Euan R. Tovey

Mite allergen content in commercial extracts and in bed dust determined by radioallergosorbent tests

Radioallergosorbent (RAST) direct binding and inhibition type assays were used to quantitate the mite (Dermatophagoides pteronyssinus) allergen content of four commercial mite extracts and a laboratory prepared extract from freeze‐dried mites. The content of mite allergen in extracts prepared from twenty samples of dust vacuumed from bedding was measured by RAST inhibition assay. The four commercial mite extracts designated A, B, C and D, and the laboratory extract, designated L, contained 52, 265, 108, 1.5 and 581 arbitrary units of allergen/ml for the direct binding assay and 128, 111, 217, ∼1 and 1083 arbitrary, but different, units of allergen/ml for the inhibition assay respectively. Qualitative differences between at least two extracts were suggested by the different slopes obtained when allergen binding of anti IgE was plotted against the volume of extract used in the direct binding assay. Differences in slope between the two extracts were also apparent when they were used in the inhibition assay. The quantities of mite allergen/gm of bed dust expressed in arbitrary units for the inhibition assay were 24 to 457 (mean 129) units. These quantities are similar to and sometimes greater than the quantity in 1 ml of mite extract and so confirm bed dust as a potent source of mite allergen. There was no significant correlation between the weight of dust, the numbers of dead and live mites and the allergen content of dust.

Effect of reagins and allergen extracts on radioallergosorbent assays for mite allergen

The reproducibility of the radicallororbent (RAST) inhibition and direct binding assays with mite allergen were investigated in the presence of heterogeneous extracts and non‐mite‐sensitive atopic sera. Both contain components similar to potential contaminants which would occur in the assay of mite allergen and dust allergen extracts. The standardized inhibition and direct binding assays employed had a day to day (n= 4) coefficient of variation [(s.d. ± 100)/mean] of 15% and 24% respectively. The inhibition assay for mite allergen was reproducible in the presence of protein concentrations of added plant, fungal, arthropod and animal extracts in excess of the protein concentrations that occur under the operational mite assay conditions. The mite inhibition assay was also reproducible in the presence of non‐mite allergen extracts, with and without additional sera containing IgE specific for the non‐mite allergens. The binding of a constant quantity of mite allergen to the activated solid phase in the direct binding assay was reproducible in the presence of added bovine serum albumin, and of a fungal or arthropod extract, representing the heterogeneous components of an allergen extract at the concentrations of total protein known to occur in the direct binding assay of mite extracts.