title: Simultaneous detection of total and allergen-specific IgE by using purified allergens in a fluorescent multiplex array.
abstract: BACKGROUND
  Testing serum samples for total and allergen-specific IgE requires separate testing for each antibody and allergen specificity.
  
  
  OBJECTIVE
  To apply fluorescent suspension array technology to allow simultaneous detection of total and allergen-specific IgE in serum in a single quantitative test.
  
  
  METHODS
  A 7-plex suspension array for the simultaneous detection of total IgE and IgE specific to Der p 1, Der p 2, Fel d 1, Can f 1, Bet v 1, and Phl p 5 was developed, using mAb or purified allergens covalently coupled to fluorescent microspheres. The multiplex array was validated by comparing total and allergen-specific IgE levels in serum from patients with allergy with results obtained by enzyme immunoassays.
  
  
  RESULTS
  There was a highly significant correlation between total IgE levels measured by multiplex array and fluorescent enzyme immunoassay (r = 0.97; P < .001; n = 63). Total and allergen-specific IgE levels also correlated with enzyme-linked and fluorescent enzyme immunoassay results (r = 0.44-0.94; n = 95 or 106). The multiplex array was reproducible (r = 0.86-0.99; mean coefficient of variance percentage, 12% to 25%). The sample volume required for a 7-plex assay was <20 microL per sample, compared with >400 microL in current immunoassays.
  
  
  CONCLUSIONS
  The multiplex array is a high-throughput system that allows simultaneous quantification of allergen-specific and total IgE.
  
  
  CONCLUSIONS
  Our results suggest that fluorescent multiplex technology will facilitate large-scale epidemiologic studies of allergic sensitization. The reduced serum volume is an advantage for pediatric studies.
authors.affiliation: Indoor Biotechnologies Inc, Charlottesville, VA 22903, USA. eking@inbio.com
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: King
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Vailes
authors.forename: Amy
authors.initials: A
authors.lastname: Tsay
authors.forename: Shama M
authors.initials: SM
authors.lastname: Satinover
authors.forename: Martin D
authors.initials: MD
authors.lastname: Chapman
mesh_terms: D000485:Allergens / Q000276:immunology*; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D005453:Fluorescence; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood*; D040081:Protein Array Analysis / Q000379:methods*; D039981:Pyroglyphidae / Q000276:immunology; D015203:Reproducibility of Results