ELISA kit for detecting airborne allergens based on IgG4 antibody

A technology of antibody detection and kit, which is applied in the field of allergen detection, can solve problems such as staying up and not being able to accurately determine allergic diseases in a timely manner, and achieve the effects of various types, improved sensitivity, and a wide range of applications

Inactive Publication Date: 2018-10-12
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the current research on airborne allergens in the existing technology stays at the allergic reaction mediated by IgE, which will lead to the inability to timely and accurately determine the occurrence of allergic diseases

Method used

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  • ELISA kit for detecting airborne allergens based on IgG4 antibody
  • ELISA kit for detecting airborne allergens based on IgG4 antibody
  • ELISA kit for detecting airborne allergens based on IgG4 antibody

Examples

Experimental program
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preparation example Construction

[0048] In the present invention, the preparation method of the detection plate coated with airborne allergens preferably includes the following steps:

[0049] 1) diluting the airborne allergen with a coating buffer to obtain the airborne allergen coating solution;

[0050] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;

[0051] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.

[0052] In the invention, the coating buffer is used to dilute the airborne allergen 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. In the present invention, the concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L. The pH...

Embodiment 1

[0088]Coating: airborne allergen coating solution diluted with dust mite, dust mite and wormwood pollen allergens to a mass concentration of 10 μg / ml with carbonate buffer solution; diluted human IgG4 protein with carbonate buffer solution to obtain Standard coating solution (see Table 1 for protein concentration). Add 0.05ml of coating solution to each reaction well of the polystyrene plate, overnight at 4°C. On the next day, the solution in the wells was discarded, and 0.35ml of washing buffer was added to each well, washed 3 times, and incubated for 0.5min each time. (referred to as washing, the same below);

[0089] Blocking: Add 0.05ml of PBS buffer solution containing 2% bovine serum albumin to each reaction well, pH: 7.2 (blocking solution), 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 dust mite allergy, serum from patients with Humulus pollen allergy (serum to be tested),...

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Abstract

The invention provides an ELISA kit for detecting airborne allergens based on an IgG4 antibody, and belongs to the technical field of the detection of airborne allergens. The ELISA kit includes the following reagents: a test plate coated with the airborne allergens; a washing buffer; a sample diluent; a standard or a positive quality control; a biotin-labeled anti-human IgG4 antibody; an enzyme-labeled avidin solution; a substrate color developing solution; and a stop solution. The ELISA kit adopts the IgG4 to detect the airborne allergens, and the ELISA kit is an ELISA kit prepared based on an indirect method, has the characteristics of high sensitivity, high accuracy and good reproducibility, and can process large quantities of samples at one time. The embodiment proves that the ELISA kit for detecting the airborne allergens based on the IgG4 antibody has a detection sensitivity of 1 ng/ml.

Description

technical field [0001] The invention relates to the technical field of allergen detection, in particular to a kit for detecting airborne allergens based on IgG4 antibodies, that is, a specific IgG4 detection kit for airborne allergens. 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, airborne allergy, drug allergy and severe allergic reactions, etc., seriously affect people's quality of life and life safety. [0003] Airborne allergens such...

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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