Background: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the
present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16
kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test.
Methods: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total
RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces
collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using
pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected
individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5)
and tuberculosis (n=5) were examined using this recombinant antigen.
Results: Recombinant protein was purified by affinity chromatography using His-Tag column. After
purification, recombinant protein was confirmed by western blot analysis using His Tag monoclonal antibody
or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values
were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for
rAgB16.
Conclusion: While the produced recombinant AgB16 kDa showed promising results in diagnosing human
hydatidosis, but more investigations should be implemented to reach an accurate gold standard.
