Open AccessEvaluation of a recombinant multiepitope antigen for diagnosis of hepatitis C virus: A lower cost alternative for antigen production
Author(s) -
Thomasini Ronaldo Luis,
Souza Hortencia Gisele Amaro,
BrunaRomero Oscar,
Totola Antonio Helvecio,
Gonçales Neiva Sellan Lopes,
Lima Cristiano Xavier,
Teixeira Mauro Martins
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22410
Subject(s) - virology , serology , antigen , recombinant dna , immunoassay , hepatitis c virus , antibody , hepatitis , western blot , medicine , biology , virus , immunology , gene , biochemistry
Background The most of the hepatitis C‐infected patients remain undiagnosed until they develop severe liver damage or submitted for serological screening. 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. 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.