Principle of Histamine Release Test (HR-Test).
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HR-Test for autoimmune urticaria |
The Histamine Release Test is performed in microtiter plates coated with glass fibers providing a solid phase for histamine binding. The glass fibers bind histamine with high affinity and selectivity. The Histamine Release Test procedure includes histamine release from peripheral whole blood basophils, in a glass fiber prepared microtiter plate, by challenging 25 µL heparinized whole blood with 25 µL buffered allergen dilutions in various concentrations for 60 minutes at 37°C. After incubation the microtiter plate is washed and incubated with SDS to remove cells and interfering substances. Histamine is determined by the fluorometric o-Phthal-di-aldehyde method. The assay is calibrated by a two point standard curve and a positive cell control (anti-IgE). Test results are either classified in classes 0,1,2 and 3 (standardized allergens) or reported as negative or positive.
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