A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein |
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Authors: | Kyoung Ju Song Zhaoshou Yang Chom-Kyu Chong Jin-Soo Kim Kyung Chan Lee Tong-Soo Kim Ho-Woo Nam |
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Affiliation: | 1.Department of Parasitology, Catholic Institute of Parasitic Disease, College of Medicine, Catholic University of Korea, Seoul 137-701, Korea.;2.Asan Biotech Institute, Asan Pharm Co Ltd, Kiheung 445-813, Korea.;3.Department of Parasitology, Inha University School of Medicine, Incheon 400-712, Korea. |
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Abstract: | Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis. |
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Keywords: | Toxoplasma gondii serodiagnosis rapid diagnostic test recombinant GST-GRA2-SAG1A ELISA intrinsically unstructured domain |
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