Characterization of Type-Common Eptiopes Encoded by Gene Segments of Hepatitis E Virus. Phase 1.
Abstract
We previously isolated two cDNA clones which encode epitopes specifically recognized by antibodies from humans and cynomolgus macaques (cyno) infected with enterically-transmitted non-A, non-B hepatitis virus (HEV). To confirm the specificity of the observed immunological reaction, we tested a total of 32 code human and monkey sera with our cloned HEV antigens using lambda gt 11 phage plaque assay. The data of this serological screening indicate that the phage assay yields appropriate results on the coded sera, confirming the utility of the two cloned HEV antigens in detecting antibodies to the virus. We have determined the entire nucleotide sequences of the two cDNA clones, 406-4-2 and 406-3-2. To develop an enzyme-linked immunosorbent assay (ELISA) for detecting HEV antibodies, we used synthetic peptides derived from the deduced amino acid sequences of clones 406-4-2 and 1L6 (Burma equivalent of 406-3-2) as antigens. Our preliminary data suggest that these synthetic peptides are not as immunoreactive in ELISA as clones 406-4-2 and 406-3-2 in lambda gt 11 phage plaque assay. We have optimized the ELISA protocol to improve this new HEV antibody assay. WE also have expressed clones 406-4-2 and 1L6 in E. coli as fusions with beta-galactosidase gene. Using these expressed HEV antigens, we generated rabbit anti-HEV antisera. The rabbit antisera may be useful for the diagnosis of hepatitis E.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 26, 1990
- Accession Number
- ADB160615
Entities
People
- Chiao-chain Huang