High-throughput in-vitro allergen screening method

Abstract

The invention discloses a high-throughput in-vitro allergen screening method, which comprises: peripheral blood drawing, separating serum, adding the serum into BSA liquid closed 96-well plates pre-coated with mouse or rabbit anti-human IgE antibody in advance to perform incubation; carrying out specific antigen-antibody reaction; after incubation, conducting washing to remove non-specifically bound components; adding 96 biotin labeled specific allergens to one of the 96-well plates respectively; adding a biotin labeled anti-IgE antibody into the other 96-well plate, performing incubation, conducting washing to remove non-specifically bound components; conducting sample adding of fluorescein labeled streptavidin to the 96-well plates, conducting inoculation, and then performing washing; then sending the 96-well plates to a high-throughput fluorescence signal detection system to carry out quantitative detection of the fluorescence intensity respectively, and using the total IgE detected by the first 96-well plate to judge allergy; and using the other 96-well plate to diagnose allergen. The method provided by the invention has an accuracy of effective detection of allergens up to 95%, and can detect more than 90 allergens at a time.

Description

technical field [0001] The invention relates to a high-throughput in vitro allergen screening method. Background technique [0002] Allergic diseases, also known as allergic diseases, refer to tissue damage or physiological functions caused by the stimulation of the body by antigenic substances (also known as allergens or allergens), such as pollen, dust, food, drugs, parasites, etc. Disorder, which is an abnormal or pathological immune response. The fundamental problem to be solved is to first determine what the allergen is? Effective prevention and treatment can only be achieved by accurately identifying the exact allergen to which the patient is allergic. At present, the main allergen screening methods include in vivo detection and in vitro detection. Among them, there are two kinds of in vivo detection methods: skin test method and spot method. The disadvantage is that the body is more painful. In vitro detection is mainly in vitro immunoassay. In vitro immunoassay ...

AI Technical Summary

Problems solved by technology

[0003] The method used by the UniCAP system of Swedish Pharmacia Company is fluorescent immunological detection, but since the UniCAP system uses patient serum to react with pre-coated and fixed allergens, the specific IgE antibody in the serum binds to the allergen It will be competed by the specific IgG antibody in the serum, so it will bias the test results and affect the specificity and accuracy

[0004] Although the products of Fooke Company in Germany can eliminate the competitive interference of specific IgG antibodies to specific IgE antibodies and improve the specificity, Fooke Company uses the enzyme-linked immunosorbent method ELISA, and the detection sensitivity is much lower than that of fluorescein detection. Therefore, it is easy to produce false negative results, resulting in missed detection