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Serum-specific ige biological activity detection method and kit used therefor

A technology of biological activity and detection method, applied in the field of serum-specific IgE biological activity detection and the kit used, can solve the problem of inability to detect serum sIgE immune activity and the like

Inactive Publication Date: 2016-06-08
TIANJIN MEDICAL UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

All in all, the serum sIgE detection method currently used clinically cannot detect the immune activity of serum sIgE, and there are certain defects

Method used

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Examples

Experimental program
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Effect test

Embodiment Construction

[0021] A kit used for the detection method of serum-specific IgE biological activity, the kit includes: ① KU812 cell line, purchased from American Type Culture Collection Center (ATCC); ② anti-human IgE antibody, purchased from ABCAM, UK Company; ③ anti-CD63-FITC labeled antibody, purchased from BD Company, USA; ④ series of allergen solutions.

[0022] To complete the said detection items of the technical invention, it is also necessary to include cell laboratory routine solutions such as cell culture medium, Hanks solution, phosphate-buffered saline, cell culture plates, and cooperate with instruments such as flow cytometers and horizontal centrifuges.

[0023] Among them, KU812 cells are suspension cells, and the RPMI-1640 medium containing more than 15% fetal bovine serum can be used. Culture plates can be 6-well, 12-well or 24-well.

[0024] Taking the detection of milk-specific IgE biological activity as an example, the detection process is as follows:

[0025](1) Prepa...

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Abstract

The invention provides a detection method for serum specificity IgE biological activity and a kit adopted by the same. The detection method comprises the following steps: adopting a KU812 cell system as indicate cells to induce expression of IgE-Fc receptors; incubating serum of a subject and the KU812 cells to facilitate the combination of sIgE to be detected in the serum and the IgE-Fc receptors on the surfaces of the KU812 cells; adding specific allergens combined with the sIgE on the surfaces of the KU812 cells in order to cross-link the receptors and activate the KU812 cells for degranulation and histamine releasing, and simultaneously promoting CD63 molecules in the cells to be transferred onto the surfaces of the cell membranes; marking anti-CD63 antibodies by fluoresceins for cell immunostaining; adopting a flow cytometry to detect the percentage of the expressing CD63 cells in order to finally determine the activated intensity of the KU812 cells. The detection method has the benefits that the detection method can be used for evaluating the risk degree of allergy when patients contact the allergens, as well as evaluating the clinical effects of specific desensitization treatment.

Description

technical field [0001] The invention relates to a serum-specific IgE biological activity detection method and a kit used therein, which are used for evaluating the serum-specific IgE pathogenic activity of allergic patients. Background technique [0002] Allergy is an abnormal immune response that occurs when the body is stimulated by certain antigens, which can be manifested as physiological dysfunction or (and) tissue cell damage. Allergies are divided into 4 types according to their mechanisms and clinical characteristics. Type I allergies are also called anaphylaxis, which are mainly mediated by immunoglobulin E (IgE) and involved in mast cells or basophils. The main characteristics of allergic reactions are as follows: ①It occurs quickly and subsides quickly; ②It is mediated by IgE, with the participation of mast cells or basophils; ③It usually causes physiological dysfunction of the body, and generally does not leave tissue damage; ④There are obvious individual differe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/582G01N33/6803G01N2333/70503
Inventor 李会强侯丽英朱黎娜林书祥常艳敏李迺昶
Owner TIANJIN MEDICAL UNIV
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