A method for identification of basophil activation and degranulation

Abstract

The invention provides a method for identifying basophil activation and degranulation, which includes the following steps: number the test samples in order, and mark the flow sample loading tubes required for the test; Add the required combination of anti-human CD123, anti-human CCR3, anti-human HLA‑DR, anti-human CD203c and / or anti-human CD63 fluorescently labeled flow cytometry antibodies; add the mixed whole blood to the labeled flow cytometry tube and keep at room temperature Incubate in the dark; add red blood cell lysis solution to each sample tube and incubate at room temperature in the dark; after the incubation, centrifuge to remove the supernatant; use a flow cytometer to detect and analyze the basophil count and average fluorescence intensity. The method of the present invention can distinguish 98-100% of basophils in peripheral blood from other types of cells without the need to separate and purify basophils, and can identify basophil activation and / or In the degranulated state, each sample to be tested requires no more than 100 μL of whole blood and has good repeatability.

Description

technical field [0001] The invention relates to the technical field of allergic disease diagnosis, in particular to an identification method for basophil activation and degranulation. 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. With the development of industrial economy and the change of ecological environment, such diseases have been increasing in recent years, becoming common and frequently-occurring diseases, which is a major problem that needs to be solved in the field of health and economic development in our country. [0003] Historically, most of the definitions and diagnostic schemes of diseases were proposed by developed countries. In 2011, He Shaoheng and Zhang Huiyun took the lead in revising and discussing the definition of allergic diseases that had been used for nea...

AI Technical Summary

Problems solved by technology

Unfortunately, to date there is no reliable and reproducible method for specifically identifying the activated, degranulated state of basophils in whole blood, leading to allergen-specific basophil priming. The test cannot be carried out

Our recent study found that if CD123, HLA-DR, and CCR3 are detected simultaneously, 98-100% of the CCR3+CD123+HLA-DR- cell population is basophils, which is the first to basically solve the specificity of basophils. sexual immune recognition, but did not address the identification of basophil activation and degranulation

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