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Journal of Clinical Microbiology, September 1999, p. 2863-2871, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Antigenic Domains of the Open Reading Frame 2-Encoded Protein of Hepatitis E Virus

Yury E. Khudyakov,1,* Elena N. Lopareva,1 Danny L. Jue,2 Tamara K. Crews,2 S. P. Thyagarajan,3 and Howard A. Fields1

Hepatitis Branch, Division of Viral and Rickettsial Diseases,1 and Biotechnology Core Facility Branch, Scientific Resources,2 National Center for Infectious Diseases, Centers for Disease Control and Prevention, U. S. Department of Health and Human Services, Atlanta, Georgia, and Department of Microbiology, University of Madras, Madras, India3

Received 9 December 1998/Returned for modification 13 April 1999/Accepted 16 June 1999

The antigenic composition of the hepatitis E virus (HEV) protein encoded by open reading frame 2 (ORF2) was determined by using synthetic peptides. Three sets of overlapping 18-, 25-, and 30-mer peptides, with each set spanning the entire ORF2 protein of the HEV Burma strain, were synthesized. All synthetic peptides were tested by enzyme immunoassay against a panel of 32 anti-HEV-positive serum specimens obtained from acutely HEV-infected persons. Six antigenic domains within the ORF2 protein were identified. Domains 1 and 6 located at the N and C termini of the ORF2 protein, respectively, contain strong immunoglobulin G (IgG) and IgM antigenic epitopes that can be efficiently modeled with peptides of different sizes. In contrast, antigenic epitopes identified within the two central domains (3 and 4) were modeled more efficiently with 30-mer peptides than with either 18- or 25-mers. Domain 2 located at amino acids (aa) 143 to 222 was modeled best with 25-mer peptides. A few 30-mer synthetic peptides derived from domain 5 identified at aa 490 to 579 demonstrated strong IgM antigenic reactivity. Several 30-mer synthetic peptides derived from domains 1, 4, and 6 immunoreacted with IgG or IgM with more than 70% of anti-HEV-positive serum specimens. Thus, the results of this study demonstrate the existence of six diagnostically relevant antigenic domains within the HEV ORF2 protein.

* Corresponding author. Mailing address: Hepatitis Branch, MS A-33, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Rd. NE, Atlanta, GA 30333. Phone: (404) 639-2335. Fax: (404) 639-1563. E-mail: yek0{at}cdc.gov.

Journal of Clinical Microbiology, September 1999, p. 2863-2871, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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