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Identification method and application of eb virus-infected lymphocyte subpopulation

A lymphocyte and EB virus technology, applied in the field of medical detection, can solve the problems of poor EB virus detection and other problems

Active Publication Date: 2018-10-23
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The first object of the present invention is to provide a kind of identification method of Epstein-Barr virus-infected lymphocyte subsets, and the second object of the present invention is to provide the application of the identification method of EB virus-infected lymphocyte subsets, to alleviate the existing technology. Existing technical problems of poor detection of EB virus

Method used

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  • Identification method and application of eb virus-infected lymphocyte subpopulation
  • Identification method and application of eb virus-infected lymphocyte subpopulation
  • Identification method and application of eb virus-infected lymphocyte subpopulation

Examples

Experimental program
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Embodiment 1

[0045] This embodiment provides a magnetic bead-based identification method for EB virus-infected lymphocyte subsets, which includes the following steps:

[0046] 1. Extract human peripheral blood mononuclear cells (HPBMC): 6mL fasting venous blood, placed in a sterile test tube with EDTA anticoagulation, diluted 1 times with PBS, and separated PBMC with lymphocyte separator (2000 turns 20 minutes), using Centrifuge time with buffer solution (MACS BSA Stock Solution (#130-091-376) and autoMACS Rinsing Solution (#130-091-222) 1:20 ratio), and place in a refrigerator at 4 degrees;

[0047] 2. Aspirate the middle tunica albuginea layer, add 10mL PBS, 1400rpm, 10min to wash and precipitate PBMC;

[0048] 3. Discard the supernatant, add 2mL red blood cell lysate (Solebold red blood cell lysate item number: Cat#R1010) for lysis, mix well, and incubate at room temperature for 5 minutes;

[0049] 4. After mixing well, draw 20μL of cells and dilute 1:3 with 2% glacial acetic acid, then take th...

Embodiment 2

[0080] This embodiment provides a flow cytometer-based identification method for EB virus-infected lymphocyte subsets, which includes the following steps:

[0081] 1. Take 6 mL of peripheral blood, anticoagulate with heparin, dilute to 12 mL with PBS, and mix;

[0082] 2. Slowly add the diluted blood along the test tube wall to the surface of the 15mL lymphocyte separation solution. Do not use too much force to avoid mixing the blood and the separation solution and maintain a clear layered state;

[0083] 3. Centrifuge at 2000 rpm for 20 minutes at 18-20°C. After centrifugation, the blood in the test tube can be clearly divided into 4 layers. The upper layer is the plasma layer, and the middle layer is the separation layer (mononuclear cells are located between the plasma layer and the separation layer) , The bottom layer is the red blood cell layer, and the red blood cell layer is the granulocyte layer;

[0084] 4. Aspirate the lymphocyte layer between the upper layer and the middle ...

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Abstract

The invention provides an identification method of an EB virus infected lymphocyte subpopulation and an application thereof and relates to the technical field of medical detection. The identification method of the EB virus infected lymphocyte subpopulation provided by the invention can accurately position the cell type of EBV infection by sorting monocytes of peripheral blood of an EBV infected patient in combination with a flow cytometry and a real-time fluorescent quantitative PCR technology, and is helpful for a clinician to formulate a therapeutic regimen for the cell type with EBV infection so as to efficiently and accurately treat diseases caused by EBV infection. In addition, by applying the identification method provided by the invention, target cells infected with EBV can be determined, preparation of related therapeutic drugs can be guided, and the targeted drugs are prepared with a clear target.

Description

Technical field [0001] The invention relates to the technical field of medical detection, in particular to a method for identifying EB virus-infected lymphocyte subgroups and its application. Background technique [0002] Epstein-Barr virus (EBV) belongs to the fourth type of human herpes virus. It was discovered by Epstein and Barr in 1964 when they were studying malignant lymphoma in children in Africa. Epstein-Barr virus is a lymphotropic virus, one of the viruses commonly infected by humans, infecting more than 90% of the world's population. The Epstein-Barr virus has both a high infection rate and a carcinogenic rate, and was designated as a Class I carcinogen by the International Cancer Research Center in 1997. [0003] At present, there are some EB virus detection kits, but most of them detect EB virus infection in whole blood, which is not targeted. [0004] In view of this, the present invention is proposed. Summary of the invention [0005] The first object of the present...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70G01N15/14G01N21/64C12R1/93
CPCC12Q1/705C12Q2600/136G01N15/14G01N21/6486
Inventor 王昭高卓王旖旎张嘉王晶石吴林
Owner BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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