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Kit for detecting milk allergen and preparation method thereof

A detection kit and technology for milk allergy, applied in the field of milk allergen detection kit and its preparation, milk allergy enzyme-linked immunoassay detection mode

Inactive Publication Date: 2012-01-25
TIANJIN MEDICAL UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, at present, the double-antigen sandwich method is only used in the detection of biologically active molecules such as human immunodeficiency virus (HIV), and there is no report on its use in the detection of food allergy

Method used

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  • Kit for detecting milk allergen and preparation method thereof
  • Kit for detecting milk allergen and preparation method thereof
  • Kit for detecting milk allergen and preparation method thereof

Examples

Experimental program
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Embodiment Construction

[0030] The present invention will be further described below in conjunction with accompanying drawings and examples.

[0031] 1. Obtaining high-purity milk allergens

[0032] β-lactoglobulin, α-lactalbumin, and casein in milk are the main allergens that cause allergic reactions. At present, these components are commercially available with high-purity labels, which can be obtained by order, or according to The purification method reported in the literature is obtained by self-extraction. Our laboratory has explored a mature purification process for extracting high-purity single allergens such as β-lactoglobulin, α-lactalbumin, and casein. Now take β-lactoglobulin as an example to introduce the extraction process.

[0033] 1.1 Materials and reagents:

[0034] 1.1.1 Required materials

[0035] Fresh raw milk; low-temperature high-speed centrifuge; water bath; vacuum pump; dextran gel (Sephadex-G200, Sephadex-DEAE-A50, American DPC Biological Company); Mini p-4 vertical gel ele...

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Abstract

The invention which discloses a kit for detecting a milk allergen and a preparation method thereof belongs to an enzyme-linked immunosorbent assay (ELISA) detection technology. The kit comprises a coloring solution A, a coloring solution B, a stop solution, and a negative and positive quality control serum for milk allergy, and the milk allergens are respectively coated with a micropore plate and a labeled horseradish peroxidase to prepare a coating antigen and an enzyme-labeled antigen. Above design in the invention which allows traditional indirect ELISA detection modes to be changed adopts a double antigen sandwich ELISA method to construct the kit for detecting the milk allergen. The kit which is used for identifying and capturing specific antibodies of milk allergy in a serum of a patient allows a result to be determined with the catalysis substrate coloration. The kit which overcomes many shortages of the traditional detection modes allows the detection background to be reduced, and false positive results to be reduced, and there is no need to predilute the serum in the detection because no detection antibody is needed, so the preparation method is simplified, the detection specificity is improved, and the reagent and operation cost is simultaneously saved.

Description

technical field [0001] The invention relates to ELISA detection technology, in particular to a milk allergy ELISA detection mode, especially using the principle of double-antigen sandwich method to coat non-milk allergens on reaction plates and mark horseradish peroxide Enzyme, a milk allergen detection kit for detecting specific antibodies in serum of patients with milk allergy and a preparation method thereof. Background technique [0002] Food allergy is a major hygienic problem today, which has a serious impact on people's health and quality of life. Research on the pathogenic mechanism, detection, and prevention of food allergy has attracted increasing attention. Milk allergy is one of the main types of food allergy, which is highly common in infants and young children, and seriously threatens the health, growth and development of children. [0003] There is a substance called β-lactoglobulin in milk, which is a strong allergen. Some people with allergies will show all...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/559
Inventor 李会强陈寅常艳敏许顺连李婵任杰
Owner TIANJIN MEDICAL UNIV
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