Cloning Of ORF2 Gene Of Hepatitis E Virus Genotype 1 And 4 And Primary Antigenicity Analysis Of Their Expressed Products

Hepatitis E virus (HEV) is the major cause of enterically transmitted non-A, non-B hepatitis. A lot of studies have been carried out on the molecular biology, pathogenicity, epidemiology, immunodiagnostic methods and vaccine of HEV. However, there are still some problems to be resolved. HEV ORF2 is putatively respected to be responsible for the synthesis of structural proteins. Most diagnostic agents and candidate vaccines are currently based on HEV ORF2. Some studies show that there are distinct genetic heterogeneity between different genotypes of HEV. However, commercial diagnostic agents are all made of the recombinant proteins or polypeptides from HEV genotype 1 and/or genotype 2. In this study, ORF2 genes of HEV genotype1 and genotype4 were cloned and expressed in E. coli. Six proteins were expressed and purified. ELISA based on these proteins were developed to detect anti-HEV antibodies from the national reference panel and 136 clinical sera samples. Tweleve monoclonal antibodies were also tested.Using RT-PCR, three overlapping cDNA fragments spanning the whole HEV...