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Protecting agent for free DNA in plasma

A protective agent and plasma technology, applied in the field of protective agent for free DNA in blood, can solve the problems of damaging cfDNA, reducing the detection rate of sequencing, etc., and achieve the effect of improving efficiency and increasing recovery efficiency

Inactive Publication Date: 2018-11-27
北京启衡星生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But these cross-linkers can damage cfDNA and reduce the detection rate of sequencing

Method used

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  • Protecting agent for free DNA in plasma
  • Protecting agent for free DNA in plasma
  • Protecting agent for free DNA in plasma

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0044] 1. Preparation of protective agent and preparation of blood collection tubes:

[0045] The technical scheme is: 50-80g / L EDTA, 100g-200g / L IDU, 200g-300g / L DU, 10g-50g / L trehalose, 5g-50g / L glyceraldehyde, 20-80g / L L of NaF, 10-25g / L of polyvinyl alcohol.

Embodiment 1

[0060] Example 1: Comparative experiment using the protective agent of the present invention and EDTA blood collection tubes and cfDNA blood collection tubes on the market

[0061] 8 cases of whole blood samples were randomly selected, and the samples were divided into four groups, which were put into the blood collection tubes containing the protective agent of the present invention and the tubes with only the anticoagulant EDTA, and domestic cfDNA blood collection tubes S and imported cfDNA blood collection tubes purchased on the market. Vascular V.

[0062] 1. The sample is placed at room temperature, and cfDNA is extracted from the blood in the three blood collection tubes on 0 day, 3 days, 7 days, 10 days, and 14 days, and then the amount of cfDNA is detected

[0063] 2. The sample is placed at 37 degrees, and cfDNA is extracted from the blood in the three blood collection tubes on 1 day, 2 days, 3 days, and 4 days, and then the amount of cfDNA is detected

[0064] 3. Th...

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Abstract

The invention relates to a protecting agent for free DNA in plasma and belongs to the biotechnical field. The protecting agent for free DNA in plasma is a sterile aqueous solution comprising the following components: a 50-600 g / L preservative, a 30-100 g / L nuclease inhibitor, a 20-200 g / L metabolic inhibitor and a 1-120 g / L anti-adsorbent. By mixing collected blood with the protecting agent in a proportion of (5-1000): 1, the free DNA of blood can be kept for 14 days in a constant temperature condition, the free DNA of blood can be kept for 4 days at 37 DEG C, and the free DNA of blood can betransported continuously for two days. By optimizing the components and the contents thereof, free nucleic acid in blood can be stored and extracted effectively, and meanwhile, nucleic acid doped intoblood cells as a result of splitting decomposition of hemocyte is avoided after blood is extracted.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a protective agent for free DNA in blood. Background technique [0002] 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 plasma free DNA (cell free DNA, cfDNA) as the research object, are booming. Due to the low content of fetal DNA and circulating tumor DNA (ctDNA) in plasma, the detection of low-frequency or ultra-low-frequency mutations is still difficult. [0003] In maternal plasma, fetuses in the first trimester of singleton pregnancy release less than 10% of the cell-free cfDNA concentration. With prolonged storage, the degradation of leukocytes and the subsequent release of maternal genomic DNA (gDNA) will seriously affect the proportion and integrity of low-level fetal cell-free DNA. This proble...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125C12Q2527/127
Inventor 柳丽萍黄秀付亮剑
Owner 北京启衡星生物科技有限公司
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