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Comparative Study
. 2025 Nov 13;15(1):39751.
doi: 10.1038/s41598-025-18601-x.

Comparison of basophil activation testing and component-resolved diagnosis in patients with cross-reactive intradermal results after anaphylactic reactions to hymenoptera venom

Affiliations
Comparative Study

Comparison of basophil activation testing and component-resolved diagnosis in patients with cross-reactive intradermal results after anaphylactic reactions to hymenoptera venom

Mohammad Hasan Bemanian et al. Sci Rep. .

Abstract

Hymenoptera venom allergy (HVA) is a significant cause of anaphylaxis, with a broad spectrum of symptoms. Accurate diagnosis is critical for selecting appropriate venom immunotherapy (VIT), yet multiple sensitizations complicate the identification of the culprit Hymenoptera. Component-resolved diagnostics (CRD) and the basophil activation test (BAT) are emerging approaches that may enhance the precision of HVA diagnosis. This study aimed to compare the outcomes of the BAT and CRD tests in patients with reaction to more than one type of Hymenoptera in intradermal test. Following 12 patient's report-based determination of the Hymenoptera types, Intradermal testing with venom extracts from honey bees, paper wasps, and yellow jackets was performed. CRD was conducted using recombinant allergens for each venom, and BAT was performed to assess basophil activation using venom extracts. The intradermal test indicated 10 triple-positive reactions (83.3%) and 2 double-positive reactions (16.7%). In addition, 7 (58.4%) individuals were triple-positive and 5 (41.6%) were double-positive in the CRD test. The BAT had 6 (50.0%) double-positive individuals and 3 (25.0%) single-positive patients. The diagnostic complexity in patients with multi-sensitivity to Hymenoptera venoms is highlighted by the results. While CRD provides a more specific identification of allergenic components, valuable information is offered by the BAT, especially when the clinical diagnosis is uncertain. As a result, relying just on one test seems to have limited diagnostic capabilities and it is more efficient to use many diagnostic tests in order to get a thorough evaluation.

Keywords: Basophil activation test; Component-resolved diagnosis; Cross reactivity; Hymenoptera venom allergy; Venom immunotherapy.

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Conflict of interest statement

Declarations. Competing interests: The authors declare no competing interests. Ethics approval: The present study was conducted according to the principles expressed in the Helsinki Declaration. The study was approved by ethics committee of the Iran University of Medical Sciences (Ethical code: IR.IUMS.FMD.REC.1400.61). Consent to participate: Informed consent for participation was obtained from all patients and/or their legal guardians included in the study.

Figures

Fig. 1
Fig. 1
Gating strategy of flow cytometry. By comparing forward scatter to side scatter, the lymphocyte/basophil fraction was initially chosen. As basophils, side scatter-low and CD203c-high cell populations were gated. Activated and degranulated basophils were ultimately selected by separating the populations of cells with low side scatter and high CD63. The activation of more than 10% of basophils was regarded as an affirmative outcome of the BAT (A: Positive control, B: Negative control, C: Honey bee, D: Paper wasp, E: Yellow jacket).
Fig. 2
Fig. 2
Results of intradermal test of patients by type of hymenoptera (HB: Honey bee, YJ: Yellow jacket, PW: Paper wasp).
Fig. 3
Fig. 3
Comparison of the outcomes of intradermal test, CRD test, and BAT in all patients (†: Patient No.6 did not respond to the positive control, so it was excluded from the results of the BAT for analysis).

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