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Multiepitope fusion protein-based ELISA for enhanced brucellosis serodiagnosis
Tiansong Zhan1, Yan Li2,3,4, Yujia Xie1
1Jiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, Jiangsu, China.
Plos Neglected Tropical Diseases
|December 3, 2025
Summary
A novel multiepitope fusion protein offers improved brucellosis diagnosis by significantly reducing cross-reactivity compared to traditional lipopolysaccharide (LPS) methods. This advancement enhances diagnostic accuracy for this widespread zoonotic disease.
Area of Science:
- Immunology and Infectious Diseases
- Diagnostic Development
- Zoonotic Disease Research
Background:
- Brucellosis affects millions globally, with current serological tests using lipopolysaccharide (LPS) antigen showing significant cross-reactivity.
- This cross-reactivity leads to inaccurate diagnoses, necessitating the development of more specific diagnostic tools.
Purpose of the Study:
- To engineer and evaluate a multiepitope fusion protein for enhanced brucellosis diagnosis.
- To assess the specificity and sensitivity of the fusion protein compared to traditional LPS-based methods.
Main Methods:
- Linear B-cell epitopes from Brucella proteins were identified using the Immune Epitope Database (IEDB).
- A multiepitope fusion protein was constructed, expressed, and purified.
- Indirect enzyme-linked immunosorbent assay (iELISA) was used to evaluate the protein with patient serum samples and cross-reactivity panels.
Main Results:
- The engineered fusion protein demonstrated high diagnostic performance with an AUC of 0.9912, sensitivity of 95.34%, and specificity of 93.65%.
- The fusion protein showed significantly lower cross-reactivity (9/283 samples) compared to LPS (41/283 samples) against non-brucellosis febrile patients.
- The developed protein offers a substantial advantage over LPS in differentiating brucellosis from other bacterial infections.
Conclusions:
- The multiepitope fusion protein shows significant potential as a superior diagnostic tool for brucellosis.
- Lower cross-reactivity compared to LPS indicates improved diagnostic accuracy.
- Further research with larger, diverse sample sets is recommended for full clinical validation.
