Molecular cloning and expression of a recombinant EpC1 protein for serodiagnosis of Echinococcus granulosus in dogs
Journal of Veterinary parasitology(2018)
Abstract
Echinococcus granulosus is considered as a significant parasitic infection with worldwide distribution. The present study was aimed to express the diagnostic antigen EpC1 in a recombinant system and set up an indirect ELISA for diagnosis of infected canines. Hydatid cysts were collected from liver and lungs of naturally infected sheep from slaughter houses and cDNA was synthesized from the protoscoleces. The EpC1 was expressed as recombinant protein in E. coli using pET28a as expression vector and purified by His-tag affinity chromatography. Expression of recombinant EpC1 was confirmed using SDS-PAGE followed by Western immunoblotting using T7-Tag monoclonal antibody and sera from naturally infected and experimentally challenged dogs. The diagnostic potential of the recombinant EpC1 was assessed against 22 dog sera using indirect ELISA. The sensitivity, specificity, positive and negative prediction values of the ELISA test were calculated as 100%, 93.3%, 88.8% and 100%, respectively. Recombinant EpC1 showed promising results for the diagnosis of echinococcosis in dogs.
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Key words
echinococcus granulosus,recombinant epc1 protein,serodiagnosis
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