Immunodiagnostic Value of Echinococcus Granulosus Recombinant B8/1 Subunit of Antigen B
Author(s):
Abstract:
Background
Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory.Objective
The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodiagnosis of human CE.Methods
The coding sequence for AgB8/1 subunit of Echinococcus granulosuswas selected from GenBank and was gene-optimized. The sequence was synthesized
and inserted into pGEX-4T-1 vector. Purification was performed with GST tag affinity
column. Diagnostic performance of the produced recombinant antigen, native antigen B
and a commercial ELISA kit were further evaluated in an ELISA system, using a panel
of sera from CE patients and controls.
Results
SDS-PAGE demonstrated that the protein of interest had a high expression level and purity after GST tag affinity purification. Western blotting verified the immunoreactivity of the produced recombinant antigen with the sera of CE patients. In an ELISA system, the sensitivity and specificity (for human CE diagnosis) of the recombinant antigen, native antigen B and commercial kit were respectively 93% and 92%, 87% and 90% and 97% and 95%.Conclusion
The produced recombinant antigen showed a high diagnostic value which can be recommended for serodiagnosis of CE in Iran and other CE-endemic areas. Utilizing the combination of other subunits of AgB8 would improve the performance value of the introduced ELISA system.Keywords:
Language:
English
Published:
Iranian journal of immunology, Volume:14 Issue: 2, Spring 2017
Pages:
111 to 122
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