An Indian strain of hepatitis E virus (HEV): cloning, sequence, and expression of structural region and antibody responses in sera from individuals from an area of high-level HEV endemicity

Abstract

Hepatitis E virus (HEV) is responsible for a majority of sporadic and epidemic viral hepatitides in India and other developing countries. Even though the genomes of four geographically distinct strains of HEV have been cloned and sequenced, the Indian strain of HEV remains largely uncharacterized. We have cloned and sequenced about 2.2 kb of the HEV genome constituting the structural region from an Indian strain of HEV. The nucleotide and amino acid sequences show a high degree of conservation with sequences from other HEV strains. Open reading frames (ORF) 2 and 3 were expressed in Escherichia coli as N-terminal hexahistidine epitope fusions. The purified proteins were then used in an immunoblot assay to evaluate the antibody status in sera from individuals from an area of high-level HEV endemicity. The anti-ORF2 antibodies were found to be nonspecific and could not be correlated to clinical disease. The immunoglobulin M anti-ORF3 was found to be specific for the presence of acute disease. The implications of these findings in HEV diagnosis and vaccine development are discussed.