Detection of Serum Antibodies to Hepatitis E Virus Based on HEV Genotype 3 ORF2 Capsid Protein Expressed in Nicotiana benthamiana.
10.3343/alm.2017.37.4.313
- Author:
Milena MAZALOVSKA
1
;
Nikola VARADINOV
;
Tsvetoslav KOYNARSKI
;
Ivan MINKOV
;
Pavel TEOHAROV
;
George P LOMONOSSOFF
;
Gergana ZAHMANOVA
Author Information
1. Department of Plant Physiology and Molecular Biology, University of Plovdiv “Paisii Hilendarski”, Plovdiv, Bulgaria. gerganaz@uni-plovdiv.bg
- Publication Type:Original Article
- Keywords:
Hepatitis E virus;
Transient expression;
ELISA;
Nicotiana benthamiana
- MeSH:
Agriculture;
Animals;
Antibodies*;
Capsid Proteins*;
Capsid*;
Chromatography;
Developing Countries;
Enzyme-Linked Immunosorbent Assay;
Genotype*;
Hepatitis E virus*;
Hepatitis E*;
Hepatitis*;
Humans;
Immunoglobulin G;
Methods;
Open Reading Frames;
Plants;
Swine;
Tobacco*
- From:Annals of Laboratory Medicine
2017;37(4):313-319
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: Hepatitis E virus (HEV) causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. There have been recent reports on the zoonotic spread of the virus, and several animal species, primarily pigs, have been recognized as reservoirs of HEV. Because of its possible spread, there is an urgent need of a method for the cost-effective production of HEV proteins that can be used as diagnostic antigens for the serological detection of anti-HEV antibodies. METHODS: The HEV open reading frame (ORF)2 protein was purified from plant tissue by using immobilized metal-anion chromatography (IMAC). The recombinant protein was used to develop an in-house ELISA for testing anti-HEV antibodies in both human and swine sera. Thirty-six serum samples collected from patients with serologically proven HEV infection with commercial kits were tested for anti-HEV IgG antibodies by using the plant-expressed protein. Forty-five serum samples collected from apparently healthy pigs in Bulgarian farms were also tested. RESULTS: We confirmed the transient expression and purification of a truncated version of the HEV genotype 3 capsid protein in Nicotiana benthamiana and its usefulness as a diagnostic antigen. ELISA showed the presence of anti-HEV IgG antibodies in 29 of the 36 human samples. The in-house ELISA showed anti-HEV IgG antibodies in 34 of the 45 pigs. CONCLUSIONS: We describe a method for the production of HEV ORF2 protein in N. benthamiana and the usefulness of this protein for the serological detection of anti-HEV antibodies in both humans and swine.
