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Method for detecting anti-anaphylaxis function of compound

A compound and anti-allergic technology, applied in biological testing, material inspection products, etc., can solve problems such as narrow field of view, large individual differences in chicken embryo vascular network, and difficult to unify standards, so as to expand the operating field of vision and facilitate the recording of results and Evaluate and observe the effects of clear indicators

Inactive Publication Date: 2012-12-19
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this method, the traditional method is to open a window in the chicken shell and directly add the test substance to the exposed chorion of the allantoic membrane. Irritation (within 5min), and this method has no record of application in the evaluation of anti-allergic properties of compounds

Method used

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  • Method for detecting anti-anaphylaxis function of compound
  • Method for detecting anti-anaphylaxis function of compound
  • Method for detecting anti-anaphylaxis function of compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Materials and Methods

[0023] Material:

[0024] 1. Eggs: specific pathogen-free (SPF) grade fertilized chicken eggs, 3 days old, purchased from Beijing Meriaton Experimental Animal Technology Co., Ltd.;

[0025] 2. Various reagents were purchased from Beijing Chemical Reagent Company and Sigma Reagent Company;

[0026] method:

[0027] 1. Disinfect chicken eggs with 75% alcohol and incubate for 84 hours in an incubator with a humidified water tray;

[0028] 2. Use a dental drill to cut a 5cm long incision on the vertical long axis of the eggshell, break the eggshell to both sides with the incision as the boundary, and move the chicken embryos into a 90mm diameter sterile glass plate, and add pre-prepared water to the plate in advance. Heat 2ml of shell-opening buffer at 37°C, the composition of the shell-opening buffer is CaCl 2 .2H 2 O: 0.265g / l; anhydrous MgSO 4 : 0.09767g / l; KCl: 0.4g / l; NaHCO 3 : 2.2g / l; NaCl: 6.8g / l; anhydrous Na 2 HPO 4 : 0.122g / l; peni...

Embodiment 2

[0043] Materials and Methods

[0044] Material:

[0045] 1. Chicken eggs: specific pathogen-free (SPF) grade fertilized chicken eggs, 3 days old, purchased from Beijing Meriaton Experimental Animal Technology Co., Ltd.; 2. Various reagents were purchased from Beijing Chemical Reagent Company and Sigma Reagent Company;

[0046] method:

[0047] 1. Disinfect chicken eggs with 75% alcohol and incubate for 90 hours in an incubator with a humidified water tray;

[0048] 2. Use a dental drill to cut a 1.5cm long incision on the vertical long axis in the center of the eggshell, split the eggshell to both sides with the incision as the boundary, and move the chicken embryos into a sterile glass plate with a diameter of 90mm. 2ml of shell-opening buffer preheated at 37°C, the composition of the shell-opening buffer is CaCl 2 .2H 2 O: 0.265g / l; anhydrous MgSO 4 : 0.09767g / l; KCl: 0.4g / l; NaHCO 3 : 2.2g / l; NaCl: 6.8g / l; anhydrous Na 2 HPO 4 : 0.122g / l; penicillin: 0.066g / l; strep...

Embodiment 3

[0063] Materials and Methods

[0064] Material:

[0065] 1. Eggs: specific pathogen-free (SPF) grade fertilized chicken eggs, 3 days old, purchased from Beijing Meriaton Experimental Animal Technology Co., Ltd.;

[0066] 2. Various reagents were purchased from Beijing Chemical Reagent Company and Sigma Reagent Company;

[0067] method:

[0068] 1. Disinfect chicken eggs with 75% alcohol and incubate for 90 hours in an incubator with a humidified water tray;

[0069] 2. Use a dental drill to cut a 3cm long incision on the vertical long axis of the eggshell, split the eggshell to both sides with the incision as the boundary, and move the chicken embryos into a sterile glass plate with a diameter of 90mm. Heat 2ml of shell-opening buffer at 37°C, the composition of the shell-opening buffer is CaCl 2 .2H 2 O: 0.265g / l; anhydrous MgSO 4 : 0.09767g / l; KCl: 0.4g / l; NaHCO 3 : 2.2g / l; NaCl: 6.8g / l; anhydrous Na 2 HPO 4 : 0.122g / l; penicillin: 0.066g / l; streptomycin: 0.154g / l; ...

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Abstract

A method for detecting antiallergic effect of compounds comprises the following steps: placing fertilized chicken eggs, which are sterilized by 75% alcohol in an incubator with humidified water pond to incubate the fertilized chicken eggs for 75-100h; opening egg shells; moving chick embryos into a sterile glass plate with diameter of 75-90mm; pre-adding buffer solution pre-heated to 37 DEG C foropening shells into the plate; selecting and placing chick embryos with un-damaged vitelline membrane into a constant-temperature incubator at 37.5 DEG C to continuously incubate the chick embryos tothe 8th to 15th day; grouping the chick embryos randomly; placing the chick embryos into the constant-temperature incubator to continuously incubate the chick embryos after adding compounds to be tested; when the chick embryos is incubated for 1h or 12h or 24h or 48h, adding materials which have stimulation to chick embryo allantoid vitelline membrane veins; assessing the changes of veins according to simulating factors; and calculating the suppression ratio of antiallergic compounds with different concentration. The method is simple, cheap and clear in observed index.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for detecting the strength of the anti-allergic effect of a compound, in particular to a method for detecting the strength of the anti-allergic effect of the compound by observing the change degree of blood vessels in the vitelline membrane of the chicken embryo allantois. Background technique [0002] In recent years, skin diseases caused by allergic substances are increasing year by year, becoming one of the common diseases in dermatology. People are paying more and more attention to the evaluation of the anti-allergic properties of compounds. The traditional evaluation of the anti-allergic properties of compounds is mainly based on animal experiments. However, under the influence of the global animal protection movement, developed countries generally carry out the following The "3R" movement with replacement, reduction and refinement as the core, in vitro replacement experi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/50
Inventor 杨晓冉董益阳邹运动王军兵郑洪艳
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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