title: Design and application of GB virus C (GBV-C) peptide microarrays for diagnosis of GBV-C/HIV-1 co-infection.
abstract: The main objectives of the design of GB virus C (GBV-C) peptide microarrays are the miniaturisation of antigen-antibody interaction assays, the simultaneous analysis of several peptide sequences and the reduction in the volume of serum required from patients since this always represents a limiting factor in studies to develop new systems for diagnosing human diseases. We herein report the design of a microarray immunoassay based on synthetic peptides derived from the GBV-C E2 protein to evaluate their diagnostic value in detecting anti-E2 antibodies in HIV-1 patients. To this end, peptide microarrays were initially prepared to identify the most relevant epitopes in the GBV-C E2 protein. Thus, 124 peptides composed of 18 amino acids covering the whole E2-protein sequence, with 15 residue overlaps, were spotted in triplicate onto γ-aminopropyl silane-functionalised adsorbent binding slides. The procedure to select the E2 protein epitopes was carried out using serum samples from HIV-1-infected patients. The samples had previously been tested for the presence or absence of GBV-C anti-E2 antibodies by means of the Abbott test. Thus, 11 specific epitopes in the GBV-C E2 protein were identified. Subsequently, peptide antigen microarrays were constructed using the E2 epitopes identified to detect GBV-C anti-E2 antibodies in the serum of HIV-1-infected patients with no known GBV-C co-infection. The 11 peptides selected identified anti-E2 GBV-C antibodies among HIV-1-infected patients, and a reactivity of 47 % was established. The potential antigenic peptides selected could be considered a useful tool for designing a new diagnostic system based on peptide microarrays to determine anti-GBV-C E2 antibodies in the serum of HIV-1-infected patients.
authors.affiliation: Unit of Synthesis and Biomedical Applications of Peptides, IQAC-CSIC, Jordi Girona, Barcelona, Spain.
authors.forename: Leticia
authors.initials: L
authors.lastname: Fernández
authors.forename: M José
authors.initials: MJ
authors.lastname: Bleda
authors.forename: M José
authors.initials: MJ
authors.lastname: Gómara
authors.forename: Isabel
authors.initials: I
authors.lastname: Haro
mesh_terms: D017502:Adenovirus E2 Proteins; D000914:Antibodies, Viral / Q000097:blood; D060085:Coinfection; D000939:Epitopes / Q000737:chemistry; D018178:Flaviviridae Infections / Q000097:blood / Q000150:complications / Q000175:diagnosis*; D029407:GB virus C / Q000302:isolation & purification*; D015658:HIV Infections / Q000097:blood / Q000150:complications* / Q000175:diagnosis; D015497:HIV-1 / Q000302:isolation & purification; D006525:Hepatitis, Viral, Human / Q000097:blood / Q000150:complications / Q000175:diagnosis*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods*; D014764:Viral Proteins / Q000737:chemistry
chemical_list: D017502:Adenovirus E2 Proteins; D000914:Antibodies, Viral; D000939:Epitopes; D014764:Viral Proteins