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. 2024 Jun 5:11:1423581.
doi: 10.3389/fvets.2024.1423581. eCollection 2024.

Comparison of the performance of SAG2, GRA6, and GRA7 for serological diagnosis of Toxoplasma gondii infection in cats

Serges Sabukunze  1 , Haorong Gu  1 , Lin Zhao  1 , Honglin Jia  1   2 , Huanping Guo  1
Affiliations

Comparison of the performance of SAG2, GRA6, and GRA7 for serological diagnosis of Toxoplasma gondii infection in cats

Serges Sabukunze et al. Front Vet Sci. .

Abstract

Toxoplasmosis is an important zoonotic disease caused by Toxoplasma gondii that can infect almost all warm-blooded animals worldwide, including humans. The high prevalence of T. gondii infection and its ability to cause serious harm to humans and animals, especially immunodeficient individuals, make it a key public health issue. Accurate diagnostic tools with high sensitivity are needed for controlling T. gondii infection. In the current study, we compared the performance of recombinant SAG2, GRA6, and GRA7 in ELISA for the serological diagnosis of T. gondii infection in cats. We further investigated the antigenicity of recombinant dense granule protein 3 (rGRA3), rGRA5, rGRA8, and rSRS29A expressed in a plant-based, cell-free expression system for detecting antibodies in T. gondii-infected cats. In summary, our data suggest that GRA7 is more sensitive than the other two antigens for the serodiagnosis of T. gondii infection in cats, and GRA3 expressed in the cell-free system is also a priming antigen in serological tests for detecting T. gondii infection in cats.

Keywords: GRA3; GRA6; GRA7; SAG2; Toxoplasma gondii; cats; serodiagnosis.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
SDS-PAGE analysis of the purified rSAG2, rGRA6, and rGRA7. (A) Purified rGRA6 fused with His. (B) Purified rSAG2 and rGRA7 fused with His. A BSA standard solution was diluted into the concentrations as indicated in the figure and used to quantify the concentration of the recombinant proteins. 1, 2000 ng/μL; 2, 1,500 ng/μL; 3, 1,000 ng/μL; and 4, 500 ng/μL.
Figure 2
Figure 2
Western blot analysis of the expression of recombinant proteins in the cell-free system. The expression of the recombinant proteins was detected with an anti-HA antibody. T is the protein without transmembrane domains.
Figure 3
Figure 3
Antigenicity analysis of the selected recombinant proteins expressed in the cell-free system. Antigenicity analysis of the selected proteins expressed in the cell-free system was detected through an ELISA. TgLys was coated as a control.
Figure 4
Figure 4
Antigenicity analysis of the CF-rGRA3 using cat serum samples. TgLys, T. gondii lysate; P: a serum from a cat experimentally infected with T. gondii; PRS: a random positive sample in TgLys-ELISA; 1, 2, 3, 4, 5, 6, and 7 represent the selected cat samples.
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