CN112958173B

CN112958173B - Microfluidic kit

Info

Publication number: CN112958173B
Application number: CN202110239424.3A
Authority: CN
Inventors: 吴玉鹏
Original assignee: Shenzhen Meimeimeichuangyi Medical Technology Co ltd
Current assignee: Shenzhen Meimeimeichuangyi Medical Technology Co ltd
Priority date: 2021-03-04
Filing date: 2021-03-04
Publication date: 2023-01-10
Anticipated expiration: 2041-03-04
Also published as: CN112958173A

Abstract

The invention discloses a microfluidic kit, which comprises a transfer pool, a sample cracking pool, a waste liquid pool, an elution pool, a reaction bin and a rotary valve, wherein the transfer pool is arranged in the sample cracking pool; the transfer pool is used for transferring liquid; the sample cracking cell is used for mixing a sample with a cracking solution; the waste liquid pool is used for storing the treated waste liquid; the elution pool is used for storing eluent; the reaction bin is provided with reactants and can perform corresponding reaction after introducing the eluted solution; the rotary valve is internally provided with a plurality of valve cores, and the different valve cores respectively communicate the transfer pool with the sample cracking pool, the waste liquid pool, the elution pool or the reaction bin in sequence when the rotary valve is rotated. The microfluidic kit can be operated orderly, has a simple integral structure and is simple to operate, so that the problem of inconvenience in molecular detection is effectively solved.

Description

Microfluidic kit

Technical Field

The invention relates to the technical field of molecular detection, in particular to a microfluidic kit.

Background

At present, molecular detection technology is applied to various aspects, such as pathogenic microorganism infectivity detection, tumor and genetic disease diagnosis, immune system disease diagnosis, prenatal screening and the like. With the current medical development, the medical field needs fast and accurate detection means more and more, so that the molecular detection has unique advantages. The mainstream technology of molecular detection is fluorescence quantitative PCR technology.

The characteristic of the current PCR technology exponential amplification template limits the further application of the fluorescent quantitative PCR technology to clinical application, and the problem is aerosol pollution caused by open consumables. And the existing PCR technology has the defects of multiple experimental steps, complex operation, PCR aerosol pollution, low automation degree, unstable result easily caused by manual operation and high difficulty in realizing multiple detections.

In summary, how to effectively solve the problem of inconvenient detection of molecules is a problem that needs to be solved by those skilled in the art.

Disclosure of Invention

In view of the above, the present invention provides a microfluidic kit, which can effectively solve the problem of inconvenient molecular detection.

In order to achieve the purpose, the invention provides the following technical scheme:

a microfluidic reagent kit comprises a transfer pool, a sample cracking pool, a waste liquid pool, an elution pool, a reaction bin and a rotary valve;

the transfer pool is used for transferring liquid;

the sample cracking cell is used for mixing a sample with a cracking solution;

the waste liquid tank is used for storing the treated waste liquid;

the elution pool is used for storing eluent;

the reaction bin is provided with reactants and can perform corresponding reaction after the eluted solution is introduced;

the rotary valve is internally provided with a plurality of valve cores, and the different valve cores respectively communicate the transfer pool with the sample cracking pool, the waste liquid pool, the elution pool or the reaction bin in sequence when the rotary valve is rotated.

Preferably, the rotary valve further comprises a valve body, and the valve core is arranged in the valve body in a penetrating manner;

the valve core is provided with an internal channel, and the direction of the internal channel of each valve core is different; the valve body is correspondingly provided with a plurality of connecting interfaces, and the orientations of the connecting interfaces are consistent.

Preferably, the internal passages are arranged in sequence at equal angles around the central axis of the valve body.

Preferably, a plurality of said spools are independently driven or ganged.

Preferably, the washing machine further comprises a washing pool, wherein the washing pool is used for storing washing liquid;

the transfer tank is communicated with the cleaning tank through the valve core.

Preferably, the microfluidic kit is provided with a plurality of reaction bins, each reaction bin is communicated with the transfer pool through different valve cores, and overflow ports of the reaction bins are communicated with overflow pools respectively so as to receive reaction liquid overflowing from the reaction bins.

Preferably, the gas transfer device further comprises a driving source, wherein a filtering tank and a safety tank which are sequentially connected in series are arranged between the driving source and the transfer tank, so that gas of the driving source sequentially passes through the filtering tank and the safety tank and enters the transfer tank;

the device also comprises magnetic beads which can flow among the transfer pool, the sample cracking pool, the cleaning pool and the elution pool through the rotary valve.

Preferably, a first shunt main body and a second shunt main body are respectively arranged on two sides of the valve body, a plurality of drainage channels communicated with the connecting interfaces on one side corresponding to the valve body are arranged on the first shunt main body, each drainage channel is respectively communicated with the sample cracking pool, the cleaning pool, the elution pool, the waste liquid pool and the reaction bin, an intercommunication channel is arranged on the second shunt main body, a plurality of channel ports on one side of the intercommunication channel are respectively communicated with the connecting interfaces on one side corresponding to the valve body, and a channel port on the other side of the intercommunication channel is communicated with the transfer pool.

Preferably, the device further comprises an air channel plate, one end of each of the transfer pool, the sample cracking pool and the elution pool is communicated with a corresponding air channel hole in the air channel plate, and the other end of each of the transfer pool, the sample cracking pool and the elution pool is communicated with the first split flow main body or the second split flow main body.

Preferably, rectangular grooves are formed in the first shunting main body and the second shunting main body, the sample cracking tank and the elution tank are arranged in the rectangular groove of the first shunting main body, a cavity between the wall of the rectangular groove cavity of the first shunting main body and the outer wall of the inner tank body forms the waste liquid tank, and a mounting opening is formed in the rectangular groove of the second shunting main body, so that the injector can be inserted into the rectangular groove through the mounting opening.

In this micro-fluidic kit, be provided with the rotary valve between transfer pond and sample lysis pond, the elution pond, the waste liquid pond, the reaction storehouse, and fixed connection between a plurality of case, so that can be through rotating rotary valve case, the intercommunication relation between adjustment transfer pond that can relax and other each pond, so that can carry out sample schizolysis in proper order, the waste liquid is got rid of, nucleic acid elution and nucleic acid reaction, regard the transfer pond as the pivot, and sample lysis pond, elution pond and reaction storehouse have been set up, so that can carry out multiple reaction, in order can carry out nucleic acid lysis, purification, amplification and detection, can realize that the sample advances the result and goes out. And the transfer pool is used as a pivot, so that the procedures can be orderly carried out, and the whole structure is simple and the operation is simple. In conclusion, the microfluidic kit can effectively solve the problem of inconvenient molecular detection.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

Fig. 1 is a schematic diagram of one-side explosion structure of a microfluidic kit according to an embodiment of the present invention;

fig. 2 is a schematic diagram of an exploded structure on the other side of the microfluidic kit according to the embodiment of the present invention;

FIG. 3 is a schematic diagram of an exploded view of a rotary valve according to an embodiment of the present invention;

FIG. 4 is a schematic diagram of an exploded structure of a reaction chamber provided in an embodiment of the present invention;

fig. 5 is a flow chart of the use of the microfluidic kit according to the embodiment of the present invention.

The drawings are numbered as follows:

the device comprises a liquid path sealing plate 1, a rotary valve 2, a second flow dividing main body 3, an elution pool 4, a filtering pool 5, a safety pool 6, a transfer pool 7, a filter 8, a gas path sealing plate 9, a gas path plate 10, an injector 11, a waste liquid filter 12, a first cleaning pool 13, a sample cracking pool 14, a second cleaning pool 15, a reaction bin 16, a first flow dividing main body 17, a waste liquid filtering pool 18, an overflow pool 19, a valve body 21, a first valve core piece 22, a second valve core piece 23, an internal channel 24, an external channel 25 and a bin sealing plate 161.

Detailed Description

The embodiment of the invention discloses a microfluidic kit which can effectively solve the problem of inconvenience in molecular detection. The micro-fluidic kit needs to be matched with a nucleic acid detection automation instrument for use, the full automation of nucleic acid detection is realized, the nucleic acid detection instrument matched with the device comprises a temperature control module, an external power module, a uniform mixing module, a magnet and other parts, the temperature control module is used for enabling nucleic acid amplification to keep a certain temperature, the external power module is used for providing power, the uniform mixing module is used for mixing, and the magnet is used for adsorbing to jointly complete the flow of nucleic acid detection.

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1 to 4, fig. 1 is a schematic diagram illustrating an exploded structure of a microfluidic kit according to an embodiment of the present invention; fig. 2 is a schematic diagram of an exploded structure on the other side of the microfluidic kit according to the embodiment of the present invention; FIG. 3 is a schematic diagram of an exploded view of a rotary valve according to an embodiment of the present invention; fig. 4 is a schematic diagram of an explosion structure of the reaction chamber according to the embodiment of the present invention.

In a particular embodiment, a microfluidic kit is provided. Specifically, the microfluidic kit mainly comprises a transfer pool 7, a sample cracking pool 14, an elution pool 4, a reaction bin 16, a waste liquid pool, an injector 11, a rotary valve 2 and the like. A washing pool is generally arranged, so that the microfluidic kit can perform nucleic acid cracking, purification, amplification and detection, and can realize sample inlet and outlet.

Wherein sample lysis bath 14 is packaged with the sample lysate when using, generally still be provided with the sample filling opening, get into sample lysis bath 14 with can injecting the sample, generally still be packaged with the magnetic bead simultaneously, with combine through magnetic bead and nucleic acid, certainly also can be the later stage and add the magnetic bead, the magnetic bead should can circulate between different pond chambeies through inside each intercommunication passageway and rotary valve 2, can pass through promptly rotary valve flow in transfer pond, sample lysis bath, washing pond and elution pond mainly follow inside circulation liquid circulation between each pond chamber, and outside this box body, at least in the transfer pond 7 outside, can adsorb in transfer pond 7 through magnetism suction pieces such as magnet, and do not follow liquid flow.

One or more reaction bins 16 may be provided, and the reaction bins are provided with reactants and can enter corresponding reactions after the eluted solution is introduced, and specifically, 1 to 100 reaction bins 16 may be provided. The reaction chambers 16 can be connected in parallel through flow channels, for example, each reaction chamber can receive the solution eluted from the transfer tank through different valve cores and the transfer tank, and different reactants can be arranged in different reaction chambers 16. Specifically, the primers and probes may be in liquid or lyophilized form, so as to realize multi-project parallel detection, and if the primers and probes are lyophilized in the reaction chamber 16, the reaction solution is injected with the eluted reverse solution after the lyophilized reaction solution is completely dissolved. The plurality of reaction compartments 16 are provided so that different primers and probes can be separately and independently placed, thereby preventing the primers and probes from interfering with each other during amplification or detection.

The rotary valve 2 mainly comprises a valve body and a valve core, wherein the valve core is arranged in the valve body in a penetrating mode. Different communication states can be realized by the rotary valve 2, a plurality of valve cores are arranged in the rotary valve 2, when the rotary valve is rotated, different valve cores can respectively communicate the transfer pool with the sample cracking pool, the waste liquid pool, the elution pool or the reaction bin in sequence, and what needs to be explained is that the rotary valve is rotated, which is used for realizing the opening and closing of the valve cores and generally refers to the rotary valve cores. Specifically, each valve core is used for communicating the transfer pool with the sample cracking pool, communicating the transfer pool with the waste liquid pool, communicating the transfer pool with the elution pool, and communicating the transfer pool with the reaction bin.

The multiple valve cores can be independently driven, namely can independently rotate without mutual interference or linkage, all the valve cores can be linked, or part of the valve cores can be linked, if the multiple valve cores are fixedly connected, the different valve cores are in a communication state by rotating different angles, namely the multiple valve cores are fixedly connected and can rotate from one end, so that the valve cores can rotate to enter the communication state in order.

The valve elements are generally provided with internal channels 24, the orientation of the internal channels of each valve element is different, in particular, for each valve element in linkage, the orientation of the internal channels 24 of each valve element is different, the valve body of the rotary valve is correspondingly provided with connecting interfaces, and the orientation of a plurality of the connecting interfaces is consistent, so that when the rotary valve is rotated to a certain angle position, only one of the valve elements is aligned with the corresponding connecting interface to be communicated. It should be noted that, how the internal passage 24 communicates with the connection ports at the two ends, specifically, the valve core generally rotates to enter a communication state when the passage ports at the two ends of the internal passage 24 align with the corresponding connection ports, and the connection ports at the two sides of the valve body are respectively connected with the corresponding tank bodies. When the valve core rotates to the state that the channel ports at the two ends of the internal channel 24 are not aligned with the corresponding connecting ports, the valve core is in a non-communication state, and the connecting ports at the two sides of the valve body are in a non-communication state. If the valve core corresponding to the sample cracking pool, the channel ports at the two ends of the internal channel 24 of the valve core are communicated with the sample connection pool and the transfer pool respectively, when the valve core rotates to the state that the channel ports at the two ends of the internal channel 24 are aligned with the corresponding connection ports, the sample cracking pool is communicated with the transfer pool through the valve core, and if the valve core rotates to the state that the channel ports at the two ends of the internal channel 24 are not aligned with the corresponding connection ports, the valve core between the sample cracking pool and the transfer pool is not communicated.

Wherein the valve of rotary valve 2 can divide into a plurality of sections controls, can realize single case or a plurality of runners of many case control, and each part of valve can be accomplished through moulding plastics, reduction cost that can be very big. For a plurality of valve cores which are relatively fixed with each other, a plurality of internal channels 24 can be sequentially arranged around the central axis of the valve body in an equally-divided angle, and the internal channels 24 are arranged at intervals of a certain angle, for example, adjacent 15 degrees, so that the rotation angle is convenient to adjust.

Wherein transfer pond 7 mainly plays the effect of transfer, and is further, can be provided with the driving source, with the transfer pond intercommunication, direct intercommunication or indirect intercommunication to can adjust transfer pond pool chamber pressure. Specifically, like the injector 11 and the air pump, an injection cavity of the injector 11 is communicated with a pool cavity of the transfer pool 7, so that the pressure of the pool cavity of the transfer pool 7 can be adjusted by pushing and pulling the plunger of the injector 11. Wherein the syringe 11 is preferably a disposable syringe, so as to effectively reduce the cost and facilitate the replacement. Specifically, can make and be provided with filter 8 and the safety pool 6 of establishing ties in proper order between driving source and the transfer pond 7 to make the gas of driving source pass through in proper order the filtering ponds the safety pool enters into to the transfer pond when effectively preventing the aerosol pollution, also can prevent that the interior fluid of kit that arouses because of the trouble from spilling over. When the device is used, under the condition that the valve of the rotary valve 2 is completely closed, the pressure can be applied to the transfer pool 7 through the plunger, so that the sample is cracked and pressurized, and the sample cracking effect is better. The transfer tank 7 is preferably a heatable transfer tank, so that ultrasonic waves or heat can be applied to the transfer tank 7 while the injector 11 pressurizes the mixed liquid in the transfer tank 7, thereby promoting sufficient lysis of the sample.

In use, a sample is first injected into the sample lysis cell 14, and magnetic beads are placed in the sample lysis cell 14, and the sample can be injected through the closable opening of the sample lysis cell 14. Then, the valve core of the rotary valve 2 is operated to rotate until the transfer cell 7 is communicated with the sample lysis cell 14, and at this time, the driving source is operated to enable the mixed solution of the sample, the lysis solution and the magnetic beads to oscillate back and forth between the sample lysis cell 14 and the transfer cell 7, so that the sample, the lysis solution and the magnetic beads are fully mixed. And then the mixed liquid is completely pumped to the transfer tank 7 by a driving source. After a certain time has elapsed, so that the internal lysis is completed. Then, a magnet is added to the bottom of the transfer well 7 to adsorb the magnetic beads in the transfer well 7. Then the valve of the rotary valve 2 is operated to connect the transfer pool 7 and the waste liquid pool, and the driving source is operated to completely empty the waste liquid after the sample is cracked to the waste liquid pool through the flow channel, and the magnetic beads combined with the nucleic acid are kept to be adsorbed at the bottom of the transfer pool 7 by the magnet in the process. Then take out the magnet to the valve of operation rotary valve 2 extremely, transfer pond 7 and elution pond 4 intercommunication, through operation driving source to make eluent and magnetic bead make a round trip to vibrate between transfer pond 7 and elution pond 4, and after the vibration is accomplished, draw through syringe 11, so that the mixed liquid of elution is extracted to transfer in the pond 7. Then, the magnetic beads are adsorbed at the bottom of the transfer tank 7 by a magnet, and the rotary valve 2 is operated to rotate to the transfer tank 7 to be communicated with the reaction chamber 16, so as to push the eluted nucleic acid to the reaction chamber 16 until the reaction chamber 16 is filled with the reaction solution. After the reaction solution is completely mixed with the primers and the probes in the reaction bin 16, a certain temperature can be applied to realize the PCR reaction in the reaction bin 16. In the microfluidic kit, a multi-way valve is arranged between the transfer pool 7 and the sample cracking pool 14, the elution pool 4, the waste liquid pool and the reaction bin 16, so that the communication relation between the transfer pool 7 and each other pool can be simply adjusted through the multi-way valve, so that sample cracking, waste liquid discharge, nucleic acid elution and nucleic acid reaction can be sequentially carried out, the transfer pool 7 is used as a pivot, and a driving source is arranged, so that liquid flow between a pool cavity and the pool cavity can be pushed, and further the procedures can be orderly carried out, the whole structure is simple, and the operation is simple. In conclusion, the microfluidic kit can effectively solve the problem of inconvenient molecular detection.

Further, one or more washing tanks are generally provided, such as a first washing tank 13 and a second washing tank 15. It has the washing liquid to encapsulate in advance in the washing pond to wash the sample schizolysis back and the magnetic bead that the nucleic acid combines, wash the case and transfer pond 7 intercommunication on the pond through rotary valve 2, when there are a plurality of washing ponds, so correspond set up a plurality of cases, with the intercommunication between different washing pond and the transfer pond of realizing respectively, so that transfer pond 7 and different washing pond intercommunication. When the device is used, after a sample is cracked, waste liquid is discharged to a waste liquid tank, then the valve 2 is rotated to enable the transfer tank 7 to be communicated with the cleaning tank, and the device is pushed and pulled by the injector 11 to enable the transfer tank to vibrate back and forth for multiple times, and then the cleaning liquid after cleaning is discharged like the waste liquid after cracking. Set up a plurality of washing ponds, can wash many times, can also realize that the passageway that transfer pond 7 links to each other effectively washs.

Further, an overflow tank 19 may be provided in communication with the reaction chamber 16 to receive the reaction liquid overflowing from the reaction chamber, and the overflow tank 19 connected to the reaction chamber 16 may be used to store the excess fluid after the reaction chamber 16 is filled with the fluid. So that the reaction chamber 16 can have a quantitative function by the arrangement of the overflow tank 19. When a plurality of reaction chambers 16 are provided, each reaction chamber 16 may have a separate overflow tank 19. The reaction chambers 16 can be in one-to-one correspondence with the overflow wells 19, so that cross contamination caused by different primer probes can be avoided. Of course, a plurality of reaction chambers 16 may share one overflow tank 19. Further, when the reaction chambers 16 are disposed in one-to-one correspondence with the overflow tanks 19, an external sensor may be disposed to detect whether each overflow tank 19 has overflow, so as to determine whether the corresponding reaction chamber 16 is filled with the reaction liquid. Namely, the reaction chambers 16 can be filled with reaction liquid by external sensors.

Wherein, the two sides of the valve body 21 are respectively provided with a connecting interface to form an external channel 25, and the connecting interfaces are arranged corresponding to the channels in the corresponding valve cores, so that the channels of different valve cores can be communicated with the connecting interfaces on the two sides when the valve cores rotate to different angles.

As mentioned above, the plurality of valve cores are fixedly connected, and specifically, two valve core members may be provided: a first valve core member 22 and a second valve core member 23, wherein the first valve core member 22 is integrated with a plurality of valve cores for respectively controlling the communication between the transfer pool and the sample cracking pool, the waste liquid pool, the first cleaning pool and the second cleaning pool, and the second valve core member is integrated with a plurality of valve cores for respectively controlling the communication between the transfer pool and the eluent pool and between the reaction bins.

Specifically, in order to facilitate the connection between each tank body and the rotary valve 2, a first diversion main body 17 and a second diversion main body 3 may be respectively disposed on two sides of the valve body 21. Wherein, be provided with a plurality of drainage channels that correspond each connection interface intercommunication of one side with valve body 21 on the first reposition of redundant personnel main part 17, each drainage channel communicates with sample pyrolysis tank 14, elution pond 4, washing pond, waste liquid pond and reaction storehouse 16 respectively, and wherein the preferred curved passage that is the drainage channel with sample pyrolysis tank 14 intercommunication can be called as the mixing flow path to play better mixed action. The second flow dividing main body 3 is provided with an intercommunicating channel, a plurality of channel ports communicated with one side of the intercommunicating channel are respectively communicated with each connecting interface on one side corresponding to the valve body 21, and the channel ports on the other side are communicated with the transfer tank 7, the specific intercommunicating channel comprises a main channel extending along the axial direction of the valve body 21, one side of the main channel close to the valve body 21 is provided with a plurality of sub channels arranged in parallel with each other so as to be respectively communicated with each connecting interface on the side of the valve body 21, the other side of the main channel is connected with a main channel port through a curve-shaped channel, and the main channel port is communicated with the transfer tank 7.

Wherein the first flow-dividing main body 17 is provided with a rectangular cavity, wherein the sample cracking tank 14, the elution tank 4 and the cleaning tank are all positioned in the rectangular cavity, and a tank cavity of the waste liquid tank is formed between the cavity wall of the rectangular cavity and the outer wall of each tank body. Wherein be provided with the intercommunication passageway on the second reposition of redundant personnel main part 3, in order being used for intercommunication between syringe 11 and the transfer pond 7, it is corresponding, when being provided with filtering ponds 5 and safety pool 6, can communicate in proper order through second reposition of redundant personnel main part 3, also can communicate through the gas circuit board, it is corresponding, can be provided with the rectangle chamber on second reposition of redundant personnel main part 3, in order can place safety pool 6, filtering ponds 5 and transfer pond 7, and be provided with the opening on second reposition of redundant personnel main part 3, so that syringe 11 can follow outside cartridge and enter into to the opening in, inside in order to enter into second reposition of redundant personnel main part 3, so that overall structure is more compact.

Furthermore, independent air paths and independent liquid paths can be respectively arranged among the pool bodies so as to effectively avoid cross contamination. Further, the device also comprises an air channel plate 10, wherein the air channel plate 10 is provided with a plurality of air channel holes so as to respectively communicate with the cell bodies. Specifically, one end of each of the sample cracking tank 14, the transfer tank 7, the waste liquid tank, the cleaning tank, and the elution tank 4 is communicated with a corresponding air passage hole in the air hole plate, and the other end is communicated with the first split flow main body 17 or the second split flow main body 3.

Specifically, a gas path sealing plate 9 and a gas path arranged on the gas path plate 10 are further arranged on one side of the gas path plate 10 away from the tank body. Correspondingly, a liquid path sealing plate 1 is further arranged to seal the intercommunicating channel and the communicating channel which are formed on the first flow dividing main body 17, and seal the drainage channel on the second flow dividing main body 3. And sealing, by a sealing plate, each channel formed in a groove shape so that each channel is opened only at a channel opening.

Can set up respectively in valve body 21 both sides and be the first reposition of redundant personnel main part 17 and the second reposition of redundant personnel main part 3 of rectangle, wherein the first side of second reposition of redundant personnel main part 3 is provided with above-mentioned intercommunication passageway and intercommunication passageway, the second side fluting setting of second reposition of redundant personnel main part 3, can specifically take out the shell setting from the second side by second reposition of redundant personnel main part 3, form the rectangular channel promptly at the second side, one side cell wall of rectangular channel sets up the installing opening, so that the front end business turn over gas port of syringe 11 can insert the vallecular cavity to the rectangular channel through the installing opening, syringe 11 sets up the one side that is close to valve body 21 in the rectangular channel vallecular cavity, and the one side of keeping away from valve body 21 in the rectangular channel vallecular cavity has set gradually filtering ponds 5 along the installing opening orientation, safety pond 6 and transfer pond 7, wherein be provided with filter 8 in the filtering ponds 5. Wherein the orientation of the control end of the valve element of the rotary valve 2 preferably coincides with the orientation of the above-mentioned openings to facilitate the operation of the syringe 11 and thus of the rotary valve 2 on the same side.

And wherein the first side of the first distribution body 17 is provided with respective tapping channels and at the side close to the valve body 21 with passage openings for the respective tapping channels. Wherein the first side of the first distribution body 17 is oriented in the same direction as the first side of the second distribution body 3, wherein the first side and the second side refer to the two sides which are oriented oppositely. Correspondingly, the second side of the first shunting body 17 is also provided with a rectangular groove, and the second side can be also provided with a shell. Wherein one side that the rectangular channel vallecular cavity is close to valve body 21 has set gradually elution pond 4, second along above-mentioned installation open-ended orientation direction, and wash pond 15, first washing pond 13, and is provided with sample pyrolysis pond 14 in one side that the valve body 21 was kept away from in the rectangular channel vallecular cavity to guarantee that the drainage channel with sample pyrolysis pond 14 intercommunication has sufficient space and forms curved passage, and the part of each cell body is got rid of in the rectangular channel vallecular cavity, forms above-mentioned waste liquid pond promptly. The first side surface portion of the first flow dividing body 17 has a projection on a side away from the valve body 21 so that the flow guiding passage communicating with the reaction chamber 16 can extend to the projection, and the reaction chamber 16 is provided on a side of the first flow dividing body 17 away from the valve body 21, and the plurality of reaction chambers 16 are arranged in series in a direction toward the mounting opening. Wherein the rectangular groove of the second flow-dividing body 3 can also be provided with an overflow tank 19. It should be noted that each tank body is preferably in a column shape, specifically, a cylinder shape.

The reaction bin 16 is L-shaped, a vertical plate on one side is provided with a connecting channel connected with the outside, a transverse plate on the other side is provided with a cavity of the reaction bin 16, and the cavity of one reaction bin 16 is correspondingly provided with two connecting channels to respectively communicate the drainage channel and the overflow tank 19. And it is preferred that the upper and lower bin sealing plates 161 of the reaction bin 16 have a thickness of between 0.1 mm and 1 mm, such as 0.5 mm. The reaction chambers 16 may be separated, i.e. a plurality of reaction chambers 16 are separated from each other, or may be connected, i.e. a plurality of reaction chambers 16 are formed by opening a component.

And wherein the liquid path sealing plate 1 seals against a first side of the first flow dividing body 17 and a first side of the second flow dividing body 3, and the gas channel plate 10 covers a second side of the first flow dividing body 17 and a second side of the second flow dividing body 3, i.e. across the rotary valve 2 to cover the rectangular groove opening of the first flow dividing body 17 and the rectangular groove opening of the second flow dividing body 3, respectively. Wherein the gas path sealing plate 9 covers the side of the gas path plate 10 away from the rotary valve 2. The width of each channel formed in the first flow-dividing body 17 and the second flow-dividing body 3 is preferably 0.2 to 3 mm, and the depth is preferably 0.2 to 1 mm. The first and second distributing

bodies

17, 3 may be manufactured by injection molding, but other parts may be manufactured by injection molding, and in particular, the assembly may be completed by welding or bonding.

In one embodiment, the microfluidic kit comprises a sample lysis cell 14, a first washing cell 13, a second washing cell 15, a waste solution cell, an elution cell 4, a transfer cell 7, a rotary valve 2, an injector 11, and a plurality of reaction chambers 16. The sample lysis tank 14 is pre-packaged with sample lysis solution and magnetic beads, the first cleaning tank 13 is pre-packaged with first cleaning solution, the second cleaning tank 15 is pre-packaged with second cleaning solution, the elution tank 4 is pre-packaged with eluent, and the reaction chambers 16 are pre-packaged with liquid or freeze-dried primers and probes. The rotary valve 2 cartridge is in a fully closed state before use. The following steps can be specifically adopted to complete the experimental scheme. Referring to fig. 5, fig. 5 is a flow chart illustrating the use of the microfluidic cartridge according to the embodiment of the present invention.

Step 100: and injecting the sample into a sample lysis cell pre-filled with lysis solution and magnetic beads.

I.e. opening the sealing plug of the sample lysis cell 14 to add a certain amount of sample. At this time, the sample lysis cell is pre-filled with a sample lysis solution. Wherein, a filter membrane is generally arranged in the sample cracking pool and is used for filtering the sample.

Step 200: and rotating the valve core to communicate the sample lysis cell with the transfer cell, and pushing the injector to enable the magnetic beads, the sample and the lysis solution to flow between the transfer cell and the sample lysis cell.

Rotate certain angle with rotary valve 2 case for sample lysis cell 14 switches on with transfer pond 7, and through push-and-pull syringe 11, make the mixed liquid of sample, lysate and magnetic bead make a round trip to vibrate between sample lysis cell 14 and transfer pond 7 this moment, through the mixed runner of schizolysis and mixed washing runner, make sample, lysate and magnetic bead intensive mixing.

Step 300: and after the magnetic beads and the mixed liquid of the sample and the lysis solution are completely sucked to the transfer pool, rotating the valve core to close.

That is, after the magnetic beads, the sample and the lysis buffer are all sucked into the transfer pool 7 through the syringe 11, the valve core of the rotary valve 2 is completely closed, so that the sample lysis pool is disconnected from the transfer pool. After the valve core is closed, the mixed liquid in the transfer tank 7 can be pressurized by the syringe 11, and ultrasonic waves or heat can be applied to the transfer tank 7 at the same time, so as to promote the sufficient lysis of the sample.

Step 400: and a magnet is added at the bottom of the transfer pool 7, then the valve core is rotated to enable the transfer pool to be communicated with the waste liquid pool, and then the valve core is closed after liquid in the transfer pool is pushed to the waste liquid pool.

Then, add the magnet in the transfer pond 7 bottom, like magnet or electro-magnet, can separate gas circuit closing plate 9 or night way closing plate and add, adsorb the magnetic bead after the schizolysis with the nucleic acid in the bottom of transfer pond 7, rotate rotary valve 2 case for transfer pond 7 links to each other with the waste liquid pond, promote syringe 11 and make the waste liquid after the sample schizolysis empty to the waste liquid pond through the runner completely, at this in-process, need keep the magnet to adsorb the magnetic bead with the combination of nucleic acid in the bottom of transfer pond 7.

Step 500: and rotating the valve core to enable the transfer pool to be communicated with the first cleaning pool, removing the magnet, pushing and pulling the injector to enable the first cleaning liquid and the magnetic beads combined with the nucleic acid to vibrate between the first cleaning pool and the transfer pool, and then pushing the cleaned solution out of the transfer pool to the waste liquid pool.

Specifically, rotate 2 spools of rotary valve for transfer pond 7 and first washing pond 13 intercommunication, 11 plungers of pull syringe make first washing liquid flow to transfer pond 7 through the miniflow, get rid of magnet, 11 plungers of pull syringe make first washing liquid vibrate between first washing pond 13 and transfer pond 7, for guaranteeing to wash thoroughly, wherein can correspond on the second reposition of redundant personnel main part 3 and be provided with the mixed washing runner. Vibrate a plurality of times after, will mix the whole extraction of liquid to transfer pond 7 through syringe 11, this moment again in 7 bottoms in transfer pond add the magnet, will adsorb to 7 bottoms in transfer pond with the magnetic bead of the combination of nucleic acid, rotate 2 spools of rotary valve for transfer pond 7 and waste liquid pond intercommunication promote syringe 11 and empty to the waste liquid pond completely with the waste liquid after the first washing, this in-process keeps magnet to adsorb in the bottom of transfer pond 7 with the magnetic bead of the combination of nucleic acid. Wherein the waste liquid tank is generally provided with a waste liquid filtering tank 205 and a waste liquid filter 128 positioned in the waste liquid filtering tank 205.

Step 600: then the valve core is rotated to make the second wash pond and transfer pond intercommunication to after wasing the completion, wash waste liquid propelling movement to the waste liquid pond in with the transfer pond.

Concretely, rotate rotary valve 2 case, make transfer pond 7 and second wash pond 15 intercommunication, pull syringe 11 makes the second washing liquid flow to transfer pond 7 through the miniflow channel, get rid of outside magnet this moment, push-and-pull syringe 11, make the second washing liquid wash pond 15 and transfer between the pond 7 vibrate at the second, for guaranteeing to wash thoroughly, the second washing is equally through mixing the washing runner, after vibrating for a plurality of times, all extract mixed liquid to transfer pond 7 through syringe 11, add the magnet again in transfer pond 7 bottom this moment, the magnetic bead that will combine with the nucleic acid adsorbs to transfer pond 7 bottom, rotate rotary valve 2 case, make transfer pond 7 and waste liquid pond intercommunication, promote syringe 11 and empty the waste liquid after the second washing to the waste liquid pond completely, this in-process will keep magnet to adsorb the bottom in rotary valve 7 with the magnetic bead of the combination of nucleic acid, the process finishes, rotate rotary valve 2 case, make 2 case be in the state of closing completely.

Step 700: and rotating the valve core to communicate the transfer tank with the elution tank, and after elution is carried out, sequentially pushing the elution waste liquid in the transfer tank to each reaction bin.

Specifically, the valve core of the rotary valve 2 is rotated to communicate the transfer pool 7 with the elution pool 4, the injector 11 is pulled to enable eluent to flow to the transfer pool 7 through the micro-channel, at the moment, an external magnet is removed, the plunger of the injector 11 is pushed and pulled to enable the eluent to vibrate between the elution pool 4 and the transfer pool 7, in order to ensure thorough elution, the elution mixed liquid passes through the mixed cleaning channel, after being vibrated for a plurality of times, the nucleic acid solution after elution is completely extracted to the transfer pool 7 through the injector 11, at the moment, the magnet is added at the bottom of the transfer pool 7 again to adsorb the magnetic beads after elution to the bottom of the transfer pool 7, the valve core of the rotary valve 2 is rotated to enable the transfer pool 7 to be communicated with the reaction bin 16, the plunger of the injector 11 is pushed to push the nucleic acid after elution to the reaction bin 16 until the reaction bin 16 is filled with the reaction liquid, the redundant reaction liquid can be discharged to the overflow pool 19, in the same operation mode, the subsequent other reaction bins 16 are sequentially filled with the reaction liquid, and the magnet is also kept to be adsorbed to the bottom of the magnetic beads after elution in the transfer pool 7;

after the three reaction chambers 16 are filled with the reaction solution, the valve core of the rotary valve 2 is rotated, the valve core of the rotary valve 2 is completely closed again, after the reaction solution is completely mixed with the primers and the probes in the reaction chambers 16, if the primers and the probes are freeze-dried in the reaction chambers 16, the freeze-dried reaction solution is completely dissolved, and then different or same temperatures are applied to the upper bottom surface and the lower bottom surface of the reaction chambers 16 at the same time to realize the PCR (polymerase chain reaction) in the reaction chambers 16, and the detection is performed on the reaction chambers 16 through an external optical path while the PCR is performed.

In the present specification, the embodiments are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A microfluidic kit is characterized by comprising a transfer pool, a sample cracking pool, a waste liquid pool, an elution pool, a reaction bin and a rotary valve;

the transfer pool is used for transferring liquid;

the sample cracking cell is used for mixing a sample with a cracking solution;

the waste liquid pool is used for storing the treated waste liquid;

the elution pool is used for storing eluent;

a plurality of valve cores are arranged in the rotary valve, and different valve cores respectively and sequentially communicate the transfer pool with the sample cracking pool, the waste liquid pool, the elution pool or the reaction bin when the rotary valve is rotated;

still include magnetic bead and syringe, the injection chamber and the transfer pond chamber intercommunication of syringe to the messenger:

when the rotary valve is rotated to enable the sample lysis cell to be communicated with the transfer cell, the injector can be pushed to enable the magnetic beads and the liquid to flow between the transfer cell and the sample lysis cell;

when the rotary valve rotates to enable the transfer pool to be communicated with the waste liquid pool, liquid in the transfer pool can be emptied to the waste liquid pool by pushing the injector;

when the rotary valve is rotated to enable the transfer pool to be communicated with the elution pool, the injector can be pushed to enable the magnetic beads and the liquid to flow between the transfer pool and the elution pool;

when the rotary valve rotates to enable the transfer pool to be communicated with the reaction bin, the liquid in the transfer pool can flow to the reaction bin by pushing the injector.

2. The microfluidic kit according to claim 1, wherein the rotary valve further comprises a valve body, and the valve core is arranged in the valve body in a penetrating manner;

3. The microfluidic kit according to claim 2, wherein the plurality of internal channels are sequentially arranged at equal angles around the central axis of the valve body.

4. The microfluidic kit of claim 3, wherein a plurality of the valve spools are independently driven or linked.

5. The microfluidic kit according to any one of claims 1 to 4, further comprising a wash reservoir for storing a wash solution;

6. The microfluidic kit according to claim 5, wherein the microfluidic kit is provided with a plurality of reaction chambers, each reaction chamber is respectively communicated with the transfer pool through a different valve core, and an overflow port of each reaction chamber is respectively communicated with an overflow pool for receiving the reaction liquid overflowing from the reaction chamber.

7. The microfluidic kit according to claim 6, wherein a filtering tank and a safety tank are arranged between the injector and the transfer tank in series in sequence, so that the gas of the injector enters the transfer tank through the filtering tank and the safety tank in sequence;

the magnetic beads can flow among the transfer pool, the sample cracking pool, the cleaning pool and the elution pool through the rotary valve.

8. The microfluidic kit according to claim 7, wherein a first flow dividing body and a second flow dividing body are respectively disposed on two sides of the valve body, the first flow dividing body is provided with a plurality of flow guiding channels communicated with the connecting ports on the corresponding side of the valve body, each flow guiding channel is respectively communicated with the sample cracking pool, the cleaning pool, the elution pool, the waste liquid pool and the reaction bin, the second flow dividing body is provided with an intercommunicating channel, a plurality of channel ports on one side of the intercommunicating channel are respectively communicated with the connecting ports on the corresponding side of the valve body, and a channel port on the other side of the intercommunicating channel is communicated with the transfer pool.

9. The microfluidic kit according to claim 8, further comprising a gas channel plate, wherein one end of each of the transfer cell, the sample lysis cell and the elution cell is communicated with a corresponding gas channel hole on the gas channel plate, and the other end of each of the transfer cell, the sample lysis cell and the elution cell is communicated with the first shunting body or the second shunting body.

10. The microfluidic kit according to claim 9, wherein the first splitter body and the second splitter body are both provided with rectangular grooves, the sample cracking tank and the elution tank are arranged in the rectangular grooves of the first splitter body, the waste liquid tank is formed by a cavity between the wall of the rectangular groove of the first splitter body and the outer wall of the inner tank body, the transfer tank is arranged in the rectangular groove of the second splitter body, and a mounting opening is arranged on one side wall of the rectangular groove of the second splitter body, so that a syringe can be inserted into the rectangular groove through the mounting opening.