A blood anticoagulant used for protecting free DNA and applications thereof

A blood anticoagulant and anticoagulant technology, applied in the field of anticoagulants, can solve the problems of plasma separation, false negative, cell rupture, etc., and achieve the effects of preventing cell rupture, stabilizing the environment, and prolonging the shelf life

Inactive Publication Date: 2015-08-12
AMOY DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although plasma DNA can be used for clinical prognosis and detection, in the clinical process, due to the limitation of experimental conditions or experimental time, it is impossible to separate the plasma immediately after the blood is obtained. DNA degradation, resulting in the inability to carry out follow-up experiments or false negative results of experiments, thus unable to provide valuable information for clinical

Method used

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  • A blood anticoagulant used for protecting free DNA and applications thereof
  • A blood anticoagulant used for protecting free DNA and applications thereof
  • A blood anticoagulant used for protecting free DNA and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The blood anticoagulant provided in this embodiment is calculated by weight to volume ratio (w / v), wherein the volume is the volume of blood. Contains the following components:

[0027]

[0028] Prepare anticoagulants according to the above components, and conduct a comparative test with common EDTA blood collection tubes. The specific steps are as follows:

[0029] 1. Use EDTA blood collection tubes and blood collection tubes added with the blood anticoagulant provided in this embodiment to collect blood respectively, and the blood collection volume of each tube is 10 mL;

[0030] 2. The collected blood is placed at room temperature, and the blood is gently mixed twice a day by inversion to simulate the transportation environment, so as to observe the changes of blood cells and plasma DNA in the transportation environment.

[0031] 3. Separate the plasma on days 0, 1, 3, 5, and 7; the step of separating the plasma is to put the blood into a centrifuge tube, centrif...

Embodiment 2

[0039] The blood anticoagulant provided in this embodiment comprises the following components by weight to volume ratio (w / v):

[0040]

[0041] Anticoagulants were prepared according to the above components, and compared with common EDTA blood collection tubes, the steps were the same as in Example 1. The results are shown in Table 2 and figure 2 shown.

[0042] Table 2 The present invention and common EDTA blood collection tube comparative test 2

[0043]

[0044] It can be seen from Table 2 that the Ct value after using the present invention for 7 days is close to the data on the 0th day, while the blood in the common EDTA blood collection tube increases with the storage time, and the difference between the Ct value and the data on the 0th day gradually increases. It can be seen that the use of the present invention can effectively guarantee the blood quality, and is beneficial to subsequent operations such as DNA extraction.

Embodiment 3

[0046] The blood anticoagulant provided in this embodiment comprises the following components by weight to volume ratio (w / v):

[0047]

[0048] Prepare anticoagulants according to the above components, and conduct a comparative test with ordinary EDTA blood collection tubes. The specific operation steps are as follows:

[0049] 1. Use EDTA blood collection tubes and blood collection tubes added with the blood anticoagulant provided in this example to collect blood respectively. The blood collection volume of each tube is 10 mL, and 2.25 μg of mutation-positive tumor cell line DNA is added to the blood;

[0050] 2. The collected blood is placed at room temperature, and the blood is gently mixed twice a day by inversion to simulate the transportation environment, so as to observe the changes of blood cells and plasma DNA in the transportation environment.

[0051] 3. Separate the plasma on days 0, 1, 5, and 7; the step of separating the plasma is to put the blood into a cent...

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PUM

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Abstract

A blood anticoagulant used for protecting free DNA is disclosed. The blood anticoagulant comprises following components by a weight-to-volume ratio (W / V): 0.1-1.5% of a formaldehyde releasing agent, 1.5-2.2 g / L of an anticoagulant, 0.014-0.035 g / L of purine, 0.01-0.15% of dihydric phosphate, 0.4-1.0% of chloride salts and 0.3-1.4% of a nuclease inhibitor. The blood anticoagulant is a blood protecting agent effectively protecting cells from breaking, retarding degradation of free DNA in plasma, providing a stable environment for the plasma and prolonging blood stability, and avoids influences on subsequent detection which are caused by blood deterioration and DNA degradation.

Description

technical field [0001] The invention relates to an anticoagulant for protecting blood samples, in particular to a blood anticoagulant for protecting free DNA. Background technique [0002] Blood collection tubes for different purposes contain different additives or addenda. It mainly includes anticoagulants and coagulants, etc. Additives can be liquid, solution dried, freeze-dried, powdered, etc. [0003] In clinical practice, because heparin can inhibit the PCR process, EDTA (including dipotassium ethylenediaminetetraacetic acid, tripotassium ethylenediaminetetraacetic acid, and disodium edetate) and sodium citrate are generally used for blood anti-inflammatory reactions. Coagulation, anticoagulant blood after centrifugation, the obtained plasma is used for follow-up research. [0004] Anticoagulated blood is centrifuged to obtain plasma. Plasma DNA has become a clinically important sample. It is mainly used for two aspects of research, one is prenatal diagnosis, and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 刘云芬林清华蔡阿莎宋庆涛郑立谋
Owner AMOY DIAGNOSTICS CO LTD
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