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Journal of Clinical Microbiology, February 2002, p. 439-445, Vol. 40, No. 2
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.2.439-445.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Sensitive Enzyme Immunoassay for Hepatitis B Virus Core-Related Antigens and Their Correlation to Virus Load

Tatsuji Kimura,1* Akinori Rokuhara,2 Yoko Sakamoto,1 Shintaro Yagi,1 Eiji Tanaka,2 Kendo Kiyosawa,2 and Noboru Maki1

R&D Division, Advanced Life Science Institute, Inc., Wako, Saitama 351-0112,1 Second Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan2

Received 24 May 2001/ Returned for modification 5 August 2001/ Accepted 12 November 2001

A sensitive enzyme immunoassay (EIA) specific for hepatitis B virus core antigen (HBcAg) and hepatitis B e antigen (HBeAg) was developed. We designated the precore/core gene products as hepatitis B virus (HBV) core-related antigens (HBcrAg). In order to detect HBcrAg even in anti-HBc/e antibody-positive specimens, the specimens were pretreated in detergents. The antibodies are inactivated by this pretreatment and, simultaneously, the antigens are released and the epitopes are exposed. The assay demonstrated 71 to 112% recovery using HBcrAg-positive sera. We observed no interference from the tested anticoagulants or blood components. When the cutoff value was tentatively set at 103 U/ml, all healthy control (HBsAg/HBV-DNA negative; n = 108) and anti-HCV antibody-positive (n = 59) sera were identified as negative. The assay showed a detection limit of 4 x 102 U/ml using recombinant antigen. Detection limits were compared in four serially diluted HBV high-titer sera. The HBcrAg assay demonstrated higher sensitivity than HBV-DNA transcription-mediated amplification (TMA) or HBeAg radio immunoassay (RIA) in the dilution test. HBcrAg concentrations correlated well with HBV-DNA TMA (r = 0.91, n = 29) and in-house real-time detection-PCR (r = 0.93, n = 47) in hepatitis B patients. On HBeAg/anti-HBe antibody seroconversion panels, the HBcrAg concentration changed in accordance with HBV-DNA levels. HBcrAg concentration provides a reflection of HBV virus load equivalent to HBV-DNA level, and the assay therefore offers a simple method for monitoring hepatitis B patients.

* Corresponding author. Mailing address: R&D Division, Advanced Life Science Institute, Inc., 2-10-23 Maruyamadai, Wako, Saitama, 351-0112, Japan. Phone: 81 48 465 2761. Fax: 81 48 465 2765. E-mail: tkimura{at}alsi-i.co.jp.

Journal of Clinical Microbiology, February 2002, p. 439-445, Vol. 40, No. 2
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.2.439-445.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wong, D. K.-H., Tanaka, Y., Lai, C.-L., Mizokami, M., Fung, J., Yuen, M.-F. (2007). Hepatitis B Virus Core-Related Antigens as Markers for Monitoring Chronic Hepatitis B Infection. J. Clin. Microbiol. 45: 3942-3947 [Abstract] [Full Text]  
  • Kimura, T., Ohno, N., Terada, N., Rokuhara, A., Matsumoto, A., Yagi, S., Tanaka, E., Kiyosawa, K., Ohno, S., Maki, N. (2005). 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. 280: 21713-21719 [Abstract] [Full Text]  
  • Kimura, T., Rokuhara, A., Matsumoto, A., Yagi, S., Tanaka, E., Kiyosawa, K., Maki, N. (2003). New Enzyme Immunoassay for Detection of Hepatitis B Virus Core Antigen (HBcAg) and Relation between Levels of HBcAg and HBV DNA. J. Clin. Microbiol. 41: 1901-1906 [Abstract] [Full Text]  


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