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Blood collection tube containing free desoxyribonucleic acid stabilizer

A deoxyribonucleic acid and blood collection tube technology, applied in the medical field, can solve the problems of being easily affected by the environment, affecting the results of cf-DNA detection and analysis, failing to realize early detection and treatment of diseases, etc., and achieving the effect of slowing down degradation

Pending Publication Date: 2022-03-01
河南驼人医疗器械研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of free DNA in blood is low, and it is easily affected by the environment after leaving the human body. Improper collection and storage of blood samples will affect the detection and analysis results of cf-DNA
[0003] At present, the sequencing equipment in the cf-DNA detection process is expensive and the sample size is usually small, which is easy to degrade. There is a situation that cf-DNA has already degraded during the inspection process. In addition, cf-DNA has very high requirements for blood collection tubes. It has the function of protecting cf-DNA from being degraded, and it needs to be stored stably for a long time
However, the common anticoagulant blood collection tubes on the market have strict requirements on storage and transportation conditions, and the storage time is limited, which can only be stored for about one day, which is not conducive to the detection of samples that need to be transported for a long time. Early detection and treatment of diseases

Method used

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  • Blood collection tube containing free desoxyribonucleic acid stabilizer
  • Blood collection tube containing free desoxyribonucleic acid stabilizer
  • Blood collection tube containing free desoxyribonucleic acid stabilizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of stabilizer:

[0023] Using a calibrated electronic balance, weigh the buffer substance Tris-HCl 6g, cyclodextrin 3g, glycine 3g; preservative imidazolidinyl urea 3g; anticoagulant ethylenediaminetetraacetic acid tripotassium (EDTA3K) 7g; metabolic inhibitor N-ethylmaleimide 3g, dextrose 3g; enzyme inhibitor β-mercaptoethanol 2g, respectively weighed and placed in a 100ml clean beaker, add 100ml deionized water to the beaker, mix and stir Evenly, set aside.

[0024] Preparation of blood collection tubes:

[0025] Dispense the prepared stabilizer into 10ml quartz blood collection tubes according to the dose of 300µl per blood collection tube, and carry out vacuum treatment with a liquid negative pressure of 10ml through a vacuum capping machine, and use 15gky irradiation to sterilize it for later use . The quartz blood collection tubes are commercially available.

[0026] Application of blood collection tube:

[0027] Use the above-mentioned blood coll...

Embodiment 2

[0034] In view of Example 1, the stabilizer of Example 1 formula can only guarantee the stability of cf-DNA in plasma within 21 days, and at the 28th day, cf-DNA is degraded, in order to solve this problem, we in Example 2 Add more kinds of nuclease inhibitors to inhibit the degradation of cf-DNA by nucleases and stabilize the content of cf-DNA.

[0035] The preparation of present embodiment 2 stabilizing agent is as follows:

[0036] Using a calibrated electronic balance, weigh the buffer substance Tris-HCl 6g, glycine 3g, cyclodextrin 3g; preservative imidazolidinyl urea 3g; anticoagulant ethylenediaminetetraacetic acid tripotassium (EDTA3K) 7g; metabolic inhibitor N-ethylmaleimide 3g, dextrose 3g; enzyme inhibitor β-mercaptoethanol 2g, aurin tricarboxylic acid ATA 2g, dithiothreitol 2g; placed in a clean beaker, and then Add 100ml of deionized water into the beaker, mix and stir evenly, and set aside for later use.

[0037] Blood collection tubes were prepared according t...

Embodiment 3

[0042] Blood collection tubes were prepared using the stabilizer formulation in Example 2, blood samples were randomly drawn, and 5 tubes were drawn for each blood sample, and a constant temperature shaker was used to simulate vehicle transportation conditions (25°C, 80r / min). Nucleic acid extraction was carried out after 0 day, 7 days, 14 days, 21 days, and 28 days of transportation respectively (the steps and reagents are the same as in Example 2).

[0043] The extracted nucleic acid samples were subjected to real-time fluorescent PCR detection, and the PCR test results are shown in Figure 5 . Similarly, common EDTA blood collection tubes were used as a control, and the test results are shown in Table 3.

[0044] table 3

[0045]

[0046] It can be seen from the above results that under the condition of simulating vehicle transportation, the blood collection tube containing stabilizer in Example 2 of the present invention can also ensure the stability of cf-DNA within ...

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Abstract

The invention discloses a blood collection tube containing a free desoxyribonucleic acid stabilizer, the blood collection tube comprises a stabilizer, a tube body and a sealing plug, the stabilizer comprises a preservative, an anticoagulant, a metabolic inhibitor, an enzyme inhibitor and a buffer substance, and the tube body of the blood collection tube is made of quartz, PET plastic and other materials. And the sealing rubber plug is made of a polypropylene material. The stabilizer in the blood collection tube can effectively prevent genomic DNA in cells from being released into plasma, can stably preserve free DNA in blood at normal temperature, effectively prolongs the storage time of blood samples at room temperature, has no influence on subsequent detection results, can be used for various medical biological monitoring institutions such as laboratories and hospitals, and has wide application prospects. Good use values and application prospects are realized.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a blood collection tube containing a free deoxyribonucleic acid stabilizer. Background technique [0002] Free DNA (cell free DNA, referred to as cf-DNA) refers to the free extracellular DNA in the blood. Early screening of DNA has become an important means of detection. It can be used not only for early screening, diagnosis, monitoring and other fields of cancer based on genetic detection technology, but also for non-invasive prenatal diagnosis, organ transplantation and parent-child Identify industries related to gene sequencing. However, the content of free DNA in blood is low, and it is easily affected by the environment after leaving the human body. Improper collection and storage of blood samples will affect the detection and analysis results of cf-DNA. [0003] At present, the sequencing equipment in the cf-DNA detection process is expensive and the sample size is usual...

Claims

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Application Information

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IPC IPC(8): C12M1/24C12Q1/6806A61B5/15
CPCB01L3/5082C12Q1/6806A61B5/150755C12Q2527/125C12Q2527/127
Inventor 程杰李燕虹冯亚普赵安琪刘志远李丹杰华静宇李菲闫立论王富利
Owner 河南驼人医疗器械研究院有限公司
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