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Journal of Clinical Microbiology, December 2001, p. 4370-4379, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4370-4379.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Sporadic Cases of Hepatitis E Virus (HEV) Infection in China Using Immunoassays Based on Recombinant Open Reading Frame 2 and 3 Polypeptides from HEV Genotype 4

Youchun Wang,1,2 Huayuan Zhang,2 Zhuo Li,3 Wenjie Gu,2 Haiyuan Lan,2 Wa Hao,3 Roger Ling,1 Hemin Li,2 and Tim J. Harrison1,*

Centre for Hepatology, Royal Free and University College Medical School, Royal Free Campus, London NW3 2PF, United Kingdom,1 and Department of Hepatitis, National Institute for the Control of Pharmaceutical and Biological Products, Temple of Heaven, Beijing 100050,2 and Beijing Institute of Hepatology, Youan Hospital, Beijing,3 People's Republic of China

Received 30 March 2001/Returned for modification 11 August 2001/Accepted 18 September 2001

We reported previously on the complete sequence of hepatitis E virus (HEV) genotype 4, isolated from patients with sporadic cases of acute HEV infection in China. At least eight HEV genotypes have now been described worldwide, and further isolates await classification. Current immunoassays for the detection of anti-HEV antibodies are based on polypeptides from genotypes 1 and 2 only and may be inadequate for the reliable detection of other genotypes. Because genotypes 1 and 4 predominate in China, we wished to investigate the antigenic reactivities of HEV genotype 4 proteins. Four overlapping regions of open reading frame 2 (ORF2) (FB5, amino acids [aa] 1 to 130; E4, aa 67 to 308; F2-2, aa 288 to 461; E5, aa 414 to 672) and the entire ORF3 product were expressed in Escherichia coli as fusion proteins. Enzyme immunoassays based on each of the five purified polypeptides were evaluated with sera from patients with sporadic cases of acute HEV infection. Individual immunoassays derived from HEV genotype 4 detected more cases of acute hepatitis E than a commercial assay. Some serum samples, which were positive for anti-HEV immunoglobulin G only by assays based on HEV genotype 4, were positive for HEV RNA by reverse transcription-PCR. Polypeptide FB5, from the N terminus of ORF2, had the greatest immunoreactivity with sera from patients with acute hepatitis E. These data indicate that the N terminus of ORF2 may provide epitopes which are highly reactive with acute-phase sera and that assays based on genotypes 1 and 2 alone may be inadequate for the detection of HEV infection in China, where sporadic cases of HEV infection are caused predominantly by HEV genotypes 4 and 1.

* Corresponding author. Mailing address: Centre for Hepatology, Royal Free and University College Medical School, Royal Free Campus, Rowland Hill St., London NW3 2PF, United Kingdom. Phone: 4420 7433 2881. Fax: 4420 7433 2852. E-mail: T.Harrison{at}rfc.ucl.ac.uk.

Journal of Clinical Microbiology, December 2001, p. 4370-4379, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4370-4379.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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