Peanut allergen specific IgE quantitative detection reagent, kit and application thereof

A technique for quantitative detection of peanut allergens, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of general specificity and sensitivity, high cost of use, and inferior sensitivity, etc., to achieve enhanced specificity and sensitivity, and dispersed Good stability and stability, the effect of increasing the potential number

Active Publication Date: 2021-01-05
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are only three quantitative detection reagents for peanut (F13) allergen-specific IgE that have obtained registration numbers, including: (1) foreign fluorescent enzyme-linked immunoassay detection reagents (Johansson SG. ImmunoCAP Specific IgE test: an objective tool for research and routine allergy diagnosis.[J].expert reviewof molecμL ar diagnostics,2004,4(3):273-279.), the main reagent components of this imported reagent are cellulose CAP caps activated by hydrogen bromide coated with allergens and The anti-human IgE antibody labeled with β-galactosidase is characterized by a large amount of CAP antigen coating, good specificity and sensitivity, but it is expensive and expensive to use; (2) Domestic ELISA capture The main components of this reagent are anti-human IgE antibody-coated microwell plates, biotinylated allergens and horseradish peroxidase-labeled streptavidin. Long time, low degree of automation, general specificity and sensitivity; (3) An invention patent (CN108333367A) applied by Shanghai Lanyi Technology Co., Ltd. discloses an enzyme immunoassay: involving a food allergy-specific IgE Detection kit and its application, in particular to a peanut allergen-specific IgE detection kit and its application
The invention mixes and incubates the sample to be tested with streptavidin-labeled magnetic particle suspension and biotin-labeled food allergen solution, and magnetically separates to remove the supernatant; then adds alkaline phosphatase-labeled anti-human IgE antibody, Mix and incubate, magnetically separate to remove the supernatant; finally add the substrate solution, measure the chemiluminescence intensity, and calculate the concentration of IgE in the sample to be tested according to the standard curve; the incubation time of this method is short (10min+10min), but the sensitivity is not as good as Fluorescent ELISA

Method used

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  • Peanut allergen specific IgE quantitative detection reagent, kit and application thereof
  • Peanut allergen specific IgE quantitative detection reagent, kit and application thereof
  • Peanut allergen specific IgE quantitative detection reagent, kit and application thereof

Examples

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preparation example Construction

[0033] Preparation of the streptavidin-coated magnetic particle suspension:

[0034]The magnetic particles coated with 10mg / mL streptavidin were subjected to magnetic separation, and the 0.1M Tris-HCl buffer (pH=7~ 8) Resuspend, wash repeatedly 4 times (5-10min / time), magnetically separate, and finally resuspend the magnetic particles in 0.1M Tris- HCl buffer solution (pH=7-8), that is, a suspension of streptavidin-coated magnetic particles with a concentration of 0.8-1.5 mg / mL can be obtained.

[0035] The peanut allergen is modified with biotin based on the hydrophilic polymer, and the structure diagram is as follows: figure 1 :

[0036] The hydrophilic polymer is poly-L-lysine, dendritic polylysine, dextran, polyacrylic acid, considering sensitivity and background background, preferably poly-L-lysine , the ratio used for the preferred labeling is hydrophilic polymer: peanut allergen is 1:180~1:210, the experimental data are shown in Table 1 and Table 2, and the samples (...

Embodiment 1

[0059] Embodiment 1 The preparation of streptavidin-coated magnetic particle suspension

[0060] The 10 mg / mL streptavidin-coated magnetic particles were magnetically separated, resuspended in 0.1M Tris-HCl buffer (pH=7) containing 1% BSA, 10% glycerol and 2% sucrose, and washed repeatedly for 4 time (5 min / time), magnetic separation, and finally the magnetic particles were resuspended in 0.1M Tris-HCl buffer solution (pH=7) containing 1% BSA, 10% glycerol and 2% sucrose, to obtain a concentration of 0.8mg / mL of streptavidin-coated magnetic particle suspension.

Embodiment 2

[0061] Embodiment 2 Preparation of streptavidin-coated magnetic particle suspension

[0062] The 10 mg / mL streptavidin-coated magnetic particles were magnetically separated, resuspended in 0.1M Tris-HCl buffer (pH=8) containing 3% BSA, 5% glycerol and 5% sucrose, and washed repeatedly for 4 time (10min / time), magnetic separation, and finally the magnetic particles were resuspended in 0.1M Tris-HCl buffer solution (pH=8) containing 3% BSA, 5% glycerol and 5% sucrose to obtain a concentration of 1.0mg / mL of streptavidin-coated magnetic particle suspension.

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Abstract

The invention relates to the field of biological detection, in particular to a peanut allergen specific IgE quantitative detection reagent, a kit and application thereof. The kit is based on a magnetic particle chemiluminescence method, used allergens are subjected to biotinylation modification based on hydrophilic polymers, and the kit is high in sensitivity, good in specificity, short in reaction time and high in automation degree.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a peanut allergen-specific IgE quantitative detection reagent, kit and application thereof. Background technique [0002] In 2013, the World Allergy Organization (WAO) pointed out in the white paper (WAO White Book on Allergy: Update 2013) on allergy (also known as anaphylaxis, Allergy): "The prevalence of allergic diseases is on the rise worldwide. Allergic diseases include allergic asthma, allergic rhinitis, eczema, urticaria, and angioedema; food allergy is a health problem that requires special attention, with a global incidence of 20%-30% (about 240-550 million people). The incidence rate is higher in young children (5%-8%) than in adults (1%-2%)". There are about 160 kinds of foods that cause allergies, among which milk, eggs, peanuts, soybeans, wheat, fish, crustaceans and tree nuts are identified by the FDA as the eight major allergic foods, and about 90% of food alle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/535G01N33/532
CPCG01N33/6854G01N33/54326G01N33/535G01N33/532G01N2333/415
Inventor 赵冬红朱宇皇韩晶李忠信刘功成
Owner AUTOBIO DIAGNOSTICS CO LTD
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