A method for constructing an animal model for detecting allergens in food and its application

A technology of animal model and construction method, which is applied in the field of medical evaluation and detection, can solve the problems of not being a food allergen, easy to produce oral tolerance, etc., and achieve the effect of reducing risk

Inactive Publication Date: 2016-08-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Intrinsic sensitization of proteins can be studied by intraperitoneal injection, but this route is not the natural route for human intake of food allergens
The advantage of oral administration is that its exposure route is most relevant to human exposure to food allergens, and its disadvantage is that it is prone to oral tolerance

Method used

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  • A method for constructing an animal model for detecting allergens in food and its application
  • A method for constructing an animal model for detecting allergens in food and its application
  • A method for constructing an animal model for detecting allergens in food and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Construction of animal models

[0104] 1. Experimental animals

[0105] SPF grade female Wistar rats weighing 40-60 g.

[0106] 2. Test substance

[0107] Human lactoferrin (rhLF), bovine lactoferrin (bLF), ovalbumin OVA, normal saline (negative control).

[0108] 3. Grouping of animals

[0109] Experimental animals were randomly divided into 4 groups according to body weight (two groups in each group), namely: rhLF gavage group / peritoneal injection group, bLF gavage group / peritoneal injection group, OVA gavage group / peritoneal injection group and normal saline gavage group group / peritoneal injection group. 8 animals per group.

[0110] 4. Dosage

[0111] rhLF and bLF: intragastric administration dose of 0.01g / kg body weight, intraperitoneal injection dose of 0.001g / kg body weight.

[0112] OVA: intragastric administration dose is 0.15g / kg body weight, intraperitoneal injection dose is 0.015g / kg body weight.

[0113] Normal saline: 8ml / kg body weight.

[0114] ...

Embodiment 2

[0121] Detection of reaction indicators

[0122] 1. Detection of specific IgE

[0123] 1) Coated

[0124] Dilute the test protein to 10 mg / L with coating buffer, select the wells to be coated on a 96-well plate, add 100 μL of coating solution to each well, and overnight at 4 °C;

[0125] 2) washing

[0126] Pour off the liquid in the wells, add 200 μL of washing solution to each well, place at room temperature for 3 minutes, shake off the washing solution, and pat it on absorbent paper several times until there is no obvious drop in the well, repeat twice;

[0127] 3) closed

[0128] Add 150 μL of blocking solution to each well, incubate for 1 h in a constant temperature incubator at 37 °C, and wash three times;

[0129] 4) Add primary antibody

[0130] Add 100 μL of serum to be tested diluted 1:50-100 to each well, make three parallel wells for each test serum, and make three negative controls on each plate. Incubate in a constant temperature incubator at 37°C for 1 hou...

Embodiment 3

[0163] Test Results and Analysis

[0164] 1. Daily situation

[0165] During the test period, the experimental animals in each group were in good condition, their nutritional status was normal, and no poisoning such as death occurred.

[0166] 2. Results of gavage group

[0167] 1) Specific IgE results

[0168] Table 1 shows the specific IgE detection results of each group in the intragastric administration group: it can be seen from the results that the OVA group was significantly higher than the bLF group on the 28th and 42nd days (P<0.05), and it was significantly higher on the 42nd day. higher than that of the normal saline control group (P<0.05). However, rhLF hardly induced IgE response in experimental animals.

[0169] Table 1 The specific IgE detection results of each group in the gavage group (mean ± standard deviation, n=8)

[0170]

[0171] Note: a indicates that there is a significant difference with the OVA group (P<0.05).

[0172] 2) Specific IgG results...

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Abstract

The invention provides a construction method of an animal model for detecting the allergen in a food, and an application of the animal model. The construction method concretely comprises the following steps: randomly dividing experiment animals into several groups, carrying out a sensitizing reaction on the experiment animals by using different test samples respectively, using one group of the test samples as negative control, dividing each group of the test samples into two subgroups, sensitizing the experiment animals through oral perfusion and lumbar injection approaches, evaluating the sensitization of the allergens by detecting the indexes comprising the specific immune proteins and the like of the animals to obtain the animal model, evaluating the sensitization of the test samples by carrying out in vitro experiments, and comparing the consistence of two evaluation results. The animal model obtained in the invention overcomes a problem that an in vitro evaluation method cannot completely stimulate the complex immune reaction of bodies to exogenous proteins. The animal model can be applied in the evaluation of the sensitization of food raw materials and various processed foods in order to fundamentally reduce the allergic risk of foods.

Description

technical field [0001] The application of the present invention relates to the technical field of medical evaluation and detection, in particular, to a method for constructing an animal model for detecting allergens in food. Background technique [0002] With the improvement of people's health awareness, food safety issues are getting more and more attention. Food safety refers to food that is non-toxic and harmless, meets proper nutritional requirements, and does not cause any acute, subacute or chronic harm to human health. Food allergy is one of allergic diseases, which may induce some serious allergic diseases and their complications. Therefore, food that can cause allergies must have food safety problems. [0003] Epidemiological survey results show that in recent years, the incidence of allergy-related diseases has been increasing year by year. In developed countries, 10% to 25% of people have experienced IgE-mediated allergic reactions, and the incidence of food all...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/40A61K38/38A61K49/00
Inventor 车会莲王静周催孙娜王翠燕贺晓云
Owner CHINA AGRI UNIV
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