ELISA kit for detecting airborne allergen based on IgA 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 high accuracy, good reproducibility, and improved sensitivity

Inactive Publication Date: 2018-09-28
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 is still focused on the allergic reaction mediated by

Method used

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

Examples

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Example Embodiment

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

[0049] 1) Dilute the air-borne allergen with the coating buffer to obtain the air-borne allergen coating solution;

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

[0051] 3) The next day, after washing the detection plate wells in the step 2) overnight, add a blocking solution, and wash again after incubation to obtain a detection plate coated with airborne allergens.

[0052] The present invention dilutes airborne allergens and human IgA protein with a coating buffer solution to obtain a coating solution. In the present invention, the dilution method is not particularly limited, and the technical solution for dilution well known to those skilled in the art may be used. In the present invention, the coating buffer is preferably a carbonate buffer. In the present...

Example Embodiment

[0087] Example 1

[0088] Coating: Dilute dust mites, Artemisia sphaerocephala pollen and Humulus humulus pollen allergens with carbonate buffer to an airborne allergen coating solution with a mass concentration of 10 μg / ml; dilute human IgA protein with carbonate buffer Obtain the standard coating solution (see Table 1 for protein concentration). Add 0.05ml of coating solution to each reaction well of the polystyrene plate and leave it at 4°C overnight. The next day, discard the solution in the wells, add 0.35ml of washing buffer to each well, wash 3 times, and incubate for 0.5 min each time. (Referred to as washing, the same below);

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

[0090] Add sample: Suspicious dust mite allergic patient serum, wormwood pollen allergic patient serum, Humulus pollen allergic patient s...

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Abstract

The invention provides an ELISA kit for detecting an airborne allergen based on an IgA antibody, and belongs to the technical field of airborne allergen detection. The kit is prepared from the following reagents: a test board coated with the airborne allergen, a washing buffer, a sample diluent, a standard substance or a positive quality control material, a biotin-labeled anti-human IgA antibody,an enzyme-labeled avidin solution, a color-substrate solution and a stop buffer. According to the ELISA kit provided by the invention, IgA is selected for detecting the airborne allergen; in addition,the ELISA kit is prepared based on an indirect method, has the characteristics of high sensitivity, high accuracy and good reproducibility, and can treat large quantities of samples at a time. The embodiment proves that the detection sensitivity of the kit provided by the invention reaches 10ng/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 IgA antibodies, that is, a specific IgA 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, food allergy, drug allergy and severe anaphylaxis, etc., seriously affect people's quality of life and life safety. [0003] Airborne allergens such as pollen pa...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/558G01N33/6854G01N2333/47
Inventor 何韶衡何萍
Owner 何韶衡
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