title: Standardization and characterization of antigens for the diagnosis of aspergillosis.
abstract: The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of  Aspergillus  species  Aspergillus fumigatus ,  Aspergillus flavus , and  Aspergillus niger  were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS-PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.
authors.affiliation: Graduate Program in Medicine, Medical Sciences, Laboratory of Pathogenic Fungi, Department of Microbiology, Institute of Basic Health Sciences, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500/Sl 210, CEP 90050-170, Porto Alegre, RS, Brazil.
authors.forename: Cheila Denise Ottonelli
authors.initials: CD
authors.lastname: Stopiglia
authors.forename: Alicia
authors.initials: A
authors.lastname: Arechavala
authors.forename: Mariana
authors.initials: M
authors.lastname: Carissimi
authors.forename: Julia Medeiros
authors.initials: JM
authors.lastname: Sorrentino
authors.forename: Valério Rodrigues
authors.initials: VR
authors.lastname: Aquino
authors.forename: Tatiane Caroline
authors.initials: TC
authors.lastname: Daboit
authors.forename: Luana
authors.initials: L
authors.lastname: Kammler
authors.forename: Ricardo
authors.initials: R
authors.lastname: Negroni
authors.forename: Maria Lúcia
authors.initials: ML
authors.lastname: Scroferneker
mesh_terms: D000946:Antigens, Fungal / Q000097:blood* / Q000276:immunology; D001228:Aspergillosis / Q000175:diagnosis* / Q000276:immunology; D001230:Aspergillus / Q000502:physiology*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D011237:Predictive Value of Tests; D000516:alpha-Amylases / Q000378:metabolism
chemical_list: D000946:Antigens, Fungal; D000516:alpha-Amylases