Measurement of Allergens Associated with Dust Mite Allergy

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Abstract
Sensitive radioimmunoassays were developed to measure the two groups of major allergens (Der I and Der II) of Dermatophagoides pteronyssinus and Dermatophagoides farinae, in which radiolabeled protein A served as a general tracer. Glass rods covalently coupled with F(ab′)2 fragments of rabbit antiserum IgG were incubated first with mite or house dust extracts, and then with affinity-purified rabbit antibodies. The bound allergen-antibody complex was detected with a 125I-labeled protein A. The two Der I allergens, Der p I and Der f I, were measured separately with the use of immunoabsorbed rabbit antibodies directed against species-specific determinants, while the two Der II allergens, Der p II and Der f II, were measured as ‘Der II’ with the use of antibodies directed against common determinants on both allergens. Each assay demonstrated consistently parallel dilution curves with mite and house dust extracts. The mean intraassay and interassay coefficient of variation for each assay ranged from 4.3 to 7.7% and from 3.6 to 9.0%, respectively. The Der p I and Der f I assays were shown to be highly species-specific so that the ratio of Der p l:Der f I would provide a good index of the distribution of the two mite species in a dust sample. The concentrations of the Der I and Der II allergens in different types of mite extract and dust samples from houses were compared using these assays. The results demonstrated that the ratio of Der I: Der II in the mite body extracts varied markedly from those in the whole culture extracts, while the ratio in individual dust samples was nearly constant and was close to that in the mite body extracts. These assays will be useful in standardizing mite and house dust extracts and for assessments of mite allergen exposure. Furthermore, the high assay sensitivity will make it possible to use these assays to measure mite allergens associated with airborne particles.

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