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Kit for measuring and diagnosing free and cell binding total IgE in human whole blood and preparation method

A kit and free-type technology, which is applied in the field of diagnostic kits and preparations for the determination of free-type and cell-bound total IgE in whole blood, can solve the problems of low detection sensitivity, avoid inaccurate data, and achieve good clinical promotion and detection convenient effect

Inactive Publication Date: 2018-01-23
SHENZHEN IMMUNOTHERAPY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The problem to be solved by the present invention is to provide a detection and diagnosis kit of whole blood total IgE (free type and cell-bound type) and its preparation and use method, so as to solve the problem that the existing IgE detection kit can only detect free type IgE in serum. limitations
Since the content of cell-bound IgE is 10-100 times that of free IgE, the present invention overcomes the problem of low detection sensitivity of existing detection kits

Method used

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  • Kit for measuring and diagnosing free and cell binding total IgE in human whole blood and preparation method
  • Kit for measuring and diagnosing free and cell binding total IgE in human whole blood and preparation method
  • Kit for measuring and diagnosing free and cell binding total IgE in human whole blood and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A diagnostic kit for measuring free and cell-bound total IgE in whole blood, comprising:

[0039] 96-well plate coated with IgE antibody.

[0040] Standard. The concentration of the standard is as follows: 480, 240, 120, 60, 30, 15 μg / mL.

[0041] Sample diluent.

[0042] Detection Antibody-HRP.

[0043] 20x wash buffer.

[0044] Chromogenic substrate A.

[0045] Chromogenic Substrate B.

[0046] stop solution.

[0047] Sealing film.

[0048] user's Guide.

[0049] Configure the above kits by the following steps:

[0050] 96-well plate coated with IgE antibody. Each well was coated with 100 ng of anti-human IgE antibody.

[0051] Standard. The concentration of the standard is as follows: 480, 240, 120, 60, 30, 15 μg / mL.

[0052] Sample diluent. 1L sample diluent contains: potassium dihydrogen phosphate (KH2PO4): 0.27g, disodium hydrogen phosphate (Na2HPO4): 1.42g, sodium chloride (NaCl): 8g, potassium chloride (KCl) 0.2g, 4g BSA, pH7.4.

[0053] Detection ...

Embodiment 2

[0056] A method for using the kit prepared in Example 1, comprising the steps of:

[0057] Take out the required strips from the aluminum foil bag after equilibrating at room temperature for 20 minutes, and seal the remaining strips with a ziplock bag and put them back at 4°C.

[0058] Set standard wells and sample wells, and add 50 μL of standard substances of different concentrations to each standard well;

[0059] Add 50 μL of the sample to be tested into the sample well; do not add to the blank well.

[0060] Except for the blank wells, add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each well of standard wells and sample wells, seal the reaction wells with plate sealing film, and incubate for 60 min in a 37°C water bath or incubator.

[0061] Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350 μL), let it stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat the plate washing...

Embodiment 3

[0065] Performance testing of the kit prepared by the present invention.

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Abstract

The invention provides a method for measuring free and cell binding total IgE in human whole blood and a diagnostic kit preparation method. The IgE is synthesized and secreted by plasma cells, most ofthe IgE and cells (such as basophilic granulocyte and mastocyte) are bound through receptors of the IgE, and only a little IgE is free in blood serum. An existing IgE detection method can only detectfree IgE in the blood serum and cannot detect the IgE bound on cell membranes. According to the method, blood pH (potential of hydrogen) is reduced, so that the IgE and the receptors of the IgE positioned on cells are dissociated, and the free and cell binding total IgE is detected by an ELISA (double-antibody one-step sandwich enzyme-linked immuno sorbent assay) method. The method overcomes theshortcoming that the existing IgE detection method can only detect the free IgE in the blood serum, and the free and cell binding total IgE can be detected.

Description

technical field [0001] The invention belongs to the field of immunoanalysis medicine, in particular to a diagnostic kit for measuring free and cell-bound total IgE in whole blood and a preparation method. Background technique [0002] IgE molecules are key molecules in the development of allergic diseases, and detection of IgE levels is very important in the diagnosis and treatment evaluation of allergic diseases. In recent years, with the accelerated development of industrialization and the rapid changes in human living environment and lifestyle, the incidence of allergic diseases has been increasing. According to the epidemiological survey results of allergic diseases published by WHO in 2006, nearly 22% (250 million people) of the total population of 1.2 billion in the 30 countries and regions surveyed suffer from IgE-mediated allergic diseases. Such as allergic rhinitis, asthma, conjunctivitis, eczema, food allergy, drug allergy, etc. According to a survey by the World...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/535
Inventor 张军方
Owner SHENZHEN IMMUNOTHERAPY BIOTECH CO LTD
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