title: Application of the dot immunobinding assay to allergy diagnosis.
abstract: The recently developed dot immunobinding assay is operationally simple and facilitates performance of multiple simultaneous assays. Here, its use as the basis for determination of total and allergen-specific IgE is established. For total IgE, the same commercially available polyclonal anti-human IgE was used on the solid phase and as a peroxidase conjugate in the liquid phase. After incubation with a chromogenic substrate, IgE was determined from the color intensity of the resulting dots with a scanning reflectance densitometer. The limit of sensitivity was 50 pg/ml of IgE. Standardized conditions gave the dynamic range 50 to 2500 IU/ml in serum. The IgE measured was not subject to interference by serum components, was labile at 56 degrees C, was soluble at 30% saturation (NH4)2SO4, and was unaffected by anti-human immunoglobulins of other specificity. Coefficients of variation were 0.05 within run, and were 0.1 between run. Comparison with data on sera obtained with the PRIST method yielded a correlation coefficient of 0.96 and a linear regression of slope 1.15. Assays for allergen-specific IgE were established with bee venom and dust mite allergens in the solid phase. The same peroxidase-conjugated antibody was used as for total IgE. Comparisons with comparable RAST assays were performed.
mesh_terms: D000918:Antibody Specificity; D001514:Bee Venoms; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis*; D007073:Immunoglobulin E / Q000032:analysis*; D007158:Immunologic Techniques; D008925:Mites; D010209:Paper; D011852:Radioallergosorbent Test; D011864:Radioimmunosorbent Test
chemical_list: D001514:Bee Venoms; D007073:Immunoglobulin E