Advances in the Molecular Diagnosis of Hepatitis B Infection: Providing Insight into the Next Generation of Disease

 

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Abstract

Hepatitis B virus (HBV) infection continues to represent a significant health threat, affecting over 400 million people worldwide. Historically, the diagnosis and treatment of chronic hepatitis B (CHB) relied in detection of the hepatitis B surface antigen (HBsAg) and more recently realtime polymerase chain reaction (PCR) analysis. The advent of novel technologies and equipment for the identification and staging of the different stages of HBV infection has resulted in dramatic changes to patient monitoring and management. Through the use of rapid, quantitative HBsAg immunoassays, it is now possible to predict the likelihood of patient response to treatment as well as the clinical course of disease. Ultradeep sequencing technologies (also known as next-generation sequencing) overcome many of the traditional limitations associated with population-based sequencing approaches, and have provided significant insight into the viral response to therapeutic intervention and the molecular pathogenesis of CHB. The authors discuss recent developments in the molecular diagnosis of HBV infection, as well as potential advantages and caveats resultant of this rapid progression of technology.

• References

• 1 Blumberg BS, Alter HJ, Visnich SA. “New” Antigen in Leukemia Sera. JAMA 1965; 191: 541-546
  CrossrefPubMedGoogle Scholar
• 2 Chen YC, Sheen IS, Chu CM, Liaw YF. Prognosis following spontaneous HBsAg seroclearance in chronic hepatitis B patients with or without concurrent infection. Gastroenterology 2002; 123 (4) 1084-1089
  CrossrefPubMedGoogle Scholar
• 3 Fattovich G, Giustina G, Sanchez-Tapias J , et al. Delayed clearance of serum HBsAg in compensated cirrhosis B: relation to interferon alpha therapy and disease prognosis. European Concerted Action on Viral Hepatitis (EUROHEP). Am J Gastroenterol 1998; 93 (6) 896-900
  CrossrefPubMedGoogle Scholar
• 4 Hsu YS, Chien RN, Yeh CT , et al. Long-term outcome after spontaneous HBeAg seroconversion in patients with chronic hepatitis B. Hepatology 2002; 35 (6) 1522-1527
  CrossrefPubMedGoogle Scholar
• 5 Stibbe W, Gerlich WH. Variable protein composition of hepatitis B surface antigen from different donors. Virology 1982; 123 (2) 436-442
  CrossrefPubMedGoogle Scholar
• 6 Seeger C, Mason WS. Hepatitis B virus biology. Microbiol Mol Biol Rev 2000; 64 (1) 51-68
  CrossrefPubMedGoogle Scholar
• 7 Vanlandschoot P, Leroux-Roels G. Viral apoptotic mimicry: an immune evasion strategy developed by the hepatitis B virus?. Trends Immunol 2003; 24 (3) 144-147
  CrossrefPubMedGoogle Scholar
• 8 Madalinski K, Burczynska B, Heermann KH, Uy A, Gerlich WH. Analysis of viral proteins in circulating immune complexes from chronic carriers of hepatitis B virus. Clin Exp Immunol 1991; 84 (3) 493-500
  PubMedGoogle Scholar
• 9 Bill CA, Summers J. Genomic DNA double-strand breaks are targets for hepadnaviral DNA integration. Proc Natl Acad Sci U S A 2004; 101 (30) 11135-11140
  CrossrefPubMedGoogle Scholar
• 10 Kimbi GC, Kramvis A, Kew MC. Integration of hepatitis B virus DNA into chromosomal DNA during acute hepatitis B. World J Gastroenterol 2005; 11 (41) 6416-6421
  PubMedGoogle Scholar
• 11 Deguchi M, Yamashita N, Kagita M , et al. Quantitation of hepatitis B surface antigen by an automated chemiluminescent microparticle immunoassay. J Virol Methods 2004; 115 (2) 217-222
  CrossrefPubMedGoogle Scholar
• 12 Mühlbacher A, Weber B, Bürgisser P , et al. Multicenter study of a new fully automated HBsAg screening assay with enhanced sensitivity for the detection of HBV mutants. Med Microbiol Immunol (Berl) 2008; 197 (1) 55-64
  PubMedGoogle Scholar
• 13 Zacher BJ, Moriconi F, Bowden S , et al. Multicenter evaluation of the Elecsys hepatitis B surface antigen quantitative assay. Clin Vaccine Immunol 2011; 18 (11) 1943-1950
  CrossrefPubMedGoogle Scholar
• 14 Sonneveld MJ, Rijckborst V, Boucher CA , et al. A comparison of two assays for quantification of hepatitis B surface antigen in patients with chronic hepatitis B. J Clin Virol 2011; 51 (3) 175-178
  CrossrefPubMedGoogle Scholar
• 15 Wursthorn K, Jaroszewicz J, Zacher BJ , et al. Correlation between the Elecsys HBsAg II assay and the Architect assay for the quantification of hepatitis B surface antigen (HBsAg) in the serum. J Clin Virol 2011; 50 (4) 292-296
  CrossrefPubMedGoogle Scholar
• 16 Nowak MA, Bonhoeffer S, Hill AM, Boehme R, Thomas HC, McDade H. Viral dynamics in hepatitis B virus infection. Proc Natl Acad Sci U S A 1996; 93 (9) 4398-4402
  CrossrefPubMedGoogle Scholar
• 17 Thompson AJ, Nguyen T, Iser D , et al. Serum hepatitis B surface antigen and hepatitis B e antigen titers: disease phase influences correlation with viral load and intrahepatic hepatitis B virus markers. Hepatology 2010; 51 (6) 1933-1944
  CrossrefPubMedGoogle Scholar
• 18 Volz T, Lutgehetmann M, Wachtler P , et al. Impaired intrahepatic hepatitis B virus productivity contributes to low viremia in most HBeAg-negative patients. Gastroenterology 2007; 133 (3) 843-852
  CrossrefPubMedGoogle Scholar
• 19 Chan HL, Wong VW, Wong GL, Tse CH, Chan HY, Sung JJ. A longitudinal study on the natural history of serum hepatitis B surface antigen changes in chronic hepatitis B. Hepatology 2010; 52 (4) 1232-1241
  CrossrefPubMedGoogle Scholar
• 20 Jaroszewicz J, Calle Serrano B, Wursthorn K , et al. Hepatitis B surface antigen (HBsAg) levels in the natural history of hepatitis B virus (HBV)-infection: a European perspective. J Hepatol 2010; 52 (4) 514-522
  CrossrefPubMedGoogle Scholar
• 21 Kim YJ, Cho HC, Choi MS , et al. The change of the quantitative HBsAg level during the natural course of chronic hepatitis B. Liver Int 2011; 31 (6) 817-823
  CrossrefPubMedGoogle Scholar
• 22 Nguyen T, Thompson AJ, Bowden S , et al. Hepatitis B surface antigen levels during the natural history of chronic hepatitis B: a perspective on Asia. J Hepatol 2010; 52 (4) 508-513
  CrossrefPubMedGoogle Scholar
• 23 Pollicino T, Amaddeo G, Restuccia A , et al. Impact of hepatitis B virus (HBV) preS/S genomic variability on HBV surface antigen and HBV DNA serum levels. Hepatology 2012; 56 (2) 434-443
  CrossrefPubMedGoogle Scholar
• 24 Brunetto MR. A new role for an old marker, HBsAg. J Hepatol 2010; 52 (4) 475-477
  CrossrefPubMedGoogle Scholar
• 25 Brunetto MR, Oliveri F, Colombatto P , et al. Hepatitis B surface antigen serum levels help to distinguish active from inactive hepatitis B virus genotype D carriers. Gastroenterology 2010; 139 (2) 483-490
  CrossrefPubMedGoogle Scholar
• 26 Chen CH, Lee CM, Wang JH, Tung HD, Hung CH, Lu SN. Correlation of quantitative assay of hepatitis B surface antigen and HBV DNA levels in asymptomatic hepatitis B virus carriers. Eur J Gastroenterol Hepatol 2004; 16 (11) 1213-1218
  CrossrefPubMedGoogle Scholar
• 27 Brunetto MR, Moriconi F, Bonino F , et al. Hepatitis B virus surface antigen levels: a guide to sustained response to peginterferon alfa-2a in HBeAg-negative chronic hepatitis B. Hepatology 2009; 49 (4) 1141-1150
  CrossrefPubMedGoogle Scholar
• 28 Moucari R, Mackiewicz V, Lada O , et al. Early serum HBsAg drop: a strong predictor of sustained virological response to pegylated interferon alfa-2a in HBeAg-negative patients. Hepatology 2009; 49 (4) 1151-1157
  CrossrefPubMedGoogle Scholar
• 29 Sonneveld MJ, Rijckborst V, Boucher CA, Hansen BE, Janssen HL. Prediction of sustained response to peginterferon alfa-2b for hepatitis B e antigen-positive chronic hepatitis B using on-treatment hepatitis B surface antigen decline. Hepatology 2010; 52 (4) 1251-1257
  CrossrefPubMedGoogle Scholar
• 30 Chan HL, Thompson A, Martinot-Peignoux M , et al. Hepatitis B surface antigen quantification: why and how to use it in 2011—a core group report. J Hepatol 2011; 55 (5) 1121-1131
  CrossrefPubMedGoogle Scholar
• 31 Chan HL, Wong VW, Chim AM, Chan HY, Wong GL, Sung JJ. Serum HBsAg quantification to predict response to peginterferon therapy of e antigen positive chronic hepatitis B. Aliment Pharmacol Ther 2010; 32 (11-12) 1323-1331
  CrossrefPubMedGoogle Scholar
• 32 Rijckborst V, Hansen BE, Cakaloglu Y , et al. Early on-treatment prediction of response to peginterferon alfa-2a for HBeAg-negative chronic hepatitis B using HBsAg and HBV DNA levels. Hepatology 2010; 52 (2) 454-461
  CrossrefPubMedGoogle Scholar
• 33 Chevaliez S, Hézode C, Bahrami S, Grare M, Pawlotsky JM. Long-term hepatitis B surface antigen (HBsAg) kinetics during nucleoside/nucleotide analogue therapy: Finite treatment duration unlikely. J Hepatol 2012; ; pii: S0168-8278(12)00909-9.
  PubMedGoogle Scholar
• 34 Beerenwinkel N, Zagordi O. Ultra-deep sequencing for the analysis of viral populations. Curr Opin Virol 2011; 1 (5) 413-418
  CrossrefPubMedGoogle Scholar
• 35 Radford AD, Chapman D, Dixon L, Chantrey J, Darby AC, Hall N. Application of next-generation sequencing technologies in virology. J Gen Virol 2012; 93 (Pt 9) 1853-1868
  CrossrefPubMedGoogle Scholar
• 36 Metzker ML. Sequencing technologies–the next generation. Nat Rev Genet 2010; 11 (1) 31-46
  CrossrefPubMedGoogle Scholar
• 37 Brockman W, Alvarez P, Young S , et al. Quality scores and SNP detection in sequencing-by-synthesis systems. Genome Res 2008; 18 (5) 763-770
  CrossrefPubMedGoogle Scholar
• 38 Schadt EE, Turner S, Kasarskis A. A window into third-generation sequencing. Hum Mol Genet 2010; 19 (R2) R227-R240
  CrossrefPubMedGoogle Scholar
• 39 Loman NJ, Misra RV, Dallman TJ , et al. Performance comparison of benchtop high-throughput sequencing platforms. Nat Biotechnol 2012; 30 (5) 434-439
  CrossrefPubMedGoogle Scholar
• 40 Margulies M, Egholm M, Altman WE , et al. Genome sequencing in microfabricated high-density picolitre reactors. Nature 2005; 437 (7057) 376-380
  PubMedGoogle Scholar
• 41 Homs M, Buti M, Quer J , et al. Ultra-deep pyrosequencing analysis of the hepatitis B virus preCore region and main catalytic motif of the viral polymerase in the same viral genome. Nucleic Acids Res 2011; 39 (19) 8457-8471
  CrossrefPubMedGoogle Scholar
• 42 Margeridon-Thermet S, Shulman NS, Ahmed A , et al. Ultra-deep pyrosequencing of hepatitis B virus quasispecies from nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI)-treated patients and NRTI-naive patients. J Infect Dis 2009; 199 (9) 1275-1285
  CrossrefPubMedGoogle Scholar
• 43 Reuman EC, Margeridon-Thermet S, Caudill HB , et al. A classification model for G-to-A hypermutation in hepatitis B virus ultra-deep pyrosequencing reads. Bioinformatics 2010; 26 (23) 2929-2932
  CrossrefPubMedGoogle Scholar
• 44 Bentley DR, Balasubramanian S, Swerdlow HP , et al. Accurate whole human genome sequencing using reversible terminator chemistry. Nature 2008; 456 (7218) 53-59
  CrossrefPubMedGoogle Scholar
• 45 Nishijima N, Marusawa H, Ueda Y , et al. Dynamics of hepatitis B virus quasispecies in association with nucleos(t)ide analogue treatment determined by ultra-deep sequencing. PLoS ONE 2012; 7 (4) e35052
  CrossrefPubMedGoogle Scholar
• 46 Ronaghi M, Uhlén M, Nyrén P. A sequencing method based on real-time pyrophosphate. Science 1998; 281 (5375) 363 , 365
  CrossrefPubMedGoogle Scholar
• 47 Lindström A, Odeberg J, Albert J. Pyrosequencing for detection of lamivudine-resistant hepatitis B virus. J Clin Microbiol 2004; 42 (10) 4788-4795
  CrossrefPubMedGoogle Scholar
• 48 Osiowy C, Giles E, Tanaka Y, Mizokami M, Minuk GY. Molecular evolution of hepatitis B virus over 25 years. J Virol 2006; 80 (21) 10307-10314
  CrossrefPubMedGoogle Scholar
• 49 Sede M, Ojeda D, Cassino L , et al. Long-term monitoring drug resistance by ultra-deep pyrosequencing in a chronic hepatitis B virus (HBV)-infected patient exposed to several unsuccessful therapy schemes. Antiviral Res 2012; 94 (2) 184-187
  CrossrefPubMedGoogle Scholar
• 50 Rodriguez-Frías F, Tabernero D, Quer J , et al. Ultra-deep pyrosequencing detects conserved genomic sites and quantifies linkage of drug-resistant amino acid changes in the hepatitis B virus genome. PLoS ONE 2012; 7 (5) e37874
  CrossrefPubMedGoogle Scholar
• 51 Wang C, Mitsuya Y, Gharizadeh B, Ronaghi M, Shafer RW. Characterization of mutation spectra with ultra-deep pyrosequencing: application to HIV-1 drug resistance. Genome Res 2007; 17 (8) 1195-1201
  CrossrefPubMedGoogle Scholar
• 52 Varghese V, Shahriar R, Rhee S-Y , et al. Minority variants associated with transmitted and acquired HIV-1 nonnucleoside reverse transcriptase inhibitor resistance: implications for the use of second-generation nonnucleoside reverse transcriptase inhibitors. J Acquir Immune Defic Syndr 2009; 52 (3) 309-315
  CrossrefPubMedGoogle Scholar
• 53 Kao J-H. Hepatitis B viral genotypes: clinical relevance and molecular characteristics. J Gastroenterol Hepatol 2002; 17 (6) 643-650
  CrossrefPubMedGoogle Scholar
• 54 Solmone M, Vincenti D, Prosperi MCF, Bruselles A, Ippolito G, Capobianchi MR. Use of massively parallel ultradeep pyrosequencing to characterize the genetic diversity of hepatitis B virus in drug-resistant and drug-naive patients and to detect minor variants in reverse transcriptase and hepatitis B S antigen. J Virol 2009; 83 (4) 1718-1726
  CrossrefPubMedGoogle Scholar
• 55 Akuta N, Kumada H. Influence of hepatitis B virus genotypes on the response to antiviral therapies. J Antimicrob Chemother 2005; 55 (2) 139-142
  CrossrefPubMedGoogle Scholar
• 56 Sugauchi F, Orito E, Ichida T , et al. Epidemiologic and virologic characteristics of hepatitis B virus genotype B having the recombination with genotype C. Gastroenterology 2003; 124 (4) 925-932
  CrossrefPubMedGoogle Scholar
• 57 Kew MC, Kramvis A, Yu MC, Arakawa K, Hodkinson J. Increased hepatocarcinogenic potential of hepatitis B virus genotype A in Bantu-speaking sub-saharan Africans. J Med Virol 2005; 75 (4) 513-521
  CrossrefPubMedGoogle Scholar
• 58 Lok AS, Akarca U, Greene S. Mutations in the pre-core region of hepatitis B virus serve to enhance the stability of the secondary structure of the pre-genome encapsidation signal. Proc Natl Acad Sci U S A 1994; 91 (9) 4077-4081
  CrossrefPubMedGoogle Scholar
• 59 Zhao J, Guo Y, Yan Z, Liang P, Zhang J, Liu Y. The natural YMDD mutations of hepatitis B virus in Western China. Scand J Infect Dis 2012; 44 (1) 44-47
  CrossrefPubMedGoogle Scholar
• 60 Ijaz S, Arnold C, Dervisevic S , et al. Dynamics of lamivudine-resistant hepatitis B virus during adefovir monotherapy versus lamivudine plus adefovir combination therapy. J Med Virol 2008; 80 (7) 1160-1170
  CrossrefPubMedGoogle Scholar
• 61 Warner N, Locarnini S. The antiviral drug selected hepatitis B virus rtA181T/sW172* mutant has a dominant negative secretion defect and alters the typical profile of viral rebound. Hepatology 2008; 48 (1) 88-98
  CrossrefPubMedGoogle Scholar
• 62 Sheldon J, Ramos B, Garcia-Samaniego J , et al. Selection of hepatitis B virus (HBV) vaccine escape mutants in HBV-infected and HBV/HIV-coinfected patients failing antiretroviral drugs with anti-HBV activity. J Acquir Immune Defic Syndr 2007; 46 (3) 279-282
  CrossrefPubMedGoogle Scholar
• 63 Lavanchy D. Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures. J Viral Hepat 2004; 11 (2) 97-107
  CrossrefPubMedGoogle Scholar
• 64 Henry M, Guétard D, Suspène R, Rusniok C, Wain-Hobson S, Vartanian JP. Genetic editing of HBV DNA by monodomain human APOBEC3 cytidine deaminases and the recombinant nature of APOBEC3G. PLoS ONE 2009; 4 (1) e4277
  CrossrefPubMedGoogle Scholar
• 65 Noguchi C, Ishino H, Tsuge M , et al. G to A hypermutation of hepatitis B virus. Hepatology 2005; 41 (3) 626-633
  CrossrefPubMedGoogle Scholar
• 66 Vartanian J-P, Henry M, Marchio A , et al. Massive APOBEC3 editing of hepatitis B viral DNA in cirrhosis. PLoS Pathog 2010; 6 (5) e1000928
  CrossrefPubMedGoogle Scholar
• 67 Pareek CS, Smoczynski R, Tretyn A. Sequencing technologies and genome sequencing. J Appl Genet 2011; 52 (4) 413-435
  CrossrefPubMedGoogle Scholar