The application of the sepharose bead immunofluorescence test for the detection of allergen-specific IgE and IgG antibodies in pollinosis.
A new application of the Sepharose bead immunofluorescence test for detection of allergen-specific IgE and IgG antibodies is described. Allergen extracts of four different grass pollens were coupled to CNBr-activated Sepharose 4B. Twenty normal and allergic sera were incubated with the allergen-coupled beads, washed and incubated with fluorescence-conjugated anti-gamma E and G globulins. After washing and staining with 0.5% trypan blue, the percentage of fluorescent beads was detected by fluorescence microscopy. In the IgG/anti IgG system, the smallest amount of IgG demonstrated was 20ng/ml; in the IgE/anti-IgE system it was 40ng/ml. Independent examination of tests gave a mean difference between observations of 4.1%. Reproducibility was also very good (var. coeff = 18%). The number of beads stained with IgE correlated well with intensity of skin reactions to the same extracts (r = 0.64; p = 5 x 10-7), the percentage of IgG stained beads being independent of skin reactivity. The Sepharose-IgE test allowed clear distinction between allergic and normal sera (p = 4 x 10-7), while the Sepharose IgG test did not distinguish between them at the time of diagnosis. However, the number of beads stained with IgG significantly increased in patients undergoing immunotherapy (p = 3.8 x 10-3). The Sepharose bead immunofluorescence test requires a very small amount of materials, is highly sensitive and easy to handle. It may be valuable in the "in vitro" diagnosis of grass pollen allergy and useful in evaluating immunotherapy.
D000485:Allergens; D000906:Antibodies / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis*; D011058:Pollen; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology*; D012685:Sepharose
D000485:Allergens; D000906:Antibodies; D007074:Immunoglobulin G; D007073:Immunoglobulin E; D012685:Sepharose