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ELISA kit for detecting airborne allergen based on IgG3 antibody

A technology of antibody detection and kit, which is applied in the field of airborne allergen-specific IgG3 detection kits, which can solve the problems of staying up and not being able to accurately determine allergic diseases in a timely manner, and achieve multiple types, improved sensitivity, and strong detection sensitivity Effect

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

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 allergen based on IgG3 antibody
  • ELISA kit for detecting airborne allergen based on IgG3 antibody
  • ELISA kit for detecting airborne allergen based on IgG3 antibody

Examples

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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 humulus pollen allergens to a mass concentration of 10 μg / ml with carbonate buffer solution; diluted human IgG3 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, discard the solution in the well, add 0.35ml washing buffer to each well, wash 3 times, and incubate 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), and normal human serum (n...

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Abstract

The invention provides an ELISA kit for detecting an airborne allergen based on an IgG3 antibody, and the ELISA kit belongs to the technical field of airborne allergen detection. The ELISA kit includes the following reagents: a detection plate coated with the airborne allergen, a washing buffer solution, a sample diluent, a standard substance or a positive quality control substance, a biotin-labelled anti-human IgG3 antibody, an enzyme-labeled avidin solution, a substrate coloring solution, and a stop solution. The ELISA kit of the invention selects the IgG3 to detect the airborne 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 one time. Embodiments of the invention prove that the detection sensitivity of the ELISA kit of the invention reaches 1 ng / ml.

Description

technical field [0001] The invention relates to the technical field of airborne allergen detection, in particular to a kit for detecting airborne allergens based on IgG3 antibodies, that is, a specific IgG3 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 aller...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/5436
Inventor 何韶衡何萍
Owner 何韶衡
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