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Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections

Mina Selseleh1, Hossein Keshavarz1, Mehdi Mohebali1, Saeedeh Shojaee1, Monavar Selseleh1, Mohammad Reza Eshragian2, Fatemeh Mansouri3, Mohammad Hossein Modarressi3
The Korean Journal of Parasitology 2012;50(3):233-238.
Published online: August 13, 2012

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Corresponding author (minaselseleh@yahoo.com)
• Received: February 29, 2012   • Revised: April 13, 2012   • Accepted: April 14, 2012

© 2012, Korean Society for Parasitology

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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Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections
Korean J Parasitol. 2012;50(3):233-238.   Published online August 13, 2012
DOI: https://doi.org/10.3347/kjp.2012.50.3.233
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Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections
Korean J Parasitol. 2012;50(3):233-238.   Published online August 13, 2012
DOI: https://doi.org/10.3347/kjp.2012.50.3.233
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Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections
Image Image
Fig. 1 SDS-PAGE analysis of rGRA7 expression using 12% acrylamide gel. (A) Lane 1, uninduced culture; Lane 2, expression after 7 hr of induction; Lane 3, molecular protein marker. (B) Lane 1, purified rGRA7 protein (29 kDa); Lane 3, molecular protein marker.
Fig. 2 Western blot analysis of the rGRA7 protein. Recombinant GRA7 protein was detected using a positive Toxoplasma gondii human sera and rabbit anti-human IgM conjugate by immunoblotting. Lane 1, induced control culture of cells lacking the GRA7 insert; Lane 2, purified rGRA7protein (29 kDa).

Fig. 1

Fig. 2

Production and Evaluation of Toxoplasma gondii Recombinant GRA7 for Serodiagnosis of Human Infections
Serum samplesa No. examined Positive (%)b
Negative (%)
Predictive valuec
rGRA7-IgG rGRA7-IgM rGRA7-IgG rGRA7-IgM Positive (%) Negative (%) Anti-Toxoplasma IgM sera 70 0 68 (96) 0 2 (4) 95/7 93/31 Anti-Toxoplasma IgG sera 74 66 (89) 0 8 (11) 0 95/6 77/1 Serum samples No. examined Absorbance (range) by rGRA7 ELISA Toxoplasma seropositive rate (%) Toxoplasma IgM positive sera 70 0.594 ± 0.292 (0.180-1/450)a 96 Toxoplasma IgG positive sera 74 0.879 ± 0.515 (0.212-2/100)b 89 Toxoplasma negative sera 30 0.177 ± 0.174 (0.046-0.650) 10 Sera from other diseases 30 0.091 ± 0.021 (0.062-0.126)c 0
Table 1. The sensitivity and specificity of rGRA7-IgM or IgG in comparison with results from commercial IgM and IgG ELISA kit

Tested sera were divided according to the results of commercial Toxoplasma IgG and IgM ELISA (Trinity, New York, USA).

Sensitivity and specificity were obtained from true positive cases/(true positive cases+ false negative cases)×100 and true negative cases/(true negative cases+false positive cases)×100.

Positive and negative predictive values were obtained from true positive cases/(true positive cases+false positive cases)×100 and true negative cases/(true negative cases+false negative cases)×100.

Table 2. Optical density of anti-Toxoplasma IgM and IgG using the rGRA7 antigen by ELISA

Mean±SD by rGRA7-IgM and IgM ELISA.

Mean±SD by rGRA7-IgG and IgG ELISA.

Sera from patients infected with malaria, Echinococcus granulosus, Fasciola hepatica, Strongyloides, Leishmania, and hepatitis B virus.

Table 1.

Table 2.

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