title: Development of a microarray-based method for allergen-specific IgE and IgG4 detection.
abstract: BACKGROUND
  sIgE 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.
  
  
  METHODS
  A 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The 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.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Guzel
authors.initials: G
authors.lastname: Feyzkhanova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Sergei
authors.initials: S
authors.lastname: Voloshin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Olga
authors.initials: O
authors.lastname: Smoldovskaya
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Alla
authors.initials: A
authors.lastname: Arefieva
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Marina
authors.initials: M
authors.lastname: Filippova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Viktor
authors.initials: V
authors.lastname: Barsky
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Pavlushkina
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Veronika
authors.initials: V
authors.lastname: Butvilovskaya
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Alexei
authors.initials: A
authors.lastname: Tikhonov
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Reznikov
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences (EIMB RAS), Vavilova str., 32, Moscow, Russia 119991.
authors.forename: Alla
authors.initials: A
authors.lastname: Rubina
keywords: Allergy diagnostics; Biochips; Immunoassay; Microarrays; sIgG4