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Background <p><i>Toxoplasma gondii</i> (<i>T. gondii</i>) a widespread zoonotic parasite infecting various host species. Since cattle serve as major intermediate hosts for <i>T. gondii</i>, giving a significant public health concern, developing effective diagnostic tools is essential. This study evaluated the serodiagnostic potential of an indirect enzyme linked immunosorbent assay (iELISA) using recombinant <i>T. gondii</i> dense granule 14 protein (<i>Tg</i>GRA14) in cattle population.</p> Methods <p>The recombinant <i>Tg</i>GRA14 used was synthesized and evaluated as previously described. The microplate was coated with the purified <i>Tg</i>GRA14 protein at 1&#xa0;µg/mL and incubated overnight. The assay utilized a 1:200 dilution of bovine sera and a 1:5000 dilution of HRP-conjugated anti-bovine IgG. The reaction was developed with 3,3’,5,5’-tetramethylbenzidine (TMB) substrate and the absorbance was measured at 450&#xa0;nm.</p> Results <p>A sensitivity of 93.30% (95% CI: 76.50–98.84) and specificity of 88.30% (95% CI: 82.61–92.36). The assay exhibited significant diagnostic potential with an area under the curve (AUC) of 0.95 and substantial agreement with the standard test (κ = 0.64). Utilizing these optimized iELISA parameters, the seroprevalence for anti-<i>T. gondii</i> IgG antibodies in the cattle population in this study was determined to be 22.93% (50/218).</p> Conclusions <p>These findings highlight the potential of the <i>Tg</i>GRA14-based iELISA as a reliable serodiagnostic tool, facilitating improved epidemiological surveillance and effective control of bovine toxoplasmosis.</p>

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Evaluation of recombinant TgGRA14 as a diagnostic antigen for detection of Toxoplasma gondii antibodies in cattle

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年8月1日

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  • Hanh Thi Ha,
  • Eukote Suwan,
  • Chanya Kengradomkij,
  • Nhung Pho Nguyen Nguyen,
  • Charoonluk Jirapattharasate,
  • Tawin Inpankaew

摘要

Background

Toxoplasma gondii (T. gondii) a widespread zoonotic parasite infecting various host species. Since cattle serve as major intermediate hosts for T. gondii, giving a significant public health concern, developing effective diagnostic tools is essential. This study evaluated the serodiagnostic potential of an indirect enzyme linked immunosorbent assay (iELISA) using recombinant T. gondii dense granule 14 protein (TgGRA14) in cattle population.

Methods

The recombinant TgGRA14 used was synthesized and evaluated as previously described. The microplate was coated with the purified TgGRA14 protein at 1 µg/mL and incubated overnight. The assay utilized a 1:200 dilution of bovine sera and a 1:5000 dilution of HRP-conjugated anti-bovine IgG. The reaction was developed with 3,3’,5,5’-tetramethylbenzidine (TMB) substrate and the absorbance was measured at 450 nm.

Results

A sensitivity of 93.30% (95% CI: 76.50–98.84) and specificity of 88.30% (95% CI: 82.61–92.36). The assay exhibited significant diagnostic potential with an area under the curve (AUC) of 0.95 and substantial agreement with the standard test (κ = 0.64). Utilizing these optimized iELISA parameters, the seroprevalence for anti-T. gondii IgG antibodies in the cattle population in this study was determined to be 22.93% (50/218).

Conclusions

These findings highlight the potential of the TgGRA14-based iELISA as a reliable serodiagnostic tool, facilitating improved epidemiological surveillance and effective control of bovine toxoplasmosis.