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Detection kit for semi-quantitative allergen screening and preparation method thereof

A technology for allergens and kits, which is applied in the field of detection kits for allergen screening and its preparation, and can solve the problems of high cost and low throughput

Pending Publication Date: 2022-07-29
NANJING LANSION BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, serum-specific IgE detection is the most efficient detection method, but a conventional experiment can only detect a single indicator, and the throughput is low and the cost is high

Method used

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  • Detection kit for semi-quantitative allergen screening and preparation method thereof
  • Detection kit for semi-quantitative allergen screening and preparation method thereof
  • Detection kit for semi-quantitative allergen screening and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0032] The preparation method of the membrane strip includes the following 4 steps:

[0033] (1) Preparation of first-level membrane strip: paste the cut nitrocellulose membrane on the double-sided tape, and press it tightly;

[0034] (2) Preparation of secondary membrane strips: Preparation of antigen coating preparation solution (0.1M, pH7.4 PB buffer, containing 2% BSA, 0.1% TX-100, 5% allergen stabilizer, 0.1% Proclin300), 19 kinds of allergens, cross-reactive carbohydrate determinants (CCD- used to judge whether it is a false positive) and quality control material (used to guide the judgment of positive results), respectively, were diluted with the corresponding coating preparation solution, using a film stripper Carry out the coating, and place it under 30% humidity for drying after completion;

[0035] (3) preparation of tertiary film strip: paste the secondary film strip on the transparent film;

[0036] (4) Use a slitter to cut the film strips according to the size,...

Embodiment 1

[0055] Example 1: Confirmation experiment of biotinylated anti-human IgE antibody working solution concentration

[0056] The house dust mites / dust mites were coated with universal coating buffer - phosphate buffer (0.1M PB, pH7.4) (antigen coating solution) at the concentrations of 0.5mg / mL, 2mg / mL and 6mg / mL. Membrane, biotinylated anti-human IgE antibody concentrations were 4 μg / mL, 10 μg / mL, 20 μg / mL to prepare working solution, SA-AP was diluted with diluent at a ratio of 1 / 5000 and prepared into enzyme-binding working solution. The gray values ​​of 3 positive samples and 3 negative samples of dust mites / dust mites, the results are aggregated as shown in Table 2 below.

[0057] Table 2. Confirmation experiment of biotinylated anti-human IgE antibody working solution concentration

[0058]

[0059] From the experimental results in Table 2 above, it can be seen that under the condition that the coating concentration of house dust mites / dust mites is unchanged, with the ...

Embodiment 2

[0060] Example 2: Confirmation experiment of working solution dilution ratio of enzyme binding solution

[0061] Dilute SA-AP according to the ratio of 1 / 2000, 1 / 5000 and 1 / 10000 to prepare an enzyme binding solution, which is matched with the above appropriate concentration of house dust mite / dust mite membrane strip and biotinylated anti-human IgE antibody working solution. In the test, the gray values ​​of 3 positive samples and 3 negative samples of house dust mites / dust mites were detected, and the results were aggregated as shown in Table 3 below.

[0062] Table 3. Confirmation experiment of working solution dilution ratio of enzyme binding solution

[0063]

[0064] From the experimental results in Table 3 above, it can be seen that in both positive and negative samples, when the SA-AP dilution ratio is 1 / 2000 or 1 / 5000, the difference in the detected grayscale values ​​is very small, indicating that SA -AP is in excess state at a concentration of 1 / 2000, while the ...

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Abstract

The invention discloses a kit for semi-quantitative allergen screening. The kit comprises a nitrocellulose membrane strip coated with 19 allergens, a biotinylated anti-human IgE antibody, streptavidin labeled by alkaline phosphatase and a substrate solution, the allergens include willow / poplar / elm, common ragweed, mugwort, dermatophagoides pteronyssinus / dermatophagoides farinae, house dust, cat hair, dog epithelium, cockroaches, penicillium notatum / cladosporium / aspergillus fumigatus / alternaria, scandent hop, egg white, milk, peanuts, soybeans, beef, mutton, cod / lobster / scallop, shrimps and crabs. 19 common allergens can be qualitatively and quantitatively detected through one-time detection, and the method has the advantages that the operation is simple, the detection efficiency is greatly improved, the production and use cost is reduced, and the popularization and application of allergen detection are facilitated.

Description

technical field [0001] The present invention relates to a detection kit, in particular to a detection kit related to allergen screening and a preparation method thereof. Background technique [0002] With the changes in the environment and people's lives in recent years, more and more people are exposed to complex living environments, the number and types of daily necessities are also increasing, and the occurrence of allergies has become more frequent. Among them, allergens can be mainly divided into two categories, one is inhalation allergens, such as dust mites, pollen, etc., which can cause allergic reactions after entering the respiratory tract through breathing, including asthma, allergic rhinitis or skin allergic reactions that cause the respiratory tract itself. ; Second, food allergens, such as milk, drugs, etc., can cause allergic reactions through the digestive tract, blood or skin contact. [0003] Common allergen detection methods include skin prick test, intra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/78
CPCG01N33/543G01N21/78
Inventor 许行尚杰弗瑞·陈徐霞飞郭金龙
Owner NANJING LANSION BIOTECH CO LTD
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