CN110343601A - A kind of micro-fluidic Rapid nucleic acid of high throughput extracts and detection device - Google Patents
A kind of micro-fluidic Rapid nucleic acid of high throughput extracts and detection device Download PDFInfo
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- CN110343601A CN110343601A CN201910500251.9A CN201910500251A CN110343601A CN 110343601 A CN110343601 A CN 110343601A CN 201910500251 A CN201910500251 A CN 201910500251A CN 110343601 A CN110343601 A CN 110343601A
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- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 65
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 65
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 65
- 238000001514 detection method Methods 0.000 title claims abstract description 59
- 239000000284 extract Substances 0.000 title claims description 37
- 239000012530 fluid Substances 0.000 claims abstract description 59
- 238000000605 extraction Methods 0.000 claims abstract description 48
- 239000000126 substance Substances 0.000 claims abstract description 13
- 230000009471 action Effects 0.000 claims abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 59
- 230000002101 lytic effect Effects 0.000 claims description 47
- 238000005336 cracking Methods 0.000 claims description 41
- 239000007788 liquid Substances 0.000 claims description 36
- 230000007704 transition Effects 0.000 claims description 32
- 239000007795 chemical reaction product Substances 0.000 claims description 27
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 24
- 238000010828 elution Methods 0.000 claims description 24
- 239000002699 waste material Substances 0.000 claims description 20
- 229910021389 graphene Inorganic materials 0.000 claims description 15
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 239000003463 adsorbent Substances 0.000 claims description 6
- 238000005192 partition Methods 0.000 claims description 6
- 238000004891 communication Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 4
- 238000000034 method Methods 0.000 abstract description 10
- 230000008569 process Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 21
- 230000000694 effects Effects 0.000 description 13
- 210000000170 cell membrane Anatomy 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 230000004907 flux Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000006166 lysate Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
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- 230000005291 magnetic effect Effects 0.000 description 3
- 238000000018 DNA microarray Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000009089 cytolysis Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 230000008676 import Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000005298 paramagnetic effect Effects 0.000 description 2
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- 238000004140 cleaning Methods 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
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- 230000007613 environmental effect Effects 0.000 description 1
- 206010016165 failure to thrive Diseases 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
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- 150000003904 phospholipids Chemical class 0.000 description 1
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- 108090000623 proteins and genes Proteins 0.000 description 1
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- 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
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Abstract
The present invention relates to a kind of micro-fluidic Rapid nucleic acid extraction of high throughput and detection devices, including scratch diskette and the extraction disk being arranged in the disk of the scratch diskette, the scratch diskette and extraction disk are disk and can be rotated centered on its own axis direction respectively, and the rotation center of the extraction disk and scratch diskette is not overlapped；At least two work pools are provided on said extracted disk, it is connected between each work pool and corresponding another work pool by fluid line able to turn on or off, said extracted disk can be positioned such that one of work pool is less than another work pool at a distance from the rotation center of scratch diskette at a distance from the rotation center of scratch diskette relative to scratch diskette, and substance can be made to pass through corresponding above-mentioned fluid line under the action of the centrifugal force of scratch diskette and circulated therebetween.Compared with prior art, the apparatus structure in the present invention is simple and convenient to operate, and is realized going on smoothly for entire operation process by centrifugal force, is realized the high efficiency extraction to nucleic acid.
Description
Technical field
It is extracted the present invention relates to molecular biology field more particularly to a kind of micro-fluidic Rapid nucleic acid of high throughput and detection fills
It sets.
Background technique
It include extraction, clone, amplification, the detection of nucleic acid with the biotechnology that nucleic acid (including DNA and RNA) is research object
And a series of technologies, the nucleic acid extraction such as sequencing are the committed steps of nucleic acids research, wherein the efficiency of nucleic acid extraction is that nucleic acid is ground
Study carefully whether successful restraining factors.
Paramagnetic particle method is a kind of currently used method for extracting nucleic acid, uses nanotechnology to superparamagnetic nano particle
Surface is improved and is modified, and superparamagnetism silica nanometer magnetic bead is prepared into, which can be in micro interface and nucleic acid molecules
Specific recognition and efficiently combination.Using the superparamagnetism of oxidation baby's nanosphere, in Chaotropi c salt (guanidine hydrochloride, different sulphur
Cyanic acid guanidine etc.) and the collective effect of externally-applied magnetic field under, can will be in blood, animal tissue, food, pathogenic microorganism equal samples
DNA and RNA are separated, which can be in clinical disease diagnosis, transfusion safety, Forensic Identification, disease control detection, nucleic acid samples
Build the multiple fields applications such as library, environmental microorganism detection, food safety detection, molecular biology research.Major part is complete certainly at present
Dynamic instrument for extracting nucleic acid is all made of above-mentioned paramagnetic particle method and carries out nucleic acid extraction, and during the extraction process, it is in spacious for extracting the test tube mouth used
Open state, and be the test tube group extracted for multisample, the test tube group extraction platform open as one, in test tube group
Test tube is in the open state, is easy to happen cross contamination during nucleic acid extraction in this way, to influence to extract as a result, and should
Kind nucleic acid-extracting apparatus volume is big, and at high cost and consumable quantity is big.In addition, in recent years, isothermal amplification technique rapid development is common
Manual nucleic acid extraction uses vibrator to mix, and generally by the way of vibrating up and down, but vibrates up and down and mix one
It is poor that cause property is mixed than Stirring, and there are problems that Aerosol Pollution.
Further, the rupture of cell wall is critical issue during nucleic acid extraction.The study found that graphene oxide can be broken
The cell membrane of bad bacterium outflows so as to cause intracellular organic matter and kills bacterium, in addition, graphene can be by bacterial cell membrane
Insertion cut, cell membrane is destroyed by the extensive direct extraction to phospholipid molecule on cell membrane, to kill thin
Bacterium.In addition, again studies have pointed out that it is that graphene can be as the incomparable sharp blade of nanoscale, directly that graphene is directly contacted with bacterium
Mechanical damage was connected to destroy bacterium membrane structure, meanwhile, there is graphene excellent electron-transport to remind, and can change easily
The point of bacterium film surface causes film surface cellular respiration, electron-transport and letter so that bacterial membrane surface potential loses stable state
Number transmission is chaotic, dead so as to cause growth failure in bacterial body.
Summary of the invention
First technical problem to be solved by this invention is to provide that a kind of structure is simple, operation side for the prior art
Just and efficiently high-throughput micro-fluidic Rapid nucleic acid extracts and detection device.
Second technical problem to be solved by this invention is to provide a kind of good airproof performance for the prior art and can solve
The certainly micro-fluidic Rapid nucleic acid extraction of the high throughput of Aerosol and detection device.
The technical scheme of the invention to solve the technical problem is: a kind of micro-fluidic Rapid nucleic acid of high throughput extracts
And detection device, which is characterized in that including scratch diskette and the extraction disk that is arranged in the disk of the scratch diskette, the scratch diskette and mention
Disk is taken to be disk and can rotate centered on own axes direction respectively, and the rotation center of the extraction disk and scratch diskette does not weigh
It is folded；At least two work pools are provided on said extracted disk, by able to turn on or off between each work pool and corresponding another work pool
Fluid line connection, said extracted disk can be positioned relative to scratch diskette, and can be made under the action of the centrifugal force of scratch diskette
Substance flows to another work pool by corresponding fluid line by one of work pool.
Preferably, the work pool includes cracking work pool, the first lytic reagent pond, the second lytic reagent pond and knot
Close pond, wherein the center point for extracting disk, above-mentioned first lytic reagent pond and the second lytic reagent pond point is arranged in above-mentioned combination pond
It is not connected to above-mentioned cracking work pool by corresponding fluid line, and the cracking work pool and above-mentioned combination pond phase fluid connect
Logical, the same side in conjunction with pond is arranged in the first lytic reagent pond, the second lytic reagent pond and cracking work pool, and first splits
The distance between solve reagent and the second lytic reagent pond and combine pond be respectively greater than cracking work pool and combine between pond away from
From.Reagent in such first lytic reagent pond and the second lytic reagent pond can separately flow into cracking work pool, work at this time
Disk stops operating, and after to sample dissociation, the centrifugation to lysate is realized by the rotation of scratch diskette, after being finally centrifuged
Supernatant is imported by the rotation of scratch diskette and is combined in pond.
Preferably, first lytic reagent pond is graphene reagent.It can promote cell membrane using graphene reagent
Rupture, consequently facilitating the extraction of nucleic acid.
Preferably, cracking work pool section it is substantially cylindrical comprising epicoele, cavity of resorption and tabula up and down every
Plate offers through-hole on the partition, and the bottom end of cavity of resorption is provided with the oar piece for stirring.It cracks the sample in work pool and splits
Solution liquid (the first lytic reagent pond and the second lytic reagent pond import the reagent for cracking work pool) rotates in cracking work pool, produces
Outside centrifugal force is given birth to, the liquid in cavity of resorption with the friction rotation speed for splitting pool wall due to slowing down, and the decrease of centrifugal force is to make
It obtains pressure difference and centrifugal force the first wave current of generation is greater than to the effect of fluid, this phenomenon is referred to as Ekman layer.Due to along edge
Centrifugal force is greater than at center, if whole liquid is rotated as a whole, internal centripetal force and outside (centripetal force)
It is associated with revolving speed, so just movement not outwardly or inwardly.It cracks in work pool, the rotation speed of bottom is slower, pressure slope
Degree generates and generates therewith second wave current inside along bottom degree, and outward, the second wave current inwardly flows the liquid flow on top along bottom
To which the liquid accumulation outside edges is at center.Due to oar piece effect, rest on the effect of the first wave current of bottom centre,
Liquid will flowed to pool wall by centrifugal force effect along the inside spiral in bottom, and reach uniformly mixed effect, Jin Erti
High lysis efficiency.
Preferably, the bottom of the cracking work pool is provided with the mode of laser that can emit laser beam to cracking work pool
Block.The effect of the laser beam issued by laser module can improve the rate of cleavage of cell membrane.
Preferably, being filled with the carbon sponge prepared by graphene in the combination pond, it is folded with and is used in the carbon sponge
Adsorb the adsorbent of nucleic acid.Have the characteristics that big, the internal aperture of surface area is big by the carbon sponge prepared by graphene, can promote in this way
Into the suction-operated for being folded in adsorbent therein, so that absorption physical efficiency preferably adsorbs nucleic acid.
Preferably, the work pool further includes end product pool and detection cell, which passes through corresponding with above-mentioned combination pond
Fluid line connection, and end product pool and detection cell configured in fluid communication, and the end product pool is respectively positioned on detection cell in conjunction with the same of pond
Side, and be located at relative to above-mentioned first lytic reagent pond, the second lytic reagent pond and cracking work pool in conjunction with the another of pond
Side, and end product pool is located at detection cell and combines between pond.The nucleic acid substances of adsorbent absorption are entered in end product pool by combination pond, and
There is end product pool to enter in detection cell, the direct detection of nucleic acid substances is able to achieve in detection cell, nucleic acid can not only be improved in this way and mentioned
The efficiency for taking and detecting, and it is avoided that pollution, and then improve the accuracy of detection of nucleic acids result.
Preferably, the work pool further includes kitchen sink, elution pool and waste liquid pool, the kitchen sink and elution pool are distinguished
With above-mentioned first lytic reagent pond, the second lytic reagent pond and the setting of cracking work pool in the same side, the kitchen sink and elution
Pond is connected to waste liquid pool by corresponding fluid line, elution pool and combination respectively and in conjunction with pond configured in fluid communication in conjunction with pond
The distance between pond is also respectively greater than the distance between the first lytic reagent pond and the second lytic reagent pond and combination pond, above-mentioned useless
Yet another side in conjunction with pond is arranged in liquid pool.So as to successfully realize to the washing during nucleic acid extraction, elution action.
Preferably, the kitchen sink and combine pond between, elution pool and combine pond between and cracking work pool and knot
Close and be respectively arranged with First Transition pond between pond, kitchen sink, elution pool and cracking work pool respectively with corresponding First Transition
Pond is connected to by corresponding fluid line, and each First Transition pond passes through corresponding fluid line respectively and is connected to in conjunction with pond, together
When kitchen sink and elution pool respectively with corresponding First Transition pond pass through return-air duct formed return-air circuit；The end product pool and inspection
Survey between pond and be respectively arranged with the second transition pond and premixing pool, end product pool and the second transition pond, the second transition pond and premixing pool with
And premixing pool passes through corresponding fluid line with detection cell respectively and is connected to.
Preferably, it is characterized in that, the extraction disk at least two, also, from overlook direction, respectively extract disk
The center of circle be located at using the center of circle of above-mentioned scratch diskette as on the same circumference in the center of circle.It is able to achieve by multiple settings for extracting disk
The big flux of nucleic acid is extracted, the big flux for being particularly suitable for nucleic acid repeats to extract, and setting by each position for extracting disk
It sets to improve and extracts quality.
Compared with the prior art, the advantages of the present invention are as follows: it is provided with scratch diskette in the present invention and extracts disk, and this is mentioned
It takes to integrate in disk and relates to the work pool of nucleic acid extraction and detection, when needing substance being oriented to another spy by one of work pool
When fixed work pool, disk and scratch diskette relative positioning will be extracted, since the two is different at a distance from the rotation center of scratch diskette, made
Scratch diskette the centrifugal force difference for both being rotated in place and applying so that substance can be flowed to by one of work pool it is another
Work pool can then be such that extraction operation goes on smoothly.As it can be seen that the apparatus structure in the present invention is simple and convenient to operate, pass through centrifugation
Power realizes going on smoothly for entire operation process, realizes the high efficiency extraction to nucleic acid.
Detailed description of the invention
Fig. 1 is that high-throughput micro-fluidic Rapid nucleic acid extracts and detection device under the first state in the embodiment of the present invention 1
Structural schematic diagram；
Fig. 2 is that high-throughput micro-fluidic Rapid nucleic acid extracts and detection device under second of state in the embodiment of the present invention 1
Structural schematic diagram；
Fig. 3 is that high-throughput micro-fluidic Rapid nucleic acid extracts and detection device under the third state in the embodiment of the present invention 1
Structural schematic diagram；
Fig. 4 is that high-throughput micro-fluidic Rapid nucleic acid extracts and detection device under the 4th kind of state in the embodiment of the present invention 1
Structural schematic diagram；
Fig. 5 is the structural schematic diagram that disk is extracted in the embodiment of the present invention 1；
Fig. 6 is the sectional view that work pool is cracked in the embodiment of the present invention 1；
Fig. 7 is the structural schematic diagram of partition in the embodiment of the present invention 1；
Fig. 8 is the structural schematic diagram of high-throughput micro-fluidic Rapid nucleic acid extraction and detection device in the embodiment of the present invention 2.
Specific embodiment
The present invention will be described in further detail below with reference to the embodiments of the drawings.
Embodiment 1:
As shown in Fig. 1~7, a kind of micro-fluidic Rapid nucleic acid of high throughput extracts and detection device, including scratch diskette 1 and setting
Extraction disk 2 in the disk of the scratch diskette 1, the scratch diskette 1 and extraction disk 2 are disk and respectively can be with own axes directions
Centered on rotate, and the rotation center O2 of the extraction disk 2 and the rotation center O1 of scratch diskette 1 are not overlapped.It is above-mentioned in the present embodiment
Extracting disk 2 is one, and the place the rotation center O1 of scratch diskette 1 is vertical equipped with the first rotary shaft (not shown), scratch diskette 1 with this first
It is rotated centered on rotary shaft, extracts and be provided with the second rotary shaft (not shown) at the rotation center O2 of disk 2, second rotary shaft is perpendicular
To being arranged in the disk of scratch diskette 1, extracts disk 2 and rotated centered on second rotary shaft.The extraction disk 2 and 1 energy of scratch diskette
Rotation respectively, also can be with 1 relative positioning of scratch diskette, so that extracting disk 2 can rotate with scratch diskette 1 by extracting disk 2.It extracts
The relative positioning of disk 2 and scratch diskette 1 can be realized by the positioning method of a variety of realizations, such as be extracted between disk 2 and scratch diskette 1
Magnetic is connected, is snapped connection.
It is provided at least two work pools on said extracted disk 2, passes through between each work pool and corresponding another work pool
Fluid line 91 able to turn on or off is connected to, and said extracted disk 2 can be positioned relative to scratch diskette 1, and can be in the centrifugal force of scratch diskette 1
Effect is lower so that substance flows to another work pool by corresponding fluid line 91 by one of work pool.Due to two work
Pond is different at a distance from the rotation center of scratch diskette 1, so that scratch diskette 1 is rotated in the centrifugal force difference applied at the two, from
And substance is enabled to flow to another work pool by one of work pool, can then extraction operation be made to go on smoothly.As it can be seen that this hair
Apparatus structure in bright is simple and convenient to operate, and is realized going on smoothly for entire operation process by centrifugal force, is realized to nucleic acid
High efficiency extraction.The valve able to turn on or off by being arranged in each fluid line 91 of each fluid line 91 is realized, is flowed in the present embodiment
The side wall of body pipeline 91 is flexible material, and valve is elastic material, so as to realize opening for fluid pipe road 91 by valve
It closes.
Specifically, in the present embodiment, above-mentioned work pool includes cracking work pool 3, the cracking of the first lytic reagent pond 41, second
Reagent 42, in conjunction with pond 5, end product pool 6 and detection cell 7, above-mentioned combination pond 5 be arranged in extract disk 2 center at, i.e., extraction disk
2 rotation center O2 is combining Chi5Chu, cracking work pool 3, the first lytic reagent pond 41 and the setting of the second lytic reagent pond 42
In the side for combining pond 5, and crack work pool 3, the first lytic reagent pond 41 and the second lytic reagent pond 42 with combine pond 5 it
Between distance successively increase, and end product pool 6 and detection cell 7 are arranged in the other side in conjunction with pond 5, and detection cell 7 and combine pond 5
Distance is greater than end product pool 6 and combines at a distance from pond 5.Above-mentioned first lytic reagent pond 41 and the second lytic reagent pond 42 respectively with it is upper
It states cracking work pool 3 to be connected to by corresponding fluid line 91, the cracking work pool 3 and above-mentioned 5 configured in fluid communication of combination pond, this
In embodiment, cracks and be provided with First Transition pond 83 between work pool 3 and combination pond 5, crack work pool 3 and combine between pond 5
And it is connected to respectively by corresponding fluid line 91 between cracking work pool 3 and combination pond 5.Pass through in conjunction with pond 5 and end product pool 6
Corresponding fluid line 91 is connected to, and the second transition pond 85 and premixing pool 86 are respectively arranged between end product pool 6 and detection cell 7, and
Between end product pool 6 and the second transition pond 85, between the second transition pond 85 and premixing pool 86 and between premixing pool 86 and detection cell 7
It is connected to respectively by corresponding fluid line 91.
Further, above-mentioned work pool further includes kitchen sink 81, elution pool 82, waste liquid pool 84 and pressure release pond 87, wherein should
Kitchen sink 81 and elution pool 82 respectively with above-mentioned first lytic reagent pond 41, the second lytic reagent pond 42 and cracking work pool 3
It is arranged in the same side, and is located at the outboard end for extracting 2 disk of disk, each kitchen sink 81 and elution pool 82 and combines between pond 5 respectively
It is provided with above-mentioned First Transition pond 83, between each kitchen sink 81 and First Transition pond 83, each elution pool 82 and First Transition pond 83
Between and each First Transition pond 83 and combine pond 5 between respectively by corresponding fluid line 91 be connected to.In addition, each kitchen sink
81 and elution pool 82 respectively between corresponding First Transition pond 83 by return-air duct 93 formed return-air circuit.Above-mentioned waste liquid pool
84 be connected tos by corresponding fluid line 91 in conjunction with pond 5, and in the present embodiment the waste liquid pool 84 be it is multiple, each adjacent gives up
It is connected to respectively by corresponding fluid line 91 between liquid pool 84, and the volume of each waste liquid pool 84 is by built-in outer incremented by successively, from
And it can preferably collect waste liquid.
In the present embodiment, above-mentioned each work pool and each fluid line 91 are the hollow structure for being arranged in and extracting in disk 2,
And the side wall of at least the above detection cell 7 is transparent, consequently facilitating the nucleic acid substances to extraction detect.This makes device
Air-tightness it is preferable, avoid the generation of Aerosol.In above-mentioned outermost waste liquid pool 84, in conjunction with pond 5 and detection
An above-mentioned pressure release pond 87 is respectively set in the side in pond 7, and each pressure release pond 87 passes through relief pipeline 92 and corresponding waste liquid pool respectively
84, it is connected in conjunction with pond 5 and detection cell 7, can guarantee the stability of whole system by the way that pressure release pond 87 is arranged.
Further, above-mentioned first lytic reagent pond 41 is graphene reagent, can promote cell membrane using graphene reagent
Rupture, consequently facilitating the extraction of nucleic acid.Crack work pool 3 section it is substantially cylindrical comprising epicoele 51, cavity of resorption 52 with
And partition 53 of the tabula between upper and lower chamber 51,52, through-hole 531 is offered on the partition 53, and the bottom end of cavity of resorption 52 is provided with
For stirring the oar piece 54 of substance in cavity of resorption 52.Crack the sample in work pool 3 and lysate (the first lytic reagent pond 41 and the
Two lytic reagent ponds 42 import the reagent of cracking work pool 3) it is rotated in cracking work pool 3, generate outside centrifugal force, cavity of resorption
Liquid in 52 is since the friction rotation speed with pool wall slows down, and the decrease of centrifugal force is so that pressure difference is big to the effect of fluid
The first wave current is generated in centrifugal force, this phenomenon is referred to as Ekman layer.Since the centrifugal force along edge is greater than at center, if will
Whole liquid rotate as a whole, then internal centripetal force is associated with external (centripetal force) with revolving speed, so just not having
Movement outwardly or inwardly.It cracks in work pool 3, the rotation speed of bottom is slower, and pressure slope is generated and generated therewith the bottom of along
The second inside wave current of degree, outward, the second wave current is inwardly flowed along bottom to the liquid of outside edges the liquid flow on top
It gathers at center.Due to oar piece 54 act on, rest on the effect of the first wave current of bottom centre, liquid will be along the inside spiral shell in bottom
Liter is screwed on, is being flowed by centrifugal force effect to pool wall, is reaching uniformly mixed effect, and then improve lysis efficiency.Crack work
The bottom for making pond 3 is provided with the laser module (not shown) that can emit laser beam to cracking work pool 3.It is issued by laser module
The effect of laser beam can improve the rate of cleavage of cell membrane.(do not show in conjunction in pond 5 filled with the carbon sponge prepared by graphene
Out), the adsorbent for adsorbing nucleic acid is folded in the carbon sponge.Due to the carbon sponge of graphene preparation have surface area it is big,
The big feature in internal aperture can promote the suction-operated for being folded in adsorbent therein in this way, so that absorption physical efficiency is preferably inhaled
Attached nucleic acid.
In the present embodiment, the relative positioning state of disk 2 and scratch diskette 1 is extracted with a variety of, to realize nucleic acid extraction process
In each operating procedure go on smoothly: the first state, cracking work pool 3 are big at a distance from the rotation center O1 of scratch diskette 1
In combination pond 5 at a distance from the rotation center O1 of scratch diskette 1, it is calculated as position one at this time, as shown in Figure 1；Second of state, waste liquid
Pond 84 is greater than the distance between the rotation center O1 in conjunction with pond 5 and scratch diskette 1 at a distance from the rotation center O1 of scratch diskette 1, this
The position Shi Jiwei two, as shown in Figure 2；The third state, end product pool 6 is greater than at a distance from the rotation center O1 of scratch diskette 1 to be combined
Pond 5 is denoted as position three, as shown in Figure 3 at a distance from the rotation center O1 of scratch diskette 1 at this time；The rotation of detection cell 7 and scratch diskette 1
The distance for turning center O1 is greater than end product pool 6 at a distance from the rotation center O1 of scratch diskette 1, position four is denoted as at this time, such as Fig. 4 institute
Show, in the present embodiment detection cell 7 have it is multiple be connected in sequence, it is above-mentioned that " detection cell 7 is at a distance from the rotation center of scratch diskette 1
Greater than end product pool 6 at a distance from the rotation center of scratch diskette 1 " refer to the rotation center of detection cell 7 and scratch diskette 1 positioned at upstream end
The distance between O1.Preferably to identify above-mentioned position one, position two, position three and position four in the present embodiment, mentioning
It takes and first positioning hole 21, second location hole 22, third location hole 23 and the 4th location hole 24 is respectively set in the disk of disk.
The course of work of the invention is as follows:
(1) extraction disk 2 is adjusted into supreme rheme and sets one, the specimen port of cracking work pool 3 will be passed through when sample to be cracked
31 are added, and the reagent in the first lytic reagent pond 41 and the second lytic reagent pond 42 is passed through scratch diskette 1 respectively by centrifugal force
Centrifugal force be added to cracking work pool 3 in, scratch diskette 1 stops operating, lytic reagent lysate sample.Start laser module, benefit
The membranolysis in sample is accelerated to cracking 3 bottom scan of work pool with the laser beam of 808nm.
(2) it extracts disk 2 and is maintained at above-mentioned position one, open between cracking work pool 3 and its corresponding First Transition pond 83
Fluid line 91, scratch diskette 1 rotate, by crack work pool 3 supernatant be centrifuged into the First Transition pond 83.Scratch diskette 1
It stops operating, closes the fluid line 91 between cracking work pool 3 and its corresponding First Transition pond 83, open the First Transition
Fluid line 91 between pond 83 and combination pond 5, scratch diskette 1 rotate, the supernatant in the First Transition pond 83 are centrifuged to combination
In pond 5.Scratch diskette 1 stops operating, and closes the First Transition pond 83 and combines the fluid line 91 between pond 5, lysate is being tied
It closes in pond 5 and suction-operated occurs with carbon sponge.It opens kitchen sink 81 and combines the fluid line 91 between pond 5, scratch diskette 1 revolves
Turn, cleaning solution in kitchen sink 81 is centrifuged to pond 5 is combined, scratch diskette 1 stops operating.It closes kitchen sink 81 and combines between pond 5
Fluid line 91, scratch diskette 1 rotate, washing combine pond 5.After several seconds, scratch diskette 1 stops operating, open combine pond 5 with and its
Fluid line 91 between the waste liquid pool 84 of connection extracts the rotation of disk 2 and adjusts supreme rheme and set two, extracts disk 2 and stop operating,
Scratch diskette 1 rotates, and the waste liquid in pond 5 will be combined to be centrifuged into waste liquid pool 84.After the completion of washing, scratch diskette 1 stops operating, and extracts
Disk 2 turns to position one, closes and combines pond 5 and the fluid line 91 between the waste liquid pool 84 of its connection.By above-mentioned steps pair
After carrying out sufficiently washing in conjunction with pond 5, opens elution pool 82 and combine the fluid line 91 between pond 5, scratch diskette 1 is rotated, will be washed
Eluent is centrifuged to pond 5 is combined in de- pond 82, and scratch diskette 1 stops operating.It closes elution pool 82 and combines the fluid hose between pond 5
Road 91, scratch diskette 1 rotate, elution of bound pond 5.After several seconds, scratch diskette 1 stops operating, open combine pond 5 with and its connect give up
Fluid line 91 between liquid pool 84 extracts the rotation of disk 2 and adjusts supreme rheme and set two, extracts disk 2 and stop operating, scratch diskette 1
Rotation will combine the waste liquid in pond 5 to be centrifuged into waste liquid pool 84.After the completion of elution, scratch diskette 1 stops operating, and extracts disk 2 and rotates
To position one, closes and combine pond 5 and the fluid line 91 between the waste liquid pool 84 of its connection.
(3) it extracts disk 2 to rotate and adjust to position three, opens the fluid line 91 combined between pond 5 and end product pool 6, work
Make the rotation of disk 1, supernatant is centrifuged into end product pool 6, scratch diskette 1 stops operating, and closes and combines between pond 5 and end product pool 6
Fluid line 91.
(4) it extracts disk 2 to rotate and adjust to position four, closes the fluid line between end product pool 6 and the second transition pond 85
91, open the fluid line 91 between the second transition pond 85 and premixing pool 86, scratch diskette 1 rotates, by the nucleic acid substances of extraction from
The heart is mixed into premix liquid pool with buffer.Scratch diskette 1 stops operating, and closes between the second transition pond 85 and premix liquid pool
Fluid line 91, open the fluid line 91 between premixing pool 86 and detection cell 7, nucleic acid substances, which enter in detection cell 7, to carry out
Detection.
Embodiment 2:
As shown in figure 8, unlike the first embodiment, extract disk 2 in the present embodiment on above-mentioned scratch diskette 1 there are two settings,
Each disk 2 that extracts is connect by the second vertically arranged rotary shaft with the disk of scratch diskette 1 respectively, also, from overlook direction,
Each center of circle for extracting disk 2 is located at using the center of circle of above-mentioned scratch diskette 1 as on the same circumference in the center of circle.Two extraction disks 2 can be right
Same sample carries out parallel extraction operation, can also carry out nucleic acid extraction to different samples respectively.It is extracted by two
The setting of disk 2 is able to achieve the big flux extraction to nucleic acid, and the big flux for being particularly suitable for nucleic acid repeats to extract, and by respectively mentioning
Extraction quality can be improved by taking the setting of the position of disk 2.
The characteristics of microfluidic chip technology, makes it carry out high speed to individual biological information, and parallel acquisition and analysis become can
Can, it is the acquisition of an important information and the processing platform of modern biotechnology science, provides technical support and behaviour for life area research
Make platform.Microfluidic chip technology is in biological gene engineering, medical diagnosis on disease and drug research, cell analysis, biology at present
The fields such as intermolecular interaction achieve significant achievement.Biological microfluidic system platform can be regarded as to a certain extent
One system platform as made of multiple and different component combinations, the system platform mainly include four most of: sampling system, control
System, biochip and analysis system processed.Microflow control technique developed it is very mature, how by nucleic acid rapidly extracting and micro-fluidic
It is the application problem of being solved place that technology, which combines,.The present invention is micro-fluidic by nucleic acid rapidly extracting and high flux biochip
Technology combines, and effectively improves the efficiency detected from nucleic acid extraction to high throughput and extracts quality.
Claims (10)
1. a kind of micro-fluidic Rapid nucleic acid of high throughput extracts and detection device, which is characterized in that including scratch diskette and be arranged at this
Extraction disk in the disk of scratch diskette, the scratch diskette and extraction disk are disk and respectively can be centered on its own axis directions
Rotation, and the rotation center of the extraction disk and scratch diskette is not overlapped；
At least two work pools are provided on said extracted disk, by able to turn on or off between each work pool and corresponding another work pool
Fluid line connection, said extracted disk can be positioned such that in the rotation of one of work pool and scratch diskette relative to scratch diskette
The distance of the heart is less than another work pool at a distance from the rotation center of scratch diskette, and can be under the action of the centrifugal force of scratch diskette
So that substance passes through corresponding above-mentioned fluid line and circulates therebetween.
2. high-throughput micro-fluidic Rapid nucleic acid as described in claim 1 extracts and detection device, which is characterized in that the work
Pond includes cracking work pool, the first lytic reagent pond, the second lytic reagent pond and combines pond, wherein above-mentioned combination pond setting
At the center for extracting disk, above-mentioned first lytic reagent pond and the second lytic reagent pond pass through phase with above-mentioned cracking work pool respectively
The fluid line connection answered, and the cracking work pool and above-mentioned combination pond configured in fluid communication, the first lytic reagent pond, second are split
The same side in conjunction with pond, and the distance between the first lytic reagent pond and combination pond is arranged in solution reagent and cracking work pool
Greater than the distance between cracking work pool and combination pond, the distance between the second lytic reagent pond and combination pond are also greater than cracking work
The distance between make pond and combine pond.
3. high-throughput micro-fluidic Rapid nucleic acid as claimed in claim 2 extracts and detection device, which is characterized in that described first
Lytic reagent pond is graphene reagent.
4. high-throughput micro-fluidic Rapid nucleic acid as claimed in claim 2 extracts and detection device, which is characterized in that the cracking
The section of work pool is substantially cylindrical comprising the partition of epicoele, cavity of resorption and tabula between upper and lower chamber, on the partition
Through-hole is offered, and the bottom end of cavity of resorption is provided with the oar piece for stirring.
5. high-throughput micro-fluidic Rapid nucleic acid as claimed in claim 2 extracts and detection device, which is characterized in that the combination
It is filled with the carbon sponge prepared by graphene in pond, the adsorbent for adsorbing nucleic acid is folded in the carbon sponge.
6. as the micro-fluidic Rapid nucleic acid extraction of the described in any item high throughputs of claim 2~5 and detection device, feature exist
In, the work pool further includes end product pool and detection cell, which is connected to above-mentioned combination pond by corresponding fluid line,
And end product pool and detection cell configured in fluid communication, and the end product pool and detection cell are respectively positioned on the same side in conjunction with pond, and relative to upper
It states the first lytic reagent pond, the second lytic reagent pond and cracking work pool and is located at the other side for combining pond, and end product pool is located at
Between detection cell and combination pond.
7. high-throughput micro-fluidic Rapid nucleic acid as claimed in claim 6 extracts and detection device, which is characterized in that the work
Pond further includes kitchen sink, elution pool and waste liquid pool, the kitchen sink and elution pool respectively with above-mentioned first lytic reagent pond, second
Lytic reagent pond and cracking work pool are arranged in the same side, and the kitchen sink and elution pool connect with pond phase fluid is combined respectively
It is logical, and be connected to waste liquid pool by corresponding fluid line in conjunction with pond, elution pool is also respectively greater than with the distance between pond is combined
The distance between first lytic reagent pond and the second lytic reagent pond and combination pond, above-mentioned waste liquid pool are arranged in conjunction with the another again of pond
Side.
8. high-throughput micro-fluidic Rapid nucleic acid as claimed in claim 7 extracts and detection device, which is characterized in that the washing
Pond and combine pond between, elution pool and combine pond between and cracking work pool and combine pond between be respectively arranged with First Transition
Pond, kitchen sink and elution pool are connected to corresponding First Transition pond by corresponding fluid line respectively, and each First Transition pond
Be connected to respectively by corresponding fluid line in conjunction with pond, at the same kitchen sink, elution pool and crack work pool respectively with it is corresponding
First Transition pond pass through return-air duct formed return-air circuit；
The second transition pond and premixing pool, end product pool and the second transition pond, are respectively arranged between the end product pool and detection cell
Two transition ponds pass through corresponding fluid line with detection cell with premixing pool and premixing pool respectively and are connected to.
9. the micro-fluidic Rapid nucleic acid of high throughput as claimed in any one of claims 1 to 5 extracts and detection device, feature exist
In, the extraction disk at least two, also, from overlook direction, each center of circle for extracting disk is located at above-mentioned work
The center of circle of disk is on the same circumference in the center of circle.
10. the micro-fluidic Rapid nucleic acid of high throughput as claimed in any one of claims 1 to 5 extracts and detection device, feature exist
In, each work pool and each fluid line be the hollow structure for being arranged in and extracting in disk, and at least the above detection cell
Side wall is transparent.
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