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CD4 (Cluster of Differentiation 4) cell chip as well as preparation method and application thereof

A CD4 cell and chip technology, applied in the field of CD4 cell chip and its preparation, can solve the problems such as the limited popularization of flow cytometer, and achieve the effects of reducing the difficulty of blood collection, short processing time and low cost

Inactive Publication Date: 2011-01-26
SHANGHAI SEMIBIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cost of flow cytometry limits its clinical popularity

Method used

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  • CD4 (Cluster of Differentiation 4) cell chip as well as preparation method and application thereof
  • CD4 (Cluster of Differentiation 4) cell chip as well as preparation method and application thereof
  • CD4 (Cluster of Differentiation 4) cell chip as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of CD4 cell chip

[0044] 1. Slide cleaning

[0045] At room temperature, soak the glass slide in chromic acid cleaning solution for 2 to 4 hours, and rinse it with process water. Soak the glass slide in 5% to 10% nitric acid aqueous solution, and heat in an oven at 85°C for 1.5 to 3.5 hours. After rinsing with process water, soak the slides in process water and store at room temperature. Since the main component of the glass slide is silicon dioxide, the surface of the slide is anionically charged after being treated with chromic acid and nitric acid aqueous solution successively.

[0046] 2. Antibody coating pre-treatment

[0047] Take out the glass slide soaked in the process water, tap off an appropriate amount of process water, and keep the slide in a wet state and soak it in 0.1-0.5% (W / V) poly-lysine solution for 30-60 minutes. After rinsing with water, centrifuge at 600-800r / min for 4 minutes to dry, and store at room temperature. Poly...

Embodiment 2

[0050] Example 2 Preparation of CD4 cell detection kit

[0051] CD4 cell detection kit contains CD4 cell chip, diluent, hydrogen peroxide solution, staining solution and counterstaining solution.

[0052] The CD4 cell chip was prepared according to Example 1.

[0053] Diluents may be prepared by dissolving the diluent tablet in an aqueous solution. Diluent tablets are commercially available, purchased from Shanghai Baokang Biological High-tech Co., Ltd., and the main components are sodium chloride, potassium chloride, potassium dihydrogen phosphate, disodium hydrogen phosphate, citric acid, Tween 20, etc., with One tablet can be dissolved in 40ml aqueous solution to make phosphate buffer solution with pH 7.0-7.4. Its specifications are 10 pieces / pack, packed in sealed aluminum foil bags, and kept in an airtight place with shading. When the kit is assembled, it is only necessary to add a label to the outside of the bag. The diluent can also be prepared by preparing the abov...

Embodiment 3

[0057] Example 3 Application of CD4 cell chip for detection (for the overall process, see figure 2 )

[0058] 1. Specific binding reaction: Add water to the wet box so that the bottom of the wet box is completely covered with water, and the place where the slides are placed should be kept dry. Take out the slide, place the slide on the wet box holder, and close the lid tightly. The diluted blood sample is dropped on the coated antibody of the CD4 cell chip for antigen-antibody reaction. React in a humid box at room temperature (20-30°C) for 40 minutes.

[0059] 2. Washing and staining: The reacted slides are pulled with diluent for 5 to 10 times, mainly to wash off non-specific binding cells and excess blood samples with diluent, and then put them into the staining solution tank (with CD4 antigen The expressed cells will be stained), after standing for 1 minute, transfer the slightly drained slide holder to fresh hydrogen peroxide working solution, and let stand for 4 minu...

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Abstract

The invention discloses a CD4 (Cluster of Differentiation 4) cell chip which comprises a solid phase support and an antibody fixed on the support, wherein the antibody is a CD4 monoclonal antibody, and the solid phase support is a slide. The invention also discloses a preparation method of the CD4 cell chip, a CD4 cell assay kit containing the CD4 cell chip and a method for applying the CD4 cell chip to detection. The invention also discloses the application of the CD4 cell chip and the CD4 cell assay kit. The CD4 cell chip has the advantages of low cost and simple and convenient operation and is especially applicable to poverty-stricken zones seriously infected by AIDS viruses.

Description

technical field [0001] The invention relates to a cell chip, in particular to a CD4 cell chip and its preparation method and application. Background technique [0002] CD4 cells refer to T lymphocytes with CD4 antigen, also known as helper T cells, which play a pivotal role in maintaining the body's cellular and humoral immunity. CD4 cells are the main fighting cells in the immune system and the main target cells for human immunodeficiency virus (HIV) infection. The main pathological process after the human body is infected with HIV is the damage of the immune system, which is mainly manifested in the loss of CD4 cells and the reduction of the absolute number. At present, CD4 cell count has been widely used to guide clinical anti-HIV treatment, and has become the gold standard for the definition of AIDS by the World Health Organization. The World Health Organization defines that when the number of CD4 cells in human peripheral blood is less than or equal to 200 / microliter,...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/552G01N15/10G01N33/576
Inventor 曹波何骏杰杨君
Owner SHANGHAI SEMIBIO TECH
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