Vol. 68, No. 3, Jun 2010: 147-152

The design of a “specific- IgE ELISA assay kit” for major cow’s milk allergens

Received: April 10, 2010    Accepted: May 19, 2010

Gholam Ali Kardar M.Sc.1
Zahra Pourpak M.D., Ph.D.2*

1- Ph.D. Student in Molecular Genetic
2- Department of Immunology

Immunology, Asthma & Allergy Research Institute, Children’s Medical Center, Tehran University of Medical Sciences

Background: The hypersensitivity to cow’s milk allergens is the most common allergies in children at the first year of life. The specific IgE evaluation is one of the important methods in diagnosis of allergic disease. The aim of this study was development of a sensitive and credible procedure for detection of cow’s milk allergens specific IgE.

Methods: The allergen discs were prepared by coating of allergens on nitrocellulose paper. After incubation of allergen discs with patients serum, anti-human IgE conjugated were used. In following optimization of any step of ELISAs test, a complete kit was designed. Efficiency of designed kits were evaluated by determination of specific IgE in normal (n= 29) and patient (n= 153) children serum samples and compared with commercial kits.

Results: The specific IgE against three allergrns involving casein, a-lactalbumin and b-lactoglobulin were measured on normal and patient children serum with designed and commercial ELISA kits. Results were demonstrated specificity of 93%, 89.7% & 82.8% and sensitivity of 86.3%, 81.3% & 89.6% respectively for casein, a-lactalbumin and b-lactoglobulin specific kits and these results were similar and comparable with commercial kits.

Conclusion: The Designed kits in comparison with the commercial kits were showed equivalent sensitivity and specificity. The designed kit stability was ultimately one month, probably due to don’t using of stabilizers for prepared allergen discs. We suggest these kits for commercial product in Iran and we hope be helpful for easier accesses for Cow’s milk allergy diagnosis and extend that for other allergens.

Keywords: Allergy, cow’s milk, ELISA, specific IgE.

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