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Peripheral blood lymphocyte subpopulation cell concentration detection kit and detection method thereof

A technology for detecting kits and lymphocytes, which can be used in biological tests, measuring devices, material inspection products, etc., and can solve the problems of cumbersome steps and large errors in cell detection.

Pending Publication Date: 2018-08-07
李小峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects of the prior art, solve the technical problems of large cell detection errors and cumbersome steps in the prior art, and provide a peripheral blood lymphocyte subset cell concentration detection kit and its detection method

Method used

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  • Peripheral blood lymphocyte subpopulation cell concentration detection kit and detection method thereof
  • Peripheral blood lymphocyte subpopulation cell concentration detection kit and detection method thereof
  • Peripheral blood lymphocyte subpopulation cell concentration detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Such as Figure 1 to Figure 3 As shown, the peripheral blood lymphocyte subset cell concentration detection kit includes an antibody kit 1 and a supporting stimulating solution kit 2, wherein: the antibody kit 1 and the stimulating solution kit 2 are both set to have a cover One side of the antibody reagent kit 1 is set as an antibody reagent area, and the other side is set as a spare reagent area 12, and the antibody reagent area is provided with six-hole reagent tanks numbered sequentially from A to F 13. Corresponding to the number of the six-hole reagent tank 13, there are reagent tubes 11 from numbers A to F representing the six antibody solutions; the stimulation solution kit 2 is placed in the spare reagent area 12 and is close to the antibody kit 1 The rear side wall of the rear side wall, in the spare reagent area 12, there are spare reagent bottles 3 for adding hemolysin, culture solution, membrane rupture solution, distilled water 10 times diluted buffer solu...

Embodiment 2

[0090] Ⅰ. Detection of cell concentration of peripheral blood lymphocyte subsets

[0091] a. Sample requirements: a 3ml EDTA anticoagulant tube with a purple cap is used as the experimental sample;

[0092] b. Reagent preparation:

[0093] Hemolysin;

[0094] Antibody A solution: Dissolve 1ml of CD3 / CD8 / CD45 / CD4 antibody reagent in 0.1% sodium azide buffer,

[0095] Among them, the mass-volume concentration of FITC-labeled CD3 antibody is 2.3 μg / mL, the mass-volume concentration of PE-labeled CD8 antibody is 1.75 μg / mL, and the mass-volume concentration of PerCP-labeled CD45 antibody is 7.5 μg / mL, The mass-volume concentration of APC-labeled CD4 antibody is 0.92 μg / mL;

[0096] Antibody B solution: Dissolve 1ml of CD3 / CD16+CD56 / CD45 / CD19 antibody reagent in 0.1% sodium azide buffer,

[0097] Among them, the mass-volume concentration of FITC-labeled CD3 antibody is 2.3 μg / mL, the mass-volume concentration of PE-labeled CD16+CD56 antibody is 2.75 μg / mL, and the mass-volume c...

example 1

[0166] Example 1: Wang Moumou, female, 27 years old, was admitted to the hospital mainly due to "intermittent facial erythema for more than 3 months, multi-joint swelling and pain for 1 month, and fever for 5 days". The diagnosis was "systemic lupus erythematosus, lupus nephritis, and neuropsychiatric lupus". According to the screening method provided in this example, the peripheral blood lymphocyte subset cells were tested: total T lymphocytes (CD3+CD19-) 198.11 / ul↓, (CD3+CD19-(%)) 51.31%↓; B lymphocytes (CD3-CD19+) 133.78 / ul, (CD3-CD19+ (%)) 37.77%↑; Th cells (CD3+CD4+) 45.29 / ul↓, (CD3+CD4+ (%)) 10.79%↓; Ts cells (CD3+CD4+) 157.45 / ul, (CD3+CD8+ (%)) 37.50%; Th / Ts (CD3+CD4+ / CD3+CD4+) 0.29↓; NK cells (CD3- / CD16+CD56+) 29.06 / ul↓, (CD3- / CD16+CD56+ (%)) 8.21%. Relative count of CD4+T cell subsets: Th1 (IFN-γ) 6.45%; Th2 (IL-4) 1.27%; Th17 (IL-17) 2.32%↑; Treg (CD25+Foxp3) 10.45%↑; Absolute count = percentage of positive cells of each subgroup × absolute number of CD4+ T cells ...

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Abstract

The invention relates to a peripheral blood lymphocyte subpopulation cell concentration detection kit and a detection method thereof, and belongs to the technical field of cell detection and screening. According to the technical scheme, the peripheral blood lymphocyte subpopulation cell concentration detection kit comprises an antibody kit and a stimulation liquid kit which is coordinately used with the antibody kit and is characterized in that a six-hole reagent tank is stored in the antibody kit, reagent tubes ranging from number A to number F and representing six antibody solutions are correspondingly arranged in the six-hole reagent tank according to the numbers of the six-hole reagent tank, and a standby reagent bottle is further stored in the antibody kit; after the detection of peripheral blood lymphocyte subpopulation cell concentration and the detection of CD4+T cell subpopulation cell concentration are carried out, samples are placed on a flow cytometer to analyze flow cytological maps, relative percentage is acquired and analyzed by MultiSET software or CellQuest software, and the cell absolute number of various subpopulations is calculated. The peripheral blood lymphocyte subpopulation cell concentration detection kit has the advantages that the technical problem that cell detection is large in errors and complicated in steps is solved, and the detection kit is capable of providing various antibodies and reagents, capable of simultaneously testing a plurality of samples during clinical application, simple in test steps, good in quality control and high in cell detection precision.

Description

technical field [0001] The invention belongs to the technical field of cell detection and screening, and in particular relates to a kit for detecting the cell concentration of peripheral blood lymphocyte subsets and a detection method thereof. Background technique [0002] The number and function of lymphocytes play a vital role in maintaining the homeostasis and balance of the autoimmune system. Peripheral blood lymphocytes can be divided into T cells, B cells, natural killer cells (NK cells), etc. Among them, each cell can be divided into different subgroups according to the differences in the antigens it expresses and its function. [0003] T cell receptor-leukocyte differentiation antigen 3 (TCR-CD3) complex is an important marker unique to T cells, so T cells are called CD3+T lymphocytes, which can reflect the cellular immune function of the body. Its increase is common in CD3+ T lymphocyte leukemia, some autoimmune diseases (such as systemic lupus erythematosus), myas...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/50
CPCG01N33/5047G01N33/533
Inventor 罗静李小峰高崇张升校赵向聪张辰吴琪梁聪聪来娜琳
Owner 李小峰
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