CN106701743A - Blood collection tube for protecting and stabilizing free DNA - Google Patents

Blood collection tube for protecting and stabilizing free DNA Download PDF

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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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CN
China
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.)
Pending
Application number
CN201611214323.6A
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Chinese (zh)
Inventor
刘淑园
陈华云
肖湘文
丁渭
张天海
黄爽
曾烨
邓金萍
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GUANGZHOU HESHI BIOTECHNOLOGY Co Ltd
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GUANGZHOU HESHI BIOTECHNOLOGY Co Ltd
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Priority to CN201611214323.6A priority Critical patent/CN106701743A/en
Publication of CN106701743A publication Critical patent/CN106701743A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor

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  • 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

A kind of protection and the heparin tube of stabilization dissociative DNA
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.
CN201611214323.6A 2016-12-26 2016-12-26 Blood collection tube for protecting and stabilizing free DNA Pending CN106701743A (en)

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)

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CN106701743A true CN106701743A (en) 2017-05-24

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Cited By (7)

* Cited by examiner, † Cited by third party
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)

* Cited by examiner, † Cited by third party
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

Patent Citations (5)

* Cited by examiner, † Cited by third party
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)

* Cited by examiner, † Cited by third party
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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Application publication date: 20170524