CN106635747A - Paper-based micro fluidic rapid nucleic acid extraction apparatus - Google Patents
Paper-based micro fluidic rapid nucleic acid extraction apparatus Download PDFInfo
- Publication number
- CN106635747A CN106635747A CN201710080604.5A CN201710080604A CN106635747A CN 106635747 A CN106635747 A CN 106635747A CN 201710080604 A CN201710080604 A CN 201710080604A CN 106635747 A CN106635747 A CN 106635747A
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- China
- Prior art keywords
- nucleic acid
- paper
- acid extraction
- filter
- top cover
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- 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
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Abstract
The invention discloses a paper-based micro fluidic rapid nucleic acid extraction apparatus. The apparatus comprises a filter and a nucleic acid extraction membrane; the filter comprises a top cover and a bottom cap, the two components have discoid structures with cambers, the top cover is provided with a convex sample adding channel, the bottom cap is provided with a convex waste liquid discharging channel, the top cover and the bottom cap are buckled together in order to form a filter cavity, and a nucleic acid extraction membrane is fixed in the middle. The apparatus is suitable for various types of samples, column passing, centrifugation, heating and other steps in the process are not needed, special apparatuses and electric power support are not needed, the whole process only takes 10-15 minutes, and the apparatus is very suitable for on-site nucleic acid template extraction, and can be directly applied to PCR, qPCR, LAMP and other different molecular diagnosis and detection experiments; especially the apparatus is combined with normal temperature nucleic acid amplification technology, in order to realize non-electric power dependence of the whole experiment process including sampling, nucleic acid extraction, amplification reaction, and result determination.
Description
Technical field
The present invention relates to a kind of nucleic acid-extracting apparatus, more particularly to a kind of paper substrate micro-fluidic Rapid nucleic acid extraction element.
Background technology
The extraction of nucleic acid is at present mainly using centrifugal column method and paramagnetic particle method extraction, not only high cost, and complex operation, consumption
When, the auxiliary facilities such as centrifuge, water bath are needed, this makes them to depart from laboratory environment, is not suitable for being applied to condition receiving
Limited region, epidemic prevention detection and field condition environment measuring.Traditional extracting method such as phenol-chloroform extraction method, extraction efficiency is high, right
Experiment condition requirement is low, but its toxicity is big, therefore seldom applies at present.
The development of Community medical treatment mode outside serious threat and institute recently as burst infectious disease, based on molecular diagnosis
The instant detection (POCT) of technology has obtained rapid development.Live PCR, constant temperature nucleic acid amplification, the detection of biochip equimolecular
Device emerges in an endless stream, and has versatility, also there is detection specific gene, but before detection they generally require loaded down with trivial details nucleic acid
Template extraction step and main equipment, hinder these technologies and really play a role in POCT.Therefore, set up a kind of easy,
Fast, portable, efficient nucleic acid-extracting apparatus are the keys for really realizing the application of detection system scene.
Traditional centrifugation column technology also carries out nucleic acid purification using film, and the principle of its film belongs to silicon adsorption method, this
Film only has stronger affinity and absorption affinity to nucleic acid.Using protease sample is entered under 56 DEG C of temperature conditionss first during operation
Row cracking, lysate then added in pillar and be centrifuged, and nucleic acid is adsorbed on film, and other impurities are thrown out of, finally with washing
De- liquid gets off Nucleic Acid Elution.The method extraction effect is good, but cracking when need heating, during nucleic acid purification need repeatedly from
The heart, time-consuming 2 hours of whole extraction process or so.Its high cost, complex operation, time-consuming, rely on laboratory environment, it is impossible to very
Good serves POCT, cannot also be applied to field and Site Detection.
The content of the invention
It is an object of the invention to provide a kind of portable for paper substrate micro-fluidic nucleic acid extraction that is quick, being independent of electric power
Device.
A kind of paper substrate micro-fluidic Rapid nucleic acid extraction element, including filter 1 and nucleic acid extraction film 2；
The filter 1 is made up of top cover 3 and bottom 4, and both are the disc-shaped structure with radian, set on top cover 3
The Loading channel 5 of evagination is equipped with, the waste liquid passing away 6 of evagination is provided with bottom 4, top cover 3 and bottom 4 are fastened to be formed and filtered
Device cavity, by nucleic acid extraction film 2 centre is fixed on；
The nucleic acid extraction film 2 is made up of the films 7 of fusion 5, cellulosic filter paper 8 and paraffin paper 9, and the films 7 of Fusion 5 are placed in fibre
The plain top of filter paper 8 of dimension, paraffin paper 9 is covered on cellulosic filter paper 8.
The films 7 of the fusion 5 are the circular membrane of diameter 2cm；The cellulosic filter paper 11 is the circular filter paper of diameter 6cm；
The paraffin paper 9 is the paraffin paper of central zone diameter 4cm circular holes.
The filter 1 is PC2805 materials.
A kind of method that paper substrate micro-fluidic Rapid nucleic acid is extracted, using above-mentioned extraction element, comprises the steps：
A, by Loading channel 5 sample solution 200-300 μ l, the μ l of lysate 500 and cleanout fluid 1ml are sequentially added, and are passed through
Liquid flowed out by waste liquid passing away 6；
B, then takes out the films 7 of fusion 5, in being placed in the EP pipes of 1.5ml, adds 300 μ l nucleic acid to carry after liquid evaporation to be cleaned
Take immersion to steep two minutes, draw extracting solution interior in another new EP pipe.
The lysate is the NaOH solution of 10mM.
The cleanout fluid is dehydrated alcohol.
The nucleic acid extraction liquid is distilled water.
Compared with prior art, the present invention has the advantages that：The present invention is carried for being independent of electric power, Rapid nucleic acid
The device for taking, supports without the need for electric power and large-scale instrument, realizes fast and efficiently nucleic acid extraction, will send out in nucleic acid POCT applications
Wave great function；The device first makes cell rupture, albuminous degeneration, nucleic acid release using alkaline lysises, reapplies the films of fusion 5
Nucleic acid is purified, whole process can complete entirely to extract without the need for loaded down with trivial details heating and centrifugal process in 10-15 minutes
Journey；Alkaline lysises application be inexpensive NaOH and dehydrated alcohol, nucleic acid extraction film cost is also very low, therefore whole extracts
Installation cost is all very low, is especially advantageous for the popularization and application in under-developed area；The application scope is wide, can for blood,
The genome of the various samples such as urine, Nasopharyngeal swabs, feces is extracted；The device small volume and less weight, is the discoid knot of diameter 6cm
Structure, is highly convenient for carrying, and the extraction element extraction efficiency is high, its extraction efficiency compared with traditional centrifugal column and paramagnetic particle method, Jing
QPCR verifies no significant difference；Such a nucleic acid-extracting apparatus and the combination of appropriate nucleic acid detection technique, for community application,
Epidemic situation examination and environment measuring etc. will all have important using value.
Description of the drawings
Fig. 1 splits schematic diagram for the device of the embodiment of the present invention；
Fig. 2 is the overall schematic of the embodiment of the present invention；
Fig. 3 is the nucleic acid extraction film schematic diagram of the embodiment of the present invention；
Fig. 4 is that same adenovirus infection Nasopharyngeal swabs specimen extracts nucleic acid point using this device and conventional centrifugal post method simultaneously
The result of qPCR detections is not carried out；
In figure, 1- filters, 2- nucleic acid extraction films, 3- top covers, 4- bottoms, 5- Loading channels, 6- waste liquid passing aways, 7-
The films of fusion 5,8- cellulosic filter papers, 9- paraffin paper.
Specific embodiment
Below in conjunction with the accompanying drawings, the specific embodiment of the present invention is described in detail, it is to be understood that the guarantor of the present invention
Shield scope is not limited by specific embodiment.
A kind of device that paper substrate micro-fluidic nucleic acid purification is combined for alkaline lysiss is present embodiments provided, as shown in figure 1, bag
Include filter 1 and nucleic acid extraction film 2；
As shown in Fig. 2 the filter 1 is made up of top cover 3 and bottom 4, both are the disc-shaped structure with radian,
The Loading channel 5 of evagination is provided with top cover 3, the waste liquid passing away 6 of evagination is provided with bottom 4, top cover 3 and bottom 4 are detained
Conjunction forms filter cavity, and nucleic acid extraction film is securely fastened to into centre；
As shown in figure 3, the nucleic acid extraction film 2 is made up of the films 7 of fusion 5, cellulosic filter paper 8 and paraffin paper 9, diameter 2cm
The paper membranes 7 of circular Fusion 5 be placed in circular fiber element filter paper 8 top of diameter 6cm, then with central zone diameter 4cm circular holes
Paraffin paper 9 cover, and using external force compress them is brought into close contact each other；
Sample solution 200-300 μ l, the μ l of NaOH solution 1000 of 10mM and dehydrated alcohol are sequentially added by Loading channel 5
1ml, the liquid of transmission is flowed out by waste liquid passing away 6.The films 7 of fusion 5 are then taken out, the EP of a new 1.5ml is placed in
Guan Zhong, adds 300 μ l distilled waters to soak two minutes after dehydrated alcohol evaporation, draws extracting solution interior in another new EP pipe.
Simultaneously nucleic acid is extracted using this device and conventional centrifugal post method to adenovirus infection Nasopharyngeal swabs specimen, and is carried out
QPCR detects that can draw from Fig. 4 results, two methods do not have significant difference.
Disclosed above is only the specific embodiment of the present invention, but, the present invention is not limited to this, any this area
What technical staff can think change should all fall into protection scope of the present invention.
Claims (7)
1. a kind of paper substrate micro-fluidic Rapid nucleic acid extraction element, it is characterised in that including filter (1) and nucleic acid extraction film (2)；
The filter (1) is made up of top cover (3) and bottom (4), and both are the disc-shaped structure with radian, top cover (3)
On be provided with the Loading channel (5) of evagination, the waste liquid passing away (6) of evagination, top cover (3) and bottom are provided with bottom (4)
(4) fasten and form filter cavity, nucleic acid extraction film (2) is fixed on into centre；
The nucleic acid extraction film (2) is made up of the films of fusion 5 (7), cellulosic filter paper (8) and paraffin paper (9), the films of Fusion 5 (7)
Cellulosic filter paper (8) top is placed in, paraffin paper (9) is covered on cellulosic filter paper (8).
2. paper substrate micro-fluidic Rapid nucleic acid extraction element according to claim 1, it is characterised in that the films of the fusion 5
(7) it is the circular membrane of diameter 2cm；The cellulosic filter paper (11) for diameter 6cm circular filter paper；The paraffin paper (9) is central authorities
Paraffin paper with diameter 4cm circular holes.
3. paper substrate micro-fluidic Rapid nucleic acid extraction element according to claim 1, it is characterised in that the filter (1)
For PC2805 materials.
4. a kind of method that paper substrate micro-fluidic Rapid nucleic acid is extracted, it is characterised in that filled using the extraction described in claim 1
Put, comprise the steps：
A, by Loading channel (5) sample solution 200-300 μ l, the μ l of lysate 500 and cleanout fluid 1ml are sequentially added, transmission
Liquid is flowed out by waste liquid passing away (6)；
B, then takes out the films of fusion 5 (7), in being placed in the EP pipes of 1.5ml, after liquid evaporation to be cleaned 300 μ l nucleic acid extraction is added
Immersion is steeped two minutes, draws extracting solution interior in another new EP pipe.
5. the method that paper substrate micro-fluidic Rapid nucleic acid according to claim 4 is extracted, it is characterised in that the lysate is
The NaOH solution of 10mM.
6. the method that paper substrate micro-fluidic Rapid nucleic acid according to claim 4 is extracted, it is characterised in that the cleanout fluid is
Dehydrated alcohol.
7. the method that paper substrate micro-fluidic Rapid nucleic acid according to claim 4 is extracted, it is characterised in that the nucleic acid extraction
Liquid is distilled water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710080604.5A CN106635747A (en) | 2017-02-15 | 2017-02-15 | Paper-based micro fluidic rapid nucleic acid extraction apparatus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710080604.5A CN106635747A (en) | 2017-02-15 | 2017-02-15 | Paper-based micro fluidic rapid nucleic acid extraction apparatus |
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| Publication Number | Publication Date |
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| CN106635747A true CN106635747A (en) | 2017-05-10 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710080604.5A Pending CN106635747A (en) | 2017-02-15 | 2017-02-15 | Paper-based micro fluidic rapid nucleic acid extraction apparatus |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108384780A (en) * | 2018-02-11 | 2018-08-10 | 中国农业科学院烟草研究所 | A kind of method and extraction element of batch DNA rapid extraction |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001070399A1 (en) * | 2000-03-22 | 2001-09-27 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Microhybridisation chamber |
| JP2004305037A (en) * | 2003-04-03 | 2004-11-04 | Fuji Photo Film Co Ltd | Method for separating and purifying nucleic acid and apparatus for separating and purifying nucleic acid |
| JP2006087424A (en) * | 2004-08-26 | 2006-04-06 | Matsushita Electric Ind Co Ltd | Filtration inspection device |
| CN101538567A (en) * | 2008-03-20 | 2009-09-23 | 杭州优思达生物技术有限公司 | Method for quickly processing filter-type micro nucleic acid clinical samples |
| WO2012007503A1 (en) * | 2010-07-14 | 2012-01-19 | Qiagen Gmbh | New storage, collection or isolation device |
| US20130089854A1 (en) * | 2010-11-25 | 2013-04-11 | Xian Weitong Bioscience Limited Company | Kit for dot immunogold directed filtration assay and use thereof |
-
2017
- 2017-02-15 CN CN201710080604.5A patent/CN106635747A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001070399A1 (en) * | 2000-03-22 | 2001-09-27 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Microhybridisation chamber |
| JP2004305037A (en) * | 2003-04-03 | 2004-11-04 | Fuji Photo Film Co Ltd | Method for separating and purifying nucleic acid and apparatus for separating and purifying nucleic acid |
| JP2006087424A (en) * | 2004-08-26 | 2006-04-06 | Matsushita Electric Ind Co Ltd | Filtration inspection device |
| CN101538567A (en) * | 2008-03-20 | 2009-09-23 | 杭州优思达生物技术有限公司 | Method for quickly processing filter-type micro nucleic acid clinical samples |
| WO2012007503A1 (en) * | 2010-07-14 | 2012-01-19 | Qiagen Gmbh | New storage, collection or isolation device |
| US20130089854A1 (en) * | 2010-11-25 | 2013-04-11 | Xian Weitong Bioscience Limited Company | Kit for dot immunogold directed filtration assay and use thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108384780A (en) * | 2018-02-11 | 2018-08-10 | 中国农业科学院烟草研究所 | A kind of method and extraction element of batch DNA rapid extraction |
| CN108384780B (en) * | 2018-02-11 | 2023-12-29 | 中国农业科学院烟草研究所 | Method and device for rapidly extracting DNA in batches |
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