Development and validation of a fluorescent microsphere immunoassay for anti-IgA

Immunohematology. 2009;25(1):24-8.

Abstract

Anti-IgA may cause anaphylactic transfusion reactions in IgA-deficient individuals. Testing for IgG anti-IgA is useful to identify persons at risk. This report describes an immunoassay for anti-IgA that uses polyclonal IgA coupled to fluorescent microspheres as an immunosorbent. Anti-IgA is detected by phycoerythrin-labeled anti-IgG. The assay is calibrated in arbitrary units by use of a serum that contains anti-IgA. Dose-response studies with sera that contain anti-IgA showed positive responses at dilutions up to 32-fold greater than the dilution used to test patients' samples. Inhibition studies with purified IgA and IgA-deficient serum showed no inhibition with IgA-deficient serum and complete inhibition with soluble IgA. Clinical tests performed in more than 90 assays had a CV of 13.6 percent for measurements of an internal positive control. The fluorescent immunoassay method is rapid, reproducible, and sensitive to low concentrations of IgG anti-IgA.

Publication types

  • Validation Study

MeSH terms

  • Anaphylaxis / blood
  • Anaphylaxis / diagnosis
  • Anaphylaxis / immunology
  • Antibodies, Anti-Idiotypic / blood*
  • Fluorescent Antibody Technique, Direct*
  • Humans
  • IgA Deficiency / blood
  • Immunoglobulin G / blood
  • Microspheres*
  • Sensitivity and Specificity

Substances

  • Antibodies, Anti-Idiotypic
  • Immunoglobulin G
  • anti-IgA