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Food anaphylactogen specificity IgG4 antibody ELISA detection kit and preparation method thereof

A detection kit and a food allergy detection technology, which is applied in the field of enzyme-linked immunosorbent assay detection kits, can solve the problems of unpopular detection methods, difficult clinical diagnosis, and difficulty for patients to make self-diagnosis

Inactive Publication Date: 2008-09-03
杭州浙大生科生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] IgG (mainly IgG4)-mediated immune food allergy has a slow onset, usually 24-120 hours after ingesting food, clinically manifested as chronic symptoms of multiple systems, it is difficult for patients to make self-diagnosis, and detection methods are not popular , causing difficulties in clinical diagnosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The food allergen-specific IgG4 antibody ELISA detection kit of the present invention comprises coated milk, eggs, meat, grains, nuts, fruits, vegetables, aquatic products, beans food allergies Original protein ELISA plate, human IgG4 standard solution, enzyme-labeled antibody, TMB substrate chromogenic solution, sample diluent, concentrated washing solution and stop reaction solution.

[0038] The above enzyme plate is 96 wells (12 strips of 8 wells or 8 strips of 12 wells).

[0039] The preparation method of the food allergen-specific IgG4 antibody ELISA detection kit of the present invention:

[0040] 1. Preparation of Food Allergen Proteins

[0041] 1.1 Crude extract of food allergen protein

[0042] 1.1.1 Milk extract

[0043] There are two methods of making milk infusion:

[0044] (1) First use centrifugation (2400 rpm) for 15 minutes, remove the upper layer of fat with a small spoon, take 400ml skimmed milk, add 5ml 1% rennet or milk jelly slices, do not stir...

Embodiment 2

[0128] Example 2: The kit is used to detect the specific IgG4 content during the desensitization treatment of allergic patients.

[0129] An 8-year-old boy who was allergic to milk when he was a baby had no allergic reaction symptoms for 15 days in a double-blind test. The milk prick fluid skin test was positive for grade IV, and the milk-specific IgE (sIgE) antibody and specific IgG4( sIgG4) antibody concentration background value. On the 16th day, oral milk desensitization preparations were gradually increased for 18 months, and the concentrations of milk-specific IgE and IgG4 antibodies in the blood were detected at the 1st, 3rd, 6th, and 18th months after the desensitization treatment. The results are listed in the table below:

[0130] desensitization treatment

[0131] The data in the above table shows that using the kit of the present invention can simply, quickly and accurately detect the sIgG4 content in the sample as a biological indicator of the curative ...

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PUM

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Abstract

The invention discloses an ELISA detection kit of food allergen specificity IgG4 antibody and preparation method thereof, using enzyme target rod coated by purified food allergen protein. The kit comprises: enzyme target rod coated by food allergen protein, human IgG2 titer, enzymic-lableled antibody, TMB substrate developing liquid, sample diluent, concentrated cleaning solution and termination reaction liquid. The specificity IgG4 antibodies in sample serum are bonded with the corresponding antigens fixed on the micropore surface of the enzyme target rod and the enzymic-lableled antibodies identify and bond with the IgG4 to form antigen-IgG4-antibody-enzyme composite. The absorbency is measured by the development reaction between the enzyme and substrate and the specificity IgG4 antibody concentration in sample serum is measured by standard curve. The allergen specificity IgG4 antibody concentration in sample serum is used as biology index of exposure state of food allergen and for judging the patient immune deviation and immune tolerance and monitoring the immunotherapy effectiveness of special food selected by patients.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay (ELISA) detection kit for semi-quantitatively detecting food allergen-specific IgG4 antibodies in human serum and a preparation method thereof. Background technique [0002] For decades, the medical community has been paying close attention to and studying food allergies and natural foods, genetically modified foods, food additives and preservatives that induce allergic diseases. Food allergic reactions are divided into IgE immune-mediated allergic reactions and non-IgE immune-mediated allergic reactions. In an IgE-mediated allergic reaction, food allergen-specific IgE antibodies stimulated by allergen components in food interact with mast cells, basophils, macrophages, lymphocytes, eosinophils, and platelets, etc. F on the surface of the cell membrane C εR1 and F C The εR2 receptor binds and releases inflammatory mediators such as histamine and tumor necrosis factor (TNFα), causing aller...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/535
Inventor 吴善东
Owner 杭州浙大生科生物技术有限公司
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