ELISA kit for detecting food allergen based on IgA antibody

A technology for food allergy and antibody detection, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of restricting the development of allergen detection in food, unsuitable for batch detection, and high detection cost, and achieves high accuracy and heavy-duty. Good performance and improved sensitivity

Inactive Publication Date: 2018-11-20
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Detect and identify allergens in food by analyzing the characteristics of allergen proteins, but special large-scale precision instruments are required, the detection cost is high, and the analysis is difficult. Special testing personnel and analysts are required.
[0005] To sum up, although there are many methods for the detection of food allergens currently studied, they all have the problems of slow detection speed, high cost, and are not suitable for batch detection, and require specific analytical instruments for machine-reading, which restricts the detection of food allergens. Development of original assays

Method used

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  • ELISA kit for detecting food allergen based on IgA antibody
  • ELISA kit for detecting food allergen based on IgA antibody
  • ELISA kit for detecting food allergen based on IgA antibody

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preparation example Construction

[0049] In the present invention, the preparation method of the detection plate coated with food allergen preferably comprises the following steps:

[0050] 1) diluting the food allergen with a coating buffer to obtain a food allergen coating solution;

[0051] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;

[0052] 3) The next day, after washing the test plate wells overnight in step 2), add blocking solution, wash again after incubation, and coat the test plate with food allergens.

[0053] The invention dilutes the food allergen and human IgA protein with the coating buffer to obtain the coating solution. In the present invention, the dilution method is not particularly limited, and a dilution technique known to those skilled in the art can be used. In the present invention, the coating buffer is preferably carbonate buffer. The concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L....

Embodiment 1

[0088] Coating: Dilute wheat, egg white and milk food allergens with carbonate buffer solution to a food allergen coating solution with a total protein concentration of 10 μg / ml; dilute human IgA protein with carbonate buffer solution to obtain a standard package Coating solution (see Table 1 for protein concentration), add 0.05ml coating solution to each reaction well of the polystyrene plate, and overnight at 4°C. On the next day, discard the solution in the wells, add 0.35ml washing buffer to each well, wash 3 times, and incubate for 30s each time (referred to as washing, the same below);

[0089] Blocking: Add 0.05ml of PBS buffer (blocking solution) containing 2% bovine serum albumin to each reaction well, pH: 7.2, incubate at 15-35°C for 120min, discard the solution in the well, and wash as above;

[0090]Add samples: serum from patients with suspected wheat allergy, serum from patients with egg allergy, serum from patients with milk allergy (test serum), normal human se...

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Abstract

The invention provides an ELISA kit for detecting a food allergen based on an IgA antibody, and the kit belongs to the technical field of food allergen detection. The kit includes the following reagents: a detection plate coated with the food allergen, a washing buffer solution, a sample diluent, a standard substance or a positive quality control substance, a biotin-labelled anti-human IgA antibody, an enzyme-labeled avidin solution, a substrate coloring solution, and a stop solution. The ELISA kit of the invention selects the IgA for the first time to detect the food allergen, and is the prepared ELISA kit based on an indirect method; and the ELISA kit has the characteristics of strong sensitivity, high accuracy and good reproducibility, and can process large quantities of samples at onetime. Embodiments of the invention prove that the detection sensitivity of the kit of the invention reaches 10 ng / ml.

Description

technical field [0001] The invention relates to the technical field of food allergen detection, in particular to a kit for detecting food allergens based on IgA antibodies, that is, a food allergen-specific IgA detection kit. Background technique [0002] The incidence of allergic diseases accounts for more than 30% of the world's total population, and is listed by the World Health Organization as one of the four major non-infectious diseases in the 21st century. In recent years, the incidence of allergic diseases has increased significantly and has become a public health issue of global concern. The epidemiological survey of allergic diseases by Guo Jia et al. shows that 22% of the total population of 1.2 billion suffer from allergic diseases. For example, allergic rhinitis, asthma, conjunctivitis, eczema, food allergy, drug allergy and severe anaphylaxis, etc., seriously affect people's quality of life and life safety. [0003] Food allergens such as eggs, peanuts, milk, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6854
Inventor 何韶衡何萍
Owner 何韶衡
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