Expression of glutathione s-transferase-fusion hepatitis E virus ORF2, 3 antigens in  and its application for diagnosis

Ri, So-Yong; Pak, Yong-Ju; Ha, Su-Gil; Ya, Hae-Gyong

doi:10.4103/kamj.kamj_21_18

Expression of glutathione s-transferase-fusion hepatitis E virus ORF2, 3 antigens in and its application for diagnosis

Authors

Abstract

Context
Hepatitis E infection, caused by the hepatitis E virus (HEV), is a common cause of acute hepatitis in developing countries with poor sanitation and hygiene.
Aims
In this paper, we expressed HEV ORF2 C-terminal 802 bp and ORF3 C-terminal 107 bp fragments and its glutathione S-transferase (GST) fusion genes in and tested the immunoreactivity of HEV antibody from sample serum with recombinant GST-HEV ORF2, 3 antigens by enzyme-linked immunosorbent assay (ELISA).
Settings and design
Eight primers designed for gene synthesis.
Materials and methods
HEV ORF2, 3 and its GST-fusion gene fragments were amplified by PCR. Expression of target genes in was induced by Isopropyl-β-D-Thio-alactopyranoside (IPTG) (0.1 mmol/l) and temperature (42°C). The detection of anti-HEV antibody with purified GST-HEV ORF2, 3 antigens was performed using the indirect ELISA method.
Results
When our ELISA was compared with the HEV-ELISA kit, the sensitivity of our assay is 97.9%, specificity is 100%, and consistency is 99.6%. The positive rate of anti-HEV antibody from the serum in acute hepatitis patients with recombinant GST-HEV ORF2, 3 antigens by our ELISA was 29.5 and 26.1%, respectively, and this result corresponded with the HEV-ELISA kit.
Conclusion
The new HEV ELIA developed in the present study is a highly specific diagnostic assay for the detection of anti-HEV antibody in serum specimens obtained from acute hepatitis patients.

Keywords