A new glass microfibre‐based histamine analysis for allergy testing in children

Abstract

A new microfibre method for allergy testing measuring histamine release from human basophil leukocytes is described. Samples of 50 μl washed blood are challenged with the suspected allergens. Released histamine is bound to microfibres and measured by a spectrofluorometrical method after removal of interfering substances by washing. The microfibre method (HR-MM) was compared to the conventional histamine release assay using the Ficoll-Hypaque gradient method (HR-FH) in 19 allergic children tested with one of three allergens. In addition, a comparison was made between the microfibre method and in vivo provocation tests, i.e. skin prick test (SPT), bronchial provocation test (BPT) and allergen specific serum IgE (RAST). It was found that the same individuals responded with histamine release to the same allergens in both histamine release assays, and the dose-response curves were almost identical. A positive correlation was found between the in vivo and in vitro tests. Thus it is concluded that the new method can provide reproducible, analytically precise (at the nanogram level) histamine release results in pediatric cases where: 1) a positive SPT does not correlate with case history; 2) BPT may be considered too hazardous or inconvenient; 3) confirmation of negative or inconclusive SPT or RAST is needed. In contrast to other histamine release assays it is a convenient diagnostic tool in children since only small amounts of blood are needed and at least 96 tests can be carried out in 21/2 h