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Food allergen specific IgE detection kit and application thereof

A technology for detecting kits and allergens, which is applied in the field of immunoanalysis, can solve the problems of poor sensitivity and cannot meet the detection requirements of trace IgE, and achieve the effects of good stability, enhanced specificity and sensitivity, and improved stability

Inactive Publication Date: 2018-07-27
AILEX TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] CN 102331495A discloses a food allergen detection kit and its preparation method, including chromogenic solution A, chromogenic solution B, stop solution, enzyme-labeled anti-IgE antibody, polystyrene microporous plate coated with biotinylated bovine serum Albumin-streptavidin, a biotinylated allergen, the kit is combined with streptavidin while the biotinylated allergen is combined with the specific antibody in the serum, realizing the complete coating and reaction Simultaneously, but the sensitivity of the horseradish peroxidase luminescence system used is poor, which cannot meet the detection requirements of trace IgE

Method used

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  • Food allergen specific IgE detection kit and application thereof
  • Food allergen specific IgE detection kit and application thereof
  • Food allergen specific IgE detection kit and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0050] The preparation of embodiment 1 kit

[0051] (1) Preparation of biotinylated peanut allergen

[0052] The peanut allergen was added to the biotin-dimethylformamide solution at a molar ratio of 1:10, purified by cross-linked Sephadex gel, and recovered by elution with 0.1M phosphate buffer (pH=7.4) , diluted to the working concentration;

[0053] (2) Preparation of alkaline phosphatase-labeled mouse anti-human IgE antibody solution

[0054] Add an equal volume of 12.8 mg / mL sodium periodate solution and 1% ethylene glycol solution to the alkaline phosphatase solution in sequence, and react in the dark at 2-8°C for 1 hour; according to the alkaline phosphatase solution: mouse anti-human IgE antibody The molar ratio is (0.5-2):1 Add mouse anti-human IgE antibody to the mixture, place it in 0.05M carbonate buffer and dialyze overnight at 4°C; take out the mixture, add Add 5 mg / mL sodium borohydride solution to 20 μL of sodium borohydride solution, and react in the dark a...

Embodiment 2

[0059] The establishment of embodiment 2 standard curve

[0060] (1) Mix 10 μL of IgE standard solutions with concentrations of 0.1IU / mL, 0.35IU / mL, 0.7IU / mL, 3.5IU / mL, 17.5IU / mL, 50IU / mL and 100IU / mL with 150 μL of 0.3mg / mL streptavidin-labeled magnetic particle suspension and 150 μL of biotinylated peanut allergen solution with a concentration of 0.3 μg / mL were thoroughly mixed, incubated at 37°C for 10 min, magnetically separated for 1 min, supernatant was removed, and washing solution was used Wash 5 times;

[0061] (2) Add 150 μL alkaline phosphatase-labeled anti-human IgE antibody with a concentration of 0.3 μg / mL, mix well, incubate at 37°C for 10 min, magnetically separate for 1 min, remove the supernatant, and wash 5 times with washing solution;

[0062] (3) Add 200 μL of AMPPD solution, mix well, and measure the chemiluminescence intensity of the solution with a Lumiray series of automatic chemiluminescence instrument;

[0063] (4) Take the chemiluminescence inten...

Embodiment 3

[0065] The determination of peanut allergen-specific IgE concentration in the sample to be tested in embodiment 3

[0066] (1) Four parts of 100 μL serum samples to be tested were fully mixed with 150 μL streptavidin-labeled magnetic particle suspension with a concentration of 1.2 mg / mL and 150 μL biotinylated peanut allergen solution with a concentration of 1.2 μg / mL, respectively, Incubate at 37°C for 10 minutes, magnetically separate for 1 minute, remove the supernatant, and wash 5 times with washing solution;

[0067] (2) Add 150 μL alkaline phosphatase-labeled anti-human IgE antibody with a concentration of 1.8 μg / mL, mix well, incubate at 37°C for 10 min, magnetically separate for 1 min, remove the supernatant, and wash 5 times with washing solution;

[0068] (3) Add 200 μL of AMPPD solution, mix well, and measure the chemiluminescence intensity of the solution with a Lumiray series of automatic chemiluminescence instrument;

[0069] (4) Calculate the concentration of p...

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Abstract

The invention provides a food allergen specific IgE detection kit and an application thereof. The kit includes magnetic micro-particles, a food allergen, an enzyme labeled anti human IgE antibody anda substrate, wherein the food allergen is modified with biotin, and the enzyme labeled anti human IgE antibody is modified with alkaline phosphatase. All the reagent components in the kit have good stability, and the kit has the advantages of long validity period, high sensitivity, good accuracy and short detection time.

Description

technical field [0001] The invention belongs to the field of immune analysis, and relates to a food allergen-specific IgE detection kit and its application, in particular to a peanut allergen-specific IgE detection kit and its application. Background technique [0002] IgE antibody-mediated type I allergic disease is one of the most common allergic diseases and a major global health problem. It is listed by the World Health Organization as one of the three major diseases to be prevented and controlled in the 21st century. The total incidence of allergic diseases in the world is about 15-30%, and about 5-10% of people in our country are harassed by allergens. [0003] After patients inhale, ingest or inject allergens or allergens containing sensitizing ingredients, the body's B cells will produce specific immunoglobulin E (IgE), thereby triggering an allergic reaction. IgE antibodies play a key role in allergic asthma, hay fever, urticaria, allergic rhinitis, eczema, conjunc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6854
Inventor 李子樵孙娱
Owner AILEX TECH GRP CO LTD
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