Blood additive

An additive, blood technology, applied in the direction of blood/immune system cells, cell culture active agents, biochemical equipment and methods, etc., can solve the problems of stimulating hemolysis, easy activation of platelets, free nucleic acid degradation, etc., to achieve strong anti-hemolysis ability. Effect

Inactive Publication Date: 2019-02-12
AMOY DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In addition, in order to achieve blood transportation at this stage, 4°C low temperature storage and transportation are mostly used. However, this solution has obvious shortcomings. First, even at a low temperature of 4°C, nucleases are still active, which will lead to the loss of free nucleic acids. Degradation, secondly, transportation at 4°C increases the cost burden, and the last and most important disadvantage is that low temperature conditions are also easy to activate platelets and stimulate hemolysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The blood additive provided by this embodiment, every 100mL comprises the following components (formulation 1):

[0050]

[0051] The present embodiment provides the following Cl-containing additives, as a control, each 100mL includes the following components:

[0052]

[0053] According to above-mentioned formula, weigh corresponding reagent respectively, be dissolved in 100mM phosphate buffer solution and be made into 100mL formula 1 additive of the present invention, each blood collection tube contains this additive 285.7 μ L, prepare Cl- blood collection tube with the same method, simultaneously with common EDTA blood collection tube for comparative test. Specific steps are as follows:

[0054] 1. Use EDTA blood collection tubes, blood collection tubes containing formula 1 additives and Cl- additives to collect blood respectively. The blood collection volume of each tube is 10mL. mix;

[0055] 2. Store at room temperature, observe the hemolysis of the blood...

Embodiment 2

[0058] The blood supplement provided by this embodiment, every 100mL comprises the following components (formulation 2):

[0059]

[0060] Additives were prepared according to the above formula, and a comparative test was carried out with common EDTA blood collection tubes. Specific steps are as follows:

[0061] 1. Use EDTA blood collection tubes, blood collection tubes containing the additive of formula 1 in Example 1, and blood collection tubes containing the additive of formula 2 in Example 2 to collect blood respectively. The blood collection volume of each tube is 10 mL, and immediately after blood collection, slowly turn the blood collection tube upside down 10 times , so that the blood and additives are fully mixed;

[0062] 2. After being transported at room temperature for 4 days, the blood collection tubes were centrifuged at 2000g for 10 minutes to separate the plasma, and the hemolysis of each blood collection tube was observed and compared.

[0063] experime...

Embodiment 3

[0065] Using the blood additive (formula 1) of Example 1, with the common EDTA blood collection tube as a control, carry out the normal temperature transportation experiment, the specific steps are as follows:

[0066] 1. The tumor cell line H1975 (EGFR T790M mutation-positive cell line) was cultured in RMPI 1640 medium (containing 10% fetal bovine serum) at 37°C and 5% CO2 for 3 days, and then transferred to a clean centrifuge tube;

[0067] 2. Centrifuge at 2000g for 10min, transfer the culture supernatant to a new clean centrifuge tube;

[0068] 3. Use the blood collection tube containing the additive of the present invention and the common EDTA blood collection tube to collect the peripheral blood of 3 volunteers respectively, each tube collects 10 mL, and immediately turn the blood collection tube upside down slowly for 10 times;

[0069] 4. Take the supernatant of the culture medium in step 2, vortex to mix, add 50 μL to each tube of 10 mL of blood in step 3, and slowly ...

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Abstract

The invention discloses a blood additive, which is prepared from 6.0 to 8.8 g / 100ml of anticoagulants, 2.0 to 8.4 g / 100ml of nuclease inhibiting agents, 4 to 20 g / 100ml of cell stabilizing agents, 0 to 0.31 g / 100ml of purine, 2 to 10 g / 100ml of metabolic inhibitors, 0.021 to 0.21 g / 100ml of apoptosis inhibitors and 0.1 to 1 g of nucleic acid protection agents. Non-Cl<-> buffer solution is 90 to 110 mM, and the pH is 7.3 to 7.5. The blood additive can be used for storing blood at the normal temperature for 14 days without influence on detection.

Description

technical field [0001] The invention relates to a blood additive. Background technique [0002] In 2015, liquid biopsy technology was selected as one of the top ten breakthrough technologies of the year, and blood has become the most important research object of liquid biopsy technology because of the free nucleic acid from tumors. The effective implementation of blood-based liquid biopsy depends on two key factors. One is the need for sufficiently sensitive detection technology, because circulating tumor DNA (circulating tumor DNA) accounts for a very low proportion of plasma cell-free DNA (cfDNA) , with the development of detection technology, the current detection sensitivity of ctDNA has reached 0.2%. Second, since in the real world, it is impossible to separate the plasma and perform ctDNA extraction and detection in the first time after blood collection, therefore, the effective preservation of blood is also very important. However, due to the rich nucleases in the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N15/10C12Q1/6851
CPCC12N5/0634C12N15/1003C12N2500/30C12N2500/32C12N2500/34C12N2500/40C12N2500/50C12N2501/998C12Q1/6851C12Q2531/113
Inventor 卢皇彬吕唯宋庆涛陈宁郑立谋
Owner AMOY DIAGNOSTICS CO LTD
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