CN205329049U - Isothermal nucleic acid increase device - Google Patents
Isothermal nucleic acid increase device Download PDFInfo
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- CN205329049U CN205329049U CN201521043284.9U CN201521043284U CN205329049U CN 205329049 U CN205329049 U CN 205329049U CN 201521043284 U CN201521043284 U CN 201521043284U CN 205329049 U CN205329049 U CN 205329049U
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Abstract
The utility model belongs to the technical field of it is biological, concretely relates to isothermal nucleic acid increase device and experimental method, including dull and stereotyped normal position nucleic acid increase chip, this chip comprises mutual laminating, the upper and lower layer carrier of slidable, and upper and lower layer carrier is the rectangle of equal size, and when the lower floor was fixed, upper carrier is friction sliding relatively. Offer various recess runner, the microcavitys that open side down on the last binding face of lower floor's carrier, offer the ascending recess discharge orifice of various openings, microcavity on the lower binding face of upper carrier to reach some and advance, export the through -hole, through quantitative application of sample and the not measuring of common mode, can carry out the valid operation to different samples simultaneously, and reduced the experiment cost. The chip can be applied to forensic detection, environment measuring, eat quality control survey etc.
Description
Technical field
This utility model belongs to biological technical field, is specifically related to a kind of flat board original position isothermal nucleic acid amplification experimental provision and experimental technique。
Background technology
Microflow control technique is one of international high and new technology Disciplinary Frontiers now, and the application in nucleic acid extraction, amplification, detection research achieves fast development, has been increasingly becoming one of most potential developing direction of nucleic acids research。It adopts micro-electromechanical technology to process the chip with micro-meter scale channel network configuration, by picoliters in chip to receiving the upgrading manipulation of microfluid and control, it is achieved biomedical and the integrated of chemical laboratory analyzes detection function。Slip chip (SlipChip) technology is exactly the one of microflow control technique, it is made up of two pieces of substrates up and down with micro structure, added primer premix, enzyme premix, DNA by upper substrate, realize liquid by slip upper substrate and shift alternately thus mixing the amplification of all reaction reagent original positions in reaction chamber at levels substrate。
Ring mediated isothermal nucleic acid amplification (loop-mediatedisothermalamplification, LAMP) is a kind of constant temperature nucleic acid amplification method。This technology utilizes 4 species-specific primers to rely on a kind of high activity strand displacement archaeal dna polymerase (about 65 DEG C) when DNA is in dynamic equilibrium to make strand displacement DNA synthesis in ceaselessly oneself's circulation。Whole reaction without thermal cycle repeatedly, and high specificity, highly sensitive, rapidly and efficiently, identify simplicity, for the fields such as prudence, ecological environment, food real-time diagnosis detect application create condition。
But domestic and international existing LAMP slip chip is still immature, is not fixed easily and operates, and slip chip is not had to mention the system and device that can realize original position amplification。Therefore, this utility model, based on the studies above background, has developed a kind of flat board original position isothermal nucleic acid amplification chip。
Utility model content
The purpose of this utility model there is provided flat board original position isothermal nucleic acid amplification chip, and this chip operation is easy, it may be achieved expand different samples simultaneously, reduces test operation intensity, and has saved the consumption of sample and reagent, reduces experimental cost。
In order to realize object above, this utility model provides a kind of flat board in-situ nucleic acid amplification chip, including upper strata carrier bonded to each other and lower floor's carrier；Upper strata carrier level can slide on lower floor's carrier；
Upper strata carrier is provided with sample feeding mouth group, polymerase injection port group, primer injection port group, the first outlet through hole, the second outlet through hole, the first groove discharge orifice group, the second groove discharge orifice group, first flow；
Lower floor's carrier is provided with sample introduction tank, the second runner group, reaction chamber group, comb teeth shape passage, the 3rd groove discharge orifice group；Reaction chamber is mounted between comb teeth shape passage increment or side, and the increment flowing into end and comb teeth shape passage of reaction chamber group is located along the same line；
When upper strata carrier and lower floor's carrier are positioned at initial position, circulate mutually in the position of sample feeding mouth group and reaction chamber group；Polymerase injection port group circulates mutually with comb teeth shape passage, the 3rd groove discharge orifice group, the second outlet through hole；Primer injection port group circulates mutually with sample introduction tank, the second runner group, the first groove discharge orifice group, first flow, the first outlet through hole；
During to reaction chamber group interpolation primer, translation upper strata carrier, make the first groove discharge orifice group circulate mutually with reaction chamber group, primer flows in reaction chamber group from the first groove discharge orifice；
During to reaction chamber group interpolation polymerase, translation upper strata carrier, make reaction chamber group circulate mutually with the second groove discharge orifice group, polymerase flows in reaction chamber group from the second groove discharge orifice group。
As preferably, upper strata carrier and lower floor's carrier are rectangle carrier。
As preferably, the first groove discharge orifice group includes the first groove discharge orifice, and the second groove discharge orifice group includes the second groove discharge orifice, and the 3rd groove discharge orifice group includes the 3rd groove discharge orifice；Reaction chamber group includes reaction chamber；Sample feeding mouth group includes sample feeding mouth；
Sample feeding mouth is arranged on same straight line equably；First groove discharge orifice is arranged on same straight line equably, primer injection port and the first groove discharge orifice are arranged on same straight line, second groove discharge orifice is arranged on same straight line equably, the position of sample feeding mouth and the first groove discharge orifice is corresponding, and the second groove discharge orifice and sample feeding mouth are staggered；3rd groove discharge orifice is arranged on same straight line equably, and reaction chamber is arranged on same straight line equably；Second runner group includes the second runner being positioned on straight line and the second runner being located on the same line with the 3rd groove discharge orifice group；
Sample feeding mouth, the first groove discharge orifice, the second groove discharge orifice are paralleled with the central horizontal axle of upper strata carrier；Reaction chamber, the 3rd groove discharge orifice are paralleled with the central horizontal axle of lower floor's carrier。
As preferably, upper strata carrier is provided with two groups of sample feeding mouth groups, polymerase injection port group, primer injection port group, the first outlet through hole, the second outlet through hole, the first groove discharge orifice group, the second groove discharge orifice group, first flow；
Two groups of sample feeding mouth groups, polymerase injection port group, primer injection port group, the first outlet through hole, the second outlet through hole, the first groove discharge orifice group, the second groove discharge orifice group, first flow are relative to the central horizontal axle axial symmetry of upper strata carrier；
Lower floor's carrier is provided with two groups of sample introduction tank, the second runner group, reaction chamber group, comb teeth shape passage, the 3rd groove discharge orifice group；
Two groups of sample introduction tank, the second runner group, reaction chamber group, comb teeth shape passage, the 3rd groove discharge orifice group are relative to the central horizontal axle axial symmetry of lower floor's carrier。
As preferably, primer injection port and the first groove discharge orifice are arranged on the central horizontal axle of upper strata carrier, and sample introduction tank is positioned on the central horizontal axle of lower floor's carrier。
As preferably, when overlooking the first groove discharge orifice, the second groove discharge orifice, the 3rd groove discharge orifice, the first groove discharge orifice is cross；Second groove discharge orifice, the 3rd groove discharge orifice are rectangle。
As preferably, the volume of each the first groove discharge orifice is identical；The volume of each the second groove discharge orifice is identical；The volume of each reaction chamber is identical。
As preferably, the volumetric ratio between reaction chamber, the first groove discharge orifice, the second groove discharge orifice is 10:6:3。
As preferably, sample feeding mouth group, polymerase injection port group, primer injection port group, the first outlet through hole, the second outlet through hole diameter be 1mm。
As preferably, first flow is T-shaped runner。
This utility model additionally provides a kind of isothermal nucleic acid amplification device, including the flat board original position isothermal nucleic acid amplification chip of square heat transfer platform and above-mentioned any one；Heat transfer platform is provided with heater；Lower floor's carrier is fixed on heat transfer platform, and lower floor's carrier is uniformly heated by heater。
As preferably, heat transfer platform includes adjacent at least 2 movable block, limit assembly and underframe；Limit assembly includes sliding axle portion and limiting section；
Lower floor's carrier is fixed on underframe, and the one end in sliding axle portion runs through that movable block and underframe are fixing to be connected；Movable block can slide in sliding axle portion level；Limiting section is located at the other end in sliding axle portion, and cross-sectional area is more than the cross-sectional area in sliding axle portion。
As preferably, each movable block is provided with at least 2 limit assemblies。
As preferably, limit assembly is iron nail。A kind of isothermal nucleic acid amplification method, make use of the isothermal nucleic acid amplification device of above-mentioned any one, comprises the following steps:
S0, upper strata carrier is moved to predeterminated position；
S1, add primer from primer injection port group, and make primer be full of the first groove discharge orifice group；
S2, translation upper strata carrier, make the first groove discharge orifice group circulate mutually with reaction chamber group, and primer flows in reaction chamber group from the first groove discharge orifice；
S3, upper strata carrier is moved back to predeterminated position；
S4, add polymerase from polymerase injection port group, and make polymerase be full of the second groove discharge orifice group；
S5, translation upper strata carrier, make reaction chamber group circulate mutually with the second groove discharge orifice group, and polymerase flows in reaction chamber group from the second groove discharge orifice group；
S6, upper strata carrier is moved back to predeterminated position；
S7, from sample feeding mouth group add sample；
S8, control heater make reactant liquor keep at the reaction temperatures。
As preferably, at two groups of sample feeding mouth groups, polymerase injection port group, primer injection port group, the first outlet through hole, the second outlet through hole, the first groove discharge orifice group, the second groove discharge orifice group, first flow relative to the central horizontal axle axial symmetry of upper strata carrier；And two groups of sample introduction tank, the second runner group, reaction chamber group, comb teeth shape passage, the 3rd groove discharge orifice group relative to the central horizontal axle axial symmetry of lower floor's carrier time, after step S3, again primer injection port group adds primer, and translate upper strata carrier, the reaction chamber group making the first groove discharge orifice group and opposite side circulates mutually, and primer flows in the reaction chamber group of opposite side from the first groove discharge orifice。
Compared with prior art, the beneficial effects of the utility model are in that:
1, flat board original position isothermal nucleic acid amplification chip of the present utility model can carry out multiple sample and carry out amplification operation simultaneously, is greatly saved the response time, has saved the consumption of sample and reagent, reduced experimental cost。
2, flat board original position isothermal nucleic acid amplification chip of the present utility model can realize smooth sliding and accurate temperature controller by Simple heater, effectively nucleic acid can be carried out original position amplification。
3, this chip reaction chamber, discharge orifice, runner volume size design by a certain percentage, it is possible to effectively control reaction system, improve amplification efficiency。
4, for part Experiment, can be detected by an unaided eye reaction by this chip。
Accompanying drawing explanation
Fig. 1 is the chip structure top view of the first embodiment of this utility model。
Fig. 2 is the chip structure top view of this utility model the second embodiment。
Fig. 3 is the movable block of platform structural representation when being positioned at initial position that conducts heat in the third embodiment of this utility model。
Fig. 4 be the third embodiment of this utility model conducts heat platform movable block slide after structural representation。
Fig. 5 is the chip structure top view that this utility model carrier at the middle and upper levels is positioned at initial position。
Chip structure top view when Fig. 6 is add primer in this utility model。
Chip structure top view when Fig. 7 is add polymerase in this utility model。
Fig. 8 adds the chip structure top view during polymerase when being utilize this utility model chip to carry out controlled trial。
Accompanying drawing labelling: 1, upper strata carrier；11, sample feeding mouth group；12, polymerase injection port group；13, primer injection port group；14, the second groove discharge orifice group；15, the first groove discharge orifice group；16, first flow；17, the first outlet through hole；18, the second outlet through hole；2, lower floor's carrier；21, reaction chamber group；22, comb teeth shape passage；23, sample introduction tank；24, the second runner group；25, the 3rd groove discharge orifice group；3, heat transfer platform；31, limit assembly；32, underframe；33, movable block。
Detailed description of the invention
For making the purpose of this utility model, technical scheme and advantage clearly, below in conjunction with accompanying drawing, each embodiment of the present utility model is explained in detail。
For solving above-mentioned technical problem, the first embodiment of the present utility model provides a kind of flat board in-situ nucleic acid amplification chip, as it is shown in figure 1, include upper strata carrier 1 bonded to each other and lower floor's carrier 2；Upper strata carrier 1 level can slide on lower floor's carrier 2；
Upper strata carrier 1 is provided with sample feeding mouth group 11, polymerase injection port group 12, primer injection port group the 13, first outlet through hole the 17, second outlet through hole the 18, first groove discharge orifice group the 15, second groove discharge orifice group 14, first flow 16；
Lower floor's carrier 2 is provided with sample introduction tank the 23, second runner group 24, reaction chamber group 21, comb teeth shape passage the 22, the 3rd groove discharge orifice group 25；Reaction chamber group 21 is located between comb teeth shape passage 22 increment or side, and the increment flowing into end and comb teeth shape passage 22 of reaction chamber group 21 is located along the same line；
When upper strata carrier 1 and lower floor's carrier 2 are positioned at initial position, circulate mutually in the position of sample feeding mouth group 11 and reaction chamber group 21；Polymerase injection port group 12 circulates with comb teeth shape passage the 22, the 3rd groove discharge orifice group the 25, second outlet through hole 18 phase；Primer injection port group 13 and sample introduction tank the 23, second runner group the 24, first groove discharge orifice group 15, the circulation of first flow the 16, first outlet through hole 17 phase；
To reaction chamber group 21 add primer time, translation upper strata carrier 1, make the first groove discharge orifice group 15 and the circulation of reaction chamber group 21 phase, primer flows in reaction chamber group 21 from the first groove discharge orifice 15；
To reaction chamber group 21 add polymerase time, translation upper strata carrier 1, make reaction chamber group 21 circulate with the second groove discharge orifice group 14 phase, polymerase flows in reaction chamber group 21 from the second groove discharge orifice group 14。
In the present embodiment, the second runner group 24 includes 6 sections of runners, and the first groove discharge orifice group 15 includes 5 the first groove discharge orifices, and the second groove discharge orifice group 14 includes 5 the second groove discharge orifices；3rd groove discharge orifice group 25 includes 10 the 3rd groove discharge orifices；Reaction chamber group 21 includes 5 reaction chambers, and reaction chamber is set to bulb-shaped；The volume of each the first groove discharge orifice is identical；The volume of each the second groove discharge orifice is identical；The volume of each reaction chamber is identical, and the volumetric ratio between reaction chamber, the first groove discharge orifice, the second groove discharge orifice is 10:6:3。
In the present embodiment, when overlooking the first groove discharge orifice, the second groove discharge orifice, the 3rd groove discharge orifice, the first groove discharge orifice is cross；Second groove discharge orifice, the 3rd groove discharge orifice are rectangle。Wherein, the first groove discharge orifice is designed to cross, has following technical effect that
1, when adding primer in the first groove discharge orifice, the first groove discharge orifice and the second runner group 24 circulate, and during mobile upper strata glass, can isolate quickly between the first groove discharge orifice and the second runner group 24。
2, when adding polymerase in reaction chamber, the first groove discharge orifice circulates with reaction chamber, the 3rd groove discharge orifice, and during mobile upper strata glass, if there is skew, polymerase is also not easy to flow in adjacent 3rd groove discharge orifice。
In the present embodiment, sample feeding mouth group 11, polymerase injection port group 12, primer injection port group the 13, first outlet through hole the 17, second outlet through hole 18 diameter be 1mm。
In the present embodiment, upper strata carrier 1, lower floor's carrier 2 select glass plate, naturally it is also possible to select other carrier boards that can be used in carrying out biotic experiment。It is positioned at the groove runner on carrier, discharge orifice and microcavity to be offered by laser engraving mode。The shape of glass plate, size are not particularly limited, however for the ease of operating and fixing, in the present embodiment, the rectangular glass plate that preferred dimension is identical。
In the present embodiment, the first groove discharge orifice group 15 includes several the first groove discharge orifices, and the second groove discharge orifice group 14 includes several the second groove discharge orifices, and the 3rd groove discharge orifice group 25 includes several the 3rd groove discharge orifices；Reaction chamber group 21 includes several reaction chambers；Sample feeding mouth group 11 includes several sample feeding mouths；
Sample feeding mouth is arranged on same straight line equably；First groove discharge orifice is arranged on same straight line equably, primer injection port 13 and the first groove discharge orifice are arranged on same straight line, second groove discharge orifice is arranged on same straight line equably, the position of sample feeding mouth and the first groove discharge orifice is corresponding, and the second groove discharge orifice and sample feeding mouth are staggered；3rd groove discharge orifice is arranged on same straight line equably, and reaction chamber is arranged on same straight line equably；Second runner group 24 includes the second runner being positioned on straight line and the second runner being located on the same line with the 3rd groove discharge orifice group 25；
Sample feeding mouth, the first groove discharge orifice, the second groove discharge orifice are paralleled with the central horizontal axle of upper strata carrier 1；Reaction chamber, the 3rd groove discharge orifice are paralleled with the central horizontal axle of lower floor carrier 2。
In the present embodiment, by sample feeding mouth, the first groove discharge orifice, the second groove discharge orifice and upper strata carrier 1 be set to parallel with central horizontal axle；Simultaneous reactions chamber, the 3rd groove discharge orifice are paralleled with the central horizontal axle of lower floor carrier 2 so that the flat board in-situ nucleic acid amplification chip outward appearance that this utility model provides seems more neat。
Second embodiment of the present utility model relates to a kind of flat board in-situ nucleic acid amplification chip, the second embodiment is the improvement of the first embodiment, as shown in Figure 2, it thes improvement is that, upper strata carrier 1 is provided with two groups of sample feeding mouth groups 11, polymerase injection port group the 12, first outlet through hole the 17, second outlet through hole the 18, second groove discharge orifice group 14；Each group of sample feeding mouth group 11, polymerase injection port group the 12, first outlet through hole the 17, second outlet through hole the 18, second groove discharge orifice group 14 are positioned at the side of the central horizontal axle of upper strata carrier 1；
Two groups of sample feeding mouth groups 11, polymerase injection port group the 12, first outlet through hole the 17, second outlet through hole the 18, second groove discharge orifice group 14, central horizontal axle axial symmetry relative to upper strata carrier 1；
Lower floor's carrier 2 is provided with two group reaction chamber groups 21, comb teeth shape passage the 22, the 3rd groove discharge orifice group 25；Each group reaction chamber group 21, comb teeth shape passage the 22, the 3rd groove discharge orifice group 25 are positioned at the side of the central horizontal axle of lower floor's carrier 2；
Two group reaction chamber groups 21, comb teeth shape passage the 22, the 3rd groove discharge orifice group 25 are relative to the central horizontal axle axial symmetry of lower floor's carrier 2。
In the present embodiment, by groove, reaction chamber, runner are disposed relative to the upper and lower layer glass axisymmetric mode of central horizontal axle, the flat board in-situ nucleic acid amplification chip that the present embodiment provides is made to possess more reaction chamber group, make it possible to carry out more group reaction experiment simultaneously, save the time。It is also preferred that the left by sample introduction tank, the second runner group, the first groove discharge orifice group, the centrally disposed trunnion axis of primer injection port group, the quantity of Pocket Machining so can be reduced。
The third embodiment of the present utility model relates to a kind of isothermal nucleic acid amplification device, including the flat board original position isothermal nucleic acid amplification chip of square heat transfer platform 3 and above-mentioned any one；Heat transfer platform 3 is provided with heater；Lower floor's carrier 2 is fixed on heat transfer platform 3, and lower floor's carrier 2 is uniformly heated by heater。
In the present embodiment, the bottom of lower floor's glass is fixed on metal fixing heat transfer platform, and heat transfer platform 3 includes adjacent at least 2 movable block 33, limit assembly 31 and underframe 32；Limit assembly 31 includes sliding axle portion and limiting section；Lower floor's carrier 2 is fixed on underframe 32, and the one end in sliding axle portion runs through that movable block 33 and underframe 32 is fixing to be connected；Movable block 33 can slide in sliding axle portion level；Limiting section is located at the other end in sliding axle portion, and cross-sectional area is more than the cross-sectional area in sliding axle portion。In the present embodiment, best, this movable block 33 is provided with four, as shown in Figure 3 and Figure 4。Namely the distance that movable block can slide determines the distance that upper strata glass can slide。In the present embodiment, limit assembly can be iron nail。In Fig. 4, give the upper strata glass state that slide in a direction wherein, when upper strata glass slides to the right, upper strata glass promotes movable block 33 to move laterally, and when movable block 33 arrives head of a nail position, movable block 33 cannot continue to slide out, and then limit the moving range of upper strata glass, fluid seepage occurs when being prevented from excessively moving upper strata glass in reaction chamber, make the amount of primer or polymerase in reaction chamber change, affect experiment effect。It is also preferred that the left each movable block is fixed on underframe by two iron nails, enable movable block when mobile with underframe edge keeping parallelism。
Meanwhile, in the present embodiment, by be arranged on by lower floor's glass can on the heat transfer platform 3 of uniformly transfer heat, it is achieved this lower floor's carrier 2 is uniformly heated by heater, makes the temperature in each reaction chamber identical, it is achieved experimental situation equivalents。
Utilize above-mentioned isothermal nucleic acid amplification device, additionally it is possible to realize a kind of isothermal nucleic acid amplification method, comprise the following steps:
S0, upper strata carrier 1 is moved to predeterminated position。
Before experiment starts, it is necessary to needing the glass plate used to be placed into initial position experiment, now, as shown in Figure 4, prepare for follow-up interpolation primer in the position of upper strata glass and lower floor's glass。
S1, add primer from primer injection port group 13, and make primer be full of the first groove discharge orifice group 15。
When adding primer premix, add from primer injection port group 13, primer premix passes sequentially through primer injection port group 13, sample introduction tank the 23, second runner group the 24, first groove discharge orifice group 15, first flow 16, and air and unnecessary primer premix in runner are discharged from the first outlet through hole 17。Identical in order to ensure the primer premixing liquid measure being added in reaction chamber, when adding, therefore, to assure that primer premix is full of each first groove discharge orifice。
S2, to reaction chamber group 21 add primer premix time, translation upper strata carrier 1, make the first groove discharge orifice group 15 and the circulation of reaction chamber group 21 phase, now, primer premix flows in reaction chamber group 21 and the 3rd groove discharge orifice group 25 from the first groove discharge orifice 15。
S3, upper strata carrier 1 is moved back to predeterminated position, prepare to add polymerase premix。
S4, add polymerase from polymerase injection port group 12, and make polymerase be full of the second groove discharge orifice group 14。
Add polymerase premix from polymerase injection port group 12, flowed in the 3rd groove discharge orifice group 25 by comb teeth shape passage 22。Air and unnecessary polymerase premix in comb teeth shape passage 22 flow out from the second outlet through hole 18。Identical in order to ensure polymerase premix addition, it is to be ensured that in the 3rd groove discharge orifice group, to be full of polymerase mix。Described polymerase premix includes polymerase, dNTP, Mg2+, reaction buffer。
S5, when adding polymerase in reaction chamber group 21, translation upper strata carrier 1, make reaction chamber group 21 circulate with the second groove discharge orifice group 14 phase, as it is shown in figure 5, polymerase flows in reaction chamber group 21 and the 3rd groove discharge orifice group 25 from the second groove discharge orifice group 14。
S6, upper strata carrier 1 is moved back to predeterminated position。
S7, add sample from sample feeding mouth group 11, make sample be directly entered in reaction chamber。
S8, control heater make reactant liquor keep at the reaction temperatures。In the present embodiment, General reactions temperature is at 60 DEG C～65 DEG C。
When carrying out feminine gender, positive control experiment, mobile upper strata glass is to the position of Fig. 8, and now, the reaction chamber group being positioned at comb teeth shape passage 22 side just will not add polymerase premix。Chip is moved back to initial position, add positive DNA sample, after the response time, the reaction chamber not adding polymerase premix is formed the negative control group without polymerase, forming the negative control group without DNA in the second runner 24 being located on the same line with the 3rd groove discharge orifice group 25, the reaction chamber adding positive DNA sample is positive controls。
In addition, if being respectively provided with axisymmetric through hole, groove or runner on upper strata glass, lower floor's glass, after step S3, again primer injection port group 13 adds primer, and translate upper strata carrier 1, reaction chamber group 21 phase making the first groove discharge orifice group 15 and opposite side circulates, and primer flows in the reaction chamber group 21 of opposite side from the first groove discharge orifice 15。
After the response time of 40 minutes, observe the turbidity of each reaction chamber, can direct qualitative reaction result。8 different DNA sample are carried out isothermal duplication by this example, it is to avoid loaded down with trivial details temperature cycles, substantially reduced the response time, reduce test operation intensity。
The respective embodiments described above are to realize specific embodiment of the utility model, it will be understood by those skilled in the art that and in actual applications, it is possible in the form and details it is done various change, without departing from spirit and scope of the present utility model。
Claims (8)
1. an isothermal nucleic acid amplification device, it is characterised in that include square heat transfer platform (3) and flat board original position isothermal nucleic acid amplification chip；
Described flat board in-situ nucleic acid amplification chip includes square upper layer carrier (1) bonded to each other and lower floor's carrier (2)；Can slide in the upper level of described lower floor carrier (2) in described upper strata carrier (1)；
Described upper strata carrier (1) is provided with sample feeding mouth group (11), polymerase injection port group (12), primer injection port group (13), the first outlet through hole (17), the second outlet through hole (18), the first groove discharge orifice group (15), the second groove discharge orifice group (14), first flow (16)；
Described lower floor carrier (2) is provided with sample introduction tank (23), the second runner (24), reaction chamber group (21), interdigitated electrode structure passage (22), the 3rd groove discharge orifice group (25)；Described reaction chamber group (21) is located between described interdigitated electrode structure passage (22) increment or side, and the increment of the inflow end of described reaction chamber group (21) and described interdigitated electrode structure passage (22) is located along the same line；
When described upper strata carrier (1) and lower floor's carrier (2) are positioned at initial position, circulate mutually in the position of described sample feeding mouth group (11) and described reaction chamber group (21)；Described polymerase injection port group (12) is circulated mutually with described interdigitated electrode structure passage (22), the 3rd groove discharge orifice group (25), the second outlet through hole (18)；Described primer injection port group (13) is circulated mutually with described sample introduction tank (23), the second runner (24), the first groove discharge orifice group (15), first flow (16), the first outlet through hole (17)；
During to described reaction chamber group (21) interpolation primer, translate described upper strata carrier (1), making described first groove discharge orifice group (15) circulate mutually with described reaction chamber group (21), primer flows in described reaction chamber group (21) from described first groove discharge orifice (15)；
During to described reaction chamber group (21) interpolation polymerase, translate described upper strata carrier (1), making described reaction chamber group (21) circulate mutually with described second groove discharge orifice group (14), polymerase flows in described reaction chamber group (21) from described second groove discharge orifice group (14)；
Described heat transfer platform (3) is provided with heater；Described lower floor carrier (2) is fixed on described heat transfer platform (3), and described lower floor carrier (2) is uniformly heated by described heater。
2. isothermal nucleic acid amplification device according to claim 1, it is characterized in that, described first groove discharge orifice group (15) includes several the first groove discharge orifices, described second groove discharge orifice group (14) includes several the second groove discharge orifices, and described 3rd groove discharge orifice group (25) includes several the 3rd groove discharge orifices；Described reaction chamber group (21) includes several reaction chambers；Described sample feeding mouth group (11) includes several sample feeding mouths；
Described sample feeding mouth is arranged on same straight line equably；Described first groove discharge orifice is arranged on same straight line equably, described primer injection port (13) and described first groove discharge orifice are arranged on same straight line, described second groove discharge orifice is arranged on same straight line equably, described sample feeding mouth is corresponding with the position of described first groove discharge orifice, and described second groove discharge orifice and described sample feeding mouth are staggered；Described 3rd groove discharge orifice is arranged on same straight line equably, and described reaction chamber is arranged on same straight line equably；Described second runner group (24) includes the second runner being positioned on straight line and the second runner being located on the same line with described 3rd groove discharge orifice group (25)；
The straight line at described sample feeding mouth place, the straight line at the first groove discharge orifice place, the straight line at the second groove discharge orifice place are paralleled with the central horizontal axle on described upper strata carrier (1)；The straight line at described reaction chamber place, the straight line at the 3rd groove discharge orifice place are paralleled with the central horizontal axle of described lower floor carrier (2)。
3. isothermal nucleic acid amplification device according to claim 2, it is characterized in that, described upper strata carrier (1) is provided with sample feeding mouth group (11) described in two groups, polymerase injection port group (12), the first outlet through hole (17), the second outlet through hole (18), the second groove discharge orifice group (14)；Each group of sample feeding mouth group (11), polymerase injection port group (12), the first outlet through hole (17), the second outlet through hole (18), the second groove discharge orifice group (14) are positioned at the side of the central horizontal axle on described upper strata carrier (1)；
Sample feeding mouth group (11) described in two groups, polymerase injection port group (12), the first outlet through hole (17), the second outlet through hole (18), the second groove discharge orifice group (14), central horizontal axle axial symmetry relative to described upper strata carrier (1)；
Described lower floor carrier (2) is provided with reaction chamber group (21) described in two groups, interdigitated electrode structure passage (22), the 3rd groove discharge orifice group (25)；Reaction chamber group (21) described in each group, interdigitated electrode structure passage (22), the 3rd groove discharge orifice group (25) are positioned at the side of the central horizontal axle of described lower floor carrier (2)；
Reaction chamber group (21) described in two groups, interdigitated electrode structure passage (22), the 3rd groove discharge orifice group (25) are relative to the central horizontal axle axial symmetry of described lower floor carrier (2)。
4. isothermal nucleic acid amplification device according to claim 3, it is characterized in that, described primer injection port (13) and described first groove discharge orifice are arranged on the central horizontal axle on described upper strata carrier (1), and described sample introduction tank is positioned on the central horizontal axle of described lower floor carrier (2)。
5. flat board in-situ nucleic acid amplification chip according to claim 2, it is characterised in that when overlooking described first groove discharge orifice, the second groove discharge orifice, the 3rd groove discharge orifice, described first groove discharge orifice is cross；Described second groove discharge orifice, the 3rd groove discharge orifice are rectangle。
6. isothermal nucleic acid amplification device according to claim 1, it is characterised in that described heat transfer platform (3) includes adjacent at least 2 movable block (33), limit assembly (31) and underframe (32)；Described limit assembly (31) includes sliding axle portion and limiting section；
Described lower floor carrier (2) is fixed on described underframe (32), and the one end in described sliding axle portion runs through that described movable block (33) and described underframe (32) are fixing to be connected；Described movable block (33) can slide in described sliding axle portion level；Described limiting section is located at the other end in described sliding axle portion, and cross-sectional area is more than the cross-sectional area in described sliding axle portion。
7. isothermal nucleic acid amplification device according to claim 6, it is characterised in that each described movable block is provided with at least 2 described limit assemblies (31)。
8. isothermal nucleic acid amplification device according to claim 6, it is characterised in that described limit assembly (31) is iron nail。
Priority Applications (1)
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105400692A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105400692A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental method |
| CN105400692B (en) * | 2015-12-15 | 2017-06-27 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental technique |
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