie, P.N. (1994) Comparison of methods for detection of hepatitis B virus DNA. J. Clin. Microbiol. 32, 2088–2091.

    PubMed  CAS  Google Scholar 

  26. Khakoo, S.I., Soni, P.N., and Dusheiko, G.M. (1996) A clinical evaluation of a new method for HBV DNA quantitation in patients with chronic hepatitis B. J. Med. Virol. 50, 112–116.

    Article  PubMed  CAS  Google Scholar 

  27. Hwang, S.J., Lee, S.D., Lu, R.H., et al. (1996) Comparison of three different hybridization assays in the quantitative measurement of serum hepatitis B virus DNA. J. Virol. Methods 62, 123–129.

    Article  PubMed  CAS  Google Scholar 

  28. Krajden, M., Minor, J., Cork, L., and Comanor, L. (1998) Multi-measurement method comparison of three commercial hepatitis B virus DNA quantification assays. J. Viral Hepat. 5, 415–422.

    Article  PubMed  CAS  Google Scholar 

  29. Ho, S.K.N., Chan, T.M., Cheng, I.K.P., and Lai, K.N. (1999) Comparison of the second-generation Digene hybrid capture assay with the branched DNA assay for measurement of hepatitis B virus DNA in serum. J. Clin. Microbiol. 37, 2461–2465.

    PubMed  CAS  Google Scholar 

  30. Pawlotsky, J.M., Bastie, A., Hezode, C., et al. (2000) Routine detection and quantification of hepatitis B virus DNA in clinical laboratories: performance of three commercial assays. J. Virol. Methods 85, 11–21.

    Article  PubMed  CAS  Google Scholar 

  31. Chan, H.L., Leung, N.W., Lau, T.C., Wong, M.L., and Sung, J.J. (2000) Comparison of three different sensitive assays for hepatitis B virus DNA in monitoring of responses to antiviral therapy. J. Clin. Microbiol. 38, 3205–3208.

    PubMed  CAS  Google Scholar 

  32. Gerlich, W.H., Heermann, K.-H., Thomssen, R., and the Eurohep Group (1995) Quantitative assays for hepatitis B virus DNA: standardization and quality control. Viral Hep. Rev. 1, 53–57.

    Google Scholar 

  33. Heermann, K.-H., Gerlich, W.H., Chudy, M., Schaefer, S., Thomssen, R., and the Eurohep Pathobiology Group (1999) Quantitative detection of hepatitis B virus DNA in two international reference plasma preparations. J. Clin. Microbiol. 37, 68–73.

    PubMed  CAS  Google Scholar 

  34. Quint, W.G., Heijtink, R.A., Schirm, J., Gerlich, W.H., and Niesters, H.G. (1995) Reliability of methods for hepatitis B virus DNA detection. J. Clin. Microbiol. 33, 225–228.

    PubMed  CAS  Google Scholar 

  35. Lindh, M., Horal, P., Dhillon, A.P., and Norkrans, G. (2000) Hepatitis B virus DNA levels, precore mutations, genotypes and histological activity in chronic hepatitis B. J. Viral Hepat. 7, 258–267.

    Article  PubMed  CAS  Google Scholar 

  36. Cacciola, I., Pollicino, T., Squadrito, G., et al. (2000) Quantification of intrahepatic hepatitis B virus (HBV) DNA in patients with chronic HBV infection. Hepatology 31, 507–512.

    Article  PubMed  CAS  Google Scholar 

  37. Otake, K., and Nishioka, K. (2000) Nucleic acid amplification testing of hepatitis B virus. Lancet 355, 1460.

    Article  PubMed  CAS  Google Scholar 

  38. Hunt, C.M., McGill, J.M., Allen, M.I., and Condreay, L.D. (2000) Clinical relevance of hepatitis B viral mutations. Hepatology 31, 1037–1044.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Internal Medicine II, University Hospital, Homburg/Saar, Germany

    ofessor of Medicine Stefan Zeuzemro (Professor of Medicine)

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  1. ofessor of Medicine Stefan Zeuzemro
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Editors and Affiliations

  1. Ribapharm Inc., Costa Mesa, CA

    Robert K. Hamatake PhD  & Johnson Y. N. Lau MD  & 

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© 2004 Humana Press Inc.

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Zeuzemro, S. (2004). Overview of Commercial HBV Assay Systems. In: Hamatake, R.K., Lau, J.Y.N. (eds) Hepatitis B and D Protocols. Methods in Molecular Medicine, vol 95. Humana Press. https://doi.org/10.1385/1-59259-669-X:3

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  • DOI: https://doi.org/10.1385/1-59259-669-X:3

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-105-9

  • Online ISBN: 978-1-59259-669-0

  • eBook Packages: Springer Protocols

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