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T lymphocyte immunophenotyping method and kit

A lymphocyte and immune typing technology, applied in the field of immunology, can solve problems such as complicated operation, unsuitable for practical application, and large demand for samples to be tested

Inactive Publication Date: 2015-02-18
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if you want to further perform immunophenotyping and quantitative analysis on cell subpopulations, you need to resort to other cellular immunotyping methods, which are complicated to operate and require a large number of samples to be tested, which is not suitable for practical applications.

Method used

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  • T lymphocyte immunophenotyping method and kit
  • T lymphocyte immunophenotyping method and kit
  • T lymphocyte immunophenotyping method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0134] Example 1 Lymphocyte Fine Immunotyping and Quantitative Analysis

[0135] Experimental Materials:

[0136] Sample to be tested: anticoagulated peripheral blood sample, derived from healthy volunteers, which is the peripheral blood of normal people.

[0137] BD Biosciences Lymphocyte Sorting Kit (cat340503), purchased from BD biosciences, wherein mixed antibody 1 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD3 (FITC) antibody, anti-CD4 (APC) antibody, anti-CD8 (PE ) antibody; mixed antibody 2 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD19 (APC) antibody, anti-CD56 and CD16 (PE) antibody, anti-CD3 (FITC) antibody. The red blood cell lysate that comes with the BD clinical kit.

[0138] Self-prepared erythrocyte lysate: every 1L of erythrocyte lysate contains NH 4 Cl8.29g, KHCO 3 1g, EDTA0.37g.

[0139] experimental method:

[0140] Take 500 μL of anticoagulated peripheral blood sample, take 200 μL of the sample to be tested, a...

Embodiment 2

[0193] Example 2 Fine Immunotyping and Quantitative Analysis of Lymphocytes

[0194] Experimental Materials:

[0195] Samples to be tested: anticoagulated peripheral blood samples, derived from venous blood samples from children who came to our hospital (with signed informed consent), and peripheral blood from children with suspected systemic lupus erythematosus (SLE).

[0196]BD Biosciences lymphocyte classification kit (cat340503), purchased from BD biosciences, wherein the mixed antibody 1 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD3 (FITC) antibody, anti-CD4 (APC) antibody, anti-CD8 (PE ) antibody; mixed antibody 2 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD19 (APC) antibody, anti-CD56 and CD16 (PE) antibody, anti-CD3 (FITC) antibody. The red blood cell lysate that comes with the BD clinical kit.

[0197] Self-prepared erythrocyte lysate: every 1L of erythrocyte lysate contains NH 4 Cl8.29g, KHCO 3 1g, EDTA0.37g.

[0198] e...

Embodiment 3

[0236] Example 3 Fine Immunotyping and Quantitative Analysis of Lymphocytes

[0237] Experimental Materials:

[0238] Samples to be tested: anticoagulant peripheral blood samples, derived from venous blood samples of children who came to our hospital (with signed informed consent), peripheral blood of children with suspected hyper-IgM syndrome.

[0239] BD Biosciences Lymphocyte Sorting Kit (cat340503), purchased from BD biosciences, wherein mixed antibody 1 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD3 (FITC) antibody, anti-CD4 (APC) antibody, anti-CD8 (PE ) antibody; mixed antibody 2 includes: fluorescently labeled Percp anti-CD45 antibody, anti-CD19 (APC) antibody, anti-CD56 and CD16 (PE) antibody, anti-CD3 (FITC) antibody. The red blood cell lysate that comes with the BD clinical kit.

[0240] Self-prepared erythrocyte lysate: every 1L of erythrocyte lysate contains NH 4 Cl8.29g, KHCO 3 1g, EDTA0.37g.

[0241] experimental method:

[0242] Take ...

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Abstract

The invention belongs to the field of immunological technique and discloses a T lymphocyte immunophenotyping method and a kit. The T lymphocyte immunophenotyping method provided by the invention comprises the following steps: taking different fluorescently-labeled antibodies and mixing the antibodies with a sample to be tested, carrying out incubation, carrying out flow cytometry detection to obtain test data, and analyzing the test data. The antibodies contain an anti-TCR alpha beta antibody, an anti-TCR gamma Delta antibody, an anti-CD3 antibody, an anti-CD4 antibody, an anti-CD8 antibody, an anti-CD27 antibody and an anti-CD45RA antibody. By the above method, more comprehensive fine T lymphocyte immunophenotyping is realized. Few samples to be tested are required. The method is simple to operate. Required time is short. And the method has high accuracy and can widely be applied in lymphocyte subpopulation immunophenotyping.

Description

technical field [0001] The invention belongs to the technical field of immunology, and in particular relates to a method and a kit for immunotyping of T lymphocytes. Background technique [0002] Lymphocytes, also known as lymphocytes, are the smallest type of white blood cells, with a diameter of 6-8 microns; they account for about 20%-30% of the number of white blood cells in the human body, with round nuclei and little cytoplasm; produced by lymphoid organs , is an important cell component of the body's immune response function, and is a type of cell line with immune recognition function. Lymphocytes are complex heterogeneous cell populations, which can be divided into different categories according to their phenotype and functional characteristics, such as T cells, B cells, NK cells, etc. These cells can be further divided into several subgroups. Lymphocytes and their subgroups cooperate and restrict each other in the process of immune response, and jointly complete the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/505G01N2333/705
Inventor 赵晓东周丽娜
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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