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In-vitro detection method of allergen in food

A detection method and in vitro detection technology, applied in the direction of biological testing, material inspection products, etc., can solve the problem of inaccurate reflection of allergic reactions to allergens

Active Publication Date: 2013-08-21
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to provide an in vitro detection method for allergens in food, so as to solve the problem that the existing evaluation methods cannot accurately reflect the allergic reaction mediated by the allergen induced by IgE

Method used

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  • In-vitro detection method of allergen in food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Embodiment 1 Animal experiment

[0117] 1. Experimental animals

[0118] 3-4 week old female BALB / c mice of SPF grade.

[0119] 2. Test substance

[0120] The tested proteins were glycinin (Glycinin, Gly), ovalbumin (OVA), potato acid phosphatase (PAP), all added with 10 μg cholera toxin adjuvant (CT, diluted in 100 μl normal saline)

[0121] 3. Animal grouping

[0122] The experimental animals were randomly divided into 5 groups according to body weight, namely: Gly+CT group, OVA+CT group, PAP+CT group, CT control group and negative control group.

[0123] 4. Dosage

[0124] The test protein was 1 mg, all adsorbed on 10 μg cholera toxin adjuvant (diluted in 100 μl normal saline), the CT control group received 10 μg cholera toxin adjuvant, and the negative control group received the same volume of normal saline.

[0125] 5. Animal handling

[0126] The experimental animals were subjected to the experiment after adaptive feeding for 1 week. Water and food were pro...

Embodiment 2

[0131] The detection of embodiment 2 specific IgE

[0132] 1) Coated

[0133] Dilute the protein to be tested to 10 mg / L with coating buffer, select wells to be coated on a 96-well plate, add 100 μl of coating solution to each well, and overnight at 4°C;

[0134] 2) washing

[0135] Pour off the liquid in the wells, add 250 μl of PBS / 0.1%BSA-Tween20 washing solution to each well, place at room temperature for 5 minutes, shake off the washing solution, pat several times on absorbent paper until there are no obvious drops in the wells, and repeat twice;

[0136] 3) closed

[0137] Add 150 μl PBS / 1%BSA to each well, incubate in a constant temperature incubator at 37°C for 1 hour, and wash three times;

[0138] 4) Add primary antibody

[0139] Add 100 μl of serum to be tested into each well, make three parallel holes for each test serum, and make three negative controls on each plate. Incubate in a constant temperature incubator at 37°C for 1 hour, and wash six times;

[014...

Embodiment 3

[0149] The detection of embodiment 3 specific IgG1

[0150] 1) Coated

[0151] Dilute the protein to be tested to 10 mg / L with coating buffer, select wells to be coated on a 96-well plate, add 100 μl of coating solution to each well, and overnight at 4°C;

[0152] 2) washing

[0153] Pour off the liquid in the wells, add 250 μl of PBS / 0.1%BSA-Tween20 washing solution to each well, place at room temperature for 5 minutes, shake off the washing solution, pat several times on absorbent paper until there are no obvious drops in the wells, and repeat twice;

[0154] 3) closed

[0155] Add 150 μl PBS / 1%BSA to each well, incubate in a constant temperature incubator at 37°C for 1 hour, and wash three times;

[0156] 4) Add primary antibody

[0157] Add 100 μl of serum to be tested into each well, make three parallel holes for each test serum, and make three negative controls on each plate. Incubate in a constant temperature incubator at 37°C for 1 hour, and wash six times;

[01...

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Abstract

The invention relates to an in-vitro detection method of allergen in food. The invitro detection method comprises the steps of: 1) establishing an IgE (Immunoglobulin E) hyperresponsiveness food allergic animal model based on an allergen immuned mouse; and 2) exciting a sensitization RBL-2H3 cell, an allergen or a rat anti-mouse IgE antibody, and detecting the release rate of beta-hexosaminidase. The invitro detection method provided by the invention is low in detection cost and high in detection sensitivity and specificity, can be used for accurately reflecting the IgE mediated anaphylactoid capability induced by the allergen and evaluating the sensitization of food raw materials and various processed foods, so as to enable the risk of food allergies to be radically reduced.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to an in vitro detection method for allergens in food. Background technique [0002] Food allergy is a type of allergic disease and may be a trigger for some serious allergic disease complications. According to epidemiological surveys, the incidence of allergies has been increasing year by year in recent years. In developed countries, 3% to 4% of adults have food allergies every year, and the incidence of children and infants is 5%. The incidence of allergic diseases in our country is also showing a rapid growth trend, which has been highly valued by the health department. Food allergy affects the health of allergic people to a considerable extent, and protecting consumers' food safety has become a serious problem for food safety management departments and food manufacturers. Therefore, food allergy has been regarded as a serious public nutrition and health problem, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 车会莲孙娜周催王静王翠燕贺晓云黄昆仑
Owner CHINA AGRI UNIV
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