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Method for detecting materials causing anaphylactoid reaction

A technology for allergic reactions and substances, applied in the field of medical testing, which can solve the problems of loss of biological information, inability to perform accurate quantitative and dynamic observation, and detection errors.

Active Publication Date: 2013-04-24
EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to detect the corresponding indicators, mast cells undergo various chemical fixation and chromogenic treatments, losing a series of biological information of degranulation in vivo, which also causes a lot of detection errors, and it is impossible to carry out accurate quantitative and dynamic observation

Method used

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  • Method for detecting materials causing anaphylactoid reaction
  • Method for detecting materials causing anaphylactoid reaction
  • Method for detecting materials causing anaphylactoid reaction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1, measuring method

[0083] Step 1, RBL-2H3 is cultivated with complete RPMI-1640 culture fluid (RPMI1640 culture fluid: RPMI1640 powder 10g, sodium pyruvate 0.11g, Hepes (hydroxyethylpiperoneethanesulfonic acid) 5.925g, NaHCO 2g plus triple distilled water 1000ml, when used, add 10% fetal bovine serum, penicillin and streptomycin (100,000 units), after three passages, inoculate the cells into a confocal culture dish (diameter 3cm, glass bottom thickness 0.17μm) and culture in a 37-degree incubator for 24 hours ;

[0084] Step 2, change the culture medium into Tyrode's solution (preparation method: NaC400mg, KCl 100mg, MgCl 2 50mg, NaHCO500mg, NaH 2 PO 4 32.5 mg, CaCl 2 100g, add distilled water to 1000ml, add glucose 1.0g before use), add Fluo4 fluorescent dye with a final concentration of 1μg / ml at the same time, incubate at 37°C for 30min;

[0085]Step 3. Move to the laser scanning confocal microscope to observe under the excitation light of 488nm...

Embodiment 2

[0087] Embodiment 2, allergic substance assay method

[0088] Step 1, RBL-2H3 is cultivated with complete RPMI-1640 culture fluid (RPMI1640 culture fluid: RPMI1640 powder 10g, sodium pyruvate 0.11g, Hepes (hydroxyethylpiperoneethanesulfonic acid) 5.925g, NaHCO 2g plus triple distilled water 1000ml, when used, add 10% fetal bovine serum, penicillin and streptomycin (100,000 units), after three passages, inoculate the cells into a confocal culture dish (diameter 3cm, glass bottom thickness 0.17μm) and culture in a 37-degree incubator for 24 hours ;

[0089] Step 2, change the culture medium into Tyrode's solution (preparation method: NaC400mg, KCl 100mg, MgCl 2 50mg, NaHCO500mg, NaH 2 PO 4 32.5 mg, CaCl 2 100g, add distilled water to 1000ml, add glucose 1.0g before use), add Fluo4 fluorescent dye with a final concentration of 1μg / ml at the same time, incubate at 37°C for 30min;

[0090] Step 3. Move to the laser scanning confocal microscope to observe under the excitatio...

Embodiment 3

[0092] Embodiment 3, Chinese medicine injection allergen determination method

[0093] Step 1, RBL-2H3 is cultivated with complete RPMI-1640 culture fluid (RPMI1640 culture fluid: RPMI1640 powder 10g, sodium pyruvate 0.11g, Hepes (hydroxyethylpiperoneethanesulfonic acid) 5.925g, NaHCO 2g plus triple distilled water 1000ml, when used, add 10% fetal bovine serum, penicillin and streptomycin (100,000 units), after three passages, inoculate the cells into a confocal culture dish (diameter 3cm, glass bottom thickness 0.17μm) and culture in a 37-degree incubator for 24 hours ;

[0094] Step 2, change the culture medium into Tyrode's solution (preparation method: NaC400mg, KCl 100mg, MgCl 2 50mg, NaHCO500mg, NaH 2 PO 4 32.5 mg, CaCl 2 100g, add distilled water to 1000ml, add glucose 1.0g before use), add Fluo4 fluorescent dye with a final concentration of 1μg / ml at the same time, incubate at 37°C for 30min;

[0095] Step 3. Move to the laser scanning confocal microscope to obs...

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Abstract

The invention belongs to the field of medicament detection, and relates to a method for detecting materials causing anaphylactoid reaction. The method comprises the following steps of: culturing cells and medicaments together; scanning images of acquired culture by a microscopic; and determining the existence of allergy substances according to change of cell morphology in images as well as the change of calcium ions in the cells.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for measuring substances that cause anaphylactoid reactions. Background technique [0002] Anaphylaxis is one of the common adverse reactions. Anaphylaxis is an abnormal immune reaction between exogenous antigenic substances and antibodies in the body. There are many substances that can induce allergic reactions, such as proteins, polypeptides, polysaccharides and other macromolecular substances with complete antigenicity; other compounds with smaller molecules can be used as haptens to combine with proteins in the body to form complete antigens, thereby causing allergic reactions. At present, there are many allergic substances involved in allergic reactions, among which medicines (traditional Chinese medicine and western medicine), food, pollen, mites, etc. occur more frequently. [0003] Acute allergic reactions include type I hypersensitivity r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 王毅王丹巧胡剑江张倩侯燕鸣雷洪涛于友华李连达
Owner EXPERIMENTAL RES CENT CHINA ACAD OF CHINESE MEDICAL SCI
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