We have previously identified two cDNA clones which encode epitopes specifically recognized by antibodies from human and cynomolgus macaques infected with enterically transmitted non-A, non-B hepatitis virus (HEV). Using the two HEV cDNA clones as probes, we have isolated two additional cDNA clones that contain inserts encoding long open reading frames. Since it is likely that the two newly isolated cDNA clones contain multiple immunogenic sequences, we propose to express both cDNA clones in E. coli or in yeast as fusion proteins. We will then use the purified recombinant proteins to develop a new HEV antibody test based on an enzyme-linked immunosorbent assay (ELISA). We will also develop two confirmatory tests for detecting HEV in human fecal specimens. One confirmatory test will be use of ELISA to capture the HEV viral antigens in clinical samples, and the other test will be based on polymerase chain reactions (PCR) to detect specific nucleotide sequences of HEV.
