CN107142194B - Multiplex test piece device and operation method thereof - Google Patents
Multiplex test piece device and operation method thereof Download PDFInfo
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- CN107142194B CN107142194B CN201610216279.6A CN201610216279A CN107142194B CN 107142194 B CN107142194 B CN 107142194B CN 201610216279 A CN201610216279 A CN 201610216279A CN 107142194 B CN107142194 B CN 107142194B
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- 238000012360 testing method Methods 0.000 title claims abstract description 146
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- 239000007924 injection Substances 0.000 claims abstract description 78
- 239000012488 sample solution Substances 0.000 claims abstract description 62
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- 239000003921 oil Substances 0.000 claims description 20
- 239000012530 fluid Substances 0.000 claims description 16
- 238000011017 operating method Methods 0.000 claims description 16
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 8
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- 239000000155 melt Substances 0.000 claims description 4
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- 229920002545 silicone oil Polymers 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract description 10
- 238000012546 transfer Methods 0.000 abstract description 2
- 238000011990 functional testing Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 41
- 238000003752 polymerase chain reaction Methods 0.000 description 15
- 238000001514 detection method Methods 0.000 description 13
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- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 1
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
Abstract
The invention provides a multiplex test piece device and an operation method thereof. The multiplex test piece device comprises a test piece and a sacrificial layer. The test strip has a plurality of reaction vessels arranged in an array, each reaction vessel having an opening and a bottom. The sacrificial layer is provided with a micro-channel, and the micro-channel is provided with an injection channel, a main channel and a tail end channel which are communicated with each other. The sacrificial layer is suitable for assembling with the test piece, wherein the main flow channel is assembled facing the opening part. Injecting a sample solution into the injection flow channel such that the sample solution flows from the injection flow channel through the main flow channel to the end flow channel, wherein the sample solution is loaded into each of the reaction vessels while flowing through the main flow channel. By means of the multi-functional test strip device and the operation method thereof of the present invention, the sample solution can be rapidly and uniformly loaded into each reaction container of the test strip, and all the reaction containers can be filled in a short time by a single back-transfer liquid operation.
Description
Technical field
The present invention relates to a kind of multiplexing test strip device and its operating methods, are detected with being applied to molecular biosciences, more particularly to
One kind is for polymerase chain reaction (PCR), the multiplexing test strip device that can fill pcr reagent in advance and its operation
Method.
Background technique
In molecular biosciences detection field, it is often necessary to carry out the inspection of a variety of project targets for a sample.For example,
It is detected by polymerase chain reaction, to examine the genotype of several single nucleotide polymorphism (SNP) in a sample, or is examined
Multiple genes show degree in one sample.(assay) is examined to form an inspection by several DNA or RNA at this point, just may require that
Set group (panel).Including at least two specific DNA primer molecules in PCR inspection in every group of testing reagent, (certain PCR are examined
In also additionally include the specific reporter probe of target (reporter probes)), this to introduction (introduction to) must correctly with
The DNA profiling mixing extracted from test sample to be carried out (sample), can verify whether specific DNA target deposits in sample
Or existing amount be how many.
Traditionally, introduction to and sample be placed in identical reaction vessel to carry out PCR.General storing mode is usually
Introduction single bottle stored by pipette to, enzyme and dNTP mixture and buffering agents and sample one by one in a manual manner
It is pipetted into reaction vessel with pipette.The most common support container is 96 porose discs.In the way of aforementioned placement, PCR detect to
The pipette pipetting of two bouts (twice) is needed less, wherein an addition sample is into reaction vessel, and another time addition
Introduction is into reaction vessel.For example, it is assumed that checking 36 targets in a sample using an inspection set group, then need
Will at least 36 times pipettings to add respective introduction into 36 different reaction vessels, in addition, it is necessary to 36 return liquid behaviour
Make to add sample to above-mentioned each reaction vessel.Such mode of operation is not only complicated to be easy error, but also must consume a large amount of people
Power.
If being pre-charged with introduction pair in individual reaction vessels, experimental implementation person only needs to add sample to filling out in advance
In the container substituted the bad for the good.For previous example, 36 targets of a sample are tested, will only need 36 times pipetting addition samples
Product are into 36 reaction vessels being pre-charged with.In addition, reaction vessel volume can be reduced simultaneously to about nanoliter (nano-
Liter in the range of), to save detection reagent usage amount.Under this improvement, common 96 porose discs as support container, sample
Formula will be changed to the titer plate of similar test piece.
However, the size and volume of the reaction vessel (also referred to as micro- well or nanometer wellhole) of titer plate are too small, it is difficult to hand
Dynamic filling introduction pair or sample used simultaneously avoid the cross contamination between adjacent reaction container simultaneously (i.e. introduction are to from one
Well dissipates to other wells), therefore, it is necessary to special microfluid ingredients to dispense technology.More specifically, introduction is provided previously to extremely
Each nanometer well and fixed introduction is in the surface of this nanometer of well.Later, user can single pipetting or by single
Microfluidic channel adds sample into each reaction vessel, and need not worry that introduction is emitted to other wells from a well, makes well and well
Between cross contamination minimize.
When subsequent progress sample detection, the sample of predetermined amount must be filled in each reaction vessel.Traditional method is to use
Sample is added in reaction well by transfer pipet(te) or needle-shaped dispenser one by one.However, as reaction vessel volume becomes smaller, between well
Apart from closer, dispenser or the path of special mechanical mechanism are designed, being sent to each reaction vessel individually is complicated and consumes
When.If to reach single pipetting or adding sample by single microfluidic channel by special test strip device design
Into each reaction vessel, manual operation needed for polymerase chain reaction prepares detection reagent will can be greatly simplified, and increase and fill out
Fill the convenience of sample.
Summary of the invention
The present invention provides a kind of multiplexing test strip device and its operating methods, are suitable for molecular biosciences and detect；In particular, with
In polymerase chain reaction；And more specifically, it is applied to real-time polymerase chain reaction.By multiplexing test strip device of the invention and its
Operating method, sample can be rapidly and uniformly loaded into each reaction vessel of test piece, with single time pipetting very short
Time in just can fill all reaction vessels.
The present invention provides a kind of multiplexing test strip device comprising test piece and sacrificial layer.Test piece has multiple reaction vessels, the
One injection hole and first row stomata.Reaction vessel is arranged as array, each reaction vessel tool opening portion and bottom.Sacrificial layer has
Fluid channel, fluid channel have interconnected injection runner, main flow paths and end runner.Sacrificial layer is suitable for assembling with test piece,
Wherein main flow paths are assembled in face of opening portion.Sample solution is injected to injection runner, so that sample solution from the first injection hole
Main flow paths are flowed through to end runner from injection runner, and wherein sample solution is loaded into each reaction and holds when flowing through main flow paths
In device.
In one embodiment of this invention, multiplexing test strip device further includes the shell to accommodate test piece and sacrificial layer, shell
Body has the second injection hole and second row stomata, wherein sequentially injecting sample solution to note from the second injection hole and the first injection hole
Air stream enter runner, so that sample solution flows through main flow paths to end runner from injection runner, wherein sample solution is flowing through main stream
When road, it is loaded into each reaction vessel.
In one embodiment of this invention, the material of shell includes Heat Conduction Material.
In one embodiment of this invention, shell includes upper cover and bottom plate, and upper cover is suitable for assembling with bottom plate, and upper cover has
To accommodate the groove of test piece and sacrificial layer, the second injection hole and second exhaust hole are located in upper cover.
In one embodiment of this invention, the material of test piece includes translucent material.
In one embodiment of this invention, translucent material includes polycarbonate or polymethyl methacrylate.
In one embodiment of this invention, the material of sacrificial layer includes wax.
The present invention provides a kind of multiplexing test strip device comprising test piece and sacrificial layer.Test piece has multiple reaction vessels, and
Reaction vessel is arranged as array, and each reaction vessel has opening portion and bottom.Sacrificial layer has fluid channel, and fluid channel has phase
Intercommunicated injection runner, main flow paths and end runner.Sacrificial layer is suitable for assembling with test piece, and wherein main flow paths are in face of opening
Portion and assemble.Sample solution is injected into injection runner, is flowed so that sample solution flows through main flow paths to end from injection runner
Road, wherein sample solution is loaded into each reaction vessel when flowing through main flow paths.
It in one embodiment of this invention, further include shell to accommodate test piece and sacrificial layer, shell has injection hole
With gas vent, wherein from injection hole inject sample solution to injection runner so that sample solution from injection runner flow through main stream
Road to end runner, wherein sample solution is loaded into each reaction vessel when flowing through main flow paths.
In one embodiment of this invention, the material of shell includes Heat Conduction Material.
In one embodiment of this invention, shell includes upper cover and bottom plate, and upper cover is suitable for assembling with bottom plate, and upper cover has
To accommodate the groove of test piece and sacrificial layer, injection hole and gas vent are located in upper cover.
In one embodiment of this invention, the material of test piece includes translucent material.
In one embodiment of this invention, translucent material includes polycarbonate or polymethyl methacrylate.
In one embodiment of this invention, the material of sacrificial layer includes wax.
The present invention provides a kind of operating method of multiplexing test strip device, including assembling comprising test piece, sacrificial layer and to hold
Set the multiplexing test strip device of the shell of test piece and sacrificial layer.Test piece has the multiple reaction vessels for being arranged as array, each reaction
Container has opening portion and bottom, and sacrificial layer has fluid channel, and fluid channel has interconnected injection runner, main flow paths and end
Runner is held, wherein main flow paths are assembled in face of opening portion.Then, sample solution is injected into injection via the injection hole of shell
Runner, so that sample solution flows through main flow paths to end runner from injection runner, wherein sample solution is flowing through main flow paths
When, it is loaded into each reaction vessel.Later, oil is injected into injection runner via the injection hole of shell, so that oil is flowed from injection
Main flow paths are flowed through to end runner, wherein oil when flowing through main flow paths, is excluded without the sample being loaded in reaction vessel in road
Solution.Next, heating so that sacrificial layer melts, the sacrificial layer after fusing is mixed with oil.
In one embodiment of this invention, oil includes mineral oil or silicone oil.
In one embodiment of this invention, the material of shell includes Heat Conduction Material.
In one embodiment of this invention, the material of test piece includes translucent material.
In one embodiment of this invention, translucent material includes polycarbonate or polymethyl methacrylate.
In one embodiment of this invention, the material of sacrificial layer includes wax.
Based on above-mentioned, the present invention provides a kind of multiplexing test strip device and its operating methods, and sample is made to flow through sacrificial layer
Main flow paths when, rapidly and uniformly be loaded into test piece each reaction vessel in, then by oil exclude without be loaded in reaction hold
Sample solution in device.In this way, all reaction vessels can be filled just in a short period of time with single time pipetting,
Therefore, experimental implementation can be simplified and save the time.
On the other hand, test piece is spaced at a distance from least about 10 μm between sacrificial layer, and sacrificial layer has certain thickness.In
In the experimentation of polymerase chain reaction, heat so that sacrificial layer melts, the sacrificial layer after fusing is mixed with oil, and then makes test piece
Have at a distance from about 600 μm between bottom plate, so that reaction is gone on smoothly.Due to being not required to excessive addition sample, test piece can be made
Specific range is maintained between bottom plate, therefore, has the function of saving sample additive amount.
To make the foregoing features and advantages of the present invention clearer and more comprehensible, special embodiment below, and it is detailed to cooperate attached drawing to make
Carefully it is described as follows.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of multiplexing test strip device according to a first embodiment of the present invention；
Fig. 2 is the structural schematic diagram of test piece according to a first embodiment of the present invention；
Fig. 3 is the diagrammatic cross-section of the reaction vessel of test piece according to a first embodiment of the present invention；
Fig. 4 is the schematic diagram that multiplexing test strip device according to a first embodiment of the present invention is applied to that sample solution loads；
Fig. 5 is the schematic diagram that multiplexing test strip device according to a second embodiment of the present invention is loaded applied to sample solution；
Fig. 6 A to Fig. 6 C is the schematic diagram of the operating method of multiplexing test strip device according to the present invention.
Appended drawing reference:
10: test piece
12,42: injection hole
14,44: gas vent
16: double stage structures
20: sacrificial layer
22: injection runner
24: main flow paths
26: end runner
28: fluid channel
30: shell
40: upper cover
50: bottom plate
60: space
70a, 70b: sample solution
80: oil
100: detection zone
102,160a, 160b, 160c, 160d, 160e, 160f: reaction vessel
102a: opening portion
102b: bottom
X: the first rank
Y: second-order
Specific embodiment
The present invention provides a kind of multiplexing test strip device and its operating method, can be widely applied to different types of reaction
It examines.Hereinafter, it is first defined and is illustrated for noun used in text in specification.
" reagent " can refer to the formula or preparation for the several ingredients of fc-specific test FC/inspection.For example, using polymerization
In the test of enzyme chain reaction, detection reagent includes a pair of of introduction, enzyme, dNTPs, fluorescence report object and salt etc..In application program
In, different introductions to and fluorescence report object may first be added in reaction vessel, be thirdly sample and enzyme, dNTP and its
The mixing of its additive is then added in reaction vessel.
" sample " refers to tested nucleic acid samples.It is mentioned from the sources such as blood, tissue, saliva for example, sample can be
The nucleic acid fragment (including DNA or RNA etc.) taken.
" analysis " or " test " can refer to same sample carry out one or more experiment or test item.For example, using PCR
300 single nucleotide polymorphism (300SNP) genotype is detected to be directed to nucleic acid samples, which includes several PCR detection projects,
By each genotype (A, T, C, G) for checking each single nucleotide polymorphism (SNP).For example, using real-time fluorescence quantitative PCR
Determine the nucleic acid amount an of particular sequence.
" sample solution " refers to the mixture or mixed solution of above-mentioned " sample " Yu mix reagent (master mix).
" reaction vessel " can refer to each pipe, each reaction tube, the hole of titer plate or the well of pipe dish, or test test piece or array
Pit/hole of plate.As described herein, " test piece ", " test piece plate ", " detection array board " or " detection plate " can refer to accommodate aforementioned anti-
Answer the same substrate or substrate of container.
When liquid volume in a reservoir reduces to a certain extent, liquid flowing in a reservoir is mainly by surface
Adhesive force and non-gravity is controlled.If the liquid volume in container only has several nanoliters, liquid surface adhesion with higher
Property can adhere on container (Na Jing), that is to say, that liquid can be considered as bonding agent stabilization and be attached to container bottom at this time
Or on chamber wall.
Preferably, reaction vessel can be test piece or detect the single reacting hole or well of array board.As front institute
It discusses, is preferably using the reaction vessel of smaller volume, size is for example differed from several nanoliters to several hundred nanoliters.
Fig. 1 is the structural schematic diagram of multiplexing test strip device according to a first embodiment of the present invention.
Fig. 1 is please referred to, multiplexing test strip device includes test piece 10, sacrificial layer 20 and shell 30, and wherein shell 30 can be used to
Accommodate test piece 10 and sacrificial layer 20.Hereinafter, it will be described in detail with initial reference to Fig. 2 and Fig. 3 for the structure of test piece 10.
Fig. 2 is the structural schematic diagram of test piece according to a first embodiment of the present invention.
Referring to figure 2., test piece 10 has a detection zone 100, and the area of detection zone 100 be, for example, 22.5 millimeters ×
22.5 millimeters.It include multiple reaction vessels 102 in detection zone 100, and reaction vessel 102 is arranged as n × n array.In addition, examination
Piece 10 is with more injection hole 12 and gas vent 14.In the present embodiment, the size of test piece 10 is, for example, 42 millimeters × 36 millimeters × 2
Millimeter.More specifically, the material of test piece 10 may include translucent material, and translucent material be, for example, polycarbonate (PC) or
Polymethyl methacrylate (PMMA).
Referring again to Fig. 2, from the point of view of the cross-sectional view (right part of Fig. 2) of test piece 10, each reaction vessel 102 have compared with
Wide opening portion 102a and relatively narrow bottom 102b.In the present embodiment, the depth d1 of each reaction vessel 102 is, for example, 100
Micron, the size of opening portion 102a is, for example, 200 microns (L1) × 185 micron (W1), and the size of bottom 102b is, for example,
106.74 microns (L2) × 91.74 micron (W2).Spacing P1 range between reaction vessel 102 is, for example, 25 microns to 40 micro-
Rice.The angle, θ of the sloped sidewall of reaction vessel 102 is, for example, 90 degree to 140 degree, more excellent preferably between 100 degree to 135 degree
It is selected between 110 degree to 120 degree.Each reaction vessel 102 can accommodate such as 2.1 nanoliters of sample solution.
Fig. 3 is the diagrammatic cross-section of the reaction vessel of test piece according to a first embodiment of the present invention.
Referring to figure 3., the reaction vessel of test piece 10 can be designed to the shapes or configure mode for having different.Citing and
Speech, reaction vessel 160a and 160b are the groove formed in test piece 10 but have no through test piece 10.It is reaction vessel 160b, anti-
Answer container 160d and reaction vessel 160f that there is sloped sidewall.Reaction vessel 160c, reaction vessel 160d, reaction vessel 160e and
Reaction vessel 160f runs through test piece 10 and there are two open ends on the top and bottom of test piece 10.Due to capillarity, sample
Liquid can be steady stagnant in reaction vessel 160c, reaction vessel 160d, reaction vessel 160e and reaction vessel 160f.Reaction vessel
160d penetrates test piece 10 and there are two open ends for tool on the top and bottom of test piece 10, and has two of inclined side wall connection
Open end.
However, structure chart shown in Fig. 2 and Fig. 3 is only used for illustrating the present embodiment, the shape of reaction vessel of the present invention
Shape, size or quantity are not limited thereto, and the cross-sectional shape of reaction vessel can also be such as circle, rectangular or polygon
Shape.
Under normal circumstances, introduction is dissolved in aqueous solvent or solution, and test piece of the invention may be designed as the interior of reaction vessel
Wall and bottom surface are hydrophilic, and are hydrophobic between each reaction vessel.Reagent or probe will only be attached to hydrophily area
Domain, that is to say, that be only adhered to the inner wall and bottom surface of reaction vessel.The size of each reaction vessel can be less than 1 milli
Rice.With this size scale, a small amount of sample liquids overflow can fill a large amount of reaction vessel in 10 seconds, significantly improve
Sample efficiency of loading.
Fig. 4 is the schematic diagram that multiplexing test strip device according to a first embodiment of the present invention is applied to that sample solution loads.It connects
, hereinafter, by referring concurrently to Fig. 1, Fig. 2 and Fig. 4 to the structure of the multiplexing test strip device of first embodiment of the invention and its
It is illustrated in the application that sample solution loads.
Fig. 1 is please referred to, sacrificial layer 20 has fluid channel 28, and fluid channel 28 has interconnected injection runner 22, master
Want runner 24 and end runner 26.In the present embodiment, the material of sacrificial layer 20 may include wax, therefore, in polymerase chain reaction
In be heated to about it is fusible at 60 DEG C.However, the present invention is not limited thereto, any fusible temperature range also can be used to be higher than
The material of room temperature to 60 DEG C, and preferably using can melting temperatur be about 60 DEG C material.More specifically, the size example of sacrificial layer 20
37.6 millimeters in this way × 39.6 millimeters × 1.3 millimeters, the depth of fluid channel 28 is, for example, 0.12 millimeter, the size of main flow paths 24
E.g. 22.5 millimeters × 22.5 millimeters.
Referring to Fig. 1 and Fig. 2, sacrificial layer 20 is suitable for assembling with test piece 10, and wherein main flow paths 24 face test piece 10
The opening portion 102a of middle reaction vessel 102 and assemble.It, can be from 12 note of injection hole of test piece 10 referring to Fig. 1, Fig. 2 and Fig. 4
Enter sample solution to injection runner 22, so that sample solution flows through main flow paths 24 to end runner 26 from injection runner 22,
Middle sample solution is loaded into the (flow direction of sample solution in each reaction vessel 102 of test piece 10 when flowing through main flow paths 24
As shown in the dotted arrow in Fig. 4).
In Fig. 2, injection hole 12 and the gas vent 14 of test piece 10 are configured with diagonal form, but the present invention not as
The configuration mode of limit, injection hole 12 and gas vent 14 also can be according to the injection runner 22 of sacrificial layer 20 and the configuration side of end runner 26
Formula and adjusted, as long as can make sample solution in flow process sufficiently extend.
As shown in Figure 1, shell 30 may include upper cover 40 and bottom plate 50, wherein upper cover 40 is suitable for assembling with bottom plate 50.More in detail
For thin, the size of upper cover 40 is, for example, 45mm × 43mm × 5mm, and the size of bottom plate 50 is, for example, 42mm × 40mm × 2mm.In
In the present embodiment, upper cover 40 can have the groove to accommodate test piece 10 and sacrificial layer 20, and injection hole 42 and gas vent 44 can
In upper cover 40.However, the structure of shell 30 is not limited thereto in the present invention, it also any other that can be used can accommodate examination
The structure of piece 10 and sacrificial layer 20.For example, shell 30 is also possible to integrated formed structure, wherein having trip and can open
It closes to be placed in test piece 10 and sacrificial layer 20.In addition, guide groove push structure can also be used to be placed in test piece 10 and sacrificial layer 20
In shell 30.
More specifically, shell 30 has the function of thermally conductive in polymerase chain reaction, and material may include Heat Conduction Material,
Heat Conduction Material is, for example, the metals such as aluminium or copper, graphite or wafer, but the present invention is not limited thereto.In addition, shell 30 has more
Test piece 10 and sacrificial layer 20 is set to be isolated from the outside world, to avoid affected effect is reacted.
In the present embodiment, shell 30 may include mark label (not shown), when multiplexing test strip device application of the invention
When being provided with instrument (such as: thermal cycler) of tag reader, tag reader can read the mark on shell 30
Indicating label, wherein mark label is, for example, hand marking, bar code or other labels, but the present invention is not limited thereto, also may be used
On demand and the tag reader used of arranging in pairs or groups selects applicable mark label.
Referring to Fig. 1, Fig. 2 and Fig. 4, can sequentially be injected from the injection hole 42 of shell 30 and the injection hole 12 of test piece 10
Sample solution is to injection runner 22, so that sample solution flows through main flow paths 24 to end runner 26 from injection runner 22, wherein
For sample solution when flowing through main flow paths 24, (flow direction of sample solution is such as in each reaction vessel 102 of loading test piece 10
Shown in dotted arrow in Fig. 4).In addition, as shown in figure 4, test piece 10 can be double with more double stage structures 16 positioned at its edge
Space 60 can be formed between stage structure 16 and sacrificial layer 20, upper cover 40 and bottom plate 50, space 60 has the work for preventing bubble from generating
With.In the present embodiment, double stage structures 16 can have the first rank x and second-order y, wherein the thickness of the first rank x and second-order y
Ratio is, for example, 1:1, and the thickness of the first rank x and second-order y is for example 1mm respectively, and overall thickness is, for example, 2mm.
In Fig. 1, the injection hole 42 and gas vent 44 of upper cover 40 are configured with diagonal form, but the present invention not as
Limit.The configuration mode of injection hole 42 and gas vent 44 be according to test piece 10 injection hole 12 and gas vent 14 configuration mode and
It is fixed.That is, as described above, can be adjusted according to the injection runner 22 of sacrificial layer 20 and the configuration mode of end runner 26, only
It wants that sample solution can be made sufficiently to extend in flow process.
Fig. 5 is the schematic diagram that multiplexing test strip device according to a second embodiment of the present invention is loaded applied to sample solution.Fig. 5
Shown in second embodiment be similar to Fig. 1, Fig. 2, Fig. 3 and first embodiment shown in Fig. 4, therefore same components are with identical label table
Show and it will not be described here.
The present embodiment and above-mentioned first embodiment the difference is that, test piece 10 does not have injection hole 12 and gas vent 14,
And test piece 10 does not have double stage structures 16 positioned at its edge.In addition, the size of sacrificial layer 20 is greater than the size of test piece 10.It lifts
For example, the size of test piece 10 can be identical as above-mentioned first embodiment, and the size of main flow paths 24 is, for example, in sacrificial layer 20
Greater than 22.5 millimeters × 22.5 millimeters.
Referring to figure 5., can from the injection hole 42 of shell 30 inject sample solution to injection runner 22 so that sample solution from
Injection runner 22 flows through main flow paths 24 to end runner 26, and wherein sample solution is loaded into test piece when flowing through main flow paths 24
In 10 each reaction vessel 102 (flow direction of sample solution is as shown in the dotted arrow in Fig. 5).
The present invention more provides a kind of operation side of multiplexing test strip device described in above-mentioned first embodiment and second embodiment
Method.Fig. 6 A to Fig. 6 C is the schematic diagram of the operating method of multiplexing test strip device according to the present invention.Then, hereinafter, will join
It is illustrated according to operating method of Fig. 6 A to Fig. 6 C to multiplexing test strip device of the invention.It should be noted that related multiplexing examination
The details that the structure and sample solution of sheet devices load is in above describing in detail, therefore it will not be described here.
Firstly, assembling multiplexing test strip device, the multiplexing test strip device of assembled completion is through such as dispensing mode fixing seal
To maintain airtight conditions, sample solution is avoided to flow outward.
Then, by pipetting (pipetting) or other applicable liquid dispensers, by sample solution via shell
Injection hole be injected into injection runner so that sample solution from injection runner flow through main flow paths to end runner.Such as Fig. 6 A institute
Show, sample solution 70a and sample solution 70b are loaded into each reaction vessel 102 when flowing through main flow paths 24.In more detail and
Speech, the total amount of adding of sample solution is, for example, 60 μ l.
Later, oil is injected into injection runner via the injection hole of shell, so that oil flows through main flow paths from injection runner
To end runner.As shown in Fig. 6 B to Fig. 6 C, oil is excluded without being loaded in reaction vessel 102 80 when flowing through main flow paths 24
Sample solution 70b.More specifically, oil is, for example, mineral oil or silicone oil.
Next, in the experimentation of polymerase chain reaction, heat so that sacrificial layer melts, the sacrificial layer after fusing with
Oil mixing, wherein the heated temperature with thawing of sacrificial layer is, for example, about 60 DEG C.It should be noted that test piece of the invention with it is sacrificial
The distance of at least about 10 μm (e.g. 10 μm to 50 μm) is spaced between domestic animal layer, and sacrificial layer has certain thickness (e.g. 550
μm to 590 μm of thickness).Therefore, when the sacrificial layer after fusing is mixed with oil, can make to have about between test piece and bottom plate
600 μm of distance, so that reaction is gone on smoothly.Due to being not required to excessive addition sample, can make to remain special between test piece and bottom plate
Therefore set a distance has the function of saving sample additive amount.
In conclusion the present invention provides a kind of multiplexing test strip device suitable for molecular biosciences detection and its operation sides
Method makes sample solution in the main flow paths for flowing through sacrificial layer, is rapidly and uniformly loaded into each reaction vessel of test piece, then
It excludes by oil without the sample solution being loaded in reaction vessel.In this way, with single time pipetting in the very short time
All reaction vessels just can be inside filled, therefore, experimental implementation can be simplified and save the time.Further, since being not required to add
Sample is measured, can make to maintain specific range between test piece and bottom plate, therefore, the present invention is with more the work for saving sample additive amount
With.
Although the present invention is disclosed as above with embodiment, however, it is not to limit the invention, any technical field
Middle those of ordinary skill, it is without departing from the spirit and scope of the present invention, therefore of the invention when can make a little change and retouching
Protection scope is subject to view appended claims confining spectrum.
Claims (18)
1. a kind of multiplexing test strip device characterized by comprising
Test piece has multiple reaction vessels, the first injection hole and first row stomata, and the reaction vessel is arranged as array, each
The reaction vessel tool opening portion and bottom；
Sacrificial layer has fluid channel, and the fluid channel has interconnected injection runner, main flow paths and end runner, and
The sacrificial layer is suitable for assembling with the test piece, wherein the main flow paths assemble, the sacrificial layer in face of the opening portion
Material include wax；And
Shell, to accommodate the test piece and the sacrificial layer, the shell has the second injection hole and second row stomata,
Wherein sequentially from second injection hole and first injection hole injection sample solution to the injection runner, so that institute
It states sample solution and flows through the main flow paths to the end runner, wherein the sample solution is flowing through from the injection runner
When the main flow paths, it is loaded into each reaction vessel.
2. multiplexing test strip device according to claim 1, which is characterized in that the material of the shell includes Heat Conduction Material.
3. multiplexing test strip device according to claim 1, which is characterized in that the shell includes upper cover and bottom plate, described
Upper cover is suitable for assembling with the bottom plate, and the upper cover has the groove to accommodate the test piece and the sacrificial layer, described
Second injection hole and the second exhaust hole are located in the upper cover.
4. multiplexing test strip device according to claim 1, which is characterized in that the shell includes mark label.
5. multiplexing test strip device according to claim 1, which is characterized in that the material of the test piece includes translucency material
Material.
6. multiplexing test strip device according to claim 5, which is characterized in that the translucent material include polycarbonate or
Polymethyl methacrylate.
7. a kind of multiplexing test strip device characterized by comprising
Test piece, has multiple reaction vessels, and the reaction vessel is arranged as array, each reaction vessel tool opening portion and bottom
Portion；
Sacrificial layer has fluid channel, and the fluid channel has interconnected injection runner, main flow paths and end runner, and
The sacrificial layer is suitable for assembling with the test piece, wherein the main flow paths assemble, the sacrificial layer in face of the opening portion
Material include wax；And
Shell, to accommodate the test piece and the sacrificial layer, the shell has injection hole and gas vent,
It wherein is injected into the injection runner from the injection hole by sample solution, so that the sample solution is flowed from the injection
The main flow paths are flowed through to the end runner, wherein the sample solution when flowing through the main flow paths, is loaded into often in road
In one reaction vessel.
8. multiplexing test strip device according to claim 7, which is characterized in that the material of the shell includes Heat Conduction Material.
9. multiplexing test strip device according to claim 7, which is characterized in that the shell includes upper cover and bottom plate, described
Upper cover is suitable for assembling with the bottom plate, and the upper cover has the groove to accommodate the test piece and the sacrificial layer, described
Injection hole and the gas vent are located in the upper cover.
10. multiplexing test strip device according to claim 7, which is characterized in that the shell includes mark label.
11. multiplexing test strip device according to claim 7, which is characterized in that the material of the test piece includes translucency material
Material.
12. multiplexing test strip device according to claim 11, which is characterized in that the translucent material includes polycarbonate
Or polymethyl methacrylate.
13. a kind of operating method of multiplexing test strip device characterized by comprising
Assemble multiplexing test strip device, the multiplexing test strip device include test piece, sacrificial layer and to accommodate the test piece with it is described
The shell of sacrificial layer, the test piece, which has, is arranged as multiple reaction vessels of array, each reaction vessel tool opening portion and
Bottom, the sacrificial layer have fluid channel, and the fluid channel has interconnected injection runner, main flow paths and end stream
Road, wherein the main flow paths are assembled in face of the opening portion, the material of the sacrificial layer includes wax；
Sample solution is injected into the injection runner via the injection hole of the shell, so that the sample solution is from the note
Air stream enter runner flows through the main flow paths to the end runner, wherein the sample solution when flowing through the main flow paths, carries
Enter in each reaction vessel；
Oil is injected into the injection runner via the injection hole of the shell, so that the oil flows through institute from the injection runner
Main flow paths are stated to the end runner, wherein the oil when flowing through the main flow paths, is excluded without being loaded in the reaction
The sample solution in container；And
It heats so that the sacrificial layer melts, the sacrificial layer after fusing is mixed with the oil.
14. the operating method of multiplexing test strip device according to claim 13, which is characterized in that the oil includes mineral oil
Or silicone oil.
15. the operating method of multiplexing test strip device according to claim 13, which is characterized in that the material packet of the shell
Include Heat Conduction Material.
16. the operating method of multiplexing test strip device according to claim 13, which is characterized in that the shell includes mark
Label.
17. the operating method of multiplexing test strip device according to claim 13, which is characterized in that the material packet of the test piece
Include translucent material.
18. the operating method of multiplexing test strip device according to claim 17, which is characterized in that the translucent material packet
Include polycarbonate or polymethyl methacrylate.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW105106095A TWI639705B (en) | 2016-03-01 | 2016-03-01 | Multiplex slide plate device and operation method thereof |
| TW105106095 | 2016-03-01 |
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| Publication Number | Publication Date |
|---|---|
| CN107142194A CN107142194A (en) | 2017-09-08 |
| CN107142194B true CN107142194B (en) | 2019-11-05 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610216279.6A Active CN107142194B (en) | 2016-03-01 | 2016-04-08 | Multiplex test piece device and operation method thereof |
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| CN (1) | CN107142194B (en) |
| TW (1) | TWI639705B (en) |
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|---|---|---|---|---|
| TWI669390B (en) * | 2018-08-08 | 2019-08-21 | 奎克生技光電股份有限公司 | Multiplex slide plate device having storage tank |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2257802A4 (en) * | 2008-02-29 | 2014-07-02 | Univ Northwestern | Barriers for facilitating biological reactions |
| US9724692B2 (en) * | 2013-06-27 | 2017-08-08 | Quark Biosciences, Inc. | Multiplex slide plate |
-
2016
- 2016-03-01 TW TW105106095A patent/TWI639705B/en active
- 2016-04-08 CN CN201610216279.6A patent/CN107142194B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| TWI639705B (en) | 2018-11-01 |
| CN107142194A (en) | 2017-09-08 |
| TW201732043A (en) | 2017-09-16 |
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