Ronaldo Luis Thomasini1, Hortencia Gisele Amaro Souza2, Oscar Bruna-Romero3, Antonio Helvecio Totola4, Neiva Sellan Lopes Gonçales5, Cristiano Xavier Lima6, Mauro Martins Teixeira7. 1. Faculdade de Medicina de Diamantina (FAMED), NEPii - Núcleo de Estudos de Patologias Infecciosas e Inflamatórias, Universidade Federal dos Vales do Jequinhonha e Mucuri, Diamantina, Minas Gerais, Brazil. 2. Bioclin-Quibasa, Belo Horizonte, Minas Gerais, Brazil. 3. Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil. 4. Universidade Federal de São João Del Rei, Ouro Branco, Minas Gerais, Brazil. 5. Hemocentro, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil. 6. Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. 7. Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Abstract
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.
BACKGROUND: The most of the hepatitis C-infectedpatients 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.
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