CN109022274A - Micro-fluidic chip, chip steerable system, micro fluidic device and method for extracting nucleic acid - Google Patents

Abstract

The invention discloses a kind of nucleic acid extraction micro-fluidic chip, chip steerable system, micro fluidic device and method for extracting nucleic acid.Micro-fluidic chip includes at least one storage chamber, at least one reagent connection control unit and at least one reaction chamber, each storage chamber is correspondingly arranged with a reaction chamber, and reagent connection control unit is provided between each storage chamber and corresponding reaction chamber to control whether the two is connected to, wherein, at least one storage chamber is packaged with the reagent for nucleic acid extraction.The present invention can simplify the operating procedure of nucleic acid extraction, conducive to the automation for realizing nucleic acid extraction, save the operating time of nucleic acid extraction, improve the efficiency of nucleic acid extraction, reduce error in operation probability.

Description

Micro-fluidic chip, chip steerable system, micro fluidic device and method for extracting nucleic acid

Technical field

The present invention relates to biomedical, diagnosis and detection technique field, in particular to a kind of micro-fluidic chips, chip manipulation System, micro fluidic device and method for extracting nucleic acid.

Background technique

The high sensitivity and specificity of detection of nucleic acids, make it occupy extremely important position in technical field of biological It sets.The premise for realizing detection of nucleic acids is to obtain highly purified nucleic acid-templated of biological sample, and existing nucleic acid extraction process is often wrapped The biological respinse step for including multiple complexity, either have been manually done, or completes by ancillary equipment, exist efficiency compared with It is low, it is easy the deficiencies of malfunctioning.

Micro-fluidic chip can be sample preparation, the mixing, separation, reaction, inspection during chemistry, biology, medical analysis The basic operation units such as survey, cell culture, sorting are transplanted on the chip of one piece of micro-meter scale, or by multiple functional modules It is integrated on a unified chip, controlled fluid is run through to the microchannel network of entire chip, building biology, chemical microreactor Or micro-system, it is automatically performed analysis overall process.It for technical field of biological, is particularly advantageous in that: by automating, flowing Sample processing time is greatly shortened in the operating mode of ability of swimming, improves detection efficiency, reduces reaction reagent and sample consumption, most Automation, low cost, the detection of intelligentized fast medical are realized eventually.It is micro-fluidic due to the outstanding advantage in terms of sample treatment Chip is by its integrated multiple microreactor, and the controllability microfluidic networks being made of micro-valve, Micropump, microchannel Sample treatment and nucleic acid automatically extract multiple steps such as related sample flow, sample mixing, nucleic acid purification, waste liquid removal and mention An ideal driving assembly is supplied.Therefore, nucleic acid extraction is realized based on microfluidic chip technology, the effect of detection of nucleic acids can be improved Rate, or construct integrated nucleic acid automatic checkout system and build up a solid foundation.

But in the prior art, most of when carrying out nucleic acid extraction using micro-fluidic chip, it is still desirable to mention nucleic acid The various reagents taken are added inside micro-fluidic chip, and it reduce the efficiency of nucleic acid extraction, increase the operation of nucleic acid extraction Error probability.

Summary of the invention

The purpose of the present invention is to provide a kind of micro-fluidic chip, chip steerable system, micro fluidic device and nucleic acid extractions Method, it is intended to conducive to the automation for realizing nucleic acid extraction process, improve the efficiency of nucleic acid extraction, the operation for reducing nucleic acid extraction goes out Wrong probability.

First aspect present invention provides a kind of nucleic acid extraction micro-fluidic chip, including at least one storage chamber, at least one A reagent connection control unit and at least one reaction chamber, each storage chamber are correspondingly arranged with a reaction chamber, and every Reagent connection control unit is provided between a storage chamber and corresponding reaction chamber both to control and whether to be connected to, In, at least one described storage chamber is packaged with the reagent for nucleic acid extraction.

Second aspect of the present invention provides a kind of nucleic acid extraction chip steerable system, and the chip steerable system is for controlling Micro-fluidic chip described in any one of according to a first aspect of the present invention executes nucleic acid extraction movement.

Third aspect present invention provides a kind of nucleic acid extraction micro fluidic device, and the micro fluidic device includes micro-fluidic core Piece and the chip steerable system that nucleic acid extraction movement is executed for controlling the micro-fluidic chip, wherein the micro-fluidic chip For the micro-fluidic chip according to any one of first aspect present invention, the chip steerable system is according to the present invention second Chip steerable system described in any one of aspect.

Fourth aspect present invention provides a kind of method for extracting nucleic acid, and the method for extracting nucleic acid utilizes third aspect present invention Any one of described in micro fluidic device extract nucleic acid, the method for extracting nucleic acid include: control it is corresponding with the storage chamber Reagent is connected to control unit, discharges the reagent at least one described storage chamber to corresponding reaction chamber.

Based on micro-fluidic chip provided by the invention, chip steerable system, micro fluidic device and method for extracting nucleic acid, due to At least one storage chamber of micro-fluidic chip is packaged with the reagent for nucleic acid extraction, then, can be by reagent in nucleic acid extraction Release is in corresponding reaction chamber, and without the specialized operations such as being measured, being injected to corresponding reagent, therefore, of the invention is micro- Fluidic chip can simplify the operating procedure of nucleic acid extraction, save the operating time of nucleic acid extraction, improve the efficiency of nucleic acid extraction, Reduce error in operation probability.

Nucleic acid extraction of the invention chip steerable system and method for extracting nucleic acid and micro-fluidic chip are having the same excellent Point.

By referring to the drawings to the detailed description of exemplary embodiment of the present invention, other feature of the invention and its Advantage will become apparent.

Detailed description of the invention

The drawings described herein are used to provide a further understanding of the present invention, constitutes part of this application, this hair Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention.In the accompanying drawings:

Fig. 1 is the structural schematic diagram of the micro fluidic device of first embodiment of the invention.

Fig. 2 is the structural schematic diagram of the chip steerable system of first embodiment of the invention.

Fig. 3 is the structural schematic diagram that Y-axis moves driving assembly in the chip steerable system of first embodiment of the invention.

Fig. 4 is the structural schematic diagram that X-axis moves driving assembly in the chip steerable system of first embodiment of the invention.

Fig. 5 is the structural schematic diagram that magnetic force applies module in the chip steerable system of first embodiment of the invention.

Fig. 6 is that the structure of the micro-fluidic chip and the vacuum suction component of chip steerable system of first embodiment of the invention is shown It is intended to.

Fig. 7 is the structural schematic diagram of the micro-fluidic chip of first embodiment of the invention.

Fig. 8 is the exploded perspective structural schematic diagram of the micro-fluidic chip of first embodiment of the invention.

Fig. 9 A and Fig. 9 B be first embodiment of the invention micro fluidic device during nucleic acid extraction magnetic bead mixed process Schematic diagram.

Figure 10 A and Figure 10 B be first embodiment of the invention micro fluidic device during nucleic acid extraction magnetic bead shifted Journey schematic diagram.

Figure 11 be first embodiment of the invention micro fluidic device during nucleic acid extraction reagent discharge Principle of Process figure.

Figure 12 A and Figure 12 B be first embodiment of the invention micro fluidic device during nucleic acid extraction hose closed Journey schematic diagram.

Figure 13 A and Figure 13 B are the partial structure diagram of the micro-fluidic chip of second embodiment of the invention.

Figure 14 is the cooperation of the micro-fluidic chip (part) of third embodiment of the invention and the solenoid valve of chip steerable system Structural schematic diagram.

Into Figure 14, each appended drawing reference respectively represents Fig. 1:

1, bottom plate, 2, Y-axis belt fixing piece, 3, Y-axis belt, 4, Y-axis sliding block, 5, Y-axis guide rail, 6, y-axis motor, 7, Y-axis Position grating, 8, Y-axis flute profile optocoupler, 9, Y-axis microswitch, 10, left magnet carrier, 11, left magnet motors, 12, left magnet cunning Block, 13, left magnet positioning grating, 14, left magnet flute profile optocoupler, 15, left magnet microswitch, 16, left side permanent magnet, 17, Right magnet carrier, 18, right magnet motors, 19, right magnet sliding block, 20, right side permanent magnet, 21, hose close motor, 22, soft Duct occlusion electric machine support, 23, closing sliding block, 24, heating film, 25, bottom magnet bracket, 26, bottom permanent magnet, 27, miniflow Control chip, 28, puncture needle tabletting, 29, reagent release motor, 30, reagent release electric machine support, 31, pinboard, 32, X-axis electricity Machine, 33, X-axis guide rail, 34, X-axis belt, 35, X-axis slide block, 36, X-axis belt fixing piece, 37, X-axis positioning grating, 38, X-axis slot Shape optocoupler, 39, X-axis microswitch, 40, chip butt joint base, 41, spring pressuring plate, 42, chip position detection microswitch, 43, suction Disc carrier, 44, sucker, 45, gas-guide tube, 46, vacuum pump, 47, shut-off valve, 48, vacuum pump fixed frame, 49, (reagent storage organization On piece) storage chamber, 50, storage chamber cover plate, 51, well, 52, puncture needle, 53, mixing chamber stomata, 54, reagent storage organization Piece, 55, puncturing structure piece, 56, connectivity structure piece, 57, accommodating blind hole, 58, isolation blind hole, 59, reagent interface channel, 60, anti- Answer structure piece, 61, mixing chamber, 62, hose, 63, elution chamber, 64, eluent storage chamber, 65, elution chamber lid piece, 66, waste liquid deposits Put structure piece, 67, waste liquid chamber, 68, waste liquid chamber stomata, 69, waste liquid bottom of chamber piece, 70, waste liquid removal channel, 71, magnetic bead, 72, rubber Rubber plug, 73, solenoid valve.

Specific embodiment

Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Below Description only actually at least one exemplary embodiment be it is illustrative, never as to the present invention and its application or make Any restrictions.Based on the embodiments of the present invention, those of ordinary skill in the art are not making creative work premise Under every other embodiment obtained, shall fall within the protection scope of the present invention.

Unless specifically stated otherwise, positioned opposite, the digital table of the component and step that otherwise illustrate in these embodiments It is not limited the scope of the invention up to formula and numerical value.Simultaneously, it should be appreciated that for ease of description, each portion shown in attached drawing The size divided not is to draw according to actual proportionate relationship.For technology, side known to person of ordinary skill in the relevant Method and equipment may be not discussed in detail, but in the appropriate case, and the technology, method and apparatus should be considered as authorizing explanation A part of book.In shown here and discussion all examples, any occurrence should be construed as merely illustratively, and Not by way of limitation.Therefore, the other examples of exemplary embodiment can have different values.It should also be noted that similar label Similar terms are indicated in following attached drawing with letter, therefore, once it is defined in a certain Xiang Yi attached drawing, then subsequent attached It does not need that it is further discussed in figure.

In the description of the present invention, it is to be understood that, components are limited using the words such as " first ", " second ", only It is merely for convenience of distinguishing corresponding components, there is no Stated otherwise such as, there is no particular meanings for above-mentioned word, therefore not It can be interpreted as limiting the scope of the invention.

In the description of the present invention, it is to be understood that, the noun of locality such as " front, rear, top, and bottom, left and right ", " it is laterally, vertical, Vertically, orientation or positional relationship indicated by level " and " top, bottom " etc. is normally based on orientation or position shown in the drawings and closes System, is merely for convenience of description of the present invention and simplification of the description, in the absence of explanation to the contrary, these nouns of locality do not indicate that It must have a particular orientation or be constructed and operated in a specific orientation with the device or element for implying signified, therefore cannot manage Solution is limiting the scope of the invention；The noun of locality " inside and outside " refers to inside and outside the profile relative to each component itself.

Fig. 1 to Figure 14 shows the structure of various embodiments of the present invention.As shown in figs. 1 to 14, the present invention provides a seed nucleus Sour extraction micro-fluidic chip 27, nucleic acid extraction chip steerable system and nucleic acid extraction micro fluidic device.

Micro-fluidic chip 27 includes at least one storage chamber, at least one reagent connection control unit and at least one reaction Chamber.Each storage chamber is correspondingly arranged with a reaction chamber, and reagent company is provided between each storage chamber and corresponding reaction chamber Logical control unit is to control whether the two is connected to.At least one storage chamber is packaged with the reagent for nucleic acid extraction.

It, then, can be by reagent in nucleic acid extraction since at least one storage chamber is packaged with the reagent for nucleic acid extraction Release is in corresponding reaction chamber, and without the specialized operations such as being measured, being injected to corresponding reagent, therefore, of the invention is micro- Fluidic chip 27 can simplify the operating procedure of nucleic acid extraction, save the operating time of nucleic acid extraction, improve the effect of nucleic acid extraction Rate reduces error in operation probability.

Preferably, at least one reaction chamber includes mixing chamber 61, at least one storage chamber include be packaged with lysate and with The lysate storage chamber and/or the washing including being packaged with cleaning solution and being correspondingly arranged with mixing chamber 61 that mixing chamber 61 is correspondingly arranged Liquid storage chamber；And/or at least one reaction chamber include elution chamber 63, at least one storage chamber include be packaged with eluent and with The eluent storage chamber 64 that elution chamber 63 is correspondingly arranged.

Due to being packaged with lysate in lysate storage chamber, when extracting nucleic acid, sample to be tested injection lysate is deposited In storage chamber, the cracking process of nucleic acid can be at least partly realized in lysate storage chamber, without being measured, being infused to lysate The specialized operations such as enter.In cracking process or after the completion, it can be connected to by controlling reagent corresponding with lysate storage chamber Control unit will crack mixed liquor and directly discharge to mixing chamber 61, carry out extracting work in next step." the cracking referred in the present invention Mixed liquor ", which refers to, is filled with the lysate of sample to be tested, regardless of whether cracking reaction is completed.

Similarly, setting is packaged with cleaning solution and the cleaning solution storage chamber being correspondingly arranged with mixing chamber 61 may not need in nucleic acid The purification step of extraction such as is measured to cleaning solution, is injected at the specialized operations, therefore can improve operating efficiency, reduces error in operation Probability.

Preferably, at least one storage chamber 49 includes two cleaning solution storage chambers, is stored not in two cleaning solution storage chambers Same cleaning solution.Under normal conditions, nucleic acid extraction needs washing operation twice, and washing operation uses different cleaning solutions twice, Different cleaning solutions is packaged in different cleaning solution storage chambers by the setting, corresponding by the cleaning solution storage chamber for controlling different Reagent connection control unit controls time and order that different cleaning solutions enters mixing chamber 61, keeps washing process easier, fast It is prompt.

Similarly, setting is packaged with eluent and can simplify with the eluent storage chamber 64 that is correspondingly arranged of elution chamber 63 and washes De- liquid metering and addition step, improve operating efficiency, reduce error in operation probability.

And setting elution chamber 63 is conducive to be directed to using micro-fluidic chip 27 in micro-fluidic chip 27 to complete elution step Amount of reagent needed for different extraction steps is different, and the reaction chamber of suitable capacity is selected to complete all operationss of nucleic acid extraction.

Preferably, micro-fluidic chip 27 further includes for sample to be tested sample-adding structure to be added to lysate storage chamber.Setting Sample-adding structure can be conducive to sample to be tested and be added in lysate.

Being loaded structure is a kind of preferred structure, but is not required in some cases.For example, if lysate stores The cavity wall of chamber is provided with the structure that can be pierced through, so that it may which specially setting is not loaded structure, and cracking is pierced through when needing to be loaded Sample to be tested can be added by piercing through the perforation that cavity wall obtains in the cavity wall of liquid storage chamber into lysate.

Sample-adding structure may include well 51 and diaphragm, and piercing through diaphragm can make lysate storage chamber pass through well 51 It is in communication with the outside.Diaphragm can be covered on well 51, also can be set between well 51 and lysate storage chamber.? Need to be when sample to be tested be added to lysate storage chamber, only need to pierce through diaphragm can carry out sample-adding operation.The material of diaphragm can be with It is metal foil, rubber membrane etc., as long as not reacting with lysate and being convenient for piercing through.

Preferably, magnetic bead 71 is packaged in mixing chamber 61.During nucleic acid extraction, the absorption cracking of magnetic bead 71 is often utilized Nucleic acid afterwards, then the magnetic bead 71 for being adsorbed with nucleic acid is washed and eluted, purification of nucleic acid template can be obtained.In mixing chamber 61 It is inside packaged with magnetic bead 71, the step of magnetic bead 71 are added to mixing chamber 61 can be saved, reduce the metering of magnetic bead 71 and step is added, Therefore, it is possible to reduce the operating procedure of nucleic acid extraction improves operating efficiency, reduces error in operation probability.

In a preferred embodiment, reagent connection control unit includes being set between storage chamber and corresponding reaction chamber Isolation structure, control storage chamber by piercing through isolation structure and be connected to reaction chamber.Pierceable isolation structure, storage chamber are set When good isolation may be implemented when not needing connection between corresponding reaction chamber, and needing to be connected to, correspondence need to be only pierced through Isolation structure, it is easy to operate.

It is further preferred that reagent connection control unit further includes that the reagent connection being set between storage chamber and reaction chamber is led to Road, isolation structure include the isolation blind hole 58 for being set to storage chamber bottom, and the bottom hole of isolation blind hole 58 is located at corresponding reagent company It connects on road.When being provided with multiple reagent connection control units, corresponding storage chamber and reaction are made by reagent interface channel 66 Chamber have connection may, recycle isolation blind hole 58 bottom hole as isolation structure, make isolation structure be easy to pierce through, and bottom hole with Outer part is opposite to be not easy to pierce through, it is possible to reduce pierces through the possibility made mistakes.

Preferably, isolation structure is pierced through for convenience, and reagent connection control unit includes the puncture for piercing through isolation structure Needle 52.The material of puncture needle 52 is preferably glass, can also use other materials not reacted with the reagent stored in storage chamber Material is made, such as stainless steel.In addition, in order to make pierce through after isolation structure between corresponding storage chamber and reaction chamber have compared with Good connected state, it is preferable that the concave portion with certain axial length can be set in the periphery of puncture needle 52.For example, puncturing The piercing end of needle 52 may include the groove extended along the length direction of puncture needle 52.

In order to accurately pierce through isolation structure, it further includes the appearance being set at the top of corresponding storage chamber that reagent, which is connected to control unit, Set blind hole 57.Puncture needle be set to accommodating blind hole 57 in and can along accommodating blind hole 57 to corresponding storage chamber internal motion to pierce Wear the bottom hole of accommodating blind hole 57.Accommodating 57 one side of blind hole can accommodate puncture needle 52, when the length of puncture needle 52 is different Waiting can prevent the position of each puncture needle 52 from mistake occurs, and have certain guiding role to puncture needle 52, can make to wear Pricker 52 accurately pierces through corresponding isolation structure, reduces the possibility for piercing through failure.In addition, each reagent connection control unit includes Corresponding puncture needle 52 can also prevent from causing cross contamination when puncturing different isolation structures using same puncture needle 52.

Preferably, at least one reagent connection control unit is different from remaining reagent connection structure of control unit and/or is arranged Height is different, so that the puncture time of the isolation structure of at least one reagent connection control unit is different from the connection control of remaining reagent The puncture time of the isolation structure in portion.For example, the length of puncture needle can be designed to difference, isolation structure and puncture needle 52 Distance is arranged different, isolation structure thickness is arranged different etc..The setting can control the storage for being respectively stored with reagent The reagent release time of chamber obtains preferable control effect with corresponding with each extraction step during nucleic acid extraction.

Certainly, puncture needle 52 is not the necessary component of micro-fluidic chip 27, such as can be used as and be worn using syringe needle Thorn tool pierces through isolation structure.

Preferably, micro-fluidic chip 27 further includes mixing chamber stomata 53, and mixing chamber stomata 53 is connected to mixing chamber 61.Setting Mixing chamber stomata 53 is conducive to mixing chamber 61 and is in communication with the outside by mixing chamber stomata 53, conducive to the cracking mixed liquor completed after absorption The reagents such as raffinate, cleaning solution release mixing chamber 61.Preferably, mixing chamber stomata 53 is closing before cleavage step completion 's.Closing mode can for example be closed using diaphragm or rubber plug etc..

Preferably, micro-fluidic chip 27 further includes internal connection control unit, and inside connection control unit is for controlling mixing chamber Whether 61 be connected to elution chamber 63.Internal connection control unit, which is arranged, can make the participation core after mixing chamber 61 completes part operation The substance (such as the magnetic bead for being adsorbed with nucleic acid) that acid extracts is directly entered elution chamber 63 through inside connection control unit and completes subsequent work Make, the control of nucleic acid extraction is made to realize automation, it is simple and convenient, save the operating time.

Preferably, internal connection control unit includes being connected to mixing chamber 61 and eluting the hose 62 between chamber 63, passes through control Whether it is connected between the cross-sectional flow area control mixing chamber 61 and elution chamber 63 of hose 62 processed.It is controlled by hose cross-sectional flow area Whether it is connected to, can be repeatedly controlled between mixing chamber 61 and elution chamber 63.And can by hose 62 by magnetic bead 71 from Mixing chamber 61 is directly transferred to elution chamber 63, simplifies the transfer process of magnetic bead 71, it is ensured that the safety of transfer process, Ke Yiti High operating efficiency reduces error in operation probability.

Preferably, the bottom of mixing chamber 61 and the bottom of elution chamber 63 can be arranged substantially in same level height, soft Pipe 62 is arranged substantially horizontally.The setting is conducive to shift magnetic bead 71 using magnetic force.It is further preferred that the water of the bottom of hose 62 Flat height and the bottom of mixing chamber 61 and the level height for the bottom for eluting chamber 63 are roughly equal.The setting is conducive to pass through hose 62 With mixing chamber 61 from elution chamber 63 in exclude in magnetic bead transfer process enter elution chamber 63 cleaning solution, without for elution chamber The 63 special cleaning solutions of setting exclude channel.

Additionally preferably, mixing chamber 61 can be set above elution chamber 63, and hose 62 is substantially vertically arranged.The setting Magnetic bead 71 is shifted into elution chamber 63 conducive to using gravity.When needed, special washing can be set for elution chamber 63 Liquid excludes channel.

In a preferred embodiment, micro-fluidic chip 27 includes the elution chamber lid for forming the part cavity wall of elution chamber 63 Piece 65, and elute chamber lid piece 65 and be provided with the gap structure to be opened/closed made of flexible material.Eluting chamber lid piece 65 can be whole It is all made of flexible material on body to be made, can also only gap structure to be opened/closed be made of flexible material.Gap structure can be opened and closed It such as can be strip crevice, cross gap, radial gap etc. various forms.

Additionally preferably, micro-fluidic chip 27 further includes waste liquid chamber 67 and waste liquid excludes control unit, and waste liquid excludes control unit For controlling whether reaction chamber is connected to waste liquid chamber 67.The setting can exclude waste liquid to waste liquid chamber 67, make micro-fluidic core Piece 27 has the function of devil liquor recovery, and it is not necessary that special waste liquid recovery apparatus is in addition arranged, waste liquid excludes simple and convenient.

Preferably, it includes that the waste liquid being set between reaction chamber and waste liquid chamber 67 removes 70 He of channel that waste liquid, which excludes control unit, Whether the pressure of pressure control portion, pressure control portion control waste liquid chamber 67 passes through waste liquid between reaction chamber and waste liquid chamber 67 to control Channel 70 is removed to be connected to.Pressure by controlling waste liquid chamber 67 can control the pressure difference that waste liquid removes 70 both ends of channel, in liquid It can control waste liquid under surface tension and the collective effect of pressure difference and remove whether channel 70 is connected to, this control mode can be micro- It is controlled outside fluidic chip 27, and can according to need and the progress on-off control of channel 70 is removed to waste liquid repeatedly, thus It is adapted to discharge the demand of waste liquid in each nucleic acid extraction step.

Preferably, pressure control portion includes the waste liquid chamber stomata 68 being connected to waste liquid chamber 67, by controlling waste liquid chamber stomata Whether 68 vacuumize or whether close the pressure that can control waste liquid chamber 67.It is very simple that waste liquid chamber stomata 68 is set, by useless The pressure that sap cavity stomata 68 controls waste liquid chamber 67 is also more convenient.

Nucleic acid extraction chip steerable system is held for coupling with aforementioned micro-fluidic chip 27 and controlling micro-fluidic chip 27 The movement of row nucleic acid extraction.It is acted using the nucleic acid extraction of chip steerable system control micro-fluidic chip 27, nucleic acid can be improved and mention The automatization level taken is further simplified operating procedure, improves operating efficiency.

Preferably, chip steerable system includes chip motion control module, chip motion control module and micro-fluidic chip 27 are drivingly connected to control the movement of micro-fluidic chip 27.Chip motion control module can pass through the fortune of control micro-fluidic chip 27 The dynamic different mixing velocities for participating in substance improved during nucleic acid extraction, improve reaction efficiency.

Chip motion control module includes the linear motion control unit for driving micro-fluidic chip 27 to move along a straight line.Linear motion Mixing efficiency during nucleic acid extraction can be improved in control unit, and the position that can also adjust micro-fluidic chip 27 makes micro-fluidic chip 27 are in suitable operating position.

Preferably, linear motion control unit includes: X-axis movement driving assembly, for driving micro-fluidic chip along horizontal X Direction movement；And/or Y-axis moves driving assembly, for driving micro-fluidic chip along the Y-direction fortune for being horizontally and perpendicularly to X-direction It is dynamic；And/or Z axis moves driving assembly, for driving micro-fluidic chip to move along vertical Z-direction.

Preferably, moving along a straight line the motion control in each direction of control unit can be by driving mechanism and guide rail slide block machine The cooperation of structure is realized.For example, in one embodiment, if simultaneously including X-axis movement driving assembly, Y-axis movement driving group Part and Z axis move driving assembly, linear motion control unit can be arranged as follows:

It may include X-axis driving mechanism, X-axis slide block 35 and X-axis guide rail 33 that X-axis, which moves driving assembly, and X-axis guide rail 33 is along X Axis direction extends, and X-axis slide block 35 is reciprocally arranged along X-axis guide rail 33, and X-axis driving mechanism and the driving of X-axis slide block 35 connect It connects, micro-fluidic chip 27 and X-axis slide block 35 are arranged relatively fixedly.Y-axis movement driving assembly includes Y-axis driving mechanism, Y-axis cunning Block 4 and Y-axis guide rail 5, Y-axis guide rail 5 extend along the y axis, and Y-axis sliding block 4 is reciprocally arranged along Y-axis guide rail 5, and Y-axis is driven Motivation structure and Y-axis sliding block 4 are drivingly connected, and X-axis guide rail 33 and Y-axis sliding block 4 are arranged relatively fixedly.Z axis moves driving assembly can To include Z axis driving mechanism, Z axis sliding block and Z axis guide rail, for Z axis guide rail along Z-direction extension, Z axis sliding block can be past along Z axis guide rail The setting of multiple movement ground, Z axis driving mechanism are connect with Z axis slider-actuated, and Y-axis guide rail 5 is opposite with Z axis sliding block to be arranged fixedly.

Each driving mechanism of linear motion control unit is preferably motor.

Preferably, chip steerable system further includes chip butt joint base 40, and chip motion control module passes through chip butt joint base 40 is detachably connected with micro-fluidic chip 27.Chip butt joint base 40 is set and is conducive to micro-fluidic chip 27 and chip motion control mould Block is accurate, rapid abutting joint, improves operating efficiency.

Preferably, chip butt joint base 40 includes slot, and the side wall of slot is fixed with spring pressuring plate 41, and micro-fluidic chip 27 is inserted Enter in slot and spring pressuring plate 41 is crimped on micro-fluidic chip 27.The chip butt joint base 40 of slot form and spring pressuring plate 41 Setting, make micro-fluidic chip 27 and chip butt joint base 40 may be implemented it is quick connect and fast quick-detach, and in spring pressuring plate 41 Under the action of pressure, micro-fluidic chip 27 can be more firmly secured in the slot of chip butt joint base 40 when connection.

Preferably, chip steerable system further includes chip position detection device.Chip position detection device is micro- for detecting Whether fluidic chip 27 docks successfully with chip butt joint base 40.Chip position detection device is, for example, that chip position detection fine motion is opened Close 42.Chip position detection microswitch 42 can be set in slot or near slot, such as can be fixedly installed on slot Rear wall.The prompt of flip chip position detection microswitch 42 is docked successfully after micro-fluidic chip 27 is inserted into slot success, favorably In the accuracy rate for guaranteeing that micro-fluidic chip 27 is docked with chip butt joint base 40.

Preferably, chip steerable system further includes that magnetic force applies module, and it includes that alternating force applies magnet that magnetic force, which applies module,. Alternating force applies magnet and is used to apply alternation magnetive attraction to the magnetic bead 71 in the reaction chamber of micro-fluidic chip 27.Magnetic force applies module The use for cooperating magnetic bead 71, can make magnetic bead 71 under the action of alternation magnetive attraction, move in cracking mixed liquor, sufficiently adsorb Crack the nucleic acid in mixed liquor.Preferably, two alternating forces that magnetic force applies that module includes relative spacing setting apply magnet, micro- Fluidic chip 27, which can move to, is located at reaction chamber between two alternating forces application magnets, while the magnetic in micro-fluidic chip 27 Pearl 71 can apply in two alternating forces to be moved back and forth between magnet.The setting is conducive to apply alternation magnetive attraction to magnetic bead 71.It hands over Variable force, which applies magnet for example, to be permanent magnet, it is of course also possible to use electromagnet.If also can use using electromagnet The energization order of electromagnet controls changes of magnetic field, is realized with this and applies alternation magnetive attraction to magnetic bead 71.

Preferably, it further includes magnet movement driving assembly that magnetic force, which applies module, and magnet movement driving assembly is handed over for driving Variable force applies magnet and moves relative to micro-fluidic chip 27.Applying magnet movement by control alternating force can make magnetic field generate change Change, to apply alternation magnetive attraction to magnetic bead 71.The movement that alternating force applies magnet can be folded with the movement of micro-fluidic chip 27 Add, to keep reaction more abundant, reaction efficiency is higher.

In one preferred embodiment, magnet movement driving assembly for drive two alternating forces apply magnets along Apply the vertical direction of the distribution arrangement of magnet with two alternating forces to move reciprocatingly.

Magnet movement driving assembly is preferably the driving assembly that moves along a straight line.For example, two alternating forces apply magnet along water Flat directional spreding, magnet movement driving assembly include applying two alternating forces that magnet is correspondingly arranged with two alternating forces respectively to apply Magnet driving mechanism, two alternating forces is added to apply magnet guide rail and two alternating forces application magnet sliding blocks, magnet guide rail is along perpendicular Histogram is connected to extension, magnet driving mechanism with corresponding magnet slider-actuated, and magnet sliding block can be past along corresponding magnet guide rail The setting of multiple movement ground, magnet are fixed on corresponding magnet sliding block.

For example, it is the left side permanent magnet 16 and right side permanent magnet being distributed in the horizontal direction that two alternating forces, which apply magnet, 20, magnet movement driving assembly include the left magnet motors 11 being correspondingly arranged with left side permanent magnet 16, left magnet sliding block 12 and Left magnet carrier 10, and include the right magnet motors 18 being correspondingly arranged with right side permanent magnet 20, right magnet sliding block 19 and the right side Magnet carrier 17, left magnet sliding block 12 are drivingly connected with left magnet motors 11, and right magnet sliding block 19 drives with right magnet motors 18 Connection, left side permanent magnet 16 and right side permanent magnet 20 are separately fixed on left magnet sliding block 12 and right magnet sliding block 19.

Preferably, it further includes bottom magnet that magnetic force, which applies module, and the reaction chamber of micro-fluidic chip 27 includes elution chamber 63, is washed De- chamber 63 can move to the top of bottom magnet, and bottom magnet is used to the magnetic bead 71 eluted in chamber 63 being adsorbed on elution chamber 63 Bottom and be in tiled state.Bottom magnet is, for example, bottom permanent magnet 26.

Preferably, chip steerable system further includes reagent release module, and reagent release module is for controlling reagent connection control Portion processed between storage chamber and corresponding reaction chamber so as to be connected to.

It include the isolation structure being set between storage chamber and corresponding reaction chamber and for piercing in reagent connection control unit In the case where the puncture needle 52 for wearing isolation structure, reagent release module preferably includes puncture needle tabletting 28.Puncture needle tabletting 28 Side positioned at the separate micro-fluidic chip 27 of puncture needle 52 and the puncture direction along puncture needle 52 are movably disposed will wear Pricker 52 is pierced into isolation structure.Control whether isolation structure pierces through with can be convenient by puncture needle tabletting 28.Reagents connect The setting of logical control unit can also effectively control the puncture time of different isolation structures.For example, if different puncture needle 52 Length is different, and the setting of isolation structure height is identical, then puncture needle tabletting 28 is pressed into longest puncture needle 52 at first, makes longest The corresponding isolation structure of puncture needle 52 be pierced, puncture needle tabletting 28 stop a period of time after, be further continued for towards push puncture The direction of needle 52 moves, and the second long corresponding isolation structure of puncture needle 52 is pierced, and so on, until all puncture needles 52 Corresponding isolation structure is pierced.In this way, can control the examination in different storage chambers by the control to puncture needle tabletting 28 Agent enters the time of corresponding reaction chamber, so that different nucleic acid extraction steps be cooperated to discharge suitable reagent, is finally completed core Acid extracts work.

Preferably, reagent release module further includes tabletting driving mechanism, and tabletting driving mechanism and puncture needle tabletting 28 drive Connection is moved with controlling puncture needle tabletting 28 along the puncture direction of puncture needle 52.Tabletting driving mechanism is arranged to be conducive to realize puncture needle The position control of tabletting 28 reduces control fault, is also conducive to the automatic control and chip steerable system of entire chip steerable system In each module cooperation.

Further, reagent release module further includes tabletting guiding mechanism, and tabletting guiding mechanism includes along puncture needle 52 The guide runner for puncturing the guide post of direction setting and being slidably matched with guide post, puncture needle tabletting 28 and the fixed company of guide runner It connects.Tabletting guiding mechanism can be such that the motion profile of puncture needle tabletting 28 determines, can more accurately control the thorn of each isolation structure Wear the time.

It include mixing chamber 61 and elution chamber 63 at least one reaction chamber of micro-fluidic chip 27, and micro-fluidic chip 27 is also In the case where being connected to control unit including inside, chip steerable system further includes the inner passage for controlling internal connection control unit Open and close module.Inner passage opening and closing module is arranged to be conducive to control mixing chamber 61 and elute the connected state of chamber 63, so that control is mixed It closes chamber 61 and elutes the substance transfer between chamber 63.

It is internal in the case where internal connection control unit includes being connected to mixing chamber 61 and eluting hose 62 between chamber 63 Channel opening and closing module includes controlling the hose control assembly of the cross-sectional flow area of hose 62.By the passage section for controlling hose 62 Product controls mixing chamber 61 and elutes the connected state between chamber 63 in which can be convenient, and can repeatedly carry out on-off control, favorably Mixing chamber 61 is connected to demand with elution chamber 63 during meeting nucleic acid extraction, is especially conducive to control mixing chamber 61 and elution Substance transfer between chamber 63.

Preferably, hose control assembly includes closing sliding block 23, closes the setting radially displaceablely along hose 62 of sliding block 23 It sets to change the cross-sectional flow area of hose 62 with the contact position of hose 62 by changing.The hose control assembly structure is simple, It is easy to control quick.Further, hose control assembly may also include closing slider-actuated mechanism, closing slider-actuated mechanism with It closes sliding block 23 to be drivingly connected, to improve the degree of automation of chip steerable system.

Preferably, chip control system further includes heating module, and heating module is for heating elution chamber 63.Heating module can With for heating eluent in the drying and elution step in purification step, conducive to the raising of nucleic acid extraction efficiency.Preferably, add Thermal modules include heating film 24.Heating film 24 is small in size, heating rate is fast, temperature control is simple.It is highly preferred that heating film 24 is solid Due to one end of the direction elution chamber 63 of closing sliding block 23.The setting can drive heating film by driving closing sliding block 23 simultaneously 24, the specialized agency of the position for control heating film 24 that no setting is required, structure is simple, easy to control.

Additionally preferably, micro-fluidic chip 27 further includes waste liquid chamber 67 and waste liquid excludes control unit, and waste liquid excludes control unit For controlling whether reaction chamber is connected to waste liquid chamber 67；Chip steerable system further includes excluding the useless of control unit for controlling waste liquid Liquid excludes control module.Can control the liquid in reaction chamber by waste liquid exclusion control module control waste liquid exclusion control unit is No inflow waste liquid chamber 67.

Excluding control unit in waste liquid includes that the waste liquid being set between mixing chamber 61 and waste liquid chamber 67 removes the feelings in channel 70 Under condition, it includes the pressure control assembly for controlling 67 pressure of waste liquid chamber that waste liquid, which excludes control module,.Pass through pressure control assembly The pressure for controlling waste liquid chamber 67 can control the pressure difference that waste liquid removes 70 both ends of channel, in the pressure difference and surface tension of liquid Under effect, it can control waste liquid and remove whether the corresponding reaction chamber in channel 70 has liquid to be discharged into waste liquid chamber 67.

In the case where micro-fluidic chip 27 includes waste liquid chamber stomata 68 being connected to waste liquid chamber 67, pressure control assembly is logical Cross the pressure of the state control waste liquid chamber 67 of control waste liquid chamber stomata 68.For example, can control whether waste liquid chamber stomata 68 is taken out very It is empty, if the pressure of the control waste liquid chamber 67 such as closing.

Preferably, pressure control assembly includes the vacuum suction component being connected to waste liquid chamber stomata 68；Alternatively, pressure controls Component includes the valve for controlling waste liquid chamber stomata 68 and being opened and closed.

Preferably, chip steerable system includes that moving parts and the movement position being correspondingly arranged with moving parts detection fill It sets.Wherein, moving parts include motor, and the corresponding movement position detection device of moving parts includes microswitch, in exercise group In part motion process, the initial position or final position of moving parts are controlled by the corresponding microswitch of moving parts, are led to Cross the speed of the movement of the motor of moving parts and the course location of time control moving parts；Alternatively, moving parts are corresponding Movement position detection device includes that microswitch, grating and flute profile optocoupler pass through moving parts in moving parts motion process Corresponding microswitch controls the initial position or final position of moving parts, passes through the corresponding grating of moving parts and flute profile The course location of optocoupler control moving parts.Wherein, in a laminar grating of strip, multiple narrow slits are integrated with, in electricity In machine motion process, by the mutual movement between narrow slit in slot optical coupling and grating, the positioning control that is achieved between different location System.

Wherein, at least one moving parts is for example including the X-axis movement driving assembly of chip motion control module, Y-axis fortune Dynamic driving assembly, magnetic force apply the magnet movement driving assembly of module, and the puncture needle tabletting 28 and tabletting of reagent release module are driven The hose control assembly etc. of motivation structure and/or inner passage opening and closing module.

The present invention also provides a kind of nucleic acid extraction micro fluidic device, including nucleic acid extraction micro-fluidic chip above-mentioned and Nucleic acid extraction above-mentioned chip steerable system.

The present invention also provides a kind of method for extracting nucleic acid, which extracts core using micro fluidic device above-mentioned Acid, method for extracting nucleic acid include: control reagent connection control unit corresponding with storage chamber, make the reagent at least one storage chamber In release to corresponding reaction chamber.

Method for extracting nucleic acid of the invention can use chip steerable system and complete nucleic acid extraction.Due to will directly be stored in Reagent in storage chamber is discharged to corresponding reaction chamber, can simplify operating procedure, improves operating efficiency, and it is general to reduce error in operation Rate.

Preferably, method for extracting nucleic acid includes: cleavage step, the cracking for being stored with lysate at least one storage chamber Sample to be tested is added in liquid storage chamber and obtains cracking mixed liquor；Crack mixed liquor transfer step, control and lysate storage chamber pair The reagent connection control unit answered, will crack mixed liquor and be discharged into the mixing chamber 61 of at least one reaction chamber；Nucleic acid absorption step Suddenly, it moves magnetic bead 71 and adsorbs the nucleic acid in cracking mixed liquor；Nucleic acid purification step, washing are adsorbed with core The magnetic bead 71 of acid；Nucleic Acid Elution on the magnetic bead 71 for being adsorbed with nucleic acid is got off to obtain purification of nucleic acid template by elution step.

Preferably, nucleic acid absorption step includes applying micro-fluidic chip 27 in two alternating forces to generate relatively between magnet Apply magnet in two alternating forces first does relatively reciprocating motion, and first direction of motion to do relatively reciprocating motion is parallel to two friendships The distribution arrangement of variable force application magnet.The setting is conducive to magnetic bead 71 and sufficiently, quickly mixes with reaction solution, improves operating efficiency.

Preferably, nucleic acid absorption step further includes generating micro-fluidic chip 27 relatively between two alternating forces apply magnet Apply magnet in two alternating forces second does relatively reciprocating motion, and the second direction to do relatively reciprocating motion is perpendicular to two alternating forces Apply the distribution arrangement of magnet.The setting similarly serves to favor magnetic bead 71 and sufficiently, quickly mixes with reaction solution, improves operating efficiency.

Preferably, nucleic acid purification step is included: and is washed to be adsorbed with nucleic acid at least once using cleaning solution in mixing chamber 61 Magnetic bead 71, so that the cleaning solution for completing last time washing is passed through the elution that is set in mixing chamber 61 and at least one reaction chamber Inside connection control unit between chamber 63 enters elution chamber 63, and washs in inside connection control unit filled with completion last time Cleaning solution, magnetic bead 71 swims across the cleaning solution in internal connection control unit under the action of gravity and/or magnetic force and enters elution Chamber 63 adsorbs magnetic bead 71 with bottom magnet later, completes from mixing chamber 61, internal connection control unit and the elution discharge of chamber 63 last The cleaning solution of once washing.This method will be in mixing chamber 61 as transport materials by the cleaning solution for completing last time washing Magnetic bead 71 is transferred to elution chamber 63, can efficiently realize transfer process of the magnetic bead 71 inside micro-fluidic chip 27, improves nucleic acid Extraction efficiency.

Preferably, nucleic acid purification step further include: after the cleaning solution that last time washing is completed in discharge, drying elution chamber The magnetic bead 71 for being adsorbed with nucleic acid in 63.Drying and processing is carried out to the magnetic bead 71 after washing, it is possible to reduce cleaning solution is to purifying core The influence of acid template.

Preferably, in elution step, reagent connection control corresponding with the eluent storage chamber 71 of micro-fluidic chip 27 is controlled Portion processed discharges the eluent in eluent storage chamber 71 to elution chamber 63, makes the movement of magnetic bead 71 to incite somebody to action in elution chamber 63 Nucleic Acid Elution on magnetic bead 71 gets off.Eluent is directly deposited in eluent storage chamber 71 and can be directly released into elution chamber 63 It is interior, it without the metering of special eluent, injection step, can simplify operating procedure, improve operating efficiency, it is general to reduce error in operation Rate.

Preferably, predetermined order and time interval discharge the reagent in each storage chamber to corresponding reaction chamber.The setting Release time can be made appropriately to cooperate with each nucleic acid extraction step, smoothly complete nucleic acid extraction.

Various embodiments of the present invention are described in detail below with reference to Fig. 1 to Figure 14.

First embodiment

Fig. 1 to Figure 12 shows the nucleic acid extraction micro fluidic device and its micro-fluidic chip of first embodiment of the invention Structure.Micro fluidic device includes nucleic acid extraction micro-fluidic chip 27 and nucleic acid extraction chip steerable system.Chip manipulation system System, which couples with micro-fluidic chip 27 and controls micro-fluidic chip 27, executes nucleic acid extraction movement.

Referring to Fig. 1, Fig. 7 and Fig. 8.In first embodiment, micro-fluidic chip 27 includes the storage set gradually from top to bottom Chamber lid piece 50, reagent storage organization piece 54, puncturing structure piece 55, connectivity structure piece 56, reaction structure piece 60, waste liquid storage configuration Piece 66 and waste liquid bottom of chamber piece 69.Additionally include elution chamber lid piece 65.

Storage chamber cover plate 50 is located at the top of reagent storage organization piece 54.Be provided on storage chamber cover plate 50 well 51, Mixing chamber stomata 53, multiple accommodating blind holes 57 and multiple puncture needles 52.Puncture needle 52 is embedded in corresponding accommodating blind hole 57.This In embodiment, puncture needle 52 is made of glass.

In the present embodiment, storage chamber cover plate 50, reagent storage organization piece 54 and the composition of puncturing structure piece 55 are deposited for reagent The single layer structure of storage.

As shown in figure 8, aforementioned single layer structure includes three storage chambers 49.Three storage chambers 49 are respectively lysate storage Chamber, the first cleaning solution storage chamber and the second cleaning solution storage chamber.Lysate is packaged in lysate storage chamber；First cleaning solution is deposited The first cleaning solution is packaged in storage chamber；The second cleaning solution is packaged in second cleaning solution storage chamber.Three storage chambers 49 are respectively provided with It is encapsulated inside reagent storage organization piece 54, and by storage chamber cover plate 50 and about 55 puncturing structure piece.In addition, the present embodiment In, a storage chamber being also provided in reaction structure piece 60, i.e. eluent storage chamber 64.Eluent storage chamber 64 and elution chamber 63 are isolated blind hole 58 unanimously with reagent connection control unit by one layer of thin-wall construction isolation, the thin-wall construction.Eluent is deposited Eluent is packaged in storage chamber 64.Eluent storage chamber 64 and elution chamber 63 are all set in reaction structure piece 60, rationally divided Match the space of storage chamber and reaction chamber, the inner flow passage of micro-fluidic chip 27 can be made shorter, it is compact-sized.Do not scheme some In the embodiment shown, each storage chamber can also be set in same storage organization piece.

Each storage chamber (three storage chambers 49 and eluent storage chamber 64 that the present embodiment includes storage organization piece 54) it is big It is small can the amount of reagent according to needed for a nucleic acid extraction setting.For example, in the present embodiment, due to the dosage of lysate and cleaning solution Larger and eluent dosage is less, and the volume ratio lysate storage chamber of eluent storage chamber 64 and two cleaning solution storage chambers are all It is small.

The shape of each storage chamber can also need to be arranged according to arrangement.Such as in the present embodiment eluent storage chamber 64 cross Section is rectangular, and the section of other three storage chambers 49 is L shape.

In the present embodiment, well 51 is connected to lysate storage chamber.Usually well 51 is coated on well 51 Diaphragm seals, when sample-adding punctures diaphragm, the splitting sample to be tested injection lysate storage chamber using liquid-transfering gun or sample needle It solves in liquid, completes sample-adding process.When sample to be tested injects lysate, it can realize in lysate storage chamber and at least partly crack Process.The present embodiment septation is made of aluminium foil, in other embodiments, diaphragm can by do not react with lysate its Its metal or nonmetallic materials are made, such as rubber membrane, resin film.

In the present embodiment, each storage chamber is correspondingly provided with a puncture needle 52.As shown in figure 8, four puncture needles 52 divide is Three groups, lysate storage chamber puncture needle 52 corresponding with the first cleaning solution storage chamber is first group and length is different, and second washes Washing the corresponding puncture needle 52 of liquid storage chamber is second group, and the corresponding puncture needle 52 of eluent storage chamber 64 is third group.Three groups are worn Difference on needle positions makes corresponding isolation structure have the different puncture time, makes the reagent in storage chamber suitable Time discharges to corresponding reaction chamber.

In the present embodiment, the length setting of first group of puncture needle 52 are as follows: puncture needle 52 corresponding with lysate storage chamber is most Long, puncture needle 52 corresponding with the first cleaning solution storage chamber takes second place.When pressing first group of puncture needle 52, with lysate storage chamber pair The puncture needle 52 answered is depressed first, is pierced its corresponding isolation structure first.

Puncturing structure piece 55 is located at the lower section of reagent storage organization piece 54, and setting is blind there are three being isolated on puncturing structure piece 55 Hole 58.In the present embodiment, isolation blind hole 58 forms isolation structure above-mentioned, and the thin-wall construction of 64 bottom of eluent storage chamber is also Blind hole 58 is isolated.Blind hole 58 is isolated in the present embodiment and storage chamber corresponds.

The thickness of the laminate structure of the bottom hole of blind hole 57 and the bottom hole of isolation blind hole 58 is accommodated convenient for the piercing of puncture needle 52.Example As the thickness is not more than 0.3mm in the present embodiment.

In first embodiment, the movement downward vertically that can use puncture needle tabletting 28 squeezes that different storage chambers are corresponding to wear Pricker 52, successively punctures accommodating blind hole 57 and isolation blind hole 58, and isolation blind hole 58 becomes through-hole then and can release corresponding storage Reagent in chamber.And needs can be reacted according to detection, certain time is stopped between the puncture of every two isolation blind hole 58, until most Puncture needle 52 afterwards is completed to puncture.

In the present embodiment, reagent connection control unit includes that the reagent connection being set between storage chamber 49 and reaction chamber is led to Road, reagent interface channel are set in connectivity structure piece 56.

Include in connectivity structure piece 56 and the one-to-one reagent interface channel 59 of storage chamber 49,59 phase of reagent interface channel It is mutually independent, it is vertical structure.

In the present embodiment, micro-fluidic chip 27 includes mixing chamber 61 and elution 63 two reaction chambers of chamber.Mixing chamber 61 is used for Magnetic bead absorption nucleic acid step and purification step during nucleic acid extraction, elution chamber 63 is for completing elution step.

Whether the hose 62 as internal connection control unit is connected to for controlling mixing chamber 61 with elution chamber 63.Mixing chamber 61, hose 62 and elution chamber 63 are all set in reaction structure piece 60.

For convenient for excluding the waste liquid in reaction chamber, waste liquid removes channel 70 and is also disposed on reaction structure piece 60, this implementation In example, waste liquid removes channel 70 and is connected to mixing chamber 61.

Hose 62 is fixed between mixing chamber 61 and elution chamber 63, for being connected to mixing chamber 61 and elution 63 two chambers of chamber Room.As shown in figure 8, mixing chamber 61 and elution chamber 63 are set to side by side in same level height.

Mixing chamber stomata 53 runs through storage chamber cover plate 50, reagent storage organization piece 54, puncturing structure piece 55 and connectivity structure Each layer of structure piece in piece 56, is connected with the mixing chamber 61 of micro-fluidic chip 27.In the present embodiment, mixing chamber stomata 53 can It is closed with rubber plug.

Hose 62 can be resiliently compressed by being made of soft rubber material.The both ends of hose 62 are separately connected mixing chamber 61 With elution chamber 63.The inside that hose 62 can be used as mixing chamber 61 when cross-sectional flow area is greater than certain value and elute between chamber 63 connects Circulation passage, it can be achieved that connecting inside this when making the cross-sectional flow area of hose 62 be less than certain value by the compressional deformation to hose 62 The closing of circulation passage.

Elution chamber lid piece 65 is set on reaction structure piece 60, and is set to 63 top of elution chamber, forms the portion of elution chamber 63 Chamber-separating wall.Elution chamber lid piece 65 is whole to be made of flexible material, and flexible material for example can be silica gel material.Elute chamber lid piece 65 Middle part be right-angled intersection narrow slit structure.Narrow slit structure is closed state usually, this to elute chamber 63 during the reaction In closed state；Chamber 63 is eluted when, the rifle point of liquid-transfering gun can after the reaction was completed protruded by the gap of narrow slit structure, it is narrow The narrow slit of crack structure can be squeezed by the rifle point of liquid-transfering gun and be opened, and take out purification of nucleic acid template after the reaction was completed by liquid-transfering gun.

Waste liquid storage configuration piece 66 is located at the lower section of reaction structure piece 60.Waste liquid chamber 67 is set to waste liquid storage configuration piece 66 In.Waste liquid bottom of chamber piece 69 is set to the bottom of waste liquid storage configuration piece 66, for closing the bottom of waste liquid chamber 67.

In first embodiment, the back upper place of waste liquid chamber 67 is provided with the waste liquid chamber stomata of the pressure for controlling waste liquid chamber 67 68.Waste liquid chamber 67 removes channel 70 by waste liquid and is connected with mixing chamber 61.Waste liquid removes channel 70 and waste liquid in the present embodiment Chamber stomata 68 constitutes waste liquid and excludes control unit.Wherein, waste liquid chamber stomata 68 constitutes the pressure control portion that waste liquid excludes control unit.

In micro-fluidic chip 27 material of each structure piece can be polycarbonate or polymethyl methacrylate materials, respectively It can be bonded by thermal bonding or chemical method between structure piece.

As shown in Figure 1, the nucleic acid extraction chip steerable system of first embodiment is coupled with micro-fluidic chip 27 to control Micro-fluidic chip executes nucleic acid extraction movement.

Chip steerable system includes that chip motion control module, reagent release module, inner passage are opened in first embodiment Die closing block, heating module, magnetic force apply module and waste liquid excludes control module.

Chip motion control module and micro-fluidic chip 27 are drivingly connected to control micro-fluidic chip movement.

In first embodiment, chip motion control module includes linear motion control unit.The linear motion control unit includes X-axis for driving micro-fluidic chip 27 to move along horizontal X-direction moves driving assembly and for driving micro-fluidic chip edge It is horizontally and perpendicularly to the Y-axis movement driving assembly of the Y-direction movement of X-direction.In the present embodiment, X-direction is the left and right in Fig. 1 Direction, Y direction are the front-rear direction in Fig. 1.

Referring to figs. 1 to Fig. 4.It includes Y-axis belt fixing piece 2, Y-axis belt 3, Y-axis sliding block 4, Y-axis that Y-axis, which moves driving assembly, Guide rail 5, y-axis motor 6, Y-axis position grating 7, Y-axis flute profile optocoupler 8 and Y-axis microswitch 9.Y-axis guide rail 5, y-axis motor 6, Y-axis Belt fixing piece 2 is fixed on bottom plate 1.Y-axis belt 3 drives Y-axis sliding block 4 in Y-axis guide rail 5 under the driving of y-axis motor 6 It moves in a straight line.Initial position is controlled by Y-axis microswitch 9 in Y-axis movement, and course location positions grating by Y-axis in Y-axis movement 7 and Y-axis flute profile optocoupler 8 control.

It includes pinboard 31, X-axis motor 32, X-axis guide rail 33, X-axis belt 34, X-axis slide block 35, X that X-axis, which moves driving assembly, Axis belt fixing piece 36, X-axis positioning grating 37, X-axis flute profile optocoupler 38 and X-axis microswitch 39.X-axis movement driving assembly passes through Pinboard 31 is integrally provided on Y-axis sliding block 4, and wherein pinboard 31 is fixed on Y-axis sliding block 4.X-axis motor 32, X-axis guide rail 33 It is fixed on pinboard 31 with X-axis belt fixing piece 36.X-axis belt 34 drives X-axis slide block 35 under the driving of X-axis motor 32 It is moved in a straight line in X-axis guide rail 33.Initial position is controlled by X-axis microswitch 39 in X-axis movement, process position in X-axis movement It sets and is controlled by X-axis positioning grating 37 and X-axis flute profile optocoupler 38.

As depicted in figs. 1 and 2, chip steerable system further includes chip butt joint base 40.Linear motion control unit passes through chip Butt joint base 40 is fixedly connected with micro-fluidic chip 27.Chip butt joint base 40 is fixed on the X-axis slide block 35 of chip motion control module On.Chip butt joint base 40 includes the slot for installing micro-fluidic chip 27, and the side wall of slot is fixed with spring pressuring plate 41.Slot Rear wall be fixed with chip position detection microswitch 42.

Chip butt joint base 40 is the interface for connecting and positioning between micro-fluidic chip 27 and chip motion control module, is one A pedestal with groove, by the cooperation of spring pressuring plate 41, make micro-fluidic chip 27 in the horizontal and vertical directions with chip pair Joint chair 40 is fitted close, and realizes the rapid abutting joint between micro-fluidic chip 27 and chip control system.Driving group is moved by Y-axis The rapid abutting joint of the cooperation of part and X-axis movement driving assembly and micro-fluidic chip 27 and chip butt joint base 41 is, it can be achieved that micro-fluidic Two-dimensional motion of the chip 27 in X-direction, Y direction.

Reagent release module is connected between storage chamber and corresponding reaction chamber for controlling reagent connection control unit with controlling.

As shown in Figure 1, Figure 2 and shown in Figure 11 A, Figure 11 B, in the present embodiment, reagent release module includes 28 He of puncture needle tabletting The tabletting driving mechanism being drivingly connected with puncture needle tabletting 28.Tabletting driving mechanism specifically includes reagent release motor 29.

In the present embodiment, reagent release module further includes reagent release electric machine support 30, reagent release positioning grating, reagent Discharge flute profile optocoupler, reagent release microswitch and tabletting guiding mechanism.Reagent discharges motor 29 and the setting of tabletting guiding mechanism In on reagent release electric machine support 30.Tabletting guiding mechanism includes the guide post being vertically arranged on reagent release electric machine support 30 With the guide runner being slidably matched with guide post, puncture needle tabletting 28 and guide runner are fixedly installed.At the beginning of puncture needle tabletting 28 Beginning, position was by reagent release microswitch control.The course location of puncture needle tabletting 28 discharges positioning grating by reagent and reagent is released Put the control of flute profile optocoupler.

During reagent release, reagent release motor 29 drives puncture needle tabletting 28 to move downward, and successively squeezes storage chamber The puncture needle 52 of top, so that puncture needle 52, which punctures it, corresponds to accommodating blind hole 57 and isolation blind hole 58, in corresponding accommodating blind hole After 57 become through-hole with isolation blind hole 58, the reagent in storage chamber is discharged to corresponding reaction chamber.In the present embodiment, by nothing Pump type drive mode, so that reagent is achieved in releasing automatically for reagent automatically into the mixing chamber 61 or elution chamber 63 of lower section Let off journey.

Chip steerable system further includes inner passage opening and closing module.Inner passage opening and closing module includes control in the present embodiment Whether the hose control assembly of the cross-sectional flow area of hose 62 is connected between mixing chamber 61 and elution chamber 63 with controlling.

Hose control assembly includes closing sliding block 23 and closing slider-actuated mechanism, and closing slider-actuated mechanism and closing are slided Block 23 is drivingly connected, and closing sliding block 23 changes the cross-sectional flow area of hose 62 along the radially movable setting of hose 62.This In embodiment, closing slider-actuated mechanism is that hose closes motor 24.

Hose closed module is fixed on pinboard 31, therefore, the opposite position of hose closed module and micro-fluidic chip 27 It sets and is easy to determining, convenient for controlling the connected state of hose 62.

In hose closed process, closing sliding block 23 upward vertical movement under the driving of hose closing motor 21, with envelope It closes sliding block 23 to move upwards, close sliding block 23 makes 62 deformation of hose to passage section face close to one end squeezing tube 62 of hose 62 Product be less than certain value, can closed mixing chamber 61 and elution chamber 63 between inside communication channel.Preferably, such as Figure 12 A and figure Shown in 12B, closing sliding block 23 includes protrusion close to one end of hose 62, and when controlling the cross-sectional flow area of hose 62, closing is slided The raised squeezing tube 62 of block 23.

The chip steerable system of the present embodiment further includes heating module, and heating module is for heating elution chamber 63.Heated mould Block specifically includes heating film 24.In the present embodiment, heating film 24 is fixed on one end of the direction elution chamber 63 of closing sliding block 23. As illustrated in figs. 12 a and 12b, it is arranged upward for the protrusion of squeezing tube 62 with heating film 24.It is upward in closing sliding block 23 While moving squeezing tube 62, heating film 24 moves up to the bottom wall of elution chamber 63 and is bonded therewith, in elution process In to magnetic bead 71 dry and elution reagent is heated.

It includes the alternation for applying magnetive attraction for the magnetic bead 71 in the reaction chamber to micro-fluidic chip 27 that magnetic force, which applies module, Power applies magnet.

As shown in Figure 1, Figure 2, shown in Fig. 5, Fig. 9 A, Fig. 9 B, Figure 10 A and Figure 10 B, in first embodiment, magnetic force applies module packet Two alternating forces for including relative spacing setting apply magnet, and the mixing chamber 61 of micro-fluidic chip 27 is located at two alternating forces and applies magnetic Between body.In the present embodiment, it is respectively the spaced left side permanent magnet 16 in left and right and right side that two alternating forces, which apply magnet, Permanent magnet 20.

It further includes magnet movement driving assembly that magnetic force, which applies module, and magnet movement driving assembly is for controlling two alternating forces Apply magnet to move back and forth along the direction vertical with two alternating forces application distribution arrangements of magnet.

In the present embodiment, it includes left magnet carrier 10, left magnet motors 11, left magnet sliding block 12, a left side that magnetic force, which applies module, Magnet position grating 13, left magnet flute profile optocoupler 14, left magnet microswitch 15, left side permanent magnet 16, right magnet carrier 17, Right magnet motors 18, right magnet sliding block 19, right magnet positioning grating, right magnet flute profile optocoupler, right magnet microswitch, right side are forever Long magnet 20.Left magnet carrier 10 and right magnet carrier 17 are separately fixed at the left and right side of bottom plate 1.Left magnet motors 11, Left magnet sliding block 12 and right magnet motors 18, right magnet sliding block 19 constitute magnet movement driving assembly.Left magnet motors 11 and a left side Magnet sliding block 12 is drivingly connected, and right magnet motors 18 are drivingly connected with right magnet sliding block 19.Left magnet sliding block 12 and right magnet are sliding The outer end face setting of block 19 is fluted, and groove is used for fixed permanent magnet.

During nucleic acid absorption, cracking mixed liquor is discharged to mixing chamber 61, it is mixed with the magnetic bead 71 in mixing chamber 61 It closes.

As shown in fig. 9 a and fig. 9b, moving driving assembly by X-axis drives micro-fluidic chip 27 reciprocal in X-direction or so Movement makes the left and right sides outer wall timesharing of mixing chamber 61 close to left side permanent magnet 16 and right side permanent magnet 20, mixing chamber 61 Inside generates a cycle changing magnetic field, the field drives magnetic bead 71 left and right in reaction reagent under permanent magnet effect It moves back and forth.

While 27 bilateral reciprocation of micro-fluidic chip, left side permanent magnet 16 and right side permanent magnet 20 are in magnet Under the driving for moving driving assembly, with the lower movement in the vertical direction of certain rate, so that magnetic bead 71 and reaction reagent is each A liquid level comes into full contact with.

The up and down motion that the bilateral reciprocation of micro-fluidic chip 27 and two alternating forces apply magnet makes magnetic bead 71 and splits Solution mixed liquor reaches well-mixed effect, and the nucleic acid after making cracking is sufficiently adsorbed by magnetic bead 71.

In washing process, magnetic bead 71 can be made to be adsorbed with the magnetic bead of nucleic acid with same or like hybrid mode washing 71。

And in elution step, it can also make to elute chamber 63 by X-axis motion control component positioned at two alternating forces application magnets Between, make the magnetic bead for being adsorbed with nucleic acid 71 complete to be sufficiently mixed and gradually with eluent with same or like hybrid mode The nucleic acid on lower magnetic bead 71 is eluted, until elution finishes.

As shown in figs. 10 a and 10b, in magnetic bead transfer process, pass through the X-axis motion control group of chip motion control module Part driving micro-fluidic chip 27 moves downward so that left side permanent magnet 16 while discharging soft close to the left side outer wall of mixing chamber 61 Pipe 62, so that the hose 62 between mixing chamber 61 and elution chamber 63 is opened, liquid reagent penetrates through mixing chamber 61 and elution chamber 63.It is logical Cross Y-axis movement driving assembly driving micro-fluidic chip 27 moved backward in Y direction, the magnetic bead 71 in mixing chamber 61 left side forever Pass through the liquid reagent in hose 62 under the driving in 16 magnetic field of long magnet, is moved in elution chamber 63.Then pass through chip motion Control module driving micro-fluidic chip 27 moves to the top of bottom permanent magnet 26, makes the bottom face of elution chamber 63 close to bottom Permanent magnet 26 elutes the bottom that the magnetic bead 71 in chamber 63 is laid in elution chamber 63 under the suction-operated of bottom permanent magnet 26 Portion excludes control module by waste liquid and exhausts cleaning solution, is achieved in the transfer process of magnetic bead 71.

In the present embodiment, it includes that waste liquid removes channel 70 and as pressure control that the waste liquid of micro-fluidic chip 27, which excludes control unit, The waste liquid chamber stomata 68 in portion processed.The waste liquid of chip steerable system excludes the pressure that control module is used to control waste liquid chamber 67.Pass through The pressure of control waste liquid chamber 67 can control the pressure difference that waste liquid removes 70 both ends of channel, so as to control mixing chamber 61 and give up Whether sap cavity 67, which can remove channel 70 by waste liquid, is connected to.

In the present embodiment, it includes the vacuum being connected to waste liquid chamber stomata 68 that the waste liquid of chip steerable system, which excludes control module, Pumping components.

As shown in fig. 6, vacuum suction component includes sucker stand 43, sucker 44, gas-guide tube 45, vacuum pump 46, shut-off valve 47 and vacuum pump fixed frame 48.

Vacuum pump fixed frame 48 is fixed on chip butt joint base 40, after sucker stand 43 is fixed on chip butt joint base 40 End, sucker 44 and the waste liquid chamber stomata 68 of micro-fluidic chip 27 are concentric.Sucker 44 is connected to by gas-guide tube 45 with vacuum pump 46. When micro-fluidic chip 27 is inserted into chip butt joint base 40, sucker 44 is bonded with the waste liquid chamber stomata 68 of horizontal structure.Shut-off valve 47 close waste liquid chamber stomata 68 in the off state, so that enclosure space is formed inside waste liquid chamber 67, so that liquid reagent Since the effect of atmospheric pressure will not leak into waste liquid chamber 67 down in mixing chamber 61 and elution chamber 63.

During waste liquid removes, shut-off valve 47 and vacuum pump 46 are opened simultaneously, and waste liquid chamber stomata 68 is opened, in waste liquid chamber 67 It is evacuated, the waste liquid in mixing chamber 61 and elution chamber 63 removes channel 70 by waste liquid under pressure difference and gravity and flow to In waste liquid chamber 67, it is achieved in waste liquid and removes process.It can accelerate the discharge of waste liquid by the pump-absorb action of vacuum pump 46.

A kind of specific operation process that nucleic acid is extracted using the micro fluidic device of first embodiment is described below.

The aluminium foil being covered on well 51 is punctured, sample to be tested is added to by well 51 and is stored with lysate Lysate storage chamber in.

Micro-fluidic chip 27 after sample-adding is inserted into chip butt joint base 40 and is allowed to fixed.At this point, sucker 44 and waste liquid chamber Stomata 68 is bonded, and shut-off valve 47 closes gas-guide tube 45 in the off state, to close waste liquid chamber stomata 68.Meanwhile hose seals It closes the driving closing sliding block 23 of motor 21 to move vertically upwards, the raised squeezing tube 62 on closing sliding block 23 is allowed to be deformed into through-flow Sectional area is almost nil, the interface channel between closed mixing chamber 61 and elution chamber 63.

Chip motion control module drive micro-fluidic chip 27 is moved to the underface of puncture needle tabletting 28, so that micro-fluidic First group of puncture needle face puncture needle tabletting 28 on chip 27.Reagent discharges motor 29 and drives puncture needle tabletting 28 vertically downward Movement squeezes puncture needle 52, and puncture needle 52 is made successively to puncture accommodating blind hole 57 corresponding with lysate storage chamber and isolation blind hole 58.Cracking mixed liquor in lysate storage chamber, which passes through corresponding reagent interface channel 59 under the effect of gravity and is discharged into, to be packaged with Magnetic bead 71 mixing chamber 61 in.

Cracking mixed liquor after the completion of cleavage step can be discharged to mixing chamber 61.This can be prevented in sample to be tested Nucleic acid other than substance adhered in the case where not cracking on magnetic bead 71 and influence the speed that magnetic bead 71 adsorbs nucleic acid. It is of course also possible to the mixed solution of the sample to be tested of unfinished cracking and lysate be discharged to mixing chamber 61, in cracking Nucleic acid is adsorbed simultaneously.

Chip motion control module driving micro-fluidic chip 27 makes mixing chamber 61 be moved to left side permanent magnet 16 and right side Between permanent magnet 20.Under the driving of X-axis movement driving assembly, micro-fluidic chip 27 is moved to the left left side permanent magnet 16 Side so that the left side outer wall of mixing chamber 61 is close to close left side permanent magnet 16, the magnetic bead 71 in lysate is permanent in left side It is moved downward under the magnetic fields of magnet 16.After magnetic bead 71 is moved fully to the inner left wall of mixing chamber 61, drive micro-fluidic Chip 27 moves right to the side of right side permanent magnet 20, so that the outer right wall of mixing chamber 61 is close to close right side permanent magnet 20, magnetic bead 71 moves right under 20 magnetic fields of right side permanent magnet, is eventually collected in the right side inner wall of mixing chamber 61.Pass through Bilateral reciprocation of the magnetic bead 71 in mixing chamber 61, so that the nucleic acid in lysate comes into full contact with magnetic bead 71, by magnetic bead 71 Absorption.

After the completion of nucleic acid absorption step, micro-fluidic chip 27 is moved to the side of right side permanent magnet 20, makes magnetic bead 71 The right side inner wall of mixing chamber 61 is gathered under the magnetic fields of right side permanent magnet 20 and is adsorbed.Start 47 He of shut-off valve Vacuum pump 46, so that the waste liquid in mixing chamber 61 removes channel 70 by waste liquid and is transferred down in waste liquid chamber 67, it is complete to waste liquid After full removal, vacuum pump 46 and shut-off valve 47 are closed, closes waste liquid chamber stomata 68.

The first cleaning solution of release is pushed using the control puncture needle tabletting 28 of reagent release module and is completed in mixing chamber 61 It is similarly operated with nucleic acid absorption step, the magnetic bead 71 of nucleic acid is adsorbed with using the washing of the first cleaning solution.Puncture needle pressure is controlled again Piece 28 pushes the second cleaning solution of release and completes similarly to operate with nucleic acid absorption in mixing chamber 61, is washed using the second cleaning solution Wash the magnetic bead 71 for being adsorbed with nucleic acid.

After the completion of the second cleaning solution is mixed with magnetic bead 71, does not start waste liquid and exclude control module, by micro-fluidic chip 27 It is moved to the side of left side permanent magnet 16, magnetic bead 71 gathers mixing chamber 61 under the magnetic fields of left side permanent magnet 16 Inner left wall.Hose closing motor 21 driving closing sliding block 23 move downward, release hose 62, mixing chamber 61 and elution chamber 63 it Between form inside communication channel, the second cleaning solution of part by hose 62 flows to elution chamber 63 and full of hose 62.

Y-axis movement driving assembly driving micro-fluidic chip 27 moves backward, so that the magnetic bead 71 in mixing chamber 61 is in left side It is transferred in elution chamber 63 under the sucking action of permanent magnet 16 by hose 62.Elution chamber 63 is completely transferred to magnetic bead 71 Afterwards, it is moved upwards using the hose closing driving closing sliding block 23 of motor 21, closed hose 62.

Chip motion control module drives micro-fluidic chip 27, so that it is eluted chamber 63 and moves to the upper of bottom permanent magnet 26 Side elutes the magnetic bead 71 in chamber 63 and is laid in the bottom of elution chamber 63 under the magneticaction of bottom permanent magnet 26 and is inhaled Draw.The hose closing driving closing sliding block 23 of motor 21 moves downward, and release hose 62 opens vacuum pump 46 and shut-off valve 47, row Fall to elute the second cleaning solution in chamber 63, hose 62 and mixing chamber 61.

Then re-closed hose 62 and waste liquid chamber stomata 68, while heating film 24 is opened, the magnetic of drying elution 63 bottom of chamber Pearl 71 avoids the second cleaning solution of 71 adsorption of magnetic bead from polluting subsequent elution process, completes purification step.

After the completion of purification step, driving reagent release module discharges eluent to elution chamber 63, and heating film 24 is in and opens State is opened, Eluant temperature is made to rise to 55 DEG C, progress is similarly operated with nucleic acid absorption, and completion magnetic bead 71 is mixed with eluent It closes, the Nucleic Acid Elution for being adsorbed on 71 surface of magnetic bead is got off to get to purification of nucleic acid template, elution step is completed.

Utilize the purification of nucleic acid template after sampling probe extraction elution.Nucleic acid is completed as a result, automatically extracts overall process.

Wherein sample to be tested is, for example, blood, the virion in saliva.Sample to be tested is carried out in micro-fluidic chip Purification of nucleic acid template is obtained after cracking, absorption, purifying and elution, purifying core needed for providing nucleic acid amplification reaction for molecular diagnosis Acid template.

Second embodiment

As shown in figure 13, the difference of second embodiment and first embodiment is: in micro-fluidic chip 27, mixing chamber 61 with Elution chamber 63 is vertical structure, and hose 62 is placed and is connected to vertically between mixing chamber 61 and elution chamber 63, and waste liquid removes channel 70 are located at the bottom of elution chamber 63, and waste liquid chamber stomata 68 is located at the top of waste liquid chamber 67.Correspondingly, the closing of chip steerable system Sliding block 23 is located at the side of micro-fluidic chip 27, and it is soft to move horizontally extruding for closing sliding block 23 under the driving of hose closing motor 21 Pipe 62 reaches closed effect.Meanwhile the heating film 24 being fixed on closing sliding block 23 fits to the side wall of elution chamber 63.

When needing to be discharged waste liquid to the waste liquid chamber 67 in mixing chamber 61, release hose 62 and waste liquid chamber stomata 68, mixing chamber Waste liquid in 61 removes channel 70 by hose 62 and waste liquid and enters waste liquid chamber 67.Elution step is completed in elution chamber 63, the After the purification process of secondary washing, the magnetic bead 71 in mixing chamber 61 is moved to the upper end of hose 62 using permanent magnet, Then release hose 62, magnetic bead 71 follow the second cleaning solution to enter in elution chamber 63 by gravity, complete magnetic bead 71 Transfer process.

Other unaccounted parts, can refer to the associated description of remaining embodiment in second embodiment.

3rd embodiment

As shown in figure 14,3rd embodiment and first embodiment the difference is that: the pressure control group of chip steerable system Part is not sucked by vacuum component, the rubber stopper 72 of solenoid valve 73 and the end for being connected to solenoid valve 73.Wherein, waste liquid chamber gas Hole 68 is arranged above waste liquid chamber 67, and solenoid valve 73 is for controlling the opening and closing of waste liquid chamber stomata 68.

Solenoid valve 73 drives the closing waste liquid chamber stomata 68 downwards of rubber stopper 72 in the on state, so that inside waste liquid chamber 67 Enclosure space is formed, the liquid reagent in reaction process in mixing chamber 61 rests on mixing chamber 61 because waste liquid chamber stomata 68 is closed It is interior without flowing downward.Solenoid valve 73 drives rubber stopper 72 to move upwards in the off state, and waste liquid chamber stomata 68 is opened, waste liquid Chamber 67 and outside air penetrate through, and the waste liquid in mixing chamber 61 is under the interaction of gravity and microchannel inner surface tension, downwards It flow in waste liquid chamber 67.Waste liquid is completed by the unlatching of waste liquid chamber stomata 68 and closed state as a result, and removes process.

Other unaccounted parts, can refer to the associated description of remaining embodiment in 3rd embodiment.

The content for description that the present invention is not limited to the above embodiments.Such as the frame sheet number of micro-fluidic chip 27, respectively Functional stomata, channel, the shape of reaction chamber, size and quantity etc. can change as needed.

In addition, it will appreciated by the skilled person that realizing all or part of the nucleic acid extraction of above-described embodiment Step may be implemented by hardware, and relevant hardware can also be instructed to complete by program, program can store in one In kind computer readable storage medium, storage medium mentioned above can be read-only memory, disk or CD etc..

Compared with prior art, above embodiments of the present invention at least one have the following effects that:

Reaction reagent is stored on micro-fluidic chip, makes reagent integrated with micro-fluidic chip composition one, disposable The reaction structure body used；In reaction process, by quickly, discharge the reaction reagent that stores in chip one by one so that all behaviour Work can be completed in a totally enclosed environment, save reagent metering and filling step, improve nucleic acid extraction efficiency, reduce behaviour Make error probability, moreover it is possible to effectively avoid the cross-infection in biomedical detect.

By the mutual cooperation between micro-fluidic chip and chip steerable system, it is sequentially completed cracking, nucleic acid absorption, pure Change, elution and etc., finally obtain purification of nucleic acid template needed for quickly providing nucleic acid amplification reaction for molecular diagnosis.

A kind of higher chip steerable system of the degree of automation is provided, using automation, the operation mode of continuous-flow type, certainly It is dynamic to complete nucleic acid extraction overall process.

Position transfer is carried out between different modules using chip motion control module driving micro-fluidic chip and function is cut It changes, space can be saved, realize portability, while the functional requirement on different dimensions can be integrated again, realize full-automatic.

It proposes based on the reagent method for releasing for puncturing pin puncture, driving puncture needle successively punctures accommodating blind hole and is isolated blind Hole allows reagent quick release by a kind of non-pump type drive mode under the collective effect of gravity and surface tension, realizes reagent Driving process.

Cyclically-varying magnetic field is generated using magnet, driving magnetic bead movement drives miniflow by chip motion driving assembly Control chip and/or magnet movement driving assembly, which drive alternating force to apply magnet, applies micro-fluidic chip and alternating force between magnet Generate relative motion, move back and forth in reaction chamber to generate cyclically-varying field drives magnetic bead, realize magnetic bead with Efficient mixing between reaction reagent.

Using the elasticity of hose, allows hose to be used as mixing chamber in the released state and elute the interface channel between chamber, benefit In completing magnetic bead transfer process, in the case where hose is in squeezed state, closed mixing chamber and elution chamber, respectively become it mutually solely Vertical reaction chamber completes each step reaction, avoids the cross contamination between reagent.

Finally it should be noted that: the above embodiments are merely illustrative of the technical scheme of the present invention and are not intended to be limiting thereof；To the greatest extent The present invention is described in detail with reference to preferred embodiments for pipe, it should be understood by those ordinary skilled in the art that: still It can modify to a specific embodiment of the invention or some technical features can be equivalently replaced；Without departing from this hair The spirit of bright technical solution should all cover within the scope of the technical scheme claimed by the invention.

Claims (43)

1. a kind of nucleic acid extraction micro-fluidic chip, which is characterized in that be connected to including at least one storage chamber, at least one reagent Control unit and at least one reaction chamber, each storage chamber is correspondingly arranged with a reaction chamber, and each storage Reagent connection control unit is provided between chamber and corresponding reaction chamber both to control and whether to be connected to, wherein it is described at least One storage chamber is packaged with the reagent for nucleic acid extraction.

2. micro-fluidic chip according to claim 1, which is characterized in that at least one described reaction chamber includes mixing chamber (61), at least one described storage chamber includes the lysate storage for being packaged with lysate and being correspondingly arranged with the mixing chamber (61) Chamber and/or the cleaning solution storage chamber including being packaged with cleaning solution and being correspondingly arranged with the mixing chamber (61)；And/or it is described extremely A few reaction chamber includes elution chamber (63), at least one described storage chamber include be packaged with eluent and with the elution chamber (63) the eluent storage chamber (64) being correspondingly arranged.

3. micro-fluidic chip according to claim 2, which is characterized in that the micro-fluidic chip further includes sample-adding structure, The sample-adding structure includes well (51) and diaphragm, and piercing through diaphragm can make the lysate storage chamber pass through the well (51) it is in communication with the outside.

4. micro-fluidic chip according to claim 1, which is characterized in that at least one described reaction chamber includes mixing chamber (61), magnetic bead (71) are packaged in the mixing chamber (61).

5. micro-fluidic chip according to claim 1, which is characterized in that the reagent connection control unit includes being set to institute State the isolation structure between storage chamber and the corresponding reaction chamber, by pierce through the isolation structure control the storage chamber with The corresponding reaction chamber connection.

6. micro-fluidic chip according to claim 5, which is characterized in that the reagent connection control unit includes being set to institute The reagent interface channel (59) between storage chamber and the corresponding reaction chamber is stated, the isolation structure includes being set to described deposit The bottom hole of the isolation blind hole (58) of storage chamber bottom, isolation blind hole (58) is located at the corresponding reagent interface channel (59) On.

7. micro-fluidic chip according to claim 5, which is characterized in that the reagent connection control unit includes for piercing through The puncture needle (52) of the isolation structure and the accommodating blind hole (57) being set at the top of corresponding storage chamber, the puncture needle (52) Be set in the accommodating blind hole (57) and can along the accommodating blind hole (57) to corresponding storage chamber internal motion to pierce through The bottom hole of accommodating blind hole (57) and the isolation structure.

8. micro-fluidic chip according to claim 5, which is characterized in that at least one described reagent is connected to control unit and its The structure of remaining reagent connection control unit is different and/or setting height is different, so that at least one described reagent connection control unit The time of piercing through of isolation structure is different from the puncture time that remaining reagent is connected to the isolation structure of control unit.

9. micro-fluidic chip according to claim 1, which is characterized in that at least one described reaction chamber includes mixing chamber (61), the micro-fluidic chip further includes mixing chamber stomata (53), and the mixing chamber stomata (53) and the mixing chamber (61) are even It is logical.

10. micro-fluidic chip according to claim 1, which is characterized in that at least one described reaction chamber includes mixing chamber (61) and elution chamber (63), the micro-fluidic chip further include internal connection control unit, and the internal connection control unit is for controlling Make whether the mixing chamber (61) is connected to the elution chamber (63).

11. micro-fluidic chip according to claim 10, which is characterized in that the internal connection control unit includes being connected to Hose (62) between the mixing chamber (61) and elution chamber (63), by the cross-sectional flow area for controlling the hose (62) It controls and whether is connected between the mixing chamber (61) and the elution chamber (63).

12. micro-fluidic chip according to claim 11, which is characterized in that the mixing chamber (61) and the elution chamber (63) it is set in same level height, the hose (62) is arranged substantially horizontally；Alternatively, the mixing chamber (61) is set to Above the elution chamber (63), the hose (62) is substantially vertically arranged.

13. micro-fluidic chip according to claim 1, which is characterized in that at least one described reaction chamber includes elution chamber (63), elution chamber lid piece (65) of the micro-fluidic chip including forming elution chamber (63) part cavity wall, and the elution Chamber lid piece (65) is provided with the gap structure to be opened/closed made of flexible material.

14. micro-fluidic chip according to claim 1, which is characterized in that the micro-fluidic chip further includes waste liquid chamber (67) and waste liquid excludes control unit, the waste liquid exclude control unit for control the reaction chamber whether with the waste liquid chamber (67) Connection.

15. micro-fluidic chip according to claim 14, which is characterized in that it includes being set to that the waste liquid, which excludes control unit, Waste liquid between the reaction chamber and the waste liquid chamber (67) removes channel (70) and pressure control portion, the pressure control portion control The pressure of the waste liquid chamber (67) is made to control between the reaction chamber and the waste liquid chamber (67) and whether to move by the waste liquid Except channel (70) are connected to.

16. micro-fluidic chip according to claim 15, which is characterized in that the pressure control portion includes and the waste liquid The waste liquid chamber stomata (68) of chamber (67) connection, by controlling whether the waste liquid chamber stomata (68) vacuumizes or whether close control The pressure of the waste liquid chamber (67).

17. a kind of nucleic acid extraction chip steerable system, which is characterized in that the chip steerable system with according to right for wanting Micro-fluidic chip described in asking any one of 1 to 16 (27) couples and controls the micro-fluidic chip (27) execution nucleic acid extraction and moves Make.

18. chip steerable system according to claim 17, which is characterized in that the chip steerable system includes chip fortune Dynamic control module, the chip motion control module and the micro-fluidic chip (27) are drivingly connected to control the micro-fluidic core Piece (27) movement.

19. chip steerable system according to claim 18, which is characterized in that the chip motion control module includes driving The linear motion control unit of the micro-fluidic chip (27) linear motion is moved, the linear motion control unit includes:

X-axis moves driving assembly, for driving the micro-fluidic chip to move along horizontal X-direction；And/or

Y-axis moves driving assembly, for driving the micro-fluidic chip to move along the Y-direction for being horizontally and perpendicularly to the X-direction； And/or

Z axis moves driving assembly, for driving the micro-fluidic chip to move along vertical Z-direction.

20. chip steerable system according to claim 18, which is characterized in that the chip steerable system further includes chip Butt joint base (40) and chip position detection device, the chip butt joint base (40) include for accommodating the micro-fluidic chip (27) Slot and the spring pressuring plate (41) that is connected on the side wall of the slot, the micro-fluidic chip (27) pass through the chip pair The slot and spring pressuring plate (41) of joint chair (40) and the chip motion control module are detachably connected, the chip position inspection Device is surveyed for detecting whether the micro-fluidic chip (27) docks successfully with the chip butt joint base (40).

21. chip steerable system according to claim 17, which is characterized in that the chip steerable system further includes magnetic force Apply module, it includes that alternating force applies magnet that the magnetic force, which applies module, and the alternating force applies magnet and is used for the miniflow The magnetic bead (71) controlled in the reaction chamber of chip (27) applies alternation magnetive attraction.

22. chip steerable system according to claim 21, which is characterized in that between the magnetic force application module includes opposite Apply magnet every two alternating forces of setting, the micro-fluidic chip (27), which can move to, makes the reaction chamber be located at described two A alternating force applies between magnet, while the magnetic bead (71) in the micro-fluidic chip (27) can be applied in described two alternating forces Add and is moved back and forth between magnet.

23. chip steerable system according to claim 21, which is characterized in that it further includes magnet that the magnetic force, which applies module, Driving assembly is moved, the magnet movement driving assembly is for driving the alternating force to apply magnet relative to the micro-fluidic core Piece (27) movement.

24. chip steerable system according to claim 22, which is characterized in that it further includes magnet that the magnetic force, which applies module, Move driving assembly, the magnet movement driving assembly for drive described two alternating forces apply magnets along with it is described two The direction that the distribution arrangement of alternating force application magnet is vertical moves back and forth；It is in the horizontal direction that described two alternating forces, which apply magnet, The left side permanent magnet (16) and right side permanent magnet (20) of distribution；The magnet movement driving assembly include with the left side forever Left magnet motors (11), left magnet sliding block (12) and the left magnet carrier (10) that long magnet (16) is correspondingly arranged, and including with Right magnet motors (18), right magnet sliding block (19) and the right magnet carrier (17) that the right side permanent magnet (20) is correspondingly arranged, The left magnet sliding block (12) and the left magnet motors (11) are drivingly connected, the right magnet sliding block (12) and the right magnet Motor (18) is drivingly connected, and the left side permanent magnet (16) and the right side permanent magnet (20) are separately fixed at the left magnetic On body sliding block (12) and the right magnet sliding block (19).

25. chip steerable system according to claim 21, which is characterized in that it further includes bottom that the magnetic force, which applies module, The reaction chamber of magnet, the micro-fluidic chip (27) includes elution chamber (63), and the elution chamber (63) can move to the bottom The top of portion's magnet, the bottom magnet are used to elute the magnetic bead (71) in chamber (63) and are adsorbed on the bottom of elution chamber (63) simultaneously In tiled state.

26. chip steerable system according to claim 17, which is characterized in that the chip steerable system further includes reagent Release module, the reagent release module is for controlling reagent connection control unit so that the storage chamber and corresponding described It is connected between reaction chamber.

27. chip steerable system according to claim 26, which is characterized in that the reagent connection control unit includes setting Isolation structure between the storage chamber and the corresponding reaction chamber and the puncture needle for piercing through the isolation structure (52), the reagent release module includes puncture needle tabletting (28), and the puncture needle tabletting (28) is located at the puncture needle (52) The side far from the micro-fluidic chip (27) and be movably disposed along the puncture direction of the puncture needle (52) with by institute It states puncture needle (52) and is pierced into the isolation structure.

28. chip steerable system according to claim 27, which is characterized in that the reagent release module further includes tabletting Driving mechanism, the tabletting driving mechanism include reagent release motor (29), reagent release electric machine support (30), and the reagent is released Electrical Discharge Machine (29) and the puncture needle tabletting (28) are respectively arranged on reagent release electric machine support (30), and the reagent is released Electrical Discharge Machine (29) and the puncture needle tabletting (28) are drivingly connected to control the puncture needle tabletting (28) along the puncture needle (52) puncture direction is mobile.

29. chip steerable system according to claim 17, which is characterized in that at least one described reaction chamber includes mixing Chamber (61) and elution chamber (63), the micro-fluidic chip (27) further include internal connection control unit, the internal connection control unit Whether it is connected to for controlling the mixing chamber (61) with the elution chamber (63), the chip steerable system further includes for manipulating The inner passage of the internal connection control unit opens and closes module.

30. chip steerable system according to claim 29, which is characterized in that the internal connection control unit includes connection Hose (62) between the mixing chamber (61) and elution chamber (63), the inner passage opening and closing module includes control institute The hose control assembly of the cross-sectional flow area of hose (62) is stated, the hose control assembly includes closing sliding block (23), the envelope Sliding block is closed to be arranged radially displaceablely along the hose (62) to change institute with the contact position of the hose (62) by changing State the cross-sectional flow area of hose (62).

31. chip steerable system according to claim 30, which is characterized in that the chip steerable system further includes being used for The heating module of elution chamber (63) is heated, the heating module includes heating film (24), and the heating film (24) is fixed on One end towards elution chamber (63) of closing sliding block (23).

32. chip steerable system according to claim 17, which is characterized in that the micro-fluidic chip (27) further includes giving up Sap cavity (67) and waste liquid exclude control unit, the waste liquid exclude control unit for control the reaction chamber whether with the waste liquid chamber (67) it is connected to, it includes that the waste liquid being set between the reaction chamber and the waste liquid chamber (67) removes that the waste liquid, which excludes control unit, Channel (70) and the waste liquid chamber stomata (68) being connected to the waste liquid chamber (67), the chip steerable system further includes for controlling The waste liquid that the waste liquid excludes control unit excludes control module, and it includes for controlling the waste liquid that the waste liquid, which excludes control module, The pressure control assembly of chamber (67) pressure, the pressure control assembly is by controlling whether the waste liquid chamber stomata (68) vacuumizes Or whether close to control the pressure of the waste liquid chamber (67).

33. chip steerable system according to claim 32, which is characterized in that the pressure control assembly include with it is described The vacuum suction component of waste liquid chamber stomata (68) connection；Alternatively, the pressure control assembly includes controlling the waste liquid chamber stomata (68) valve being opened and closed.

34. chip steerable system described in any one of 7 to 33 according to claim 1, the chip steerable system includes movement Component and the movement position detection device being correspondingly arranged with the moving parts, wherein

The moving parts include motor, and the corresponding movement position detection device of the moving parts includes microswitch, in institute It states in moving parts motion process, initial position or the termination of moving parts is controlled by the corresponding microswitch of moving parts Position controls the course location of moving parts by the movement velocity of the motor of moving parts and time；Alternatively,

The corresponding movement position detection device of the moving parts includes microswitch, grating and flute profile optocoupler, in the movement During component movement, the initial position or final position of moving parts are controlled by the corresponding microswitch of moving parts, is led to Cross the course location of the corresponding grating of moving parts and flute profile optocoupler control moving parts.

35. a kind of nucleic acid extraction micro fluidic device, the micro fluidic device includes micro-fluidic chip and described micro- for controlling Fluidic chip executes the chip steerable system of nucleic acid extraction movement, which is characterized in that the micro-fluidic chip is to be wanted according to right Micro-fluidic chip described in asking any one of 1 to 16, the chip steerable system are according to any one of claim 17 to 34 The chip steerable system.

36. a kind of method for extracting nucleic acid, which is characterized in that the method for extracting nucleic acid utilizes micro-fluidic described in claim 35 Device extracts nucleic acid, and the method for extracting nucleic acid includes: control reagent connection control unit corresponding with the storage chamber, makes described Reagent at least one storage chamber is discharged to corresponding reaction chamber.

37. method for extracting nucleic acid according to claim 36, which is characterized in that the method for extracting nucleic acid includes:

Cleavage step is added sample to be tested into the lysate storage chamber for being stored with lysate of at least one storage chamber and obtains To cracking mixed liquor；

Mixed liquor transfer step is cracked, reagent corresponding with the lysate storage chamber is controlled and is connected to control unit, will be cracked mixed Liquid is closed to be discharged into the mixing chamber (61) of at least one reaction chamber；

Nucleic acid absorption step moves magnetic bead (71) in the mixing chamber (61) and adsorbs the nucleic acid in cracking mixed liquor；

Nucleic acid purification step, washing are adsorbed with the magnetic bead (71) of nucleic acid；

Nucleic Acid Elution on the magnetic bead (71) for being adsorbed with nucleic acid is got off to obtain purification of nucleic acid template by elution step.

38. the method for extracting nucleic acid according to claim 37, which is characterized in that the nucleic acid absorption step is described including making Micro-fluidic chip (27) applies to generate between magnet in two alternating forces applies the first of magnet relative to described two alternating forces It does relatively reciprocating motion, described first direction of motion to do relatively reciprocating motion is parallel to the distribution that described two alternating forces apply magnet Direction.

39. the method for extracting nucleic acid according to claim 38, which is characterized in that the nucleic acid absorption step further includes making institute It states micro-fluidic chip (27) and is generated between described two alternating forces apply magnet and apply magnet relative to described two alternating forces Second does relatively reciprocating motion, and the described second direction to do relatively reciprocating motion applies the distribution of magnet perpendicular to described two alternating forces Direction.

40. the method for extracting nucleic acid according to claim 37, which is characterized in that the nucleic acid purification step includes: in institute It states in mixing chamber (61) and washs the magnetic bead (71) for being adsorbed with nucleic acid at least once using cleaning solution, make to complete to wash for the last time The cleaning solution washed passes through the inside that is set between the mixing chamber (61) and the elution chamber (63) of at least one reaction chamber It is connected to control unit and enters the elution chamber (63), and filled with the institute for completing last time washing in the internal connection control unit State cleaning solution, the magnetic bead (71) swum across under the action of gravity and/or magnetic force in the internal connection control unit described in wash It washs liquid and enters the elution chamber (63), the magnetic bead is adsorbed by the bottom magnet of described elution chamber (63) bottom later (71), and last time is completed from the mixing chamber (61), the internal connection control unit and the elution chamber (63) discharge to wash The cleaning solution washed.

41. method for extracting nucleic acid according to claim 40, which is characterized in that the nucleic acid purification step further include: After the cleaning solution of last time washing is completed in discharge, the magnetic bead for being adsorbed with nucleic acid in elution chamber (63) is dried (71)。

42. the method for extracting nucleic acid according to claim 37, which is characterized in that in the elution step, control with it is described The corresponding reagent of the eluent storage chamber (71) of at least one storage chamber is connected to control unit, will be in the eluent storage chamber (64) Eluent discharge in the elution chamber (63) at least one reaction chamber, make in elution chamber (63) magnetic bead (71) movement with Nucleic Acid Elution on the magnetic bead (71) is got off.

43. method for extracting nucleic acid according to claim 36, which is characterized in that predetermined order and time interval are to correspondence Reaction chamber discharge the reagent in each storage chamber.