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Circulation free DNA and blood cell preservation medium

A medium and protective agent technology, applied in the field of biological preparations, can solve the problems of high cost, inability to obtain effects, high price, etc., and achieve the effect of ensuring accuracy, obvious protective effect, and reducing carcinogenic risk.

Inactive Publication Date: 2019-07-12
江苏芯超生物科技(集团)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The two products meet the requirements of users to varying degrees, but because of their high prices and single functions, users cannot get better results while bearing high costs.

Method used

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  • Circulation free DNA and blood cell preservation medium
  • Circulation free DNA and blood cell preservation medium
  • Circulation free DNA and blood cell preservation medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The storage medium contains the following components:

[0033] 1) Caprolactone and imidazolidinyl urea are used as protective agents;

[0034] 2) tripotassium ethylenediaminetetraacetic acid dihydrate as a chelating stabilizer;

[0035] 3) tunicamycin and aurin tricarboxylic acid are used as metabolic inhibitors;

[0036] 4) Glyceraldehyde is used as an enzyme inhibitor;

[0037] 5) Polyvinyl alcohol and trehalose are used as anti-adsorption agents;

[0038] 6) Glycine-phosphate as buffer.

[0039] The storage medium is 16g / L ethylenediaminetetraacetic acid tripotassium dihydrate, 24g / L caprolactin, 16g / L imidazolidinyl urea, 33.6g / L tunicamycin, 101.6g / L aurintricarboxylic acid , 80g / L polyvinyl alcohol, 120g / L trehalose, 120g / L glyceraldehyde, 8×glycine-phosphate buffer (50g / L glycine) configuration, the final pH is 7.4, and storage medium and blood samples or products by 1 : 7 blends.

[0040] Use conventional EDTA anticoagulant tubes to collect blood. After bl...

Embodiment 2

[0043] Preservation medium is formulated according to the dose in Table 1, and the final pH is 7.4, which is mixed with the preservation medium and blood samples or products at a ratio of 1:7.

[0044]

[0045]

[0046] The blood of non-pregnant women was collected with conventional EDTA anticoagulant tubes, and then divided into 21 tubes. 2 mL of blood was added to each tube, and the same amount of male white blood cell suspension was added to each tube of blood. Among them, 571.4 μL of the storage medium 1 and 2 configured in the above Table 1 were added to 14 tubes, 7 tubes of each storage medium, and the other 7 tubes were not added with the storage medium, and they were all stored at room temperature (25-28°C). Day, 2 days, 3 days, 4 days, 7 days, 10 days and 14 days, a total of 7 time points. At each time point, a tube of blood without protective medium and a tube of blood added with various preservation media were taken out, and the plasma was separated by centrif...

Embodiment 3

[0054] Blood was collected in conventional EDTA anticoagulant tubes, and divided into 21 tubes, and 2 mL was added to each tube. Among them, 285.6 μL of storage media 1 and 2 in Example 2 were added to 14 tubes, 7 tubes of each storage medium, and the other 7 tubes were stored at room temperature (25-28° C.) without adding storage media. 2 days, 3 days, 4 days, 7 days, 10 days and 14 days, a total of 7 time points. At each time point, a tube of blood without a protective medium and a tube of blood added with a preservation medium in Example 1 of the present invention were taken out for detection and analysis of cell activity. The instrument used for the detection is CellometerAUTO2000 automatic double fluorescence cell viability counting analyzer from Nexcelom Company of the United States. The composite dye AO / PI was used to adjust its concentration to 2×, and the preserved one was diluted 10 times with 1×PBS solution, and compounded with 2× After the dyes were mixed at 1:1, ...

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Abstract

The invention relates to a circulation free DNA and blood cell preservation medium which comprises the following components: 1) one or more protection agents; 2) one or more chelation stabilizers; 3)one or more metabolic inhibitors; 4) one or more enzyme inhibitors; 5) one or more anti-adsorption agents; 6) a buffer solution. The preservation medium has a remarkable circulation free DNA and cellprotection effect, is capable of preserving a blood preparation or a biological sample for a long time at a normal temperature and is free of free aldehyde substance, anaphylactic reactions and toxicreactions of human bodies in contact with liquid samples and biological samples with the preservation medium are reduced, potential carcinogenic risks of the preservation medium are reduced, the completeness of nucleic acid in a product preparation and a biological sample can be maintained, accuracy and scientificity of nucleic acid substance detection can be ensured, and detection results approximate to reality can be provided.

Description

technical field [0001] The invention discloses a preservation medium, specifically a circulation free DNA and blood cell preservation medium, and belongs to the technical field of biological preparations. Background technique [0002] Because of its convenient sampling and simple operation, blood is currently the most widely used sample in life science research and clinical diagnosis, and blood products are also widely used in clinical practice. Traditional anticoagulant tubes such as EDTA anticoagulant tubes, heparin anticoagulant tubes, etc. have the effect of preserving blood samples, but the storage time is short. Even if they are stored at 4°C for 12 hours, the cell activity will be lost by as much as 30-40%. This makes the sample unable to meet the requirements in some more demanding experiments. [0003] As a branch of in vitro diagnosis, liquid biopsy is used to detect circulating tumor cells (circulating tumor cell, CTC), circulating tumor DNA (circulating tumor DN...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N15/10
CPCA01N1/0226C12N15/10
Inventor 张涛张海天
Owner 江苏芯超生物科技(集团)有限公司
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