RÉSUMÉ
Acute hepatitis C virus (HCV) infection is usually asymptomatic, therefore, early diagnosis is rare. It may remain undiagnosed in individuals who progress to chronic infection, often until serious liver damage has developed. To incorporate the diagnosis of this viral disease in a multiple-diagnostic assay, we first analyzed by immunoinformatics the HCV subtype 1a polyprotein (specifically Core, E2, NS3, NS5A proteins) to select antigenic peptides to be tested initially by the Pepscan technique. Next, we performed the immunodiagnosis of HCV infection, using the Multiple Antigen Blot Assay (MABA). In 22 patients' sera included in this study, a 20-mer linear peptide belonging to the N-terminus of the worldwide conserved Core protein showed 100% sensitivity and specificity; other sequences showed different levels of antibody recognition. The use of MABA in combination with synthetic peptides as a source of multiple, specific, and nonexpensive antigens for other infectious diseases could represent a rapid, integrated, and inexpensive diagnostic methodology.
Sujet(s)
Hepacivirus/immunologie , Hépatite C/diagnostic , Tests immunologiques/méthodes , Peptides/immunologie , Protéines virales non structurales/immunologie , Maladie aigüe , Antigènes viraux/immunologie , Hepacivirus/isolement et purification , Hépatite C/sang , Hépatite C/immunologie , Anticorps de l'hépatite C/sang , Humains , Immunotransfert/méthodes , Peptides/synthèse chimique , Protéines du core viral/immunologie , Protéines virales non structurales/isolement et purificationRÉSUMÉ
This technique is based on the sensitization of different antigens in a single nitrocellulose strip, which react when exposed to an immune serum and thereafter with the appropriate peroxidase conjugate and the corresponding substrate. Signals in those reactive spots are recorded as black squares in a negative photographic film, using a chemiluminiscent substrate or as blue spots when a precipitable colorimetric substrate is used. This technique allows the simultaneous demonstration of antigenicity of different antigens (peptides, recombinant molecules, and crude preparations), with a high sensitivity and specificity. Its major value is based on its versatility, since it is possible to rapidly evaluate and to compare various antigenic preparations and to use it for diagnosis of different infectious, allergic and autoimmune diseases, at a low cost.