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Research Article| Volume 75, ISSUE 2, P302-310, August 2021

Clinical efficacy of a novel, high-sensitivity HBcrAg assay in the management of chronic hepatitis B and HBV reactivation

Published:March 21, 2021DOI:https://doi.org/10.1016/j.jhep.2021.02.017

      Highlights

      • A high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) has been developed.
      • iTACT-HBcrAg was about 10-fold more sensitive than the conventional HBcrAg assay.
      • HBcrAg could be detected prior to HBV reactivation in most patients.
      • iTACT-HBcrAg should be used as a surrogate marker for disease progression and treatment response.

      Background & Aims

      A fully automated, novel high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) has been developed. We demonstrate the clinical utility of iTACT-HBcrAg for monitoring chronic hepatitis B (CHB) and for the early detection of HBV reactivation.

      Methods

      After fundamental assessments, the clinical performance of iTACT-HBcrAg was compared with other HBV markers. i) Serial sera, available from 161 HBeAg-negative patients with CHB and persistently undetectable HBV DNA, were measured by iTACT-HBcrAg and a conventional HBcrAg assay (G-HBcrAg). ii) Serial sera from 13 HBV-reactivated patients were measured by iTACT-HBcrAg and an ultra-high-sensitivity HBsAg immune complex transfer-chemiluminescent enzyme immunoassay (lower limit of detection; 0.0005 IU/ml, ICT-CLEIA) to compare HBV DNA detection. iii) To elucidate the various HBcrAg components detected by iTACT-HBcrAg, OptiPrep density gradient centrifugation analysis was performed on sera obtained before and after HBV reactivation.

      Results

      The analytical performance of iTACT-HBcrAg was satisfactory. The sensitivity of iTACT-HBcrAg (2.1 Log U/ml) was approximately 10-fold greater than that of G-HBcrAg (2.8 Log U/ml). i) HBcrAg was detectable in the sera of 97.5% (157/161) of patients with CHB by iTACT-HBcrAg, of whom 75.2% (121/161) had ≥2.8 Log U/ml HBcrAg and 22.4% (36/161) had 2.1–2.8 Log U/ml HBcrAg, which was undetectable by G-HBcrAg. ii) 9 and 2 of 13 HBV-reactivated patients were HBcrAg-positive by iTACT-HBcrAg before and at HBV DNA positivity, respectively; 7 and 4 were HBcrAg-positive by iTACT-HBcrAg before and at HBsAg-positivity by ICT-CLEIA, respectively. iii) The HBcrAg detected by iTACT-HBcrAg before HBV reactivation was contained in empty particles (22 KDa precore protein).

      Conclusions

      iTACT-HBcrAg could be used to better monitor responses to anti-HBV treatments in HBeAg-negative patients and for the early detection of HBV reactivation.

      Lay summary

      A fully automated, novel high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) has been developed. iTACT-HBcrAg can be used to monitor HBeAg-negative patients with chronic hepatitis B, as well as for the early detection of HBV reactivation. iTACT-HBcrAg could be used as a general marker of disease progression and treatment response.

      Graphical abstract

      Keywords

      Linked Article

      • Reply to: “Understanding HBcrAg components improves the interpretation of clinical HBcrAg assay results”
        Journal of HepatologyVol. 75Issue 4
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          We thank Hong and Hu1 for their interest in our article2 and we also thank the Journal of Hepatology for the opportunity to reply to their thoughtful discussion on the nature of HBcrAg. We appreciate their questions, but the discussion does not influence our major conclusion that the novel high-sensitivity HBcrAg quantitative assay, iTACT-HBcrAg, could be useful to monitor responses in HBeAg-negative patients and detect HBV reactivation.
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      • Understanding HBcrAg components improves the interpretation of clinical HBcrAg assay results
        Journal of HepatologyVol. 75Issue 4
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          We read with great interest the paper by Inoue et al. on the development of a novel high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) and its application for monitoring hepatitis B e antigen (HBeAg)-negative patients and HBV reactivation.1 Without a doubt, this high-sensitivity HBcrAg assay improves the sensitivity of the current HBcrAg assay and is timely for the ongoing pursuit of an HBV cure. However, a few important issues need to be clarified.
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      References

        • World Health Organization (WHO)
        Hepatitis B factsheet. WHO web site.
        • Lai C.L.
        • Yuen M.F.
        The natural history and treatment of chronic hepatitis B: a critical evaluation of standard treatment criteria and end points.
        Ann Intern Med. 2007; 147: 58-61
        • Lai C.L.
        • Wong D.
        • Ip P.
        • Kopaniszen M.
        • Seto W.K.
        • Fung J.
        • et al.
        Reduction of covalently closed circular DNA with long-term nucleos(t)ide analogue treatment in chronic hepatitis B.
        J Hepatol. 2017; 66: 275-281
        • Inoue T.
        • Tanaka Y.
        The role of hepatitis B core-related antigen.
        Genes. 2019; 10: 357
        • Mak L.Y.
        • Wong D.K.
        • Cheung K.S.
        • Seto W.K.
        • Lai C.L.
        • Yuen M.F.
        Review article: hepatitis B core-related antigen (HBcrAg): an emerging marker for chronic hepatitis B virus infection.
        Aliment Pharmacol Ther. 2018; 47: 43-54
        • Rokuhara A.
        • Tanaka E.
        • Matsumoto A.
        • Kimura T.
        • Yamaura T.
        • Orii K.
        • et al.
        Clinical evaluation of a new enzyme immunoassay for hepatitis B virus core-related antigen; a marker distinct from viral DNA for monitoring lamivudine treatment.
        J Viral Hepat. 2003; 10: 324-330
        • Wong D.K.
        • Seto W.K.
        • Cheung K.S.
        • Chong C.K.
        • Huang F.Y.
        • Fung J.
        • et al.
        Hepatitis B virus core-related antigen as a surrogate marker for covalently closed circular DNA.
        Liver Int. 2017; 37: 995-1001
      1. EASL 2017 Clinical Practice Guidelines on the management of hepatitis B virus infection.
        J Hepatol. 2017; 67: 370-398
        • Mak L.Y.
        • Seto W.K.
        • Fung J.
        • Yuen M.F.
        New biomarkers of chronic hepatitis B.
        Gut Liver. 2019; 13: 589-595
        • Kumada T.
        • Toyoda H.
        • Tada T.
        • Kiriyama S.
        • Tanikawa M.
        • Hisanaga Y.
        • et al.
        Effect of nucleos(t)ide analogue therapy on hepatocarcinogenesis in chronic hepatitis B patients: a propensity score analysis.
        J Hepatol. 2013; 58: 427-433
        • Hosaka T.
        • Suzuki F.
        • Kobayashi M.
        • Fujiyama S.
        • Kawamura Y.
        • Sezaki H.
        • et al.
        Impact of hepatitis B core-related antigen on the incidence of hepatocellular carcinoma in patients treated with nucleos(t)ide analogues.
        Aliment Pharmacol Ther. 2019; 49: 457-471
        • Suzuki Y.
        • Maekawa S.
        • Komatsu N.
        • Sato M.
        • Tatsumi A.
        • Miura M.
        • et al.
        Hepatitis B virus (HBV)-infected patients with low hepatitis B surface antigen and high hepatitis B core-related antigen titers have a high risk of HBV-related hepatocellular carcinoma.
        Hepatol Res. 2019; 49: 51-63
        • Shinkai N.
        • Matsuura K.
        • Sugauchi F.
        • Watanabe T.
        • Murakami S.
        • Iio E.
        • et al.
        Application of a newly developed high-sensitivity HBsAg chemiluminescent enzyme immunoassay for hepatitis B patients with HBsAg seroclearance.
        J Clin Microbiol. 2013; 51: 3484-3491
        • Kusumoto S.
        • Tanaka Y.
        • Suzuki R.
        • Watanabe T.
        • Nakata M.
        • Sakai R.
        • et al.
        Ultra-high sensitivity HBsAg assay can diagnose HBV reactivation following rituximab-based therapy in patients with lymphoma.
        J Hepatol. 2020; 73: 285-293
        • Shinkai N.
        • Kusumoto S.
        • Murakami S.
        • Ogawa S.
        • Ri M.
        • Matsui T.
        • et al.
        Novel monitoring of hepatitis B reactivation based on ultra-high sensitive hepatitis B surface antigen assay.
        Liver Int. 2017; 37: 1138-1147
        • Kimura T.
        • Rokuhara A.
        • Sakamoto Y.
        • Yagi S.
        • Tanaka E.
        • Kiyosawa K.
        • et al.
        Sensitive enzyme immunoassay for hepatitis B virus core-related antigens and their correlation to virus load.
        J Clin Microbiol. 2002; 40: 439-445
        • Kimura T.
        • Rokuhara A.
        • Matsumoto A.
        • Yagi S.
        • Tanaka E.
        • Kiyosawa K.
        • et al.
        New enzyme immunoassay for detection of hepatitis B virus core antigen (HBcAg) and relation between levels of HBcAg and HBV DNA.
        J Clin Microbiol. 2003; 41: 1901-1906
        • Kimura T.
        • Ohno N.
        • Terada N.
        • Rokuhara A.
        • Matsumoto A.
        • Yagi S.
        • et al.
        Hepatitis B virus DNA-negative dane particles lack core protein but contain a 22-kDa precore protein without C-terminal arginine-rich domain.
        J Biol Chem. 2005; 280: 21713-21719
        • Aguiar J.
        • Garcia G.
        • Leon Y.
        • Canales E.
        • Angel Silva J.
        • Gell O.
        • et al.
        High functional stability of a low-cost HBV DNA qPCR primer pair and plasmid standard.
        Euroasian J Hepatogastroenterol. 2016; 6: 19-24
        • Wong D.K.
        • Tanaka Y.
        • Lai C.L.
        • Mizokami M.
        • Fung J.
        • Yuen M.F.
        Hepatitis B virus core-related antigens as markers for monitoring chronic hepatitis B infection.
        J Clin Microbiol. 2007; 45: 3942-3947
        • Rokuhara A.
        • Sun X.
        • Tanaka E.
        • Kimura T.
        • Matsumoto A.
        • Yao D.
        • et al.
        Hepatitis B virus core and core-related antigen quantitation in Chinese patients with chronic genotype B and C hepatitis B virus infection.
        J Gastroenterol Hepatol. 2005; 20: 1726-1730
        • Wong D.K.
        • Seto W.K.
        • Cheung K.S.
        • Chong C.K.
        • Huang F.Y.
        • Fung J.
        • et al.
        Hepatitis B virus core-related antigen as a surrogate marker for covalently closed circular DNA.
        Liver Int. 2017; 37: 995-1001
        • Chuaypen N.
        • Posuwan N.
        • Payungporn S.
        • Tanaka Y.
        • Shinkai N.
        • Poovorawan Y.
        • et al.
        Serum hepatitis B core-related antigen as a treatment predictor of pegylated interferon in patients with HBeAg-positive chronic hepatitis B.
        Liver Int. 2016; 36: 827-836
        • Seto W.K.
        • Wong D.K.
        • Fung J.
        • Huang F.Y.
        • Liu K.S.
        • Lai C.L.
        • et al.
        Linearized hepatitis B surface antigen and hepatitis B core-related antigen in the natural history of chronic hepatitis B.
        Clin Microbiol Infect. 2014; 20: 1173-1180
        • Seto W.K.
        • Tanaka Y.
        • Wong D.K.
        • Lai C.L.
        • Shinkai N.
        • Yuen J.C.
        • et al.
        Evidence of serologic activity in chronic hepatitis B after surface antigen (HBsAg) seroclearance documented by conventional HBsAg assay.
        Hepatol Int. 2012; 7: 98-105
        • Corcuera A.
        • Stolle K.
        • Hillmer S.
        • Seitz S.
        • Lee J.Y.
        • Bartenschlager R.
        • et al.
        Novel non-heteroarylpyrimidine (HAP) capsid assembly modifiers have a different mode of action from HAPs in vitro.
        Antivir Res. 2018; 158: 135-142
        • Seto W.K.
        • Wong D.K.
        • Chan T.S.
        • Hwang Y.Y.
        • Fung J.
        • Liu K.S.
        • et al.
        Association of hepatitis B core-related antigen with hepatitis B virus reactivation in occult viral carriers undergoing high-risk immunosuppressive therapy.
        Am J Gastroenterol. 2016; 111: 1788-1795
        • Iossa D.
        • Vitrone M.
        • Liotti A.
        • Portella G.
        • Durante-Mangoni E.
        Hepatitis B core-related antigen to detect hepatitis B virus (HBV) reactivation in heart transplant recipients with past HBV infection: a pilot study.
        Clin Transpl. 2019; 33e13574
        • Ning X.
        • Luckenbaugh L.
        • Liu K.
        • Bruss V.
        • Sureau C.
        • Hu J.
        Common and distinct capsid and surface protein requirements for secretion of complete and genome-free hepatitis B virions.
        J Virol. 2018; 92 (e00272-18)
        • Yaginuma K.
        • Shirakata Y.
        • Kobayashi M.
        • Koike K.
        Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA.
        Proc Natl Acad Sci U S A. 1987; 84: 2678-2682
        • Yoshida K.
        • Desbiolles A.
        • Feldman S.F.
        • Ahn S.H.
        • Alidjinou E.K.
        • Atsukawa M.
        • et al.
        Assay for hepatitis B core-related antigen identify patients with high viral load: systematic review and meta-analysis of individual participant data.
        Clin Gastroenterol Hepatol. 2021; 19: 46-60.e8