Expression of glutathione s-transferase-fusion hepatitis E virus ORF2, 3 antigens in and its application for diagnosis | ||||
| Kasr Al Ainy Medical Journal | ||||
| Volume 24, Issue 3, September 2018 PDF (1.23 MB) | ||||
| DOI: 10.4103/kamj.kamj_21_18 | ||||
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| Authors | ||||
| So-Yong Ri; Yong-Ju Pak; Su-Gil Ha; Hae-Gyong Ya | ||||
| 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 | ||||
| hepatitis E virus ORF2; 3 genes; Expression vector; enzyme-linked immunosorbent assay | ||||
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