Gastroenterology
Volume 134, Issue 2 , Pages 405-415 , February 2008

Virologic Monitoring of Hepatitis B Virus Therapy in Clinical Trials and Practice: Recommendations for a Standardized Approach

  • Jean–Michel Pawlotsky

      Affiliations

    • French National Reference Center for Viral Hepatitis B, C and delta, Department of Virology, Henri Mondor Hospital, University of Paris XII, Créteil, France
    • INSERM U841, Créteil, France
    • Corresponding Author InformationAddress requests for reprints to: Jean-Michel Pawlotsky, MD, PhD, Department of Virology, Hopital Henri Mondor, 51 Avenue du Maréchal de Lattre de Tassigny, 94010 Créteil, France. fax: (33) 1-4981-4831.
  • ,
  • Geoffrey Dusheiko

      Affiliations

    • Centre for Hepatology, Royal Free and University College School of Medicine, London, United Kingdom
  • ,
  • Angelos Hatzakis

      Affiliations

    • Department of Hygiene, Epidemiology and Medical Statistics, Athens University Medical School, Athens, Greece
  • ,
  • Daryl Lau

      Affiliations

    • Liver Center, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts
  • ,
  • George Lau

      Affiliations

    • Department of Medicine, Queen Mary Hospital, University of Hong Kong, Hong Kong SAR, China
  • ,
  • T. Jake Liang

      Affiliations

    • Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland
  • ,
  • Stephen Locarnini

      Affiliations

    • Victorian Infectious Diseases Reference Laboratory, North Melbourne, Victoria, Australia
  • ,
  • Paul Martin

      Affiliations

    • Division of Liver Diseases and Recanati-Miller Transplant Institute, Mount Sinai School of Medicine, New York, New York
  • ,
  • Douglas D. Richman

      Affiliations

    • VA San Diego Healthcare System and University of California San Diego, La Jolla, California
  • ,
  • Fabien Zoulim

      Affiliations

    • INSERM U871, Hospices Civils de Lyon, Department of Liver Diseases, Université Lyon 1, Lyon, France

Received 16 February 2007 ,Accepted 1 November 2007.

References 

  1. Lee WM. Hepatitis B virus infection. N Engl J Med. 1997;337:1733–1745
  2. Ganem D, Prince AM. Hepatitis B virus infection—natural history and clinical consequences. N Engl J Med. 2004;350:1118–1129
  3. Zoulim F. Mechanism of viral persistence and resistance to nucleoside and nucleotide analogs in chronic hepatitis B virus infection. Antiviral Res. 2004;64:1–15
  4. Shaw T, Bartholomeusz A, Locarnini S. HBV drug resistance: mechanisms, detection and interpretation. J Hepatol. 2006;44:593–606
  5. Liaw YF, Sung JJ, Chow WC, et al. Lamivudine for patients with chronic hepatitis B and advanced liver disease. N Engl J Med. 2004;351:1521–1531
  6. Perrillo RP, Schiff ER, Davis GL, et al. A randomized, controlled trial of interferon alfa-2b alone and after prednisone withdrawal for the treatment of chronic hepatitis B. N Engl J Med. 1990;323:295–301
  7. Manesis EK, Hadziyannis SJ. Interferon alpha treatment and retreatment of hepatitis B e antigen-negative chronic hepatitis B. Gastroenterology. 2001;121:101–109
  8. Lai CL, Chien RN, Leung NW, et al. A one-year trial of lamivudine for chronic hepatitis B. N Engl J Med. 1998;339:61–68
  9. Dienstag JL, Schiff ER, Wright TL, et al. Lamivudine as initial treatment for chronic hepatitis B in the United States. N Engl J Med. 1999;341:1256–1263
  10. Leung NW, Lai CL, Chang TT, et al. Extended lamivudine treatment in patients with chronic hepatitis B enhances hepatitis B e antigen seroconversion rates: results after 3 years of therapy. Hepatology. 2001;33:1527–1532
  11. Cooksley WG, Piratvisuth T, Lee SD, et al. Peginterferon alpha-2a (40 kDa): an advance in the treatment of hepatitis B e antigen-positive chronic hepatitis B. J Viral Hepat. 2003;10:298–305
  12. Lau GK, Piratvisuth T, Luo KX, et al. Peginterferon alfa-2a, lamivudine, and the combination for HBeAg-positive chronic hepatitis B. N Engl J Med. 2005;352:2682–2695
  13. Marcellin P, Lau GK, Bonino F, et al. Peginterferon alfa-2a alone, lamivudine alone, and the two in combination in patients with HBeAg-negative chronic hepatitis B. N Engl J Med. 2004;351:1206–1217
  14. Lai CL, Gane E, Hsu CW, et al. Two-year results from the GLOBE trial in patients with hepatitis B: greater clinical and antiviral efficacy for telbivudine (LdT) versus lamivudine. Hepatology. 2006;44(Suppl 1):222A
  15. Chang TT, Gish RG, de Man R, et al. A comparison of entecavir and lamivudine for HBeAg-positive chronic hepatitis B. N Engl J Med. 2006;354:1001–1010
  16. Chang TT, Gish RJ, de Man RA, et al. Entecavir is superior to lamivudine for the treatment of HBeAg-positive chronic hepatitis B: results of phase III study ETV-022 in nucleoside-naive patients. Hepatology. 2004;40(Suppl 1):193A
  17. Marcellin P, Chang TT, Lim SG, et al. Adefovir dipivoxil for the treatment of hepatitis B e antigen-positive chronic hepatitis B. N Engl J Med. 2003;348:808–816
  18. Hadziyannis SJ, Tassopoulos NC, Heathcote EJ, et al. Long-term therapy with adefovir dipivoxil for HBeAg-negative chronic hepatitis B for up to 5 years. Gastroenterology. 2006;131:1743–1751
  19. Hadziyannis SJ, Tassopoulos NC, Heathcote EJ, et al. Adefovir dipivoxil for the treatment of hepatitis B e antigen-negative chronic hepatitis B. N Engl J Med. 2003;348:800–807
  20. Chen CJ, Yang HI, Su J, et al. Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level. JAMA. 2006;295:65–73
  21. Iloeje UH, Yang HI, Su J, et al. Predicting cirrhosis risk based on the level of circulating hepatitis B viral load. Gastroenterology. 2006;130:678–686
  22. Yu MW, Yeh SH, Chen PJ, et al. Hepatitis B virus genotype and DNA level and hepatocellular carcinoma: a prospective study in men. J Natl Cancer Inst. 2005;97:265–272
  23. Wong DK, Cheung AM, O’Rourke K, et al. Effect of alpha-interferon treatment in patients with hepatitis B e antigen-positive chronic hepatitis B (A meta-analysis). Ann Intern Med. 1993;119:312–323
  24. Mommeja-Marin H, Mondou E, Blum MR, et al. Serum HBV DNA as a marker of efficacy during therapy for chronic HBV infection: analysis and review of the literature. Hepatology. 2003;37:1309–1319
  25. Dienstag JL, Goldin RD, Heathcote EJ, et al. Histological outcome during long-term lamivudine therapy. Gastroenterology. 2003;124:105–117
  26. Colonno RJ, Rose RE, Pokornowski K, et al. Assessment at three years shows high barrier to resistance is maintained in entecavir-treated nucleoside naive patients while resistance emergence increases over time in lamivudine refractory patients. Hepatology. 2006;44(Suppl 2):229A
  27. Di Bisceglie A, Lai CL, Gane E, et al. Telbivudine GLOBE trial: maximal early HBV suppression is predictive of optimal two-year efficacy in nucleoside-treated hepatitis B patients. Hepatology. 2006;44(Suppl 1):230A
  28. Yuen MF, Sablon E, Hui CK, et al. Factors associated with hepatitis B virus DNA breakthrough in patients receiving prolonged lamivudine therapy. Hepatology. 2001;34:785–791
  29. Ide T, Kumashiro R, Koga Y, et al. A real-time quantitative polymerase chain reaction method for hepatitis B virus in patients with chronic hepatitis B treated with lamivudine. Am J Gastroenterol. 2003;98:2048–2051
  30. Werle-Lapostolle B, Bowden S, Locarnini S, et al. Persistence of cccDNA during the natural history of chronic hepatitis B and decline during adefovir dipivoxil therapy. Gastroenterology. 2004;126:1750–1758
  31. Sung JJ, Wong ML, Bowden S, et al. Intrahepatic hepatitis B virus covalently closed circular DNA can be a predictor of sustained response to therapy. Gastroenterology. 2005;128:1890–1897
  32. Wursthorn K, Lutgehetmann M, Dandri M, et al. Peginterferon alpha-2b plus adefovir induce strong cccDNA decline and HBsAg reduction in patients with chronic hepatitis B. Hepatology. 2006;44:675–684
  33. Laras A, Koskinas J, Dimou E, et al. Intrahepatic levels and replicative activity of covalently closed circular hepatitis B virus DNA in chronically infected patients. Hepatology. 2006;44:694–702
  34. Zoulim F. Assessment of treatment efficacy in HBV infection and disease. J Hepatol. 2006;44:S95–S99
  35. Yuen MF, Wong DK, Sum SS, et al. Effect of lamivudine therapy on the serum covalently closed-circular (ccc) DNA of chronic hepatitis B infection. Am J Gastroenterol. 2005;100:1099–1103
  36. Wong DK, Yuen MF, Yuan H, et al. Quantitation of covalently closed circular hepatitis B virus DNA in chronic hepatitis B patients. Hepatology. 2004;40:727–737
  37. Zoulim F. New insight on hepatitis B virus persistence from the study of intrahepatic viral cccDNA. J Hepatol. 2005;42:302–308
  38. Hui CK, Bowden S, Jackson K, et al. Clinical significance of intrahepatic hepatitis B virus covalently closed circular DNA in chronic hepatitis B patients who received cytotoxic chemotherapy. Blood. 2005;105:2616–2617
  39. Kidd-Ljunggren K, Miyakawa Y, Kidd AH. Genetic variability in hepatitis B viruses. J Gen Virol. 2002;83:1267–1280
  40. Arauz-Ruiz P, Norder H, Robertson BH, et al. Genotype H: a new Amerindian genotype of hepatitis B virus revealed in Central America. J Gen Virol. 2002;83:2059–2073
  41. Chu CM, Liaw YF. Genotype C hepatitis B virus infection is associated with a higher risk of reactivation of hepatitis B and progression to cirrhosis than genotype B: a longitudinal study of hepatitis B e antigen-positive patients with normal aminotransferase levels at baseline. J Hepatol. 2005;43:411–417
  42. Orito E, Ichida T, Sakugawa H, et al. Geographic distribution of hepatitis B virus (HBV) genotype in patients with chronic HBV infection in Japan. Hepatology. 2001;34:590–594
  43. Grandjacques C, Pradat P, Stuyver L, et al. Rapid detection of genotypes and mutations in the pre-core promoter and the pre-core region of hepatitis B virus genome: correlation with viral persistence and disease severity. J Hepatol. 2000;33:430–439
  44. Funk ML, Rosenberg DM, Lok AS. Worldwide epidemiology of HBeAg-negative chronic hepatitis B and associated precore and core promoter variants. J Viral Hepat. 2002;9:52–61
  45. Livingston SE, Simonetti JP, McMahon BJ, et al. Hepatitis B virus genotypes in Alaska native people with hepatocellular carcinoma: preponderance of genotype F. J Infect Dis. 2007;195:5–11
  46. Janssen HL, van Zonneveld M, Senturk H, et al. Pegylated interferon alfa-2b alone or in combination with lamivudine for HBeAg-positive chronic hepatitis B: a randomised trial. Lancet. 2005;365:123–129
  47. Flink HJ, van Zonneveld M, Hansen BE, et al. Treatment with peginterferon alpha-2b for HBeAg-positive chronic hepatitis B: HBsAg loss is associated with HBV genotype. Am J Gastroenterol. 2006;101:297–303
  48. Nafa S, Ahmed S, Tavan D, et al. Early detection of viral resistance by determination of hepatitis B virus polymerase mutations in patients treated by lamivudine for chronic hepatitis B. Hepatology. 2000;32:1078–1088
  49. Gintowt AA, Germer JJ, Mitchell PS, et al. Evaluation of the MagNA Pure LC used with the Trugene HBV Genotyping Kit. J Clin Virol. 2005;34:155–157
  50. Hong SP, Kim NK, Hwang SG, et al. Detection of hepatitis B virus YMDD variants using mass spectrometric analysis of oligonucleotide fragments. J Hepatol. 2004;40:837–844
  51. Hussain M, Fung S, Libbrecht E, et al. Sensitive line probe assay that simultaneously detects mutations conveying resistance to lamivudine and adefovir. J Clin Microbiol. 2006;44:1094–1097
  52. Kim HS, Han KH, Ahn SH, et al. Evaluation of methods for monitoring drug resistance in chronic hepatitis B patients during lamivudine therapy based on mass spectrometry and reverse hybridization. Antivir Ther. 2005;10:441–449
  53. Osiowy C, Villeneuve JP, Heathcote EJ, et al. Detection of rtN236T and rtA181V/T mutations associated with resistance to adefovir dipivoxil in samples from patients with chronic hepatitis B virus infection by the INNO-LiPA HBV DR line probe assay (version 2). J Clin Microbiol. 2006;44:1994–1997
  54. Pallier C, Rodriguez C, Brillet R, et al. Complex dynamics of hepatitis B virus resistance to adefovir dipivoxil. J Hepatol. 2007;46(Suppl 1):S26
  55. Sertoz RY, Erensoy S, Pas S, et al. Comparison of sequence analysis and INNO-LiPA HBV DR line probe assay in patients with chronic hepatitis B. J Chemother. 2005;17:514–520
  56. Pallier C, Castera L, Soulier A, et al. Dynamics of hepatitis B virus resistance to lamivudine. J Virol. 2006;80:643–653
  57. Saldanha J, Gerlich W, Lelie N, et al. An international collaborative study to establish a World Health Organization international standard for hepatitis B virus DNA nucleic acid amplification techniques. Vox Sang. 2001;80:63–71
  58. Pawlotsky JM. Hepatitis B virus (HBV) DNA assays (methods and practical use) and viral kinetics. J Hepatol. 2003;39(Suppl 1):S31–S35
  59. Pawlotsky JM. Molecular diagnosis of viral hepatitis. Gastroenterology. 2002;122:1554–1568
  60. Gordillo RM, Gutierrez J, Casal M. Evaluation of the Cobas TaqMan 48 real-time PCR system for quantitation of hepatitis B virus DNA. J Clin Microbiol. 2005;43:3504–3507
  61. Hochberger S, Althof D, Gallegos de Schrott R, et al. Fully automated quantitation of hepatitis B virus (HBV) DNA in human plasma by the Cobas AmpliPrep/Cobas TaqMan system. J Clin Virol. 2006;35:373–380
  62. Laperche S, Thibault V, Bouchardeau F, et al. Expertise of laboratories in viral load quantification, genotyping, and precore mutant determination for hepatitis B virus in a multicenter study. J Clin Microbiol. 2006;44:3600–3607
  63. Lole KS, Arankalle VA. Quantitation of hepatitis B virus DNA by real-time PCR using internal amplification control and dual TaqMan MGB probes. J Virol Methods. 2006;135:83–90
  64. Ronsin C, Pillet A, Bali C, et al. Evaluation of the Cobas AmpliPrep-total nucleic acid isolation-Cobas TaqMan hepatitis B virus (HBV) quantitative test and comparison to the Versant HBV DNA 3.0 assay. J Clin Microbiol. 2006;44:1390–1399
  65. Sum SS, Wong DK, Yuen JC, et al. Comparison of the Cobas TaqMan HBV test with the COBAS Amplicor monitor test for measurement of hepatitis B virus DNA in serum. J Med Virol. 2005;77:486–490
  66. Punia P, Cane P, Teo CG, et al. Quantitation of hepatitis B lamivudine resistant mutants by real-time amplification refractory mutation system PCR. J Hepatol. 2004;40:986–992
  67. Pawlotsky JM. The concept of hepatitis B virus mutant escape. J Clin Virol. 2005;34(Suppl 1):S125–S129
  68. Villet S, Ollivet A, Pichoud C, et al. Stepwise process for the development of entecavir resistance in a chronic hepatitis B virus infected patient. J Hepatol. 2006;46:531–538
  69. Villet S, Pichoud C, Villeneuve JP, et al. Selection of a multiple drug-resistant hepatitis B virus strain in a liver-transplanted patient. Gastroenterology. 2006;131:1253–1261
  70. Chin R, Shaw T, Torresi J, et al. In vitro susceptibilities of wild-type or drug-resistant hepatitis B virus to (-)-beta-D-2,6-diaminopurine dioxolane and 2′-fluoro-5-methyl-beta-L-arabinofuranosyluracil. Antimicrob Agents Chemother. 2001;45:2495–2501
  71. Delaney WET, Edwards R, Colledge D, et al. Cross-resistance testing of antihepadnaviral compounds using novel recombinant baculoviruses which encode drug-resistant strains of hepatitis B virus. Antimicrob Agents Chemother. 2001;45:1705–1713
  72. Delaney WET, Edwards R, Colledge D, et al. Phenylpropenamide derivatives AT-61 and AT-130 inhibit replication of wild-type and lamivudine-resistant strains of hepatitis B virus in vitro. Antimicrob Agents Chemother. 2002;46:3057–3060
  73. Durantel D, Carrouee-Durantel S, Werle-Lapostolle B, et al. A new strategy for studying in vitro the drug susceptibility of clinical isolates of human hepatitis B virus. Hepatology. 2004;40:855–864
  74. Walters KA, Tipples GA, Allen MI, et al. Generation of stable cell lines expressing lamivudine-resistant hepatitis B virus for antiviral-compound screening. Antimicrob Agents Chemother. 2003;47:1936–1942
  75. Ying C, De Clercq E, Neyts J. Lamivudine, adefovir and tenofovir exhibit long-lasting anti-hepatitis B virus activity in cell culture. J Viral Hepat. 2000;7:79–83
  76. Shaw T, Edwards R, Sozzi T, et al. Large variation in the replication efficiencies of HBV mutants: implications for phenotypic assays for antiviral drug resistance. Hepatology. 2006;44(Suppl 1):252A
  77. Lai CL, Yuen MF. The natural history and treatment of chronic hepatitis B: a critical evaluation of standard treatment criteria and endpoints. Ann Intern Med. 2007;147:58–61
  78. Degertekin B, Lok AS. When to start and stop hepatitis B treatment: can one set of criteria apply to all patients regardless of age at infection?. Ann Intern Med. 2007;147:62–64
  79. Lok AS, Zoulim F, Locarnini S, et al. Antiviral drug-resistant HBV: standardization of nomenclature and assays and recommendations for management. Hepatology. 2007;46:254–265
  80. Pichoud C, Seigneres B, Wang Z, et al. Transient selection of a hepatitis B virus polymerase gene mutant associated with a decreased replication capacity and famciclovir resistance. Hepatology. 1999;29:230–237
  81. Locarnini S, Hatzakis A, Heathcote J, et al. Management of antiviral resistance in patients with chronic hepatitis B. Antivir Ther. 2004;9:679–693
  82. Delaney WET, Ray AS, Yang H, et al. Intracellular metabolism and in vitro activity of tenofovir against hepatitis B virus. Antimicrob Agents Chemother. 2006;50:2471–2477
  83. Haas DW, Smeaton LM, Shafer RW, et al. Pharmacogenetics of long-term responses to antiretroviral regimens containing efavirenz and/or nelfinavir: an Adult Aids Clinical Trials Group Study. J Infect Dis. 2005;192:1931–1942
  84. Joerger M, Bosch TM, Doodeman VD, et al. Novel deoxycytidine kinase gene polymorphisms: a population screening study in Caucasian healthy volunteers. Eur J Clin Pharmacol. 2006;62:681–684
  85. Richman DD. The impact of drug resistance on the effectiveness of chemotherapy for chronic hepatitis B. Hepatology. 2000;32:866–867
  86. Lai CL, Dienstag J, Schiff E, et al. Prevalence and clinical correlates of YMDD variants during lamivudine therapy for patients with chronic hepatitis B. Clin Infect Dis. 2003;36:687–696
  87. Lok AS, Lai CL, Leung N, et al. Long-term safety of lamivudine treatment in patients with chronic hepatitis B. Gastroenterology. 2003;125:1714–1722
  88. Colonno RJ, Rose RE, Baldick CJ, et al. High barrier to resistance results in no emergence of entecavir resistance in nucleoside-naïve subjects during the first two years of therapy. J Hepatol. 2006;44(Suppl 2):S182
  89. Tenney DJ, Levine SM, Rose RE, et al. Clinical emergence of entecavir-resistant hepatitis B virus requires additional substitutions in virus already resistant to lamivudine. Antimicrob Agents Chemother. 2004;48:3498–3507
  90. Tenney DJ, Rose RE, Baldick CJ, et al. Two-year entecavir resistance in lamivudine refractory HBV patients reveals different clinical outcomes depending on the resistance substitutions present. Antimicrob Agents Chemother. 2007;51:902–911
  91. Cane PA, Mutimer D, Ratcliffe D, et al. Analysis of hepatitis B virus quasispecies changes during emergence and reversion of lamivudine resistance in liver transplantation. Antivir Ther. 1999;4:7–14

 Supported in part by the VIRGIL European Network of Excellence on Antiviral Drug Resistance, by a grant (LSHM-CT-2004-503359) from the Priority 1 “Life Sciences, Genomics and Biotechnology for Health” program in the 6th Framework Program of the European Union, and by the French National Agency for AIDS and Viral Hepatitis Research (ANRS) (J.M.P. and F.Z.); in part by the Intramural Research Program of the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health (T.J.L.); and in part by the Extramural Research Program of the National Institutes of Health (RO-1: AIO60449 to S.L.).

 Jean-Michel Pawlotsky acted as a consultant/advisor for Roche (Basel, Switzerland), Gilead (Foster City, CA), Bristol-Myers Squibb (Princeton, NJ), Idenix (Cambridge, MA), and Novartis (Basel, Switzerland), and received research support from Roche and Gilead; Geoffrey Dusheiko received consultancy fees from Glaxo SmithKline (London, UK), Roche, Novartis, Schering Plough (Kenilworth, NJ), Idenix, Gilead Sciences, and Bristol-Myers Squibb, and received research support from the same groups; Daryl Lau acted as a consultant/advisor and has received research grants from Roche, Gilead, Bristol-Myers Squibb, and Novartis; George Lau is a Consultant for Roche and Novartis; Stephen Locarnini has received royalties and is a patent holder for Melbourne Health (Parkville, Victoria, Australia), has received consulting fees from Evivar (Melbourne, Victoria, Australia), Gilead, Pharmasset, and Bristol-Myers Squibb, has received research support from Evivar and Gilead, and has ownership interests in Pharmasset (Princeton, NJ); Paul Martin is a consultant and speaker for Bristol-Myers Squibb, Gilead, Idenix, and Roche; Douglas D. Richman is a consultant for Bristol-Myers Squibb, Gilead, Idenix, and Roche; and Fabien Zoulim is a consultant and has received speaker fees from Gilead, Bristol-Myers Squibb, Idenix, and Novartis.

PII: S0016-5085(07)02113-0

doi: 10.1053/j.gastro.2007.11.036

Gastroenterology
Volume 134, Issue 2 , Pages 405-415 , February 2008