Abstract
BackgroundsIgE and sIgG4 detection is necessary for more accurate and effective type I hypersensitivity diagnostics and the estimation of disease development. Typically, the analyses of these antibodies are performed separately with the help of various specialized systems. The aim of this study was to develop a microarray-based method for the simultaneous quantitative detection of sIgE and sIgG4 to the most common allergens in a single sample.MethodsA quantitative method for the simultaneous detection of sIgE and sIgG4 was developed based on the technology of hydrogel microchips previously designed at Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS). The microarray contained gel pads with immobilized allergens and gel pads that allow for the obtaining of sIgE and sIgG4 internal calibration curves for each allergen during the assay. The possibility of the simultaneous detection of sIgE and sIgG4 was developed using the corresponding Cy5 and Cy3 fluorescent dyes.ResultsThe multiplex immunoassay method using hydrogel microarrays developed in this study allowed the quantitative detection of sIgE and sIgG4 to 31 allergens from different groups in a single assay. A comparison of the microarray with the existing plate-based analogues (i.e., ALLERG-O-LIQ and sIgG4 ELISA) was performed by analysing 152 blood serum samples and by evaluating Pearson correlation coefficients, ROC analysis, and Passing-Bablok linear regression results.ConclusionThe implementation of this method in allergy diagnostics will provide the possibility of simultaneously performing primary patient screening and obtaining additional information concerning the severity of the allergies and the choice of an appropriate therapy.
Highlights
SIgE and specific immunoglobulin G4 (sIgG4) detection is necessary for more accurate and effective type I hypersensitivity diagnostics and the estimation of disease development
The allergens were chosen with consideration of the frequencies of allergen reactions in Central Russia, which mainly corresponds to the frequencies in Central and Northern Europe [20]
The structure of the microarray was enlarged with gel pads with immobilized immunoglobulins E and G4, which were used to plot an internal calibration curve
Summary
SIgE and sIgG4 detection is necessary for more accurate and effective type I hypersensitivity diagnostics and the estimation of disease development. The aim of this study was to develop a microarray-based method for the simultaneous quantitative detection of sIgE and sIgG4 to the most common allergens in a single sample. The manifestation of allergy symptoms can be suppressed by the presence of immunoglobulin G, which acts as a “blocking antibody”. According to the number of studies sIgG4 level as well as the ratio sIgE/sIgG4 is associated with likelihood of allergic symptoms reporting [6, 7] and can improve the prediction of tolerance to some allergens [8]. SIgG4 detection together with sIgE measurement gives more inclusive information for in vitro analysis interpretation
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