Evaluation of a recombinant multiepitope antigen for diagnosis of hepatitis C virus: A lower cost alternative for antigen production

评估重组多表位抗原对丙型肝炎病毒诊断的作用:一种低成本的抗原生产替代方法

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作者:Ronaldo Luis Thomasini, Hortencia Gisele Amaro Souza, Oscar Bruna-Romero, Antonio Helvecio Totola, Neiva Sellan Lopes Gonçales, Cristiano Xavier Lima, Mauro Martins Teixeira

Background

The most of the hepatitis C-infected patients remain undiagnosed until they develop severe liver damage or submitted for serological screening.

Conclusion

The multiepitope protein used here has a lower cost compared to production of each antigen separately and could be an alternative for the serological diagnosis of hepatitis C.

Objective

To evaluate a recombinant multiepitope protein for detection of IgG anti-hepatitis C virus. Method: A synthetic gene was cloned, expressed in Escherichia coli, and the recombinant protein was purified. Human serum panel consisted of 88 positives (20 HCV genotyped) and 376 negatives for hepatitis C, 6 positives for human acquired immunodeficiency virus, 6 syphilis positives, 6 hepatitis B positives were tested by IgG antihepatitis C virus using the protein by enzyme-linked immunosorbent assay. In addition, 20 positive (all genotyped samples) and 20 negative samples were also tested by immunoblot and dot blot assays.

Results

Positive hepatitis C sera were strongly reactive against the protein by immunoblot assay. In the dot blot assay, positive sera were reactive until 1:1000 dilution and there were no false positive results in the hepatitis C negative sera. In the enzyme-linked immunosorbent assay, positive and negative sera had significant discrimination. No cross-reaction was observed in samples positive for syphilis; human acquired immunodeficiency virus and hepatitis B. All 20 genotyped samples were positive by the three methods.

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