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Whole blood free DNA preservative, vacuum blood collection tube and preparation method of vacuum blood collection tube

A technology for vacuum blood collection tubes and preservatives, applied in the fields of blood sample preservation and biomedicine, can solve the problems of plasma cfDNA being easily degraded by nucleases, genomic DNA contamination, and requiring cold chain preservation, etc., so as to facilitate long-distance transportation and inhibit nucleic acid Enzymatic activity, the effect of avoiding lysis of blood cells

Active Publication Date: 2021-02-12
SOUTH CHINA UNIV OF TECH
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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 provide a whole blood cfDNA protective agent, the corresponding vacuum blood collection tube and its preparation method, aiming to solve the problems of nuclease degradation, genomic DNA contamination, short storage time and need for refrigeration in the existing plasma cfDNA storage. Chain preservation and other issues

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  • Whole blood free DNA preservative, vacuum blood collection tube and preparation method of vacuum blood collection tube

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Embodiment 1

[0061] A method for preparing a whole blood cfDNA preservation agent and a whole blood free DNA vacuum blood collection tube, comprising the following steps:

[0062] (1) Preparation of whole blood cfDNA preservative: mix the following components by weight: 50 parts of anticoagulant sodium citrate, 10 parts of ribozyme inhibitor ammonium sulfate, 10 parts of DNA stabilizer glucose, 10 parts Protein cross-linker glutaraldehyde, 20 parts reactive cell stabilizer HAuCl 4 , 9900 parts of RNase-free purified water; obtain whole blood cfDNA preservative; the total amount of each component is preferably 10000 parts;

[0063] (2) Filter the prepared cfDNA preservative with a 220nm filter membrane and set aside;

[0064] (3) Add the filtered cfDNA preservative into the 3mL blood collection tube according to the preservative volume: blood collection tube volume ratio of 1:30, and pump it into positive air and negative pressure, and the vacuum negative pressure is greater than 16kg / cm ...

Embodiment 2

[0067] A method for preparing a whole blood cfDNA preservation agent and a whole blood free DNA vacuum blood collection tube, comprising the following steps:

[0068] (1) Preparation of whole blood cfDNA preservative: mix the following components by weight: 100 parts of anticoagulant tripotassium edetate, 10 parts of ribozyme inhibitor guanidine isothiocyanate, 10 parts DNA stabilizer trehalose, 10 parts of protein cross-linking agent 1,2-ethanedithiol, 20 parts of reactive cell stabilizer TEOS, 9900 parts of RNase-free purified water to obtain whole blood cfDNA preservative;

[0069] (2) Filter the prepared cfDNA preservative with a 220nm filter membrane and set aside;

[0070] (3) Add the filtered cfDNA preservative into the 3mL blood collection tube according to the volume of the preservative: the volume ratio of the blood collection tube is 1:150, and pump it into a vacuum negative pressure, and the vacuum negative pressure is greater than 16kg / cm 2 , to obtain whole bloo...

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Abstract

The invention belongs to the technical field of biological medicines, and discloses a whole blood free DNA preservative, a vacuum blood collection tube and a preparation method of the vacuum blood collection tube. The whole blood free DNA preservative comprises the following components in parts by weight: 2-200 parts of an anticoagulant, 1-200 parts of a ribozyme inhibitor, 5-200 parts of a DNA stabilizer, 1-1000 parts of a protein cross-linking agent, 1-1000 parts of a reactive cell stabilizer and 5000-10000 parts of RNase-free purified water. The vacuum blood collection tube contains the whole blood free DNA preservative. The invention further discloses the preparation method of the vacuum blood collection tube. All the components in the preservative cooperate with one another and achieve a synergistic effect, the activity of nuclease in blood plasma is efficiently inhibited, and cfDNA is prevented from being degraded by nuclease in the preservation and transportation process; meanwhile, the cell stability is enhanced, and the interference on cfDNA detection caused by blood cell lysis and genome DNA release is avoided.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the technical field of blood sample preservation, in particular to a whole blood free DNA preservative, a vacuum blood collection tube and a preparation method thereof. Background technique [0002] Cell free DNA (cfDNA) refers to a type of DNA free from cells. The cfDNA in the blood of healthy people mainly comes from apoptotic cells, while tumor cells and fetuses of pregnant women release cfDNA into the blood. In recent years, with the rapid development of sequencing technology and the promotion of a series of favorable national policies, the non-invasive prenatal diagnosis and tumor liquid biopsy industries, which use cfDNA in plasma as the research object, are booming. However, the content of cfDNA in plasma is very small, which is easily degraded by nucleases in the blood, and easily polluted by genomic DNA released by leukocyte lysis, which seriously affects the detection effect. I...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12M1/24
CPCC12Q1/6806C12Q2527/125C12Q2527/127
Inventor 朱伟周亮雷琪
Owner SOUTH CHINA UNIV OF TECH