Allergenic specific IgE antibody immunoassay detection kit and preparation method thereof

A detection kit and immunoassay technology, applied in the direction of chemical reaction of materials for analysis, analysis of materials, measurement devices, etc., can solve the problems of difficulty in providing sensitivity, low accuracy, long time consumption, etc. Small background interference, high accuracy, easy to use effects

Inactive Publication Date: 2010-04-21
深圳市博卡生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The content of specific IgE antibodies in serum is very low, and it is difficult for ELISA detection kits to provide high enough s...

Method used

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  • Allergenic specific IgE antibody immunoassay detection kit and preparation method thereof
  • Allergenic specific IgE antibody immunoassay detection kit and preparation method thereof
  • Allergenic specific IgE antibody immunoassay detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Dust mite allergen-specific IgE antibody immunoassay detection kit

[0037] D. farinae allergen-specific IgE antibody immunoassay kit, including: (1) superparamagnetic microbeads coupled with D. farinae allergen protein, both of which are 100 μg D. farinae allergen protein / mg superparamagnetic microbead (2) superparamagnetic microbeads coupled with mouse anti-human IgE antibody, the two are coupled and mixed at a ratio of 10 μg goat anti-human IgE antibody / mg superparamagnetic microbeads; (3) human IgE antibody standard, derived from human serum, purified by affinity chromatography with a concentration of 100IU / mL (refer to the international reference standard for human IgE published by the World Health Organization); (4) rabbit anti-human IgE antibody labeled with alkaline phosphatase; And (5) alkaline phosphatase chemiluminescence substrate 3-(2′-helixadamantane)-4-methoxy-4-(3″-o-oxyacylphenyl)-1,2-dioxane Butane (AMPPD).

[0038] Wherein, the interior o...

Embodiment 2

[0039] Example 2: Mango allergen-specific IgE antibody immunoassay detection kit

[0040] Mango allergen-specific IgE antibody immunoassay detection kit, including: (1) magnetic microbeads coupled with mango allergen protein, the two are coupled and mixed at a ratio of 300 μg mango allergen protein / mg magnetic microbead; ( 2) Magnetic beads coupled with rabbit anti-human IgE antibody, the two are coupled and mixed at a ratio of 40 μg rabbit anti-human IgE antibody / mg magnetic beads; (3) Human IgE antibody standard, derived from human serum, tested The concentration after purification by affinity chromatography is 100IU / mL (refer to the international reference standard for human IgE published by the World Health Organization); (4) Goat anti-human IgE antibody labeled with alkaline phosphatase; (5) Alkaline phosphatase chemiluminescent substrate 3-(2′-spiroadamantane)-4-methoxy-4-(3″-o-oxyacylphenyl)-1,2-dioxetane (AMPPD); and (6) black plastic Opaque microplates.

[0041] Suc...

Embodiment 3

[0042] Example 3: Honeybee Allergen-Specific IgE Antibody Immunoassay Detection Kit

[0043] Honeybee allergen-specific IgE antibody immunoassay detection kit, including: (1) superparamagnetic microbeads coupled with bee allergen protein, the two are coupled at a ratio of 200 μg bee allergen protein / mg superparamagnetic microbeads (2) Superparamagnetic microbeads coupled with goat anti-human IgE antibody, the two are coupled and mixed at a ratio of 30 μg rabbit anti-human IgE antibody / mg superparamagnetic microbeads; (3) Human IgE antibody standard The product is derived from human serum, and the concentration after purification by affinity chromatography is 100IU / mL (refer to the international reference standard for human IgE published by the World Health Organization); (4) mouse anti-human IgE antibody labeled with alkaline phosphatase; (5) Alkaline phosphatase chemiluminescence substrate 3-(2′-helicadamantane)-4-methoxy-4-(3″-o-oxyacylphenyl)-1,2-dioxetane (AMPPD ); (6) ce...

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PUM

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Abstract

The invention relates to an immunoassay detection kit, and provides an allergenic specific IgE antibody immunoassay detection kit and a preparation method thereof. The detection kit comprises magnetic microspheres coupled with allergenic protein, magnetic microspheres coupled with anti-human IgE antibody, a human IgE antibody standard substance, alkali phosphatase labeled anti-human IgE antibody and an alkali phosphatase chemical luminous substrate AMPPD. The preparation method for the detection kit comprises the following steps: coupling the allergenic protein to the magnetic microspheres, coupling the anti-human IgE antibody to the magnetic microspheres, preparing the human IgE antibody standard substance, preparing the alkali phosphatase labeled anti-human IgE antibody, preparing the alkali phosphatase chemical luminous substrate AMPPD and packing the combination. The detection kit has the advantages of high sensitivity, high accuracy, quick response, convenient usage, low dosage and low cost, can assist in precisely and quantitatively measuring sensitive allergene, and guides clinical anaphylactic reaction treatment.

Description

technical field [0001] The invention relates to an immunoassay detection kit, in particular to an allergen-specific IgE antibody immunoassay detection kit and a preparation method thereof. Background technique [0002] Anaphylaxis detection is an important prerequisite for the treatment of anaphylaxis. Specific IgE antibodies in the patient's serum can be detected to determine what allergen is sensitizing the patient and the degree of sensitization. [0003] For many years, enzyme-linked immunosorbent assay (ELISA) has been commonly used to detect specific IgE antibodies in serum. The corresponding ELISA detection kit includes a microtiter plate coated with allergen protein, human IgE standard, enzyme-labeled antibody and substrate chromogenic solution. The content of specific IgE antibodies in serum is very low, and it is difficult for ELISA detection kits to provide high enough sensitivity, especially for the detection of small or trace amounts of specific IgE antibodies,...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76
Inventor 雷均平黄志坚邱于斌何素华薛晓英赵振军和水祥
Owner 深圳市博卡生物技术有限公司
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