Journal of clinical laboratory analysis · 2018
Evaluation of a recombinant multiepitope antigen for diagnosis of hepatitis C virus: A lower cost alternative for antigen production.
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Thomasini Ronaldo Luis, Souza Hortencia Gisele Amaro, Bruna-Romero Oscar, Totola Antonio Helvecio, Gonçales Neiva Sellan Lopes, Lima Cristiano Xavier, Teixeira Mauro Martins
Аннотация
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.
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.