Development of flow injection system for detection of allergen using fluorochrome.

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Many immunoassay methods have been developed for sensing or detecting allergens. The skin tests including prick test and scratch test are typical methods. However, the these tests are dangerous because of anaphylaxis by serum antibodies. Therefore, a safe, simple and rapid method for detecting allergic reaction is required in the clinical and food industry fields. We have developed an automated flow immunoassay system using an ion-exchange column for the detection of food allergens. Antigen-antibody complexes were subsequently separated from unreacted analytical substrate by ion-exchange column, on the basis of the difference in isoelectric point, In this study, the allergen detection system equipped with an immunoreaction column, an ion-exchange column and a detector was constructed. A linear relationship was obtained between the fluorescence intensity and allergen concentration in the range of 0.01 2.0 mg/ml. The correlation coefficient was 0.994 within this range. This method was faster (10 min) and simple to use, and gives higher precision (<2%) than the conventional ELISA using microtiter plates. Reuse of free fluorochrome conjugated IgE was possible. We also constructed a novel automated flow immunoassay system for quantification of anti-double-stranded (anti-ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus. The assay yielded a linear relationship between signal and concentration of anti-dsDNA antibody in the range of 0 300 mug/ml. This technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. These simple and rapid immunoassay methods can be used for the continuous detection of allergen.