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Novel and highly sensitive immunoassay for total hepatitis B surface antigen, including that complexed with hepatitis B surface antibody

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
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Abstract

Background

Hepatitis B surface antigen (HBsAg) is used as a clinical marker of hepatitis B virus (HBV) infection. However, conventional HBsAg assays have so far failed to accurately detect HBsAg in blood because of interference by patient-derived antibodies against HBsAg (HBsAb).

Methods

We developed a novel, fully automated assay system that can detect total HBsAg in blood, including antigens complexed with HBsAb. The immunoassay inactivates HBsAb via a simple pretreatment step to dissociate the HBsAg molecule from HBsAg–HBsAb complexes and thereby estimate total HBsAg. Accordingly, the test has been termed the “immunoassay for total antigen including complex via pretreatment (iTACT)-HBsAg.”

Results

The recovery rate of HBsAg in the presence of HBsAb was greater than 87 % at a cutoff value set at 5.0 mIU/mL on the basis of data from 545 healthy controls. Analyses using serial serum samples from 25 HBV carriers who became negative for HBsAg during follow-up showed that the iTACT-HBsAg could detect HBsAg over a period of years despite a loss in detection by conventional assays and was able to detect HBsAg in 39 (53 %) of 73 samples with HBsAb.

Conclusions

The new iTACT-HBsAg assay appears to detect total HBsAg with high sensitivity, even in the presence of HBsAb, and may useful in identifying subclinical or occult HBV carriers.

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Abbreviations

ALP:

Alkaline phosphatase

CLEIA:

Chemiluminescent enzyme immunoassay

HBcAb:

Hepatitis B core antibody

HBsAb:

Hepatitis B surface antibody

HBsAg:

Hepatitis B surface antigen

HBV:

Hepatitis B virus

HCV:

Hepatitis C virus

iTACT:

Immunoassay for total antigen including complex via pretreatment

References

  1. World Health Organization. Hepatitis B. Fact sheet no 204. http://www.who.int/mediacentre/factsheets/fs204/en/. Accessed 7 Jan 2016.

  2. Liaw YF, Chu CM. Hepatitis B virus infection. Lancet. 2009;373:582–92.

    Article  CAS  PubMed  Google Scholar 

  3. Lozano R, Naghavi M, Foreman K, et al. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet. 2012;380:2095–128.

    Article  PubMed  Google Scholar 

  4. Hoofnagle JH, Doo E, Liang TJ, et al. Management of hepatitis B: summary of a clinical research workshop. Hepatology. 2007;45:1056–75.

    Article  CAS  PubMed  Google Scholar 

  5. Rehermann B, Ferrari C, Pasquinelli C, et al. The hepatitis B virus persists for decades after patients’ recovery from acute viral hepatitis despite active maintenance of a cytotoxic T-lymphocyte response. Nat Med. 1996;2:1104–8.

    Article  CAS  PubMed  Google Scholar 

  6. Seto WK, Tanaka Y, Wong DK, 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.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Marusawa H, Uemoto S, Hijikata M, et al. Latent hepatitis B virus infection in healthy individuals with antibodies to hepatitis B core antigen. Hepatology. 2000;31:488–95.

    Article  CAS  PubMed  Google Scholar 

  8. Umemura T, Tanaka E, Kiyosawa K, et al. Mortality secondary to fulminant hepatic failure in patients with prior resolution of hepatitis B virus infection in Japan. Clin Infect Dis. 2008;47:e52–6.

    Article  PubMed  Google Scholar 

  9. Kusumoto S, Tanaka Y, Suzuki R, et al. Monitoring of hepatitis B virus (HBV) DNA and risk of HBV reactivation in B-cell lymphoma: a prospective observational study. Clin Infect Dis. 2015;61:719–29.

    Article  CAS  PubMed  Google Scholar 

  10. Drafting Committee for Hepatitis Management Guidelines, Japan Society of Hepatology. JSH guidelines for the management of hepatitis B virus infection. Hepatol Res. 2014;44(Suppl S1):1–58.

  11. Terrault NA, Bzowej NH, Chang KM, et al. AASLD guidelines for treatment of chronic hepatitis B. Hepatology. 2016;63:261–83.

    Article  PubMed  Google Scholar 

  12. Yeo W, Johnson PJ. Diagnosis, prevention and management of hepatitis B virus reactivation during anticancer therapy. Hepatology. 2006;43:209–20.

    Article  CAS  PubMed  Google Scholar 

  13. Shinkai N, Matsuura K, Sugauchi F, et al. Application of a newly developed high-sensitivity HBsAg chemiluminescent enzyme immunoassay for hepatitis B patients with HBsAg seroclearance. J Clin Microbiol. 2013;51:3484–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Deguchi M, Yamashita N, Kagita M, et al. Quantitation of hepatitis B surface antigen by an automated chemiluminescent microparticle immunoassay. J Virol Methods. 2004;115:217–22.

    Article  CAS  PubMed  Google Scholar 

  15. Takiwaki M, Uji Y, Kitajima I. Evaluation of HBs antigen chemiluminescence enzyme immunoassay for chronic hepatitis B patients. J Anal Bio Sci. 2012;35:127–32.

    CAS  Google Scholar 

  16. Nishizono I, Iida S, Suzuki N, et al. Rapid and sensitive chemiluminescent enzyme immunoassay for measuring tumor markers. Clin Chem. 1991;37:1639–44.

    CAS  PubMed  Google Scholar 

  17. Yoshitake S, Imagawa M, Ishikawa E, et al. Mild and efficient conjugation of rabbit Fab’ and horseradish peroxidase using a maleimide compound and its use for enzyme immunoassay. J Biochem. 1982;92:1413–24.

    Article  CAS  PubMed  Google Scholar 

  18. Sugauchi F, Mizokami M, Orito E, et al. A novel variant genotype C of hepatitis B virus identified in isolates from Australian Aborigines: complete genome sequence and phylogenetic relatedness. J Gen Virol. 2001;82:883–92.

    Article  CAS  PubMed  Google Scholar 

  19. Dimeski G. Interference testing. Clin Biochem Rev. 2008;29(Suppl 1):S43–8.

    PubMed  PubMed Central  Google Scholar 

  20. Carman WF. The clinical significance of surface antigen variants of hepatitis B virus. J Viral Hepat. 1997;4(Suppl 1):11–20.

    Article  PubMed  Google Scholar 

  21. Anh-Tuan N, Novak E, Hollan SR. Hepatitis B surface antigen circulating immune complexes (HBsAg-CICs) in patients with bleeding disorders. Vox Sang. 1981;40:12–6.

    Article  CAS  PubMed  Google Scholar 

  22. Sugiura K, Hasumura Y, Takeuchi J. Significance of circulating HBs antigen-antibody immune complexes in patients with HBs antigen-positive liver disease. Gastroenterol Jpn. 1982;17:241–5.

    Article  CAS  PubMed  Google Scholar 

  23. Madalinski K, Burczynska B, Heermann KH, et al. Analysis of viral proteins in circulating immune complexes from chronic carriers of hepatitis B virus. Clin Exp Immunol. 1991;84:493–500.

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Colson P, Borentain P, Motte A, et al. Clinical and virological significance of the co-existence of HBsAg and anti-HBs antibodies in hepatitis B chronic carriers. Virology. 2007;367:30–40.

    Article  CAS  PubMed  Google Scholar 

  25. Zhang JM, Xu Y, Wang XY, et al. Coexistence of hepatitis B surface antigen (HBsAg) and heterologous subtype-specific antibodies to HBsAg among patients with chronic hepatitis B virus infection. Clin Infect Dis. 2007;44:1161–9.

    Article  CAS  PubMed  Google Scholar 

  26. Aoyagi K, Ohue C, Iida K, et al. Development of a simple and highly sensitive enzyme immunoassay for hepatitis C virus core antigen. J Clin Microbiol. 1999;37:1802–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Tanaka E, Ohue C, Aoyagi K, et al. Evaluation of a new enzyme immunoassay for hepatitis C virus (HCV) core antigen with clinical sensitivity approximating that of genomic amplification of HCV RNA. Hepatology. 2000;32:388–93.

    Article  CAS  PubMed  Google Scholar 

  28. Kimura T, Rokuhara A, Sakamoto Y, et al. Sensitive enzyme immunoassay for hepatitis B virus core-related antigens and their correlation to virus load. J Clin Microbiol. 2002;40:439–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This research was supported in part by research grants (15fk0210035h0001 and 15fk0310012h0004) from the Japan Agency for Medical Research and Development. We thank Trevor Ralph for his English editorial assistance.

Author information

Authors and Affiliations

  1. Department of Medicine, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto, 390-8621, Japan

    Akihiro Matsumoto & Eiji Tanaka

  2. New Product Development Department, Fujirebio Inc., Hachioji, Tokyo, Japan

    Masayasu Imaizumi, Kazushige Moriyama & Katsumi Aoyagi

  3. Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan

    Yasuhito Tanaka

  4. Division of Hepatobiliary and Pancreatic Disease, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan

    Shuhei Nishiguchi

  5. Clinical Research Center, National Hospital Organization, Nagasaki Medical Center, Omura, Nagasaki, Japan

    Hiroshi Yatsuhashi

  6. Research and Development Department, Advanced Life Science Institute Inc., Wako, Saitama, Japan

    Takehiko Ishida & Katsumi Aoyagi

Authors
  1. Akihiro Matsumoto
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  2. Masayasu Imaizumi
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  3. Yasuhito Tanaka
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  4. Shuhei Nishiguchi
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  5. Hiroshi Yatsuhashi
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  6. Takehiko Ishida
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  7. Kazushige Moriyama
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  8. Katsumi Aoyagi
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  9. Eiji Tanaka
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Corresponding author

Correspondence to Eiji Tanaka.

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Conflict of interest

Masayasu Imaizumi, Kazushige Moriyama, and Katsumi Aoyagi are employees of Fujirebio, Inc. Takehiko Ishida and Katsumi Aoyagi are employees of Advanced Life Science Institute, Inc. Yasuhito Tanaka received lecture fees from Fujirebio, Inc., Chugai Pharmaceutical Co. Ltd, Bristol-Myers K.K, and GlaxoSmithKline K.K. Yasuhito Tanaka received a research grant from Chugai Pharmaceutical Co. Ltd. Hiroshi Yastuhashi received commercial research funding from Chugai Pharmaceutical Co. Ltd.

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Matsumoto, A., Imaizumi, M., Tanaka, Y. et al. Novel and highly sensitive immunoassay for total hepatitis B surface antigen, including that complexed with hepatitis B surface antibody. J Gastroenterol 52, 376–384 (2017). https://doi.org/10.1007/s00535-016-1244-7

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  • DOI: https://doi.org/10.1007/s00535-016-1244-7

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