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Clinical & Experimental Allergy

Assessment of recombinant dog allergens Can f 1 and Can f 2 for the diagnosis of dog allergy

Authors


Tuomas Virtanen, Department of Clinical Microbiology, University of Kuopio, PO Box 1627, FIN-70211 Kuopio, Finland.
E-mail: Tuomas.Virtanen@uku.fi

Summary

Background The use of recombinant allergens for the diagnosis and immunotherapy of allergy may offer several advantages over allergen extracts.

Objective To produce recombinant dog allergens Can f 1 and Can f 2 in Pichia pastoris yeast and to assess their suitability for the diagnosis of dog allergy.

Methods Clinically diagnosed dog-allergic patients' and healthy non-atopic dog owners' reactivities against recombinant Can f 1 and Can f 2 and commercial dog epithelial extract were studied by a panel of methods including skin prick test (SPT), ELISA and IgE immunoblotting.

Results Recombinant Can f 1 and Can f 2 were found immunologically functional: they bound dog-allergic patients' IgE in immunoblotting and inhibited specifically the binding of IgE to their natural counterparts in the dog allergen extract. Moreover, patients' IgE reactivity in immunoblotting to natural Can f 1 and their SPT with the recombinant allergen were perfectly concordant (φ coefficient 1.0, P<0.001). The concordance was slightly lower with recombinant Can f 2 (φ coefficient 0.92, P<0.001). A lower number of dog-allergic patients, 52%, reacted against Can f 1 than previously reported. About one-third of the patients reacted to Can f 2. In immunoblotting, the highest prevalence of reactivity, 60%, was directed to an 18 kDa component. Aminoterminal sequencing showed this to be a previously unidentified allergenic protein.

Conclusions The recombinant allergens can be used reliably to identify Can f 1 and Can f 2-sensitized individuals. However, the two allergens are insufficient as reagents for diagnosing dog allergy.

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