In vitro hymenoptera venom allergy diagnosis: improved by screening for cross‐reactive carbohydrate determinants and reciprocal inhibition

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Abstract
Background:  Immunoglobulin (Ig) E‐double positivity for honeybee (HB) and yellow jacket (YJ) venom causes diagnostic difficulties concerning therapeutical strategies. The aim of this study was to clarify the cause and relation of the cross‐reactivity in patients with insect venom allergy.Methods:  For this purpose, 147 patients with suspected stinging insect allergy and CAP‐FEIA‐double positivity were investigated for specific sIgE to additional cross‐reactive carbohydrate determinant (CCD)‐containing allergens: timothy grass pollen, rape pollen, natural rubber latex (NRL), bromelain, and horseradish peroxidase (HRP). Sera with sIgE to NRL were further investigated with the commercially available recombinant latex allergens. Reciprocal inhibition assays with both venoms and HRP were performed.Results:  About 36 of 147 (24.5%) patients had sIgE to both venoms only. However, 111 of 147 (75.5%) additionally reacted to CCD‐carrying allergens. 89 of 111 CCD‐reactive sera had NRL‐sIgE. In cases where inhibition experiments were performed, the NRL‐sIgE binding was completely abolished in the presence of HRP. Only nine of 61 sera were positive for at least one recombinant latex allergen; all of them were negative in history and NRL‐skin prick test. In 43 sera containing sIgE to CCD, HRP inhibition revealed unequivocal results: In 28 of 43 (65%) an HRP‐inhibition >70% of sIgE to one venom occurred, pointing out the relevant venom. In three of 43 sIgE proved to be entirely CCD‐specific.Conclusions:  Our data indicate that in cases of IgE positivity to both insect venoms supplementary screening tests with at least one CCD‐containing allergen should be performed; HRP being a suitable tool for this test. In addition, subsequent reciprocal inhibition is an essential diagnostic method to specify cross‐reacting sIgE results.