title: [Development of an antigen 'sandwich' enzyme immunoassay for the detection of antibodies against HIV-2 by using a biotinylated synthetic peptide of gp36 protein].
abstract: BACKGROUND
  Among the several existing methods for the detection of antibodies to HIV, the 'sandwich' ELISA is currently the most used. This study aims to assess a biotinylated monomeric synthetic peptide of the glycoprotein trans-membrane gp36 from HIV-2, in a sandwich assay, for the detection of antibodies against this HIV-2 protein.
  
  
  METHODS
  To perform the assay, plates coated with recombinant protein gp36 at 0.5μg/mL and synthetic peptide gp36(5) at 1μg/mL were used. The concentration of the biotinylated synthetic peptide (gp36(5)-B) used was 0.1μg/mL prepared with a Tris-BSA-NaCl buffer solution and the Streptavidin- Alkaline Phosphatase conjugate diluted 1:30000 prepared with a PBS-Sucrose-BSA solution. Positive serum samples to antibodies against HIV-1 and HIV-2 viruses (88 and 34, respectively) were tested, with 483 negative samples from blood donors and 96 serum samples to assess the analytical specificity. All the samples were tested using the UMELISA HIV 1+2 RECOMBINANT assay, and all positives were confirmed using a DAHIV-BLOT assay.
  
  
  RESULTS
  Thirty four samples with antibodies against HIV-2 were assessed as positive for both coating variants. The highest specificity was obtained with the variant using the synthetic peptide gp36(5) in its coating. The antigen 'sandwich' assay developed by using gp36(5)-B enables the detection of antibodies against gp36 protein of HIV-2.
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba. Electronic address: iqhbsag1@cie.sld.cu.
authors.forename: Aurora
authors.initials: A
authors.lastname: Delahanty-Fernández
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Dunia Clara
authors.initials: DC
authors.lastname: Bequer-Ariza
authors.affiliation: Laboratorio de Síntesis de Péptidos, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Milenen
authors.initials: M
authors.lastname: Hernández-Marín
authors.affiliation: Laboratorio de Síntesis de Péptidos, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Orlando
authors.initials: O
authors.lastname: Zulueta-Rodríguez
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Lilliam
authors.initials: L
authors.lastname: Pozo-Peña
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Idialis
authors.initials: I
authors.lastname: Hernández-Spengler
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Grisell
authors.initials: G
authors.lastname: Ramos-Martínez
authors.affiliation: Laboratorio Retrovirus, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Marcos Antonio
authors.initials: MA
authors.lastname: Valdespino-Díaz
authors.affiliation: Laboratorio de Hepatitis, Centro de InmunoEnsayo, La Habana, Cuba.
authors.forename: Julio
authors.initials: J
authors.lastname: Ventura-Paz
mesh_terms: D000469:Alkaline Phosphatase; D019921:Biotinylation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005188:False Negative Reactions; D005189:False Positive Reactions; D015483:HIV Antibodies / Q000097:blood*; D015658:HIV Infections / Q000175:diagnosis*; D015498:HIV-2 / Q000276:immunology*; D006801:Humans; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D012680:Sensitivity and Specificity; D019809:Streptavidin; D054299:env Gene Products, Human Immunodeficiency Virus / Q000276:immunology*
chemical_list: D015483:HIV Antibodies; D010446:Peptide Fragments; D054299:env Gene Products, Human Immunodeficiency Virus; C059763:env protein gp36, Human immunodeficiency virus; D019809:Streptavidin; D000469:Alkaline Phosphatase
keywords: Esptretavidina/biotina; HIV; Péptidos sintéticos; Sandwich; Streptavidin/biotin; Synthetics peptides; VIH; gp36