ELISA kit for detecting airborne allergen based on IgM antibody

An antibody detection and kit technology, which is applied in the field of allergen detection, can solve the problems of inability to timely and accurately determine allergic diseases and stay, and achieve the effect of improving sensitivity, various types and wide application range.

Inactive Publication Date: 2018-09-28
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 IgM antibody
  • ELISA kit for detecting airborne allergen based on IgM antibody
  • ELISA kit for detecting airborne allergen based on IgM 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 a coating solution for the airborne allergen;

[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 test plate wells overnight in step 2), add blocking solution, and wash again after incubation to obtain a test plate coated with airborne allergens.

[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 carbo...

Embodiment 1

[0088] Coating: Dilute Humulus japonicus pollen and Artemisia grandis pollen allergens with carbonate buffer solution to airborne allergen coating solution with a mass concentration of 10 μg / ml; dilute human IgM protein with carbonate buffer solution to obtain standard products 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: sera from patients with suspected humulus pollen allergy, sera from patients with wormwood pollen allerg...

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Abstract

The invention provides an ELISA kit for detecting an airborne allergen based on an IgM 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 IgM antibody,an enzyme-labeled avidin solution, a color-substrate solution and a stop buffer. According to the ELISA kit provided by the invention, IgM 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 IgM antibodies, that is, a specific IgM 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 a...

Claims

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Application Information

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