ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting airborne allergen based on IgG2 antigen

A technology of antibody detection and kit, which is applied in the field of airborne allergen-specific IgG2 detection kit, which can solve the problems of inability to timely and accurately determine allergic diseases and retention, and achieve high accuracy, variety, and reproducibility Good results

Inactive Publication Date: 2018-11-23
JINZHOU MEDICAL UNIV
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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 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 (Enzyme Linked Immunosorbent Assay) kit for detecting airborne allergen based on IgG2 antigen
  • ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting airborne allergen based on IgG2 antigen
  • ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting airborne allergen based on IgG2 antigen

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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, more pr...

Embodiment 1

[0088]Coating: Dilute dust mite, Artemisia grandis pollen and Humulus pollen allergens with carbonate buffer solution to the airborne allergen coating solution with a mass concentration of 10 μg / ml; dilute human IgG2 protein with carbonate buffer solution Obtain standard coating solution (protein concentration is shown in Table 1). 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 Artemisia grandis pollen allergy, serum from patient...

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Abstract

The invention provides an ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting an airborne allergen based on an IgG2 antigen, and belongs to the field of airborne allergen detection. The kit consists of a detection board coated with the airborne allergen, a washing buffer, a sample diluent, a standard substance or a positive quality control product, a biotin-labeled anti-human IgG2 antibody, an enzyme-labeled avidin solution, a substrate developing solution and a stop solution. IgG2 is used for detecting the airborne allergen, the ELISA kit is prepared based on an indirect method and has the characteristics of high detection sensitivity, high accuracy and high reproducibility, and a large batch of samples can be treated through at one time. As proved by the embodiment, the kit provided by the invention is up to 0.5ng / ml in the detection sensitivity.

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 IgG2 antibodies, that is, a specific IgG2 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 a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/558
CPCG01N33/535G01N33/558
Inventor 何韶衡张慧云
Owner JINZHOU MEDICAL UNIV
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