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J Immunol Methods ; 323(2): 123-31, 2007 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-17490674

RESUMO

The concentration of mould-specific IgG antibodies in serum may objectively indicate mould exposure and can help identifying exposed individuals. Although inhaled spores probably are the most important source of mould exposure, the commonly used methods for detecting mould-specific IgG antibodies are based on extracts from all mould components, with only low contribution from spores. We have developed a flow cytometric method using surface antigens on mould spores for quantifying mould-specific IgG antibodies in serum. Flow cytometric results were evaluated by comparison with ImmunoCap and ELISA measurements. The flow cytometric assay showed a broad linear dose-dependency and correlated moderately to strongly (r=0.41-0.97) with ImmunoCap and ELISA measurements. The IgG antibody binding was studied in detail by immunolabelling in scanning electron microscopy (SEM), revealing that morphology and IgG antibody binding differed among spores, both within and between mould strains. Germination studies by flow cytometry and SEM showed that IgG antibody binding to mould spores was altered during germination due to loss of coat. The present spore based antibody assay are simple and suitable for quantification of mould-specific IgG antibodies in serum, and includes specificity to other and possibly more relevant antigens than existing methods.


Assuntos
Antígenos de Fungos/imunologia , Citometria de Fluxo/métodos , Imunoglobulina G/sangue , Microscopia Eletrônica de Varredura/métodos , Fungos Mitospóricos/imunologia , Esporos Fúngicos/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Fungos Mitospóricos/ultraestrutura , Esporos Fúngicos/ultraestrutura
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