Use of a novel hepatitis C virus (HCV) major-epitope chimeric polypeptide for diagnosis of HCV infection

David Y. Chien, Phillip Arcangel, Angelica Medina-Selby, Doris Coit, Mark Baumeister, Steve Nguyen, Carlos George-Nascimento, Alexander Gyenes, George Kuo, Pablo Valenzuela

Resultado de la investigación: Contribución a una revistaArtículo

14 Citas (Scopus)

Resumen

The genome of hepatitis C virus (HCV) consists of seven functional regions: the core, E1, E2/NS1, NS2, NS3, NS4, and NS5 regions. The U.S. Food and Drug Administration-licensed 2.0G immunoassay for the detection of anti- HCV uses proteins from the core, NS3, and NS4 regions (McHutchinson et al., Hepatology 15:19-25, 1992). The 3.0G enzyme-linked immunosorbent assay includes the protein from the NS5 region (Uyttendaele et al., Vox Sang. 66:122-129, 1994). The necessity of detecting antibodies to viral envelope proteins (E1 and E2) and to different genotype samples has been demonstrated previously (Chien et al., Lancet 342:933, 1993; Lok et al., Hepatology 18:497-502, 1993). In this study we have attempted to improve the sensitivity of the anti-HCV assay by developing a single multiple-epitope fusion antigen (MEFA; MEFA-6) which incorporates all of the major immunodominant epitopes from the seven functional regions of the HCV genome. A nucleic acid sequence consisting of proteins from the viral core, E1, E2, NS3, NS4, and NS5 regions and different subtype-specific regions of the NS4 region was constructed, cloned, and expressed in yeast. The epitopes present on this antigen can be detected by epitope-specific monoclonal and polyclonal antibodies. In a competition assay, the MEFA-6 protein competed with 83 to 96% of genotype- specific antibodies from HCV genotype-specific peptides. This recombinant antigen was subsequently used to design an anti-HCV chemiluminescent immunoassay. We designed our assay using a monoclonal anti-human immunoglobulin G antibody bound to the solid phase. Because MEFA-6 is fused with human superoxide dismutase (h-SOD), we used an anti-human superoxide dismutase, dimethyl acridinium ester-labeled monoclonal antibody for detection. Our results indicate that MEFA-6 exposes all of the major immunogenic epitopes. Its excellent sensitivity and specificity for the detection of clinical seroconversion are demonstrated by this assay.

Idioma originalInglés
Páginas (desde-hasta)1393-1397
Número de páginas5
PublicaciónJournal of Clinical Microbiology
Volumen37
N.º5
EstadoPublicada - 29 abr 1999

Áreas temáticas de ASJC Scopus

  • Microbiología (médica)

Huella Profundice en los temas de investigación de 'Use of a novel hepatitis C virus (HCV) major-epitope chimeric polypeptide for diagnosis of HCV infection'. En conjunto forman una huella única.

  • Citar esto

    Chien, D. Y., Arcangel, P., Medina-Selby, A., Coit, D., Baumeister, M., Nguyen, S., George-Nascimento, C., Gyenes, A., Kuo, G., & Valenzuela, P. (1999). Use of a novel hepatitis C virus (HCV) major-epitope chimeric polypeptide for diagnosis of HCV infection. Journal of Clinical Microbiology, 37(5), 1393-1397.