Original Article

The Diagnostic Importance of Recombinant Allergen IgE Testing in Patients with Hymenoptera Venom Allergy: Comparison of Two Methods


Adults with systemic anaphylactic reactions (SAR) to insect sting show often multiple-positivity of serum-specific IgE (sIgE) to Hymenoptera venoms. Unnecessary long-lasting venom-specific immunotherapies (VIT) in false-positive patients increase the risk of recurrent SAR. This report aims to analyze the diagnostic importance of recombinant allergen IgE testing in patients with SAR to Hymenoptera sting.
In 82 patients we measured sIgE to honeybee venom (HBV), wasp venom (WV) and hornet venom (HV) extracts, recombinant phospholipase A2 from HBV (sIgE-rApi m1), recombinant antigen 5 from WV (sIgE-rVes v5), and cross-reactive carbohydrate determinants-CCD-bromelain by ImmunoCAP. We analyzed the correlation of ImmunoCAP and Immunoblot for HBV and WV extracts, rApi m1, and rVes v5 in 39/82 patients. According to the history of the culprit insect, we compared sensitivity and specificity between the two methods.
The severity of the SAR does not depend on the sIgE level to venom extracts and recombinant allergens. Fifty-one percent of the patients had a multiple-positivity to HBV/WV or HBV/WV/HV extracts. Severe SAR and CCD-sIgE were more frequent in multiple-positive than single-positive patients. CCD-sIgE were more frequent in HBV allergic patients than WV and HV allergic patients. There was a significant correlation between levels of sIgE to venom extracts and recombinant allergens measured by ImmunoCAP and Immunoblot. ImmunoCAP has higher sensitivity and specificity than Immunoblot for diagnosis of SAR to Hymenoptera venoms.
IgE testing to recombinant CCD-free allergens is necessary for the adequate selection of long-lasting VIT, especially in patients with multiple sensitivities to venom extracts.

1. Ruëff F, Przybilla B, Bilo´ MB, Müller U, Scheipl F, Aberer W, et al. Predictors of severe systemic anaphylactic reactions in patients with Hymenoptera venom allergy: importance of baseline serum tryptase-a study of the European Academy of Allergology and Clinical Immunology Interest Group on Insect Venom Hypersensitivity. J Allergy Clin Immunol. 2009;124(5):1047-54.
2. Müller UR, Johansen N, Peterson AB, Fromberg-Neilsen J, Haeberli G. Hymenoptera venom allergy: Analysis of double positivity of honey bee and vespula venom by estimation of IgE antibodies to species-specific major allergens Api m1 and Ves v5. Allergy. 2009;64(4):543-8.
3. Blank S, Biló MB, Ollert M. Component-resolved diagnostics to direct in venom immunotherapy: Important steps towards precision medicine. Clin Exp Allergy. 2018;48(4):354–64.
4. Biló MB. Anaphylaxis caused by Hymenoptera stings: from epidemiology to treatment. Allergy. 2011;95(2):35-7.
5. Ollert M, Blank S. Anaphylaxis to Insect Venom Allergens: Role of Molecular Diagnostics. Curr Allergy Asthma Rep. 2015;15(5):26.
6. King TP, Kochoumian L, Lam T. Immunochemical observations of antigen 5, a major venom allergen of hornets, yellowjackets and wasps. MolImmunol. 1987;24(8):857–64.
7. Schrautzer C, Bokanovic D, Hemmer W, Lang R, Hawranek T, Schwarz I, et al. Sensitivity and specificity of Hymenoptera allergen components depend on the diagnostic assay employed. J Allergy Clin Immunol. 2016;137(5):1603-5.
8. Hofman SC, Pfender N, Weckesser S, Huss-Marp J, Jakob T. Added value of IgE detection to rApi m1 and rVes v5 in patients with Hymenoptera venom allergy. J Allergy Clin Immunol. 2011;127(1):265-7.
9. Monsalve RI, Vega A, Marqués L, Miranda A, Fernández J, Soriano V, et al. Componenet-resolved diagnosis of vespid venom allergic individuals: phospholipases and antigen 5s are necessary to identify Vespula and Polistes sensitization. Allergy. 2012;67(4):528-36.
10. Korosec P, Valenta R, Mittermann I, Celesnik N, Erzen R, Zidarn M, et al. Low sensitivity of commercially available rApi m1 for diagnosis of honey bee venom allergy. J Allergy Clin Immunol. 2011;128(3):671-3.
11. Müller U, Schmid-Grendelmeier P, Hausmann O, Helbling A. IgE to recombinant allergens Api m 1, Ves v 1, and Ves v 5 distinguish double sensitization in venom allergy. Allergy. 2012;67(8):1069‒73.
12. Šelb J, Kogovšek R, Šilar M, Košnik M, Korošec P. Improved recombinant Api m1- and Ves v5-based IgE testing to dissect bee and yellow jacket allergy and their correlations with the severity of the sting reaction. Clin Exp Allergy. 2016;46(4):621-30.
13. Köhler J, Blank S, Müller S, Bantleon F, Frick M, Huss-Marp J, et al. Component resolution reveals additional major allergens in patients with honeybee venom allergy. J Allergy Clin Immunol. 2014;133(5):383-9.
14. Mueller HL. Diagnosis and treatment of insect sensitivity. Asthma Res 1966; 3 (4): 331-3.
15. Krishna MT, Ewan PW, Diwakar L, Durham SR, Frew AJ, Leech SC, et al. Diagnosis and management of hymenoptera venom allergy: British Society for Allergy and Clinical Immunology (BSACI) guidelines. Clin Exp Allergy. 2011;41(9):1201-20.
16. Hoffman DR. Allergens in Hymenoptera venom. XXV: The amono acids equences of antigen 5 molecules and the structural basis of antigenic cross-reactivity. J Allergy Clin Immunol.1993;92(5):707-16.
17. Fehr D, Micalleto S, Moehr T, Schmid-Grendelmeier P. Risk factors for severe systemic sting reactions in wasp (Vespula spp.) and honeybee (Apismellifera) venom allergic patients. ClinTransl Allergy. 2019;11(2):9-54.
18. Sturm GJ, Varga E-M, Roberts G, Mosbech H, Bilo MB, Akdis CA, et al. EAACI guidelines on allergen immunotherapy: Hymenoptera venom allergy. Allergy. 2018;73(4):744–64.
19. Eberlein B, Krischan L, Darsow U, Ollert M, Ring J. Double positivity to bee and wasp venom: improved diagnostic procedure by recombinant allergen-based IgE testing and basophil activation test including data about cross-reactive carbohydrate determinants. J Allergy Clin Immunol. 2012;130(1):155‒61.
20. Jovanovic D, Peric-Popadic A, Andrejevic S, Jovanovic I, Bonaci-Nikolic B. Triple IgE-positivity to hornet, wasp and bee venom in the patient with SAR: diagnostic and therapeutic approach. Vojnosanit Pregl. 2019;76(2):839-42.
21. Jin C, Focke M, Leonard R, Jarisch R, Altmann F, Hemmer W. Reassessing the role of hyaluronidase in yellow jacket venom allergy. J Allergy Clin Immunol. 2010;125(1):184–90.
22. Blank S, et al. Identification, recombinant expression and characterization of the 100 kDa high molecular weight Hymenoptera venom allergens Api m5 and Ves v3. J Immunol.2010;184(9):5403-13.
23. Watanabe M, Hirata H, Arima M, Hayashi Y, Chibana K, Yoshida N, et al. Measurement of Hymenoptera venom specific IgE by the IMMULITE 3gAllergy in subjects with negative or positive result by ImmunoCAP. Asia Pac Allergy. 2012; 2(3):195-202.
24. Hemmer W, Focke M, Kolarich D, Wilson B I, Almann F, Wöhrl S, et al. Antibody binding to venom carbohydrates is a frequent cause for double positivity to honey bee and yellow jacket venom in patients with stinging-insect allergy. J Allergy Clin Immunol.2001;108(6):1045-52.
25. King TP, Spangfort MD. Structure and biology of stinging insect venom allergens. Int Arch Allergy Immunol. 2000;123(8):99-106.
26. Sturm GJ, Biló MB, Bonadonna P, Hemmer W, Caruso B, Bokanovic D, Aberer W. Ves v 5 can establish the diagnosis in patients without detectable specific IgE to wasp venom and a possible north-south difference in Api m 1 sensitization in Europe. J Allergy Clin Immunol.2012;130(3):818-9.
27. Korošec P, Valenta R, Mittermann I, Čelsnik N, Šilar M, Zidarn M, Košnik M . High sensitivity of CAP-FEIA rVes v5 and rVes v1 for diagnosis of Vespula venom allergy. J Allergy ClinImmunol. 2012;129(3):1406-8.
28. King TP, Lu G, Gonzales M, Qian N, SoldatovaL.Yellow jacket venom allergens, hyaluronidase and phospholipase: sequence similarity and antigenic cross-reactivity with hornet and wasp homologs and possible implications for clinical allergy. J Allergy Clin Immunol. 1996;98(4):588-600.
29. Yoshida N, Hirata H, Watanabe M, Sugiyama K, Arima M, Fukushima Y, Ishii Y. Improved sensitivity to venom specific-immunoglobulin E by spiking with the allergen component in Japanese patients suspected of Hymenoptera venom allergy. Allergol Int. 2015;64(3):248–52.
30. Sturm GJ, Hemmer W, Hawranek T, Lang R, Ollert M, Spillner E, et al. Detection of IgE to recombinant Api m1 and rVes v5 is valuable but not sufficient to distinguish bee from wasp venom allergy. J Allergy Clin Immunol. 2011;128 (1):247-8.
IssueVol 20 No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijaai.v20i4.6951
Anaphylaxis Hymenoptera Honeybee venoms Immunoblotting Wasp venoms

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How to Cite
Jovanovic D, Peric-Popadic A, Andrejevic S, Stojanovic M, Bonaci-Nikolic B. The Diagnostic Importance of Recombinant Allergen IgE Testing in Patients with Hymenoptera Venom Allergy: Comparison of Two Methods. Iran J Allergy Asthma Immunol. 2021;20(4):413-422.