Development of Recombinant Multi-Epitope Hepatitis C Virus (HCV) Protein for Hepatitis C Diagnosis.
- 1Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Rio de Janeiro 21040-900, Brazil.
- 2Laboratory of Biotechnology and Structural Bioengineering, Federal University of Rio de Janeiro, Rio de Janeiro 21941-901, Brazil.
- 3Recombinant Technology Laboratory, Institute of Technology on Immunobiologicals, Rio de Janeiro 21040-900, Brazil.
Abstract
Hepatitis C virus (HCV) infection poses a major challenge to global public health, highlighting the need for accessible and reliable diagnostic tools to support large-scale screening and early detection. In this study, a recombinant multi-epitope HCV protein was designed, expressed, and evaluated as a diagnostic antigen for the detection of anti-HCV antibodies. The synthetic gene was constructed using immunodominant regions (core, NS3, NS4, NS5A, and NS5B) representative of HCV genotypes 1a, 1b, and 3a. The recombinant protein was expressed in Escherichia coli and purified by affinity chromatography, yielding a soluble protein with an approximate molecular weight of 43 kDa and antigenicity confirmed by Western blot analysis. An enzyme-linked immunosorbent assay (ELISA) was developed in-house using the recombinant protein as the capture antibody. Diagnostic performance was evaluated using a panel of 260 human serum samples, including 127 anti-HCV reactive and 133 anti-HCV non-reactive specimens, and receiver operating characteristic (ROC) curve analysis. The assay demonstrated excellent performance with an area under the curve (AUC) of 0.968, sensitivity of 86.6%, specificity of 98.5%, and overall accuracy of 92.7%. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis. The assay demonstrated excellent performance in serum samples, with an area under the curve (AUC) of 0.968, a sensitivity of 86.6%, and a specificity of 98.5%. These results demonstrate that the recombinant multi-epitope antigen represents a robust and scalable biomolecular platform for HCV serodiagnosis, contributing to strategies that expand access to hepatitis C screening.
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BibTeX
@article{deodato2026development,
title = {{Development of Recombinant Multi-Epitope Hepatitis C Virus (HCV) Protein for Hepatitis C Diagnosis.}},
author = {Raíssa Martins Deodato and Mayara Torquato Lima da Silva and Diego Allonso and Erik Machado Ferreira and Julia Trece Marques Possas and Jurema Corrêa da Mota and Livia Melo Villar},
journal = {Viruses},
year = {2026},
month = sep,
doi = {10.3390/v18101049},
url = {https://europepmc.org/article/MED/42943969},
note = {CC BY},
}
RIS
TY - JOUR
TI - Development of Recombinant Multi-Epitope Hepatitis C Virus (HCV) Protein for Hepatitis C Diagnosis.
AU - Raíssa Martins Deodato
AU - Mayara Torquato Lima da Silva
AU - Diego Allonso
AU - Erik Machado Ferreira
AU - Julia Trece Marques Possas
AU - Jurema Corrêa da Mota
AU - Livia Melo Villar
PY - 2026
DA - 2026/09/22
JO - Viruses
DO - 10.3390/v18101049
UR - https://europepmc.org/article/MED/42943969
AB - Hepatitis C virus (HCV) infection poses a major challenge to global public health, highlighting the need for accessible and reliable diagnostic tools to support large-scale screening and early detection. In this study, a recombinant multi-epitope HCV protein was designed, expressed, and evaluated as a diagnostic antigen for the detection of anti-HCV antibodies. The synthetic gene was constructed using immunodominant regions (core, NS3, NS4, NS5A, and NS5B) representative of HCV genotypes 1a, 1b, and 3a. The recombinant protein was expressed in Escherichia coli and purified by affinity chromatography, yielding a soluble protein with an approximate molecular weight of 43 kDa and antigenicity confirmed by Western blot analysis. An enzyme-linked immunosorbent assay (ELISA) was developed in-house using the recombinant protein as the capture antibody. Diagnostic performance was evaluated using a panel of 260 human serum samples, including 127 anti-HCV reactive and 133 anti-HCV non-reactive specimens, and receiver operating characteristic (ROC) curve analysis. The assay demonstrated excellent performance with an area under the curve (AUC) of 0.968, sensitivity of 86.6%, specificity of 98.5%, and overall accuracy of 92.7%. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis. The assay demonstrated excellent performance in serum samples, with an area under the curve (AUC) of 0.968, a sensitivity of 86.6%, and a specificity of 98.5%. These results demonstrate that the recombinant multi-epitope antigen represents a robust and scalable biomolecular platform for HCV serodiagnosis, contributing to strategies that expand access to hepatitis C screening.
SN - 1999-4915
ER -