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Veterinary journal (London, England : 1997)2013; 198(1); 141-147; doi: 10.1016/j.tvjl.2013.06.010

Evaluation of an IgE ELISA with Culicoides spp. extracts and recombinant salivary antigens for diagnosis of insect bite hypersensitivity in Warmblood horses.

Abstract: Insect bite hypersensitivity (IBH) in horses represents an immunoglobulin E (IgE)-mediated hypersensitivity to salivary antigens from biting midges (Culicoides spp.). The aim of this study was to evaluate and compare the performances of IgE ELISAs using recombinant Culicoides spp. Obsoletus group salivary gland antigens or crude whole body extracts ('ObsWBE'), C. nubeculosus recombinant proteins (Culn1, 3, 4, 5, 7, 8 and 10) and Obsoletus group recombinant proteins (Culo1 and 2). IgE levels were measured in plasma of 343 Warmblood horses classified as IBH-affected (n=167) and IBH-unaffected (n=176) according to the owners' descriptions. IBH-affected horses were subdivided based on the severity of their clinical signs at sampling and whether or not their IBH history was considered to be classical. The accuracies of the tests increased when clinical signs at sampling were more pronounced or when the IBH history could be considered as classical. A combination of IgE levels against the three best performing Culicoides spp. recombinant proteins (Culn4, Culo1 and Culo2) and ObsWBE resulted in the best performing test. When IBH-affected horses showing a classical history of the disease and severe clinical signs were compared with IBH-unaffected horses, the Youden's index at the optimal cut-off for the three tests in combination was 0.67. This optimal cut-off had a sensitivity of 70%, a specificity of 97% and a total accuracy of 92%. The performance of the IgE ELISA was affected by the severity of IBH clinical signs at sampling and was improved when IgE levels against several recombinant proteins were combined.
Publication Date: 2013-07-25 PubMed ID: 23891138DOI: 10.1016/j.tvjl.2013.06.010Google Scholar: Lookup
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  • Evaluation Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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The research article investigates the efficiency of an IgE ELISA using Culicoides spp. extracts and recombinant salivary antigens as a diagnostic tool for insect bite hypersensitivity (IBH) in Warmblood horses. The best results were obtained when the test combined IgE levels against three best performing Culicoides spp. recombinant proteins (Culn4, Culo1, and Culo2) and ObsWBE, especially when the horses showed a classical history of the disease and severe clinical signs of IBH.

Understanding Insect Bite Hypersensitivity

  • Insect bite hypersensitivity (IBH) is a common condition in horses that is caused by an allergic reaction to the saliva of biting insects, specifically Culicoides spp.
  • It’s an immunoglobulin E (IgE) mediated hypersensitivity, which means the horses’ immune systems overreact to the antigens in the insect saliva, leading to allergic symptoms.

Objective of the Study

  • The aim of this research was to evaluate how effective the IgE ELISA tests can be in diagnosing IBH in horses.
  • The researchers tested two kinds of ELISA: one using recombinant (engineered) Culicoides spp. Obsoletus group salivary gland antigens and another using crude whole body extracts (‘ObsWBE’), in combination with various C. nubeculosus and Obsoletus group recombinant proteins.

Methodology and Findings

  • The study involved 343 Warmblood horses, some affected by, and others unaffected by, IBH as defined by their owners.
  • IgE levels in each horse were measured and compared using the different variations of the ELISA tests.
  • The best performing test was a combination of IgE levels against the three best performing Culicoides spp. recombinant proteins (Culn4, Culo1, and Culo2) and ObsWBE.
  • The performance of the IgE ELISA was improved when it was used on horses showing severe clinical signs of IBH and a classical history of the disease.

Conclusion

  • The study found that the IgE ELISA tests were effective in diagnosing IBH in horses, especially when used in combination and on horses with a classical history and more severe signs of the disease.
  • The optimal cut-off point for the tests was found to have a sensitivity of 70%, a specificity of 97%, and a total accuracy of 92%.

Cite This Article

APA
Peeters LM, Janssens S, Goddeeris BM, De Keyser K, Wilson AD, Kaufmann C, Schaffartzik A, Marti E, Buys N. (2013). Evaluation of an IgE ELISA with Culicoides spp. extracts and recombinant salivary antigens for diagnosis of insect bite hypersensitivity in Warmblood horses. Vet J, 198(1), 141-147. https://doi.org/10.1016/j.tvjl.2013.06.010

Publication

ISSN: 1532-2971
NlmUniqueID: 9706281
Country: England
Language: English
Volume: 198
Issue: 1
Pages: 141-147
PII: S1090-0233(13)00294-3

Researcher Affiliations

Peeters, L M
  • Department of Biosystems, Katholieke Universiteit Leuven, BE-3001 Heverlee, Belgium. Electronic address: liesbet.peeters01@gmail.com.
Janssens, S
    Goddeeris, B M
      De Keyser, K
        Wilson, A D
          Kaufmann, C
            Schaffartzik, A
              Marti, E
                Buys, N

                  MeSH Terms

                  • Allergens / genetics
                  • Allergens / immunology
                  • Animals
                  • Ceratopogonidae / physiology
                  • Enzyme-Linked Immunosorbent Assay / methods
                  • Enzyme-Linked Immunosorbent Assay / veterinary
                  • Horse Diseases / diagnosis
                  • Horse Diseases / immunology
                  • Horses
                  • Hypersensitivity / diagnosis
                  • Hypersensitivity / immunology
                  • Hypersensitivity / veterinary
                  • Immunoglobulin E / blood
                  • Insect Bites and Stings / diagnosis
                  • Insect Bites and Stings / immunology
                  • Insect Bites and Stings / veterinary
                  • Insect Proteins / genetics
                  • Insect Proteins / immunology
                  • Molecular Sequence Data
                  • Recombinant Proteins / genetics
                  • Recombinant Proteins / immunology
                  • Salivary Glands / metabolism
                  • Sequence Analysis, DNA / veterinary

                  Grant Funding

                  • BBS/E/I/00001445 / Biotechnology and Biological Sciences Research Council
                  • BBS/E/I/00001701 / Biotechnology and Biological Sciences Research Council

                  Citations

                  This article has been cited 12 times.
                  1. Pessoa VC, Branco-Ferreira M, Jónsdóttir S, Marti E, Tilley P. Comparison of Skin Prick Tests (SPT), Intradermal Tests (IDT) and In Vitro Tests in the Characterization of Insect Bite Hypersensitivity (IBH) in a Population of Lusitano Horses: Contribution for Future Implementation of SPT in IBH Diagnosis.. Animals (Basel) 2023 Aug 28;13(17).
                    doi: 10.3390/ani13172733pubmed: 37684997google scholar: lookup
                  2. Söderroos D, Ignell R, Haubro Andersen P, Bergvall K, Riihimäki M. The Effect of Insect Bite Hypersensitivity on Movement Activity and Behaviour of the Horse.. Animals (Basel) 2023 Apr 8;13(8).
                    doi: 10.3390/ani13081283pubmed: 37106846google scholar: lookup
                  3. Cvitas I, Oberhaensli S, Leeb T, Marti E. Equine keratinocytes in the pathogenesis of insect bite hypersensitivity: Just another brick in the wall?. PLoS One 2022;17(8):e0266263.
                    doi: 10.1371/journal.pone.0266263pubmed: 35913947google scholar: lookup
                  4. Birras J, White SJ, Jonsdottir S, Novotny EN, Ziegler A, Wilson AD, Frey R, Torsteinsdottir S, Alcocer M, Marti E. First clinical expression of equine insect bite hypersensitivity is associated with co-sensitization to multiple Culicoides allergens.. PLoS One 2021;16(11):e0257819.
                    doi: 10.1371/journal.pone.0257819pubmed: 34780496google scholar: lookup
                  5. Novotny EN, White SJ, Wilson AD, Stefánsdóttir SB, Tijhaar E, Jonsdóttir S, Frey R, Reiche D, Rose H, Rhyner C, Schüpbach-Regula G, Torsteinsdóttir S, Alcocer M, Marti E. Component-resolved microarray analysis of IgE sensitization profiles to Culicoides recombinant allergens in horses with insect bite hypersensitivity.. Allergy 2021 Apr;76(4):1147-1157.
                    doi: 10.1111/all.14556pubmed: 32780483google scholar: lookup
                  6. Raza F, Ivanek R, Freer H, Reiche D, Rose H, Torsteinsdóttir S, Svansson V, Björnsdóttir S, Wagner B. Cul o 2 specific IgG3/5 antibodies predicted Culicoides hypersensitivity in a group imported Icelandic horses.. BMC Vet Res 2020 Aug 10;16(1):283.
                    doi: 10.1186/s12917-020-02499-wpubmed: 32778104google scholar: lookup
                  7. Olomski F, Fettelschoss V, Jonsdottir S, Birkmann K, Thoms F, Marti E, Bachmann MF, Kündig TM, Fettelschoss-Gabriel A. Interleukin 31 in insect bite hypersensitivity-Alleviating clinical symptoms by active vaccination against itch.. Allergy 2020 Apr;75(4):862-871.
                    doi: 10.1111/all.14145pubmed: 31816097google scholar: lookup
                  8. Wilkołek P, Szczepanik M, Sitkowski W, Rodzik B, Pluta M, Taszkun I, Gołyński M. Evaluation of multiple allergen simultaneous (sIgE) testing compared to intradermal testing in the etiological diagnosis of atopic dermatitis in horses.. J Vet Sci 2019 Nov;20(6):e60.
                    doi: 10.4142/jvs.2019.20.e60pubmed: 31775187google scholar: lookup
                  9. François L, Hoskens H, Velie BD, Stinckens A, Tinel S, Lamberigts C, Peeters L, Savelkoul HFJ, Tijhaar E, Lindgren G, Janssens S, Ducro BJ, Buys N, Schurink AA. Genomic Regions Associated with IgE Levels against Culicoides spp. Antigens in Three Horse Breeds.. Genes (Basel) 2019 Aug 8;10(8).
                    doi: 10.3390/genes10080597pubmed: 31398914google scholar: lookup
                  10. Knickelbein KE, Luethy D, Thomasy SM, Reilly C, Strom AR, Lassaline ME. Equine eosinophilic keratoconjunctivitis in California: retrospective study of 47 eyes from 29 cases (1993-2017).. Vet Ophthalmol 2019 Jul;22(4):510-519.
                    doi: 10.1111/vop.12621pubmed: 30706617google scholar: lookup
                  11. Verdon M, Lanz S, Rhyner C, Gerber V, Marti E. Allergen-specific immunoglobulin E in sera of horses affected with insect bite hypersensitivity, severe equine asthma or both conditions.. J Vet Intern Med 2019 Jan;33(1):266-274.
                    doi: 10.1111/jvim.15355pubmed: 30520523google scholar: lookup
                  12. Mueller RS, Janda J, Jensen-Jarolim E, Rhyner C, Marti E. Allergens in veterinary medicine.. Allergy 2016 Jan;71(1):27-35.
                    doi: 10.1111/all.12726pubmed: 26280544google scholar: lookup