CLONING, EXPRESSION & CHARACTERISATION OF THE HEPATITIS E VIRUS CAPSID PROTEIN OF GENOTYPES 1-4 FOR SERODIAGNOSTIC

ARCE, L.; STELLBERGER, T.; BAIKER, A.; VIZOSO PINTO, M. G.

Córdoba

Congreso; 52nd Annual Meeting Argentine Society of Biochemistry and Molecular Biology; 2016

The National Institutes of Health classified Hepatitis E as an emerging disease because: Hepatitis E Virus (HEV) is the major cause of acute hepatitis in developing countries, there is an increasing number of HEV infections in industrialized countries associated to live stock and, chronic HEV infections are being reported in immunocompromised patients (HIV, transplant recipients, etc). HEV transmission by blood transfusions has also been recently reported. There are scarce epidemiological data about HEV occurrence and prevalence in Argentina, and diagnostic tests are needed. Open reading frame 2 (ORF2) of HEV genotypes (GT) 1 to 4 were recombinatorially cloned into a customized, N-terminally His-tagged bacterial expression vector and analysed for recombinant protein expression and purification. We optimized expression under native and denaturing conditions. Purified proteins were blotted onto a nitrocellulose membrane (dot-blot) and examined for their antigenic potential by serum profiling experiments. Although the four established GTs of HEV belong to a single serotype, the serum of a patient infected with HEV ORF2 GT3 reacted with more intensity with ORF2 GT3. We provide a prototype of immunoassay for complementary confirmation of HEV seropositivity as detected in screening assays such as ELISA and plan to test a panel of sera from Argentinean blood donors.