A spray-type cell-free dna sample storage tube and its application

A technology for preserving tubes and samples, which is applied in specific-purpose bioreactors/fermenters, laboratory containers, and microbial determination/inspection, etc. Waste of environment and other problems, to achieve wide application prospects and market promotion value, prevent genomic DNA pollution, and reduce the probability of tube wall damage

Active Publication Date: 2021-11-16
北京健为医学检验实验室有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, glass tubes are inconvenient to transport and are easily broken during transportation, resulting in waste of samples and environmental pollution. Therefore, in recent years, PET materials have gradually replaced glass tubes and are widely used in clinical medicine due to their advantages of light weight, pressure resistance, non-fragility, and easy transportation. use
However, the preservation tube made of PET material has poor airtightness and water retention. The PET body fluid sample storage tube made of PET tube combined with a large volume of liquid preservation solution (about 1ml) will gradually volatilize the preservation solution in the preservation tube at room temperature. After that, the solution matrix (such as water) will evaporate completely, and the crystallization of the effective components in the preservation solution will be formed at the bottom of the tube.
During use of this type of storage tube, after adding blood and other body fluids, invert and mix according to the routine operating procedures. The crystalline components are not easy to dissolve completely, which increases the difficulty of operation during use, and the effect of sample preservation will also be affected.
Moreover, due to the different volumes of storage solution remaining in storage tubes with different storage periods, the total volume of sample solutions in different sample tubes after blood collection varies from high to low. When subsequent sampling is performed according to a fixed operation process, the actual sampling consistency cannot be achieved. Guaranteed, resulting in the stability of the sample test results cannot be effectively guaranteed
In addition, the matrix of the sample preservation solution volatilizes, the total volume of the sample solution becomes smaller after sampling, the concentration of effective preservation components in the sample solution becomes higher, and the osmotic pressure of the solution changes, which may cause cell rupture in the sample, resulting in the release of gDNA, resulting in genomic nucleic acid contamination
[0004] In order to solve the above problems, the present invention provides a free DNA sample collection and storage tube made of PET material. The optimized preservative is attached to the tube wall in the form of atomization, which solves the disadvantage of poor water retention performance of PET tubes and greatly prolongs the sample storage tube. The shelf life is long, and it is also conducive to the full mixing of the collected body fluid samples and the free DNA protective agent. The nuclease stabilizes the cfDNA in it, and effectively avoids the release of the genomic DNA of the nucleated cells in the sample, reducing the background noise of the genomic DNA

Method used

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  • A spray-type cell-free dna sample storage tube and its application
  • A spray-type cell-free dna sample storage tube and its application
  • A spray-type cell-free dna sample storage tube and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation of cfDNA storage tubes

[0024] Preparation of cfDNA sample preservative 1: weigh ethylenediaminetetraacetic acid dipotassium salt (EDTA·K 2 2H 2 O) 80g, glycine 30g, imidazolidinyl urea 150g, add to 900ml sterile deionized water, after completely dissolved, dilute to 1L.

[0025]Preparation of cfDNA sample preservative 2: weigh ethylenediaminetetraacetic acid dipotassium salt (EDTA·K 2 2H 2 O) 80g, glycine 30g, imidazolidinyl urea 150g, cell and membrane stabilizing peptide 60mg, add to 900ml sterile deionized water, after completely dissolved, dilute to 1L.

[0026] cfDNA preservation tube 1 uses the formula of preservative 1, and cfDNA preservation tube 2 uses the formula of preservative 2.

[0027] Atomization and sampling of preservatives: A precision sprayer is used to atomize and add samples of preservatives. The pressure valves of the nozzles must be fully opened, the pressure should be controlled at 3-4kg, and the adjustment of the me...

Embodiment 2

[0030] Embodiment 2: Disposable vacuum cell-free DNA sample preservation tube strength test

[0031] (1) Experimental materials:

[0032] Disposable vacuum cell-free DNA sample storage tubes provided by the present invention: brand new, 10 pieces;

[0033] (2) Reagents and instruments:

[0034] Water, room temperature 20℃~25℃;

[0035] The swing arm centrifuge can make the bottom of the container withstand 3000gn (longitudinal) centrifugal acceleration for 10min (gn =9.80665m / s2).

[0036] (3) Test method:

[0037] Add nominal volume of water into 10 sample preservation tubes by injecting water with blood collection needles to ensure that the preservation tubes are properly supported and fully balanced in the centrifuge tubes, start the centrifuge, and make the bottom of the preservation tubes withstand a longitudinal centrifugal acceleration of 3000gn for 10 minutes (not including the specified time before acceleration). After centrifugation is complete, remove the blood...

Embodiment 3

[0040] Example 3: Aspirated Volume Detection of Disposable Vacuum cfDNA Sample Preservation Tubes

[0041] Select the environment of 101.3kPa (0 meters above sea level), 96.4kPa (about 400 meters above sea level), and 91.7kPa (about 800 meters above sea level) to carry out the vacuum blood collection tube suction volume test. Numbers 1-10 are disposable vacuum cell-free DNA sample preservation tubes (5mL tubes) provided by the present invention. Note: This part refers to and conforms to the pharmaceutical industry standard of the People's Republic of China "Disposable human venous blood sample collection container (YY0314-2007)" "Appendix NA recommended ambient atmosphere calibration method judgment standard: vacuum degree (blood collection volume) in its nominal blood collection volume within ±10% of the amount”.

[0042] Table 1: Test results of vacuum suction volume

[0043]

[0044] The test results in the expected use area are shown in Table 1. In an environment belo...

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Abstract

The invention relates to a spray-type free DNA sample preservation tube and its application in blood free DNA preservation. The present invention uses a cfDNA sample preservation tube made of PET material, and a specially designed preservation agent is attached to the inner wall. The cfDNA sample storage tube of the present invention solves the disadvantage of poor water retention performance of PET tubes, greatly prolongs the shelf life of the sample storage tube, and is conducive to fully mixing the collected body fluid samples with the cfDNA protective agent, effectively inhibiting the nuclease in the body fluid samples, and stabilizing The cfDNA in it can effectively prevent the release of the genomic DNA of nucleated cells in the sample.

Description

technical field [0001] The invention mainly relates to the field of biological material storage devices, in particular to a spray-type free DNA sample storage tube and its application in blood free DNA storage. Background technique [0002] Circulating cell free DNA (cfDNA) is a kind of extracellular DNA present in human body fluids (including peripheral blood, pleural and peritoneal fluid, cerebrospinal fluid, etc.), mainly derived from apoptotic cells. The study found that both tumor cells and fetuses in pregnant women release cell-free DNA into bodily fluids. Moreover, with the continuous improvement of the requirements for precise diagnosis and treatment of malignant tumors, liquid biopsy technology represented by circulating free DNA (cfDNA) has become a research hotspot at home and abroad in recent years. In recent years, cfDNA in body fluids, especially peripheral blood, has been used more and more clinically, and has become an important marker for tumor screening di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/24C12M1/00C12Q1/6806
CPCB01L3/5082C12Q1/6806C12Q2527/127C12Q2527/125
Inventor 殷剑峰王春香
Owner 北京健为医学检验实验室有限公司
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