CN106701743A - Blood collection tube for protecting and stabilizing free DNA - Google Patents
Blood collection tube for protecting and stabilizing free DNA Download PDFInfo
- Publication number
- CN106701743A CN106701743A CN201611214323.6A CN201611214323A CN106701743A CN 106701743 A CN106701743 A CN 106701743A CN 201611214323 A CN201611214323 A CN 201611214323A CN 106701743 A CN106701743 A CN 106701743A
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- Prior art keywords
- heparin tube
- blood
- blood collection
- collection tube
- tube according
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 239000008280 blood Substances 0.000 title claims abstract description 27
- 210000004369 blood Anatomy 0.000 title claims abstract description 27
- 230000000087 stabilizing effect Effects 0.000 title abstract 2
- 238000012360 testing method Methods 0.000 claims abstract description 12
- 239000003223 protective agent Substances 0.000 claims abstract description 6
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 31
- 229960002897 heparin Drugs 0.000 claims description 31
- 229920000669 heparin Polymers 0.000 claims description 31
- 230000006641 stabilisation Effects 0.000 claims description 11
- 238000011105 stabilization Methods 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 6
- 229920001971 elastomer Polymers 0.000 claims description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- -1 Formic acid alcohol ester Chemical class 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 2
- MGSKVZWGBWPBTF-UHFFFAOYSA-N aebsf Chemical compound NCCC1=CC=C(S(F)(=O)=O)C=C1 MGSKVZWGBWPBTF-UHFFFAOYSA-N 0.000 claims description 2
- 229920005549 butyl rubber Polymers 0.000 claims description 2
- 230000002209 hydrophobic effect Effects 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- JZBRFIUYUGTUGG-UHFFFAOYSA-J tetrapotassium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [K+].[K+].[K+].[K+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O JZBRFIUYUGTUGG-UHFFFAOYSA-J 0.000 claims description 2
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims 3
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims 2
- 235000019253 formic acid Nutrition 0.000 claims 1
- 239000002775 capsule Substances 0.000 abstract 1
- 238000007789 sealing Methods 0.000 abstract 1
- 108020004707 nucleic acids Proteins 0.000 description 9
- 150000007523 nucleic acids Chemical class 0.000 description 9
- 102000039446 nucleic acids Human genes 0.000 description 9
- 238000001514 detection method Methods 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000000605 extraction Methods 0.000 description 3
- 230000023555 blood coagulation Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 244000144725 Amygdalus communis Species 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a blood collection tube for protecting and stabilizing free DNA. The blood collection tube comprises a test tube, a sealing capsule and a protective agent. After blood is collected into the blood collection tube, the blood is shaken up and down ten times to be sufficiently and evenly mixed and then is preserved at room temperature. The blood free DNA can be protected for as long as 14 days at normal temperature, and the blood collection tube can be used for protecting and storing the blood free DNA.
Description
Technical field
The present invention relates to a kind of protection and the heparin tube of stabilization dissociative DNA.
Background technology
Free nucleic acid is earliest to be found by Mandel and Metais in nineteen forty-seven, but special due to lacking high sensitivity and height
The experimental technique of property, causes to be made slow progress within the longer term with disease associated research about dissociative DNA in blood.Until
The appearance and application of dissociative DNA technology, PCR detection techniques are efficiently separated, the research in this field was obtained at nearest more than 20 years
To relatively developing rapidly.After dissociative DNA can contain the mutation of DNA of tumor cell homologous genes in self-discovery blood, using molecular biosciences
Learning to do section increasingly increases the interest for circulating the research of free nucleic acid.But these free DNA in conventional heparin tube very
Easily just degraded disappears, therefore the heparin tube of a kind of protection and stabilization dissociative DNA is in demand.
At present, various heparin tubes are there are on the market, and all simply being played in these heparin tubes prevents blood clotting
Effect, without play a part of protect free nucleic acid.In CC, due to the limitation of condition or time, it is impossible to
Very first time separated plasma after blood is obtained, the cell rupture and prolonged unprotect exposed state in blood can cause blood
In the DNA degradation that dissociates, cause that the false negative of subsequent experimental or experimental result cannot be carried out, so as to cannot provide valuable for clinical
The information of value.
In view of this, the heparin tube of a kind of protection and stabilization dissociative DNA is invented herein, to reduce what subsequent survey was detected
Interference.
The content of the invention
A kind of heparin tube it is an object of the invention to manufacture protection and stabilization dissociative DNA, the heparin tube can not only be prevented
Blood clotting, and can protect and stablize wherein free DNA.The heparin tube is by test tube, seal rubber plug and protective agent group
Into.By blood collection to Guan Zhonghou, turning upside down 10 times, it is uniform to be sufficiently mixed, room temperature preservation.So that the dissociative DNA in blood
Can preserve at normal temperatures 14 days.
Main technical points of the invention are as follows：A kind of protection and the heparin tube of stabilization dissociative DNA, described heparin tube
It is to be fabricated by by PET's material of hydrophobic property, described seal rubber plug is manufactured by butyl rubber material
Form, described protective agent is formed by the dissolving of the material such as fourth EDTA-3K, IDU, glycine, bestain, AEBSF, concentration
Respectively 50g/L, 550g/L, 30g/L, 1mg/L, 0.172g/L, 0.24g/L.
In the application, by blood collection to Guan Zhonghou, turning upside down 10 times, it is uniform to be sufficiently mixed, room temperature preservation for the present invention.
Fig. 1 is composition structural representation of the invention.1 refers to test tube, and 2 refer to seal rubber plug, and 3 refer to protective agent.
Fig. 2 is that whole blood sample carries out 0,3,7,14 in the heparin tube of a kind of protection of the present invention and stabilization dissociative DNA
The testing result of its rear stability.
Fig. 3 is whole blood sample in conventional EDTA-Na20,3,7,14 days testing results of stability are carried out in heparin tube.
Specific implementation method
Following case study on implementation is only used for explaining the present invention, rather than the limitation present invention.
Example：
Blood sample is randomly selected simultaneously with heparin tube of the present invention and routine EDTA-Na2 heparin tubes, each blood sample extracts 4 pipes respectively, often
Temperature place 0 day, 3 days, 7 days, after 14 days carry out nucleic acid extraction（The QIAamp DNA Blood Mini Kit of Qiagen companies
(50)-kit of article No. 51104）, enter the real-time PCR detection of pedestrian's β-ACTB genes as sample with the nucleic acid for extracting.
Testing result：
It is placed in conventional EDTA-Na2The whole blood sample of the heparin tube of heparin tube and protection of the present invention and stabilization dissociative DNA,
After room temperature different time, nucleic acid extraction is carried out respectively, the nucleic acid of extraction enters the real-time fluorescence PCR of pedestrian's β-ACTB genes
Detection, the Ct values such as table 1 of its testing result.
Table 1
As shown in Table 1, the heparin tube of a kind of protection of the present invention and stabilization dissociative DNA can effectively be protected in 14 days
With stablize blood middle reaches from DNA, only extremely slight degraded；And routine EDTA-Na2The stability of heparin tube is excessively poor,
Degraded situation is extremely serious.
Fig. 2 and Fig. 3 is the real-time PCR detection result figure of the corresponding people β-ACTB genes of table 1, and wherein Fig. 2 is for herein
A kind of described protection and the heparin tube of stabilization dissociative DNA carry out 0,3,7,14 days testing results of stability, as seen from the figure,
In the detection time of 0 ~ 14 day, all detection samples are all correctly detected, and are had good stability, and the free nucleic acid in blood is obtained
It is effectively protected；Fig. 3 is routine EDTA-Na20,3,7,14 days testing results of stability, testing result are carried out in heparin tube
It has been shown that, conventional EDTA-Na2The stability of the free nucleic acid in heparin tube is excessively poor, and degraded situation is extremely serious.
Claims (6)
1. it is a kind of protect and stabilization dissociative DNA heparin tube, it is characterised in that the heparin tube be by test tube, seal rubber plug and
Protective agent composition.
2. the heparin tube according to claim 1, it is characterised in that described test tube is by the poly- to benzene two of hydrophobic property
Formic acid alcohol ester material is fabricated by.
3. the heparin tube according to claim 1, it is characterised in that described seal rubber plug is by butyl rubber material system
Make and form.
4. the heparin tube according to claim 1, it is characterised in that described protective agent be by fourth EDTA-3K, IDU,
The materials such as glycine, bestain, AEBSF dissolving, concentration be respectively 50g/L, 550g/L, 30g/L, 1mg/L,
0.172g/L、0.24g/L。
5. the heparin tube according to claim 1, it is characterised in that the negative pressure of vacuum of described heparin tube is 10ml blood
Amount.
6. the heparin tube according to claim 1 ~ 4, it is characterised in that in the application, by blood collection to Guan Zhonghou, up and down
It is sufficiently mixed uniform, room temperature preservation for reverse 10 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611214323.6A CN106701743A (en) | 2016-12-26 | 2016-12-26 | Blood collection tube for protecting and stabilizing free DNA |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611214323.6A CN106701743A (en) | 2016-12-26 | 2016-12-26 | Blood collection tube for protecting and stabilizing free DNA |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106701743A true CN106701743A (en) | 2017-05-24 |
Family
ID=58903282
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611214323.6A Pending CN106701743A (en) | 2016-12-26 | 2016-12-26 | Blood collection tube for protecting and stabilizing free DNA |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106701743A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107083382A (en) * | 2017-05-25 | 2017-08-22 | 广州奇辉生物科技有限公司 | A kind of blood preseration agent and its application for being used to protect dissociative DNA |
| CN108261332A (en) * | 2018-01-22 | 2018-07-10 | 无锡百泰克生物技术有限公司 | A kind of blood rna preserves pipe |
| CN108279307A (en) * | 2018-01-30 | 2018-07-13 | 郑州安图生物工程股份有限公司 | Protection and the heparin tube for stablizing gastrin-releasing peptide precursor in serum or blood plasma |
| CN109055353A (en) * | 2018-07-17 | 2018-12-21 | 无锡百泰克生物技术有限公司 | A kind of protective agent of dissociative DNA in blood |
| CN110747193A (en) * | 2019-09-25 | 2020-02-04 | 广州市达瑞生物技术股份有限公司 | Lung cancer polygene joint detection kit quality control product based on second-generation sequencing platform |
| RU198611U1 (en) * | 2019-07-10 | 2020-07-21 | Линд Виталий Викторович | Blood sample preparation device for liquid biopsy with recombinant protein |
| CN113265443A (en) * | 2021-05-21 | 2021-08-17 | 深圳逗点医疗科技有限公司 | Blood free DNA protection reagent, protection method and blood collection tube |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010134246A1 (en) * | 2009-05-20 | 2010-11-25 | オリンパス株式会社 | Method for preparation of nucleic acid-containing sample |
| CN103789202A (en) * | 2014-01-26 | 2014-05-14 | 付士明 | Container for collecting nucleic acid preserved at normal temperature |
| CN104673623A (en) * | 2015-02-05 | 2015-06-03 | 广州新诚生物科技有限公司 | Blood sample collecting device |
| CN204897890U (en) * | 2015-02-05 | 2015-12-23 | 广州新诚生物科技有限公司 | Novel blood sample is collected device |
| CN105985904A (en) * | 2015-02-26 | 2016-10-05 | 付士明 | Circulating free DNA vacuum collection tube |
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2016
- 2016-12-26 CN CN201611214323.6A patent/CN106701743A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010134246A1 (en) * | 2009-05-20 | 2010-11-25 | オリンパス株式会社 | Method for preparation of nucleic acid-containing sample |
| CN103789202A (en) * | 2014-01-26 | 2014-05-14 | 付士明 | Container for collecting nucleic acid preserved at normal temperature |
| CN104673623A (en) * | 2015-02-05 | 2015-06-03 | 广州新诚生物科技有限公司 | Blood sample collecting device |
| CN204897890U (en) * | 2015-02-05 | 2015-12-23 | 广州新诚生物科技有限公司 | Novel blood sample is collected device |
| CN105985904A (en) * | 2015-02-26 | 2016-10-05 | 付士明 | Circulating free DNA vacuum collection tube |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107083382A (en) * | 2017-05-25 | 2017-08-22 | 广州奇辉生物科技有限公司 | A kind of blood preseration agent and its application for being used to protect dissociative DNA |
| CN107083382B (en) * | 2017-05-25 | 2018-07-31 | 广州奇辉生物科技有限公司 | A kind of blood preseration agent and its application for protecting dissociative DNA |
| CN108261332A (en) * | 2018-01-22 | 2018-07-10 | 无锡百泰克生物技术有限公司 | A kind of blood rna preserves pipe |
| CN108279307A (en) * | 2018-01-30 | 2018-07-13 | 郑州安图生物工程股份有限公司 | Protection and the heparin tube for stablizing gastrin-releasing peptide precursor in serum or blood plasma |
| CN108279307B (en) * | 2018-01-30 | 2021-01-26 | 郑州安图生物工程股份有限公司 | Blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma |
| CN109055353A (en) * | 2018-07-17 | 2018-12-21 | 无锡百泰克生物技术有限公司 | A kind of protective agent of dissociative DNA in blood |
| RU198611U1 (en) * | 2019-07-10 | 2020-07-21 | Линд Виталий Викторович | Blood sample preparation device for liquid biopsy with recombinant protein |
| CN110747193A (en) * | 2019-09-25 | 2020-02-04 | 广州市达瑞生物技术股份有限公司 | Lung cancer polygene joint detection kit quality control product based on second-generation sequencing platform |
| CN110747193B (en) * | 2019-09-25 | 2021-08-31 | 广州市达瑞生物技术股份有限公司 | Lung cancer polygene joint detection kit quality control product based on second-generation sequencing platform |
| CN113265443A (en) * | 2021-05-21 | 2021-08-17 | 深圳逗点医疗科技有限公司 | Blood free DNA protection reagent, protection method and blood collection tube |
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|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170524 |