Direct Allergy Test (DAT) – New method for detection of antigenantibody reactivity
Journal Title: IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) - Year 2018, Vol 13, Issue 6
Abstract
An alternative for the tests of immunodetection diagnoses is proposed in this work, based on the elaboration of a qualitative method that can aid in the identification of antigen-antibody immunochemical reactivity in rapid diagnoses of allergies. For this purpose, the allergen-containing Dermatophagoides farinae mite extract was immobilized with glutaraldehyde on reusable support and blocking steps, addition of antibody containing serum to the allergens present in the mite extract. The reactivity response was obtained after colorimetric revelation. The appearance of the purple color signaled the reactivity between antigen-antibody on the plate. The colored area was photographed and the reactivity was represented by the intensity of color in pixels using the gel analyzer software. The method was evaluated by repeatability tests, negative assay and variation in protein content. From the proposed method a positive reactivity response was verified for the serum of individuals sensitized by mite. The method showed better performance for application of the protein load of 500 μg of the mite extract, 0.50% (v/v) glutaraldehyde and 0.25% (w/v) blocking. Repeated analyzes performed on different days showed that the method is reproducible being in agreement when compared to the immunoblotting technique. The results obtained for the development of the antigen-antibody detection method showed its promising potential as an alternative aid in the screening of antigen-antibody allergic reactions. The qualitative method, however, does not aim at replacing traditional techniques, but it provides speed and cost reduction, as it eliminates the electrophoretic separation step, dispenses standard purified proteins, and reuses the support, besides providing less reagent consumption.
Authors and Affiliations
Francisca das C. S. Silva, Patrícia R. Pereira, Luciana D. de C. Girão, Anderson B. Matos, Maria C. Queiroz
Keywords
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- EP ID EP422643
- DOI 10.9790/3008-1306022834.
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