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https://www.arca.fiocruz.br/handle/icict/5536
AN IMPROVED ANTI-C3/IGG ELISA FOR QUANTIFICATION OF SOLUBLE IMMUNE COMPLEXES
Complemento C3/imunologia
Ensaio de Imunoadsorção Enzimática/métodos
Imunoglobulina G/imunologia
Humanos
Sensibilidade e Especificidade
Affilliation
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Escola Bahiana de Medicina e Saúde Pública. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Escola Bahiana de Medicina e Saúde Pública. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Abstract
A semi-quantitative ELISA for complement-fixing, IgG-containing immune complexes (IC) is described. The assay is based on the insolubilization of IC by polyethyleneglycol, their capture by solid-phase anti-C3 antibodies, reaction with peroxidase-labeled anti-IgG antibodies and incubation with a chromogenic peroxidase substrate. It was markedly improved by the use of a single-step procedure which simultaneously washed and precipitated the insolubilized immune complexes. Intra-assay and inter-assay coefficients of variation were lower than 8.6 and 14.7%, respectively. As expected, higher levels of circulating immune complexes, in relation to healthy individuals, were found in patients with American visceral leishmaniasis (AVL), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), with prevalences comparable to those described in the literature. The ELISA can be quickly assembled from reagents and plasticware widely available commercially, detects immune complexes fulfilling three different criteria and is more sensitive than a previously published method based on the same principles (detection limit for complement-sensitized aggregated IgG of 2 microg ml(-1) as compared with a detection limit above 16 microg ml(-1)).
DeCS
Complexo Antígeno-Anticorpo/análiseComplemento C3/imunologia
Ensaio de Imunoadsorção Enzimática/métodos
Imunoglobulina G/imunologia
Humanos
Sensibilidade e Especificidade
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