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Clinical and Diagnostic Laboratory Immunology, May 2002, p. 639-648, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.639-648.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantitation of Immunoglobulin to Hepatitis E Virus by Enzyme Immunoassay

Bruce L. Innis,1* Jitvimol Seriwatana,1 Robin A. Robinson,2 Mrigendra P. Shrestha,3 Patrice O. Yarbough,4,{dagger} Charles F. Longer,1,{ddagger} Robert M. Scott,3 David W. Vaughn,5,§ and Khin Saw Aye Myint5

Department of Virus Diseases, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910,1 Novavax, Inc., Rockville, Maryland 20852,2 Walter Reed/AFRIMS Research Unit—Nepal, Kathmandu, Nepal,3 Genelabs Technologies, Inc., Redwood City, California 94063,4 Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand5

Received 4 December 2001/ Returned for modification 17 January 2002/ Accepted 25 January 2002

We developed a quantitative enzyme immunoassay (EIA) for antibody to hepatitis E virus (HEV) by using truncated HEV capsid protein expressed in the baculovirus system to improve seroepidemiology, to contribute to hepatitis E diagnosis, and to enable vaccine evaluations. Five antigen lots were characterized; we used a reference antiserum to standardize antigen potency. We defined Walter Reed antibody units (WR U) with a reference antiserum by using the four-parameter logistic model, established other reference pools as assay standards, and determined the conversion factor: 1 WR U/ml = 0.125 World Health Organization unit (WHO U) per ml. The EIA performed consistently; median intra- and intertest coefficients of variation were 9 and 12%, respectively. The accurate minimum detection limit with serum diluted 1:1,000 was 5.6 WR U/ml; the test could detect reliably a fourfold antibody change. In six people followed from health to onset of hepatitis E, the geometric mean antibody level rose from 7.1 WR U/ml to 1,924.6 WR U/ml. We used the presence of 56- and 180-kDa bands by Western blotting as a confirmatory test and to define true-negative and -positive serum specimens. A receiver-operating characteristics plot identified 30 WR U/ml as an optimum cut-point (sensitivity, 86%; specificity, 89%). The EIA detected antibody more sensitively than a commercially available test. The EIA was transferred to another laboratory, where four operators matched reference laboratory results for a panel of unknowns. Quantitation of antibody to HEV and confirmation of its specificity by Western blotting make HEV serology more meaningful.

* Corresponding author. Mailing address: GlaxoSmithKline, 1250 S. Collegeville Rd. (mail code UP 4330), Collegeville, PA 19426-0989. Phone: (610) 917-6142. Fax: (610) 917-4287. E-mail: bruce.2.innis{at}gsk.com

{dagger} Present address: Tanox, Inc., Houston, TX 77025.

{ddagger} Present address: Chattanooga Unit, University of Tennessee College of Medicine, Chattanooga, TN 37403.

§ Present address: Department of Virus Diseases, Walter Reed Army Institute of Research, Silver Spring, MD 20910.

Clinical and Diagnostic Laboratory Immunology, May 2002, p. 639-648, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.639-648.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chen, H. Y., Lu, Y., Howard, T., Anderson, D., Fong, P. Y., Hu, W.-P., Chia, C. P., Guan, M. (2005). Comparison of a New Immunochromatographic Test to Enzyme-Linked Immunosorbent Assay for Rapid Detection of Immunoglobulin M Antibodies to Hepatitis E Virus in Human Sera. CVI 12: 593-598 [Abstract] [Full Text]  
  • Seriwatana, J., Shrestha, M. P., Scott, R. M., Tsarev, S. A., Vaughn, D. W., Myint, K. S. A., Innis, B. L. (2002). Clinical and Epidemiological Relevance of Quantitating Hepatitis E Virus-Specific Immunoglobulin M. CVI 9: 1072-1078 [Abstract] [Full Text]  


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