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Original Article

A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein

Kyoung Ju Song1,†, Zhaoshou Yang1,†, Chom-Kyu Chong2, Jin-Soo Kim2, Kyung Chan Lee2, Tong-Soo Kim3, Ho-Woo Nam1
The Korean Journal of Parasitology 2013;51(5):503-510.
Published online: October 31, 2013

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Corresponding author (howoo@catholic.ac.kr)

†The first two authors equally contributed to this paper.

• Received: July 8, 2013   • Revised: August 30, 2013   • Accepted: September 2, 2013

© 2013, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein
Korean J Parasitol. 2013;51(5):503-510.   Published online October 31, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.5.503
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A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein
Korean J Parasitol. 2013;51(5):503-510.   Published online October 31, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.5.503
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A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein
Image Image Image Image
Fig. 1 Amino acid sequence and structure of GST-GRA2-SAG1A. (A) Amino acid sequence of GST-GRA2-SAG1A. (B) Schematic diagram of the structure of GST-GRA2-SAG1A containing the intrinsically unstructured disorder region.
Fig. 2 Expression, purification, solubility, and antigenicity of recombinant GST-GRA2-SAG1A proteins. (A) Expression and purification of rGST-GRA2-SAG1A proteins in E. coli BL21(DE3). Cell lysates were separated on a 12% SDS-PAGE gel. The protein was stained with Coomassie blue. Lane M, dual protein marker; lane 1, sonicates of E. coli/GST/GRA2-SAG1A before induction; lane 2, sonicates of E. coli/GST/GRA2-SAG1A after induction with 1 mM IPTG; lane 3, purified rGST-GRA2-SAG1A protein; lane 4, purified rGST-SAG1A protein. (B) Solubility of rGST-GRA2-SAG1A proteins in E. coli. Lane M, dual protein marker; lane 1, whole cell lysates of E. coli/-GST-SAG1A after induction; lane 2, soluble part of E. coli/GST-SAG1A after induction; lane 3, insoluble part of E. coli GST-SAG1A after induction; lane 4, whole cell lysates of E. coli/GST-GRA2-SAG1A after induction; lane 5, soluble part of E. coli/GST-GRA2-SAG1A after induction; lane 6, insoluble part of E. coli/GST-GRA2-SAG1A after induction. Coomassie blue stain (B-1) and western blot with anti-GST antibody (B-2) (asterisk indicates molecular weight of insoluble part of rGST-GRA2-SAG1A). (C) Antigenicity of rGST-GRA2-SAG1A proteins. Proteins were stained with Coomassie blue. Lane M, dual protein marker; lane 1, cell lysates of T. gondii RH strain; lane 2, purified rGST-SAG1A protein; lane 3, purified rGST-GRA2-SAG1A protein (C-1). Western blot with toxoplasmosis patients sera (C-2/C-3).
Fig. 3 Comparison of ELISA and TgRDT of Korean sample. (A) Interpretation of TgRDT. Normal or negative: only 1 red upper band. Positive: + means weak, ++ mild, +++ strong antigen reactivity indicated as band. (B) Comparison of ELISA and TgRDT. Serum ELISA index of 0.25 was determined as the cut-off point.
Fig. 4 Seroprevalence of Uganda sample. (A) Analysis of TgRDT result. The result was analyzed depend on age and sex. (B) Comparison of ELISA and TgRDT. Serum ELISA index of 0.25 was determined as the cut-off point.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein
ELISA TgRDT Negative ( < 0.25) 0/33 (0%) Positive ( ≥ 0.25) 33/34 (97.1%)
Table 1. Comparison of ELISA positive and negative group to TgRDT

Table 1.

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