METHOD TO ASSESS HYPERSENSITIVITY TO HISTAMINE RELEASE FROM WHOLE BLOOD LEUKOCYTES

摘要

FIELD: medicine.;SUBSTANCE: method includes venous blood taking to vials with heparin, blood centrifugation, plasma replacement with buffer. The resuspended blood cells aliquotes are incubated with the tested substances in a microplate, as well as with anti-IgE as a positive control and with buffer as a negative control. After microplate centrifugation, thehistamine level is determined in the microplate holes supernatant using the method of reversed-phase high-performance liquid chromatography in combination with a tandem mass spectrometric detector. Levels of histamine, obtained for each of the substances being tested, are compared to positive and negative controls. With increased levels of histamine in the sample compared to the negative control in the presence of a positive control reaction, hypersensitivity to the substance being tested is revealed.;EFFECT: application of this method allows to diagnose sensitisation to an allergen, and also to monitor the effectiveness of antiallergic treatment by direct quantitative determination of histamine released from blood leukocytes by high-performance liquid chromatography and mass spectrometric analysis.;5 dwg, 1 tbl, 4 ex