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. 2020 Sep 22;20(1):694.
doi: 10.1186/s12879-020-05414-8.

Evaluation of toxoplasmosis in pregnant women using dot-immunogold-silver staining with recombinant Toxoplasma gondii peroxiredoxin protein

Zhuan-Zhuan Liu  1 , Xue-Yan Li  1 , Lin-Lin Fu  1 , Fei Yuan  1 , Ren-Xian Tang  1 , Yi-Sheng Liu  1 , Kui-Yang Zheng  2
Affiliations

Evaluation of toxoplasmosis in pregnant women using dot-immunogold-silver staining with recombinant Toxoplasma gondii peroxiredoxin protein

Zhuan-Zhuan Liu et al. BMC Infect Dis. .

Abstract

Background: Toxoplasma gondii infection endangers human health and affects animal husbandry. Serological detection is the main method used for epidemiological investigations and diagnosis of toxoplasmosis. The key to effective diagnosis of toxoplasmosis is the use of a standardized antigen and a specific and sensitive detection method. Peroxiredoxin is an antigenic protein and vaccine candidate antigen of T. gondii that has not yet been exploited for diagnostic application.

Methods: In this study, recombinant T. gondii peroxiredoxin protein (rTgPrx) was prepared and used in dot-immunogold-silver staining (Dot-IGSS) to detect IgG antibodies in serum from mice and pregnant women. The rTgPrx-Dot-IGSS method was established and optimized using mouse serum. Furthermore, serum samples from pregnant women were analyzed by rTgPrx-Dot-IGSS.

Results: Forty serum samples from mice infected with T. gondii and twenty negative serum samples were analyzed. The sensitivity and specificity of rTgPrx-Dot-IGSS were 97.5 and 100%, respectively, equivalent to those of a commercial ELISA kit for anti-Toxoplasma IgG antibody. Furthermore, 540 serum samples from pregnant women were screened with a commercial ELISA kit. Eighty-three positive and 60 negative serum samples were analyzed by rTgPrx-Dot-IGSS. The positive rate was 95.18%, comparable to that obtained with the commercial ELISA kit.

Conclusions: The Dot-IGSS method with rTgPrx as an antigen might be useful for diagnosing T. gondii infection in individuals.

Keywords: Dot-IGSS; Peroxiredoxin; Pregnant women; Serum; Toxoplasma gondii.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Detection of rTgPrx by SDS-PAGE and Western blotting. a: rTgPrx was expressed in E. coli, purified using GST affinity chromatography, analyzed by SDS-PAGE on 12% gels, and stained with Coomassie brilliant blue. M: marker. Lane 1: The molecular weight of rTgPrx digested by PreScission Protease was 25 kDa. Lane 2: GST-tagged rTgPrx was purified by GST affinity chromatography and had a molecular weight of 51 kDa. Lane 3: SDS-PAGE showed that unpurified rTgPrx was expressed in E. coli. b: The immunoreactivity of rTgPrx was detected by Western blotting. M: marker. Lane 1: rTgPrx was reacted with rabbit anti-T.gondii serum. Lane 2: rTgPrx was reacted with negative serum
See this image and copyright information in PMC

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