CN116139954A - Microfluidic chip for multichannel isothermal amplification CRISPR detection - Google Patents
Microfluidic chip for multichannel isothermal amplification CRISPR detection Download PDFInfo
- Publication number
- CN116139954A CN116139954A CN202310154380.3A CN202310154380A CN116139954A CN 116139954 A CN116139954 A CN 116139954A CN 202310154380 A CN202310154380 A CN 202310154380A CN 116139954 A CN116139954 A CN 116139954A
- Authority
- CN
- China
- Prior art keywords
- cavity
- branch
- isothermal amplification
- chip body
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502723—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by venting arrangements
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502738—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by integrated valves
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a microfluidic chip for detecting CRISPR (serial enhanced surface plasmon resonance) by multichannel isothermal amplification, which relates to the field of microfluidic chips and comprises the following components: the device comprises a first liquid path, a second liquid path, a first valve, a second valve and a cover plate. When the microfluidic chip provided by the invention is used for isothermal amplification CRISPR detection, the first valve and the second valve can separate isothermal amplification reaction from CRISPR detection, and the isothermal amplification reaction and the CRISPR detection are independently completed, so that the problem of low detection efficiency caused by simultaneous implementation of isothermal amplification reaction and CRISPR detection is effectively avoided. In addition, when the microfluidic chip provided by the invention is used for isothermal amplification CRISPR detection, isothermal amplification reaction and CRISPR detection are both carried out in a totally-enclosed pollution-free internal environment, an operator can realize liquid transfer by pushing a piston, liquid uncapping transfer in the traditional two-step operation is not needed, and the risk of aerosol pollution caused by the entry of a nucleic acid template into the environment is reduced.
Description
Technical Field
The invention relates to the field of microfluidic chips, in particular to a microfluidic chip for detecting CRISPR (crisp short amplification) by multichannel isothermal amplification.
Background
Isothermal amplification CIRSPR detection: CRISPR means "clustered regularly interspaced short palindromic repeats", capable of "recognizing sequences" and "cleaving", cas13, cas12, etc. have the property of recognizing and cleaving target genes while indifferently cleaving DNA/RNA, and DNA/RNA can be designed to be a reporter group labeled with a fluorescent reporter gene and a quencher gene, and Cas protein recognizes and cleaves target genes while cleaving fluorescent reporter genes, and the number of fluorescence released by the reporter gene is identical to the number of target genes, i.e., target genes can be indirectly detected by detecting the number of fluorescence.
Isothermal amplification techniques maintain the reaction process of nucleic acid template amplification at a constant temperature without the need for a thermal cycling process. Combining CRISPR with isothermal amplification can further improve detection sensitivity. The combination of conventional isothermal amplification and CRISPR techniques is typically performed in EP tubes (plastic centrifuge tubes), although the detection results are better, the uncapping operation of the two-step reaction may increase the risk of aerosol contamination, the one-pot reaction which integrates the two-step reaction into one EP tube makes the operation simpler and reduces aerosol contamination, but the CRISPR cis and trans cleavage characteristics may suppress the equivalent Wen Kuozeng reaction, while its cleavage may lead to a sharp decrease in the number of nucleic acid templates, resulting in a low detection efficiency.
Therefore, how to ensure the detection efficiency on the basis of reducing the aerosol pollution is a problem to be solved by the person skilled in the art.
Disclosure of Invention
In order to solve the technical problems, the invention provides the microfluidic chip for detecting the multichannel isothermal amplification CRISPR, which can prevent the overflow of aerosol generated in the CRISPR and isothermal amplification reaction processes and has high detection efficiency.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a microfluidic chip for detecting CRISPR by multichannel isothermal amplification, which is characterized by comprising the following components:
the chip body is provided with a cavity in a recessed mode, a liquid injection channel is arranged in the center of the chip body, a flow path is arranged in the chip body, the flow path comprises a first liquid path communicated with the liquid injection channel, a second liquid path communicated with the first liquid path and an exhaust channel communicated with the second liquid path, the first liquid path and the second liquid path are arranged at intervals along the circumferential direction of the chip body, the first liquid path comprises a first reaction cavity, the second liquid path comprises a second reaction cavity, and a first annular slideway and a second annular slideway are further arranged at the bottom end of the cavity;
the plugging structure is provided with a gas discharging passage communicated with the gas discharging passage, the plugging structure can be slidably arranged in the gas discharging passage, the gas discharging passage can be communicated with the outside atmosphere when the plugging structure is in a first position, and the gas discharging passage is isolated from the outside atmosphere when the plugging structure is in a second position;
the cover plate can be rotatably arranged at one end of the chip body, where the cavity is arranged, and can move towards a direction close to or far away from the chip body, a limit structure is arranged at one end of the cover plate, close to the cavity, and when the blocking structure is positioned at the second position, the limit structure is propped against the blocking structure;
the valve bank comprises a first valve and a second valve, the first valve comprises a first concave cavity, a first elastic membrane and a first ball plunger, the second valve comprises a second concave cavity, a second elastic membrane and a second ball plunger, the first concave cavity and the second concave cavity are both arranged at the bottom end of the cavity, the first concave cavity is arranged on a first flow path, the second concave cavity is arranged on a second flow path, the first concave cavity is arranged between the first reaction cavity and the liquid injection channel, the second concave cavity is arranged between the first reaction cavity and the second reaction cavity, the first elastic membrane seals the first concave cavity, the second elastic membrane seals the second concave cavity, the first ball plunger and the second ball plunger are both arranged on the cover plate, one end of the first ball plunger, which is provided with the first ball, slides along the first annular slide way, one end of the second plunger is provided with the second ball, and can be inserted into the second concave cavity and can be blocked by the second plunger when the second ball plunger is arranged in the first position and the second slide way;
and the cross section shape of the piston is matched with that of the liquid injection channel.
Optionally, the number of the flow paths is multiple, all the flow paths are uniformly arranged along the circumferential direction of the chip body, the number of the first valves and the second valves is multiple, one flow path corresponds to one first valve and one second valve, and all the first valves and all the second valves are uniformly arranged along the circumferential direction of the chip body.
Optionally, a plurality of first positioning pits are uniformly arranged in the circumferential direction of the first annular slideway, a plurality of second positioning pits are uniformly arranged in the circumferential direction of the second annular slideway, one first positioning pit is arranged between any two adjacent first cavities, one second positioning pit is arranged between any two adjacent second cavities, the included angle between the first positioning pit and the adjacent first cavity is equal to the included angle between the second positioning pit and the adjacent second cavity, the shape of the first positioning pit is matched with the shape of the first ball head, and the shape of the second positioning pit is matched with the shape of the second ball head.
Optionally, the liquid injection channel is disposed along a height direction of the chip body, the first liquid path includes a first branch, a second branch, a third branch, a fourth branch and a fifth branch, the first branch and the second branch are collinear and are all disposed along a radial direction of the chip body, the third branch, the fourth branch and the fifth branch are parallel to each other and are all disposed along the height direction of the chip body, and the first branch, the third branch, the first cavity, the fourth branch, the second branch and the fifth branch are sequentially communicated, and the second branch has the first reaction cavity; the second liquid way includes first branch road, second branch road, third branch road, fourth branch road and fifth branch road, first branch road second branch road and third branch road are parallel to each other, and all follow the direction of height of the chip body sets up, fourth branch road with fifth branch road collineation, and all follow the radial setting of the chip body, first branch road fourth branch road second cavity third branch road and fifth branch road communicate in proper order, fifth branch road has the second reaction chamber, first branch road is linked together through third branch road with fifth branch road, fifth branch road with exhaust passage is linked together.
Optionally, the chip body includes a main body, a first blocking film and a second blocking film, where the cavity, the liquid injection channel, the flow path, the first cavity, the second cavity, the first annular slide and the second annular slide are all disposed on the main body, and the first branch, the second branch, the fourth branch and the fifth branch penetrate through one end of the main body far away from the cover plate, the first blocking film is disposed on one end of the main body far away from the cover plate, and the first blocking film blocks the first branch, the second branch, the fourth branch and the fifth branch;
the third liquid path comprises a third concave cavity and an arc-shaped groove, the third concave cavity is communicated with one end of the fifth branch, the diameter of the third concave cavity is larger than that of the fifth branch, one end of the arc-shaped groove is communicated with the third concave cavity, the other end of the arc-shaped groove is communicated with the first branch, an included angle of a liquid outlet end of the first reaction cavity is 30-60 degrees, an included angle of a liquid inlet end of the first reaction cavity is 10-20 degrees, and the second blocking film is used for sealing the third concave cavity and the arc-shaped groove.
Optionally, the fifth branch further includes a partial pressure cavity, and the second reaction cavity, the partial pressure cavity, and the exhaust channel are sequentially communicated.
Optionally, the area ratio of the first reaction chamber to the second reaction chamber is 2:1, depth ratio of 4:3, the area of the second reaction cavity is equal to that of the partial pressure cavity, and the depth ratio is 1:2.
optionally, two first arc slopes matched with the first ball head shape are circumferentially arranged at intervals of the first concave cavity, two second arc slopes matched with the second ball head shape are circumferentially arranged at intervals of the second concave cavity, the arc centers of the two first arc slopes coincide, the arc centers of the two first arc slopes are located on the axis of the fourth branch, the arc centers of the two second arc slopes coincide, and the arc centers of the two second arc slopes are located on the axis of the third branch;
the bottom of the cavity is also provided with a third cavity and a fourth cavity, the shape of the third cavity is matched with that of the first elastic membrane, the third cavity is used for accommodating the first elastic membrane, the shape of the fourth cavity is matched with that of the second elastic membrane, and the fourth cavity is used for accommodating the second elastic membrane.
Optionally, the microfluidic chip for detecting the multichannel isothermal amplification CRISPR further comprises a plurality of connection assemblies arranged along the circumferential direction of the chip body, the connection assemblies comprise screws, first nuts and second nuts, the screws can be detachably arranged on the chip body, the first nuts and the second nuts are in threaded connection with the screws, a plurality of arc holes are formed in the cover plate, the screws are in one-to-one correspondence with the arc holes, each screw penetrates through each corresponding arc hole and is in sliding connection with each corresponding arc hole, so that the cover plate and the chip body can rotate relatively, the first nuts and the second nuts are respectively arranged at two ends of the cover plate, and the two ends of the cover plate are respectively abutted to the first nuts and the second nuts.
Optionally, a plurality of bumps for supporting the cover plate are circumferentially arranged at one end of the chip body, which is close to the cover plate.
Compared with the prior art, the invention has the following technical effects:
when isothermal amplification reaction or CRISPR detection is required, the first valve cuts off the first liquid path, so that the first reaction cavity is separated from the liquid injection channel, and the second valve cuts off the second liquid path, so that the first reaction cavity is separated from the second reaction cavity. Thus, isothermal amplification reaction and CRISPR detection are separated from each other and are independently completed, and the problem of low detection efficiency caused by simultaneous operation of isothermal amplification reaction and CRISPR detection is effectively avoided. Compared with the one-pot reaction of isothermal amplification reaction and CRISPR detection integrated in one EP tube, the microfluidic chip for multi-channel isothermal amplification CRISPR detection provided by the embodiment is used for isothermal amplification CRISPR detection, and the detection efficiency is higher.
In addition, the first liquid path and the second liquid path are both arranged in the chip body, when isothermal amplification reaction or CRISPR detection is carried out, when the blocking structure is at the second position, the exhaust channel is in a closed state, so that isothermal amplification reaction and CRISPR detection are carried out in a totally-closed pollution-free internal environment, in a specific use process, an operator only needs to push the piston to convey sample lysate and a sample to be detected after the isothermal amplification reaction in the first reaction cavity to the second reaction cavity for CRISPR detection, liquid uncovering transfer during the traditional two-step operation is not needed, and the risk of aerosol pollution caused by the entering of a nucleic acid template into the environment is reduced.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic structural diagram of a microfluidic chip for CRISPR detection by multichannel isothermal amplification provided in an embodiment of the present invention;
fig. 2 is a perspective view of a microfluidic chip body for CRISPR detection for multichannel isothermal amplification provided in an embodiment of the present invention;
fig. 3 is a bottom view of a microfluidic chip body for multichannel isothermal amplification CRISPR detection provided in an embodiment of the present invention;
fig. 4 is a schematic diagram of a first ball plunger, a second ball plunger, and a cover plate of a microfluidic chip for multichannel isothermal amplification CRISPR detection according to an embodiment of the present invention;
fig. 5 is a schematic diagram of a flow path arrangement mode of a microfluidic chip for detecting CRISPR by multichannel isothermal amplification according to an embodiment of the present invention
Fig. 6 is a first cross-sectional view of a microfluidic chip body for multichannel isothermal amplification CRISPR detection provided in an embodiment of the present invention;
fig. 7 is a second cross-sectional view of a microfluidic chip body for multichannel isothermal amplification CRISPR detection provided in an embodiment of the present invention;
fig. 8 is a schematic structural diagram of a plugging structure of a microfluidic chip for detecting CRISPR by multichannel isothermal amplification, which is provided in an embodiment of the present invention;
fig. 9 is a schematic structural diagram of a microfluidic chip piston for CRISPR detection by multichannel isothermal amplification provided in an embodiment of the present invention.
Reference numerals illustrate: 100. microfluidic chip for multichannel isothermal amplification CRISPR detection; 1. a main body; 2. a flow path; 201. a first shunt; 202. a second shunt; 203. a third shunt; 204. a fourth shunt; 205. a fifth shunt; 206. a first branch; 207. a second branch; 208. a third branch; 209. a fourth branch; 210. a fifth branch; 211. a third cavity; 212. an arc-shaped groove; 213. a pressure dividing chamber; 214. a first reaction chamber; 215. a second reaction chamber; 216. an exhaust passage; 3. a cavity; 4. a liquid injection channel; 5. a first annular slide; 6. a second annular slide; 7. a bleed passage; 8. a plugging structure; 9. a cover plate; 901. an arc-shaped hole; 10. a first elastic film; 11. a first ball plunger; 12. a connecting plate; 13. a second ball plunger; 14. a first cavity; 15. a second cavity; 16. a piston; 17. a push rod; 18. a first locating recess; 19. a second locating recess; 20. a first circular arc ramp; 21. a second circular arc ramp; 22. a screw; 23. a first nut; 24. a second nut; 25. a bump; 26. a limit structure; 27. a first blocking film.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention aims to provide a microfluidic chip for detecting the multi-channel isothermal amplification CRISPR, which can prevent the overflow of aerosol generated in the CRISPR and isothermal amplification reaction processes and has high detection efficiency.
In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention will be rendered by reference to the appended drawings and appended detailed description.
Referring to fig. 1 to 9, a microfluidic chip 100 for CRISPR detection with multichannel isothermal amplification provided in this embodiment includes: chip body, plugging structure 8, apron 9, valves and piston 16.
As shown in fig. 2, one end of the chip body is concavely formed with a cavity 3, and the center of the chip body is provided with a liquid injection channel 4, and when in use, the sample lysate and the sample to be tested are injected into the liquid injection channel 4.
Referring to fig. 5 to 7, a flow path 2 is provided inside the chip body, and the flow path 2 includes a first liquid path communicating with the liquid injection passage 4, a second liquid path communicating with the first liquid path, and an exhaust passage 216 communicating with the second liquid path. The first liquid way and the second liquid way set up along the circumference interval of chip piece, and first liquid way includes first reaction chamber 214, and the second liquid way includes second reaction chamber 215, and the bottom of cavity 3 still is provided with first annular slide 5 and second annular slide 6. The first reaction chamber 214 is used for carrying out isothermal amplification reaction, the second reaction chamber 215 is used for carrying out CRISPR detection, the first reaction chamber 214 is pre-stored with a freeze-dried ball generating target sequence of components such as a primer, an enzyme, a buffer solution, magnesium ions and the like, and the second reaction chamber 215 is pre-stored with a freeze-dried ball containing components such as dNTP, an enzyme, RNA, magnesium ions and the like.
As shown in fig. 8, the blocking structure 8 is provided with a gas release passage 7 communicating with the gas release passage 216, the blocking structure 8 is slidably disposed in the gas release passage 216, the gas release passage 7 is capable of communicating with the outside atmosphere when the blocking structure 8 is in the first position, the gas release passage 7 is isolated from the outside atmosphere when the blocking structure 8 is in the second position, and the gas release passage 216 is closed. As shown in fig. 7, when the sample lysate and the sample to be measured are injected into the interior of the injection channel 4, the exhaust channel 216 is kept in communication with the outside atmosphere. After the sample lysate and the sample to be tested are injected, the exhaust channel 216 is closed. Further, the air release passage 7 is an air release groove.
As shown in fig. 1 and 4, the cover plate 9 can be rotatably disposed at one end of the chip body where the cavity 3 is disposed, and the cover plate 9 can move toward a direction approaching or separating from the chip body, and one end of the cover plate 9 approaching the cavity 3 is provided with a limiting structure 26, when the plugging structure 8 is in the second position, the limiting structure 26 abuts against the plugging structure 8, so that the exhaust channel 216 maintains a closed state. Specifically, in this embodiment, the limiting structure 26 is a limiting boss.
Referring to fig. 2-7, the valve set includes a first valve and a second valve, the first valve includes a first cavity 14, a first elastic membrane 10 and a first ball plunger 11, the second valve includes a second cavity 15, a second elastic membrane and a second ball plunger 13, the first cavity 14 and the second cavity 15 are all disposed at the bottom end of the cavity 3, the first cavity 14 is disposed on the first flow path 2, the second cavity 15 is disposed on the second flow path 2, the first cavity 14 is disposed between the first reaction cavity 214 and the liquid injection channel 4, the second cavity 15 is disposed between the first reaction cavity 214 and the second reaction cavity 215, the first cavity 14 is sealed by the first elastic membrane 10, the second cavity 15 is sealed by the second elastic membrane, the top ends of the first cavity 14 and the second cavity 15 are all open, and the sealing means that the top ends of the first cavity 14 and the second cavity 15 are sealed by the first elastic membrane 10 and the second elastic membrane respectively, so that the first cavity 14 and the second cavity 15 are all a sealed cavity. The first ball plunger 11 and the second ball plunger 13 are both disposed on the cover plate 9. In the rotation process of the cover plate 9, one end of the first ball plunger 11, provided with a first ball, slides along the first annular slide way 5, one end of the second ball plunger 13, provided with a second ball, slides along the second annular slide way 6, and when the position of the first ball plunger 11 is opposite to that of the first cavity 14, the first ball can be embedded into the first cavity 14 and cut off the first liquid path, and at the moment, the first cavity 14 is disconnected from the liquid injection channel 4. When the position of the second ball plunger 13 is opposite to the position of the second cavity 15, the second ball can be embedded into the second cavity 15 and cut off the second liquid path, and at this time, the first reaction cavity 214 and the second reaction cavity 215 are disconnected. It should be noted that, the first ball plunger 11 and the second ball plunger 13 belong to ball plungers, the ball plungers can retract after being stressed, and can automatically reset after being removed, and the detailed structure of the ball plungers belongs to the prior art, and is not described herein.
When the sample lysate and the sample to be detected need to be injected into the liquid injection channel 4 and the sample lysate and the sample to be detected after the isothermal amplification reaction in the first reaction chamber 214 need to be conveyed to the second reaction chamber 215 for CRISPR detection, the first valve and the second valve are both in an open state, at this time, the first ball is separated from the first cavity 14, and the second ball is separated from the second cavity 15. As shown in fig. 6, when isothermal amplification reaction or CRISPR detection is required, the first valve and the second valve are both in a closed state, and at this time, the first ball is embedded in the first cavity 14, and the second ball is embedded in the second cavity 15. In addition, the positions of the first ball head and the second ball head are changed by rotating the cover plate 9 to realize the switching of the opening and closing states of the first valve and the second valve, and it should be noted that in the embodiment, the opening and the closing of the first valve and the second valve are performed simultaneously.
Referring to fig. 9, the cross-sectional shape of the piston 16 matches the cross-sectional shape of the liquid injection passage 4. The piston 16 is arranged in the liquid injection channel 4, and the piston 16 moves along the axial direction of the liquid injection channel 4, so that the sample lysate and the sample to be measured in the liquid injection channel 4 are driven to move. Further, a push rod 17 for driving the piston 16 to move is provided on the piston 16.
The whole reaction process is as follows:
injecting sample pyrolysis liquid and a sample to be detected into the liquid injection channel 4, filling the sample pyrolysis liquid and the sample to be detected into the first reaction cavity 214, closing the first valve and the second valve, arranging the piston 16 in the liquid injection channel 4, performing isothermal amplification reaction, opening the first valve and the second valve after the reaction is finished, pushing the piston 16 to drive the sample pyrolysis liquid and the sample to be detected which are subjected to isothermal amplification reaction into the second reaction cavity 215, closing the first valve and the second valve, and performing CRISPR detection.
In a specific use process, when isothermal amplification reaction or CRISPR detection is required, the first valve cuts off the first liquid path, so that the first reaction cavity 214 is separated from the liquid injection channel 4, and the second valve cuts off the second liquid path, so that the first reaction cavity 214 is separated from the second reaction cavity 215. Thus, isothermal amplification reaction and CRISPR detection are separated from each other and are independently completed, and the problem of low detection efficiency caused by simultaneous operation of isothermal amplification reaction and CRISPR detection is effectively avoided. Compared with the one-pot reaction of isothermal amplification reaction and CRISPR detection integrated in one EP tube, the microfluidic chip for multi-channel isothermal amplification CRISPR detection provided by the embodiment is used for isothermal amplification CRISPR detection, and the detection efficiency is higher.
In addition, the first liquid path and the second liquid path are both arranged inside the chip body, when isothermal amplification reaction or CRISPR detection is performed, when the blocking structure 8 is at the second position, the exhaust channel 216 is in a closed state, so that the isothermal amplification reaction and the CRISPR detection are performed in a totally-closed pollution-free internal environment, and in a specific use process, an operator only needs to push the piston 16 to convey sample lysate and a sample to be detected after the isothermal amplification reaction in the first reaction cavity 214 to the second reaction cavity 215 for CRISPR detection, liquid cover opening transfer during the traditional two-step operation is not needed, and the risk of aerosol pollution caused by the entering of a nucleic acid template into the environment is reduced.
As shown in fig. 2 and 8, in this embodiment, the number of the flow paths 2 is plural, all the flow paths 2 are uniformly arranged along the circumferential direction of the chip body, the number of the first valves and the second valves are plural, one flow path 2 corresponds to one first valve and one second valve, and all the first valves and all the second valves are uniformly arranged along the circumferential direction of the chip body. By providing the flow path 2, the first valve and the second valve in a plurality, the microfluidic chip 100 for multi-channel isothermal amplification CRISPR detection can perform isothermal amplification and CRISPR detection of a plurality of pathogens or mutants on the same sample. In a specific use process, isothermal amplification and CRISPR detection of different pathogens or mutant strains are respectively carried out in different flow paths 2.
Further, the number of the air release passages 7 and the blocking structures 8 is plural, the two are in one-to-one correspondence, the air release passages 7 are in one-to-one correspondence with the flow paths 2, all the air release passages 7 are uniformly arranged along the circumferential direction of the chip body, and all the blocking structures 8 are connected to the same connecting plate 12.
In this embodiment, as shown in fig. 2, a plurality of first positioning pits 18 are uniformly arranged in the circumferential direction of the first annular slide 5, a plurality of second positioning pits 19 are uniformly arranged in the circumferential direction of the second annular slide 6, one first positioning pit 18 is arranged between any two adjacent first cavities 14, one second positioning pit 19 is arranged between any two adjacent second cavities 15, and the included angle between the first positioning pit 18 and the first cavity 14 adjacent thereto is equal to the included angle between the second positioning pit 19 and the second cavity 15 adjacent thereto, the shape of the first positioning pit 18 is matched with the shape of the first ball, and the shape of the second positioning pit 19 is matched with the shape of the second ball.
When both the first valve and the second valve are opened, the first ball and the second ball are respectively embedded into the first positioning pit 18 and the second positioning pit 19, and the first positioning pit 18 and the second positioning pit 19 respectively position the first ball and the second ball.
In addition, in a specific use process, when the first valve and the second valve are required to be switched from the open state to the closed state, the two states can be switched only by rotating the cover plate 9 by a certain angle (the included angle between the first positioning pit 18 and the first concave cavity 14 adjacent to the first positioning pit), and when the first valve is switched from the open state to the closed state and the second valve is switched from the closed state to the open state, the rotating directions of the cover plate 9 are opposite.
Referring to fig. 6-7, in this embodiment, the liquid injection channel 4 is disposed along the height direction of the chip body, the first liquid path includes a first branch 201, a second branch 202, a third branch 203, a fourth branch 204 and a fifth branch 205, the first branch 201 and the second branch 202 are collinear, and are disposed along the radial direction of the chip body, the third branch 203, the fourth branch 204 and the fifth branch 205 are parallel to each other, and are disposed along the height direction of the chip body, the first branch 201, the third branch 203, the first cavity 14, the fourth branch 204, the second branch 202 and the fifth branch 205 are sequentially communicated, and the second branch 202 has a first reaction cavity 214; the second liquid path includes a first branch 206, a second branch 207, a third branch 208, a fourth branch 209 and a fifth branch 210, where the first branch 206, the second branch 207 and the third branch 208 are parallel to each other and all set along the height direction of the chip body, the fourth branch 209 and the fifth branch 210 are collinear and all set along the radial direction of the chip body, the first branch 206, the fourth branch 209, the second branch 207, the second cavity 15, the third branch 208 and the fifth branch 210 are sequentially communicated, the fifth branch 210 has a second reaction chamber 215, the first branch 206 is communicated with the fifth branch 205 through the third liquid path, and the fifth branch 210 is communicated with the exhaust channel 216.
Further, as shown in fig. 7, when the sample lysate and the sample to be measured are injected into the interior of the injection channel 4, the exhaust channel 216 remains in communication with the outside atmosphere. When the fifth branch 205 is filled with the sample lysate and the sample to be tested, the vent channel 216 is closed, and the height of the fifth branch 205 is smaller than the height of the filling channel 4. In addition, according to the principle of the communicating vessel, the height of the liquid in the liquid injection passage 4 is equal to the height of the liquid in the fifth branch 205 during the liquid injection.
In this embodiment, as shown in fig. 6, the chip body includes a main body 1, a first blocking film 27 and a second blocking film, the cavity 3, the liquid injection channel 4, the flow path 2, the first cavity 14, the second cavity 15, the first annular slide 5 and the second annular slide 6 are all disposed on the main body 1, the first shunt 201, the second shunt 202, the fourth shunt 209 and the fifth shunt 210 all penetrate through one end of the main body 1 far away from the cover plate 9, the first blocking film 27 is disposed at one end of the main body 1 far away from the cover plate 9, and the first blocking film 27 blocks the first shunt 201, the second shunt 202, the fourth shunt 209 and the fifth shunt 210. The third liquid path comprises a third cavity 211 and an arc-shaped groove 212, the third cavity 211 is communicated with one end of the fifth branch 205, in order to destroy capillary force, the diameter of the third cavity 211 is larger than that of the fifth branch 205, one end of the arc-shaped groove 212 is communicated with the third cavity 211, the other end of the arc-shaped groove 212 is communicated with the first branch 206, an included angle of a liquid inlet end of the first reaction cavity 214 is 10-20 degrees, an included angle of a liquid outlet end of the first reaction cavity 214 is 30-60 degrees, and the second blocking film is used for blocking the third cavity 211 and the arc-shaped groove 212. As shown in fig. 3, in this embodiment, specifically, the included angle α of the liquid outlet end of the first reaction chamber 214 is 30 °, and the included angle β of the liquid inlet end of the first reaction chamber 214 is 15 °.
Specifically, the ends of the first blocking film 27 and the second blocking film, which are close to the main body 1, are both provided with pressure sensitive adhesives, and the first blocking film 27 and the second blocking film can be fixed on the main body 1 by applying pressure.
In this embodiment, as shown in fig. 7, the fifth branch 210 further includes a partial pressure chamber 213, and the second reaction chamber 215, the partial pressure chamber 213, and the exhaust channel 216 are sequentially connected. When the sample lysate and the sample to be detected after the isothermal amplification reaction in the first reaction chamber 214 are conveyed to the second reaction chamber 215, the exhaust channel 216 is in a closed state, and the gas in the second reaction chamber 215 enters the partial pressure chamber 213, so that the excessive pressure in the flow path 2 can be avoided by arranging the partial pressure chamber 213.
Specifically, in this embodiment, the area ratio of the first reaction chamber 214 to the second reaction chamber 215 is 2:1, depth ratio of 4:3, the area of the second reaction chamber 215 is equal to that of the partial pressure chamber 213, and the depth ratio is 1:2, the larger volume of the pressure dividing chamber 213 relative to the second reaction chamber 215 is advantageous for sharing the pressure inside the liquid path.
In this embodiment, as shown in fig. 2, two first arc slopes 20 matching the first ball head shape are circumferentially spaced apart from the first cavity 14, two second arc slopes 21 matching the second ball head shape are circumferentially spaced apart from the second cavity 15, the arc centers of the two first arc slopes 20 coincide, the arc centers of the two first arc slopes 20 are located on the axis of the fourth branch 204, the arc centers of the two second arc slopes 21 coincide, and the arc centers of the two second arc slopes 21 are located on the axis of the third branch 208. By providing the first circular arc slope 20 and the second circular arc slope 21, the first ball and the second ball are more convenient to move.
In this embodiment, as shown in fig. 2 and 6, the bottom end of the cavity 3 is further provided with a third cavity 211 and a fourth cavity, the shape of the third cavity 211 is matched with the shape of the first elastic membrane 10, the third cavity 211 is used for accommodating the first elastic membrane 10, the shape of the fourth cavity is matched with the shape of the second elastic membrane, and the fourth cavity is used for accommodating the second elastic membrane. So set up, first elastic membrane 10 and second elastic membrane hold respectively in third cavity 211 and fourth cavity, first elastic membrane 10 and second elastic membrane imbeds cavity 3 bottom like this for cavity 3 bottom is leveled. In a specific use process, when the first ball head and the second ball head are respectively embedded into the first positioning concave pit 18 and the second positioning concave pit 19, the shapes of the first elastic film 10 and the second elastic film are changed.
As shown in fig. 1, in this embodiment, the microfluidic chip 100 for detecting CRISPR by multichannel isothermal amplification further includes a plurality of connection assemblies disposed along a circumferential direction of the chip body, the connection assemblies include a screw 22, a first nut 23 and a second nut 24, the screw 22 is detachably disposed on the chip body, the first nut 23 and the second nut 24 are in threaded connection with the screw 22, a plurality of arc holes 901 are disposed on the cover 9, the screw 22 is in one-to-one correspondence with the arc holes 901, each screw 22 passes through each corresponding arc hole 901 and is in sliding connection with each corresponding arc hole 901, so that the cover 9 and the chip body can rotate relatively, the first nut 23 and the second nut 24 are disposed at two ends of the cover 9, and two ends of the cover 9 are respectively abutted against the first nut 23 and the second nut 24.
In a specific use process, the height of the cover plate 9 can be adjusted by rotating the first nut 23 and the second nut 24, so that the cover plate 9 moves towards a direction approaching or separating from the chip body.
In this embodiment, as shown in fig. 2, a plurality of bumps 25 for supporting the cover plate 9 are circumferentially arranged at one end of the chip body close to the cover plate 9. By supporting the cover plate 9 on the bump 25, the contact area between the cover plate 9 and the chip body becomes small, and thus the friction between the cover plate 9 and the chip body becomes small.
The principles and embodiments of the present invention have been described in this specification with reference to specific examples, the description of which is only for the purpose of aiding in understanding the method of the present invention and its core ideas; also, it is within the scope of the present invention to be modified by those of ordinary skill in the art in light of the present teachings. In view of the foregoing, this description should not be construed as limiting the invention.
Claims (10)
1. A microfluidic chip for multichannel isothermal amplification CRISPR detection, comprising:
the chip body is provided with a cavity in a recessed mode, a liquid injection channel is arranged in the center of the chip body, a flow path is arranged in the chip body, the flow path comprises a first liquid path communicated with the liquid injection channel, a second liquid path communicated with the first liquid path and an exhaust channel communicated with the second liquid path, the first liquid path and the second liquid path are arranged at intervals along the circumferential direction of the chip body, the first liquid path comprises a first reaction cavity, the second liquid path comprises a second reaction cavity, and a first annular slideway and a second annular slideway are further arranged at the bottom end of the cavity;
the plugging structure is provided with a gas discharging passage communicated with the gas discharging passage, the plugging structure can be slidably arranged in the gas discharging passage, the gas discharging passage can be communicated with the outside atmosphere when the plugging structure is in a first position, and the gas discharging passage is isolated from the outside atmosphere when the plugging structure is in a second position;
the cover plate can be rotatably arranged at one end of the chip body, where the cavity is arranged, and can move towards a direction close to or far away from the chip body, a limit structure is arranged at one end of the cover plate, close to the cavity, and when the blocking structure is positioned at the second position, the limit structure is propped against the blocking structure;
the valve bank comprises a first valve and a second valve, the first valve comprises a first concave cavity, a first elastic membrane and a first ball plunger, the second valve comprises a second concave cavity, a second elastic membrane and a second ball plunger, the first concave cavity and the second concave cavity are both arranged at the bottom end of the cavity, the first concave cavity is arranged on a first flow path, the second concave cavity is arranged on a second flow path, the first concave cavity is arranged between the first reaction cavity and the liquid injection channel, the second concave cavity is arranged between the first reaction cavity and the second reaction cavity, the first elastic membrane seals the first concave cavity, the second elastic membrane seals the second concave cavity, the first ball plunger and the second ball plunger are both arranged on the cover plate, one end of the first ball plunger, which is provided with the first ball, slides along the first annular slide way, one end of the second plunger is provided with the second ball, and can be inserted into the second concave cavity and can be blocked by the second plunger when the second ball plunger is arranged in the first position and the second slide way;
and the cross section shape of the piston is matched with that of the liquid injection channel.
2. The microfluidic chip for detecting multichannel isothermal amplification CRISPR according to claim 1, wherein the number of flow paths is plural, all the flow paths are uniformly arranged along the circumferential direction of the chip body, the number of the first valves and the second valves is plural, one flow path corresponds to one first valve and one second valve, and all the first valves and all the second valves are uniformly arranged along the circumferential direction of the chip body.
3. The microfluidic chip for detecting multichannel isothermal amplification CRISPR according to claim 1, wherein a plurality of first positioning pits are uniformly arranged in the circumferential direction of the first annular slide, a plurality of second positioning pits are uniformly arranged in the circumferential direction of the second annular slide, one first positioning pit is arranged between any two adjacent first cavities, one second positioning pit is arranged between any two adjacent second cavities, an included angle between the first positioning pit and the adjacent first cavities is equal to an included angle between the second positioning pit and the adjacent second cavities, the shape of the first positioning pit is matched with that of the first ball head, and the shape of the second positioning pit is matched with that of the second ball head.
4. The microfluidic chip for multi-channel isothermal amplification CRISPR detection according to claim 1, wherein the liquid injection channel is arranged along a height direction of the chip body, the first liquid channel comprises a first branch, a second branch, a third branch, a fourth branch and a fifth branch, the first branch and the second branch are collinear and are arranged along a radial direction of the chip body, the third branch, the fourth branch and the fifth branch are parallel to each other and are arranged along the height direction of the chip body, and the first branch, the third branch, the first concave cavity, the fourth branch, the second branch and the fifth branch are sequentially communicated, and the second branch has the first reaction cavity; the second liquid way includes first branch road, second branch road, third branch road, fourth branch road and fifth branch road, first branch road second branch road and third branch road are parallel to each other, and all follow the direction of height of the chip body sets up, fourth branch road with fifth branch road collineation, and all follow the radial setting of the chip body, first branch road fourth branch road second cavity third branch road and fifth branch road communicate in proper order, fifth branch road has the second reaction chamber, first branch road is linked together through third branch road with fifth branch road, fifth branch road with exhaust passage is linked together.
5. The microfluidic chip for multichannel isothermal amplification CRISPR detection according to claim 4, wherein the chip body comprises a main body, a first blocking film and a second blocking film, the cavity, the liquid injection channel, the flow path, the first cavity, the second cavity, the first annular slide and the second annular slide are all arranged on the main body, the first branch, the second branch, the fourth branch and the fifth branch penetrate through one end of the main body far away from the cover plate, the first blocking film is arranged at one end of the main body far away from the cover plate, and the first blocking film blocks the first branch, the second branch, the fourth branch and the fifth branch;
the third liquid path comprises a third concave cavity and an arc-shaped groove, the third concave cavity is communicated with one end of the fifth branch, the diameter of the third concave cavity is larger than that of the fifth branch, one end of the arc-shaped groove is communicated with the third concave cavity, the other end of the arc-shaped groove is communicated with the first branch, an included angle of a liquid inlet end of the first reaction cavity is 10-20 degrees, an included angle of a liquid outlet end of the first reaction cavity is 30-60 degrees, and the second blocking film is used for sealing the third concave cavity and the arc-shaped groove.
6. The microfluidic chip for multichannel isothermal amplification CRISPR detection according to claim 4, wherein the fifth branch further comprises a partial pressure cavity, and the second reaction cavity, the partial pressure cavity and the exhaust channel are sequentially communicated.
7. The microfluidic chip for multichannel isothermal amplification CRISPR detection according to claim 6, wherein the area ratio of the first reaction chamber to the second reaction chamber is 2:1, depth ratio of 4:3, the area of the second reaction cavity is equal to that of the partial pressure cavity, and the depth ratio is 1:2.
8. the microfluidic chip for detecting the multichannel isothermal amplification CRISPR according to claim 4, wherein two first circular arc slopes matched with the first ball head shape are arranged at intervals in the circumferential direction of the first concave cavity, two second circular arc slopes matched with the second ball head shape are arranged at intervals in the circumferential direction of the second concave cavity, the arc centers of the two first circular arc slopes coincide, the arc centers of the two first circular arc slopes are positioned on the axis of the fourth branch, the arc centers of the two second circular arc slopes coincide, and the arc centers of the two second circular arc slopes are positioned on the axis of the third branch;
the bottom of the cavity is also provided with a third cavity and a fourth cavity, the shape of the third cavity is matched with that of the first elastic membrane, the third cavity is used for accommodating the first elastic membrane, the shape of the fourth cavity is matched with that of the second elastic membrane, and the fourth cavity is used for accommodating the second elastic membrane.
9. The microfluidic chip for detecting multichannel isothermal amplification CRISPR according to claim 1, further comprising a plurality of connection assemblies arranged along the periphery of the chip body, wherein the connection assemblies comprise screws, first nuts and second nuts, the screws can be detachably arranged on the chip body, the first nuts and the second nuts are in threaded connection with the screws, a plurality of arc holes are formed in the cover plate, the screws are in one-to-one correspondence with the arc holes, each screw penetrates through each corresponding arc hole and is in sliding connection with each corresponding arc hole so that the cover plate and the chip body can rotate relatively, the first nuts and the second nuts are respectively arranged at two ends of the cover plate, and the two ends of the cover plate are respectively propped against the first nuts and the second nuts.
10. The microfluidic chip for multichannel isothermal amplification CRISPR detection according to claim 1, wherein a plurality of bumps for supporting the cover plate are circumferentially arranged at one end of the chip body close to the cover plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310154380.3A CN116139954B (en) | 2023-02-20 | 2023-02-20 | Microfluidic chip for multichannel isothermal amplification CRISPR detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310154380.3A CN116139954B (en) | 2023-02-20 | 2023-02-20 | Microfluidic chip for multichannel isothermal amplification CRISPR detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116139954A true CN116139954A (en) | 2023-05-23 |
| CN116139954B CN116139954B (en) | 2023-08-22 |
Family
ID=86356052
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310154380.3A Active CN116139954B (en) | 2023-02-20 | 2023-02-20 | Microfluidic chip for multichannel isothermal amplification CRISPR detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116139954B (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100252116A1 (en) * | 2009-04-03 | 2010-10-07 | Integrated Nano-Technologies, Inc. | Multi-chamber rotating valve |
| US20110070654A1 (en) * | 2009-09-23 | 2011-03-24 | Ecolab Inc. | Valve analytical system |
| US20110297866A1 (en) * | 2009-01-21 | 2011-12-08 | Thinxxs Microtechnology Ag | Valve, in particular for a component in microfluid technology |
| KR20120048735A (en) * | 2010-11-04 | 2012-05-16 | 도병록 | Multiway valve and system for extracting regenerative cells using the same |
| US20150308578A1 (en) * | 2010-10-07 | 2015-10-29 | Vanderbilt University | Normally closed microvalve and applications of the same |
| CN108192816A (en) * | 2018-02-05 | 2018-06-22 | 宁波东夏生物科技有限公司 | A kind of sample automatically processes and nucleic acid amplifier and application method |
| US20200326316A1 (en) * | 2018-01-23 | 2020-10-15 | Ldetek Inc. | Valve assembly for a gas chromatograph |
| CN113846004A (en) * | 2021-10-29 | 2021-12-28 | 广东粤港澳大湾区国家纳米科技创新研究院 | Rotary type nucleic acid amplification product detection device and detection method |
| WO2022100270A1 (en) * | 2020-11-16 | 2022-05-19 | 北京迅识科技有限公司 | Method for testing target nucleic acid in sample |
| CN114713304A (en) * | 2022-05-10 | 2022-07-08 | 杭州霆科生物科技有限公司 | Totally-enclosed micro-fluidic chip for nucleic acid amplification detection |
| CN115305183A (en) * | 2022-08-10 | 2022-11-08 | 合肥中科易康达生物医学有限公司 | Centrifugal microfluidic chip integrating isothermal amplification and CRISPR/Cas nucleic acid detection and method |
| CN115466656A (en) * | 2022-08-31 | 2022-12-13 | 广东凯普科技智造有限公司 | Microfluidic nucleic acid extraction detection box and use method thereof |
-
2023
- 2023-02-20 CN CN202310154380.3A patent/CN116139954B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110297866A1 (en) * | 2009-01-21 | 2011-12-08 | Thinxxs Microtechnology Ag | Valve, in particular for a component in microfluid technology |
| US20100252116A1 (en) * | 2009-04-03 | 2010-10-07 | Integrated Nano-Technologies, Inc. | Multi-chamber rotating valve |
| US20110070654A1 (en) * | 2009-09-23 | 2011-03-24 | Ecolab Inc. | Valve analytical system |
| US20150308578A1 (en) * | 2010-10-07 | 2015-10-29 | Vanderbilt University | Normally closed microvalve and applications of the same |
| KR20120048735A (en) * | 2010-11-04 | 2012-05-16 | 도병록 | Multiway valve and system for extracting regenerative cells using the same |
| US20200326316A1 (en) * | 2018-01-23 | 2020-10-15 | Ldetek Inc. | Valve assembly for a gas chromatograph |
| CN108192816A (en) * | 2018-02-05 | 2018-06-22 | 宁波东夏生物科技有限公司 | A kind of sample automatically processes and nucleic acid amplifier and application method |
| WO2022100270A1 (en) * | 2020-11-16 | 2022-05-19 | 北京迅识科技有限公司 | Method for testing target nucleic acid in sample |
| CN113846004A (en) * | 2021-10-29 | 2021-12-28 | 广东粤港澳大湾区国家纳米科技创新研究院 | Rotary type nucleic acid amplification product detection device and detection method |
| CN114713304A (en) * | 2022-05-10 | 2022-07-08 | 杭州霆科生物科技有限公司 | Totally-enclosed micro-fluidic chip for nucleic acid amplification detection |
| CN115305183A (en) * | 2022-08-10 | 2022-11-08 | 合肥中科易康达生物医学有限公司 | Centrifugal microfluidic chip integrating isothermal amplification and CRISPR/Cas nucleic acid detection and method |
| CN115466656A (en) * | 2022-08-31 | 2022-12-13 | 广东凯普科技智造有限公司 | Microfluidic nucleic acid extraction detection box and use method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 任冀峰等: ""类微血管的直型微通道中对细胞速度的三维仿真分析"", 《医用生物力学》, vol. 36, no. 8, pages 110 * |
| 伍辉: ""LAMP结合CRISPR技术用于致病微生物集成检测的研究"", 中国博士论文全文数据库（电子期刊）, no. 2, pages 52 - 61 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116139954B (en) | 2023-08-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20210322980A1 (en) | Whole-process biological detection device | |
| WO2020001030A2 (en) | Nucleic acid integrated detection method and detection reagent tube | |
| WO2022253145A1 (en) | Blood testing microfluidic chip | |
| WO2023000654A1 (en) | Genetic test kit and genetic test apparatus | |
| CN1308681C (en) | Slide cassette for fluidic injection | |
| CN111909835A (en) | Closed micro-fluidic nucleic acid detection card box | |
| TWI411779B (en) | Microfluidic bio-chip and automatic reaction detection system thereof | |
| WO2022253146A1 (en) | Platelet detection microfluidic chip | |
| CN116139954B (en) | Microfluidic chip for multichannel isothermal amplification CRISPR detection | |
| CN113186098B (en) | Integrated lower-opening nucleic acid quick-lifting test tube, quick-lifting detection device and method | |
| CN113817601A (en) | Sample treatment and detection device | |
| CN115703991A (en) | Micro amplification instrument | |
| US20220008925A1 (en) | Rotary integrated micro-device for gene diagnosis | |
| CN212316088U (en) | Totally-enclosed multi-index nucleic acid detection device | |
| CN207210427U (en) | A kind of rotary detection of nucleic acids cartridge | |
| CN113146313A (en) | Pneumatic piston driving rotary centering clamp | |
| CN113492024B (en) | Microfluidic chip with self-driving unit, microfluidic method and application of microfluidic chip | |
| CN112877197B (en) | Summer and autumn tea detection system in compound white spirit fermented grains | |
| CN214458048U (en) | Low-cost autonomous liquid suction type nucleic acid detection card box based on magnetic rod method | |
| CN209260070U (en) | A kind of Novel free instrument nucleic acid on-site detection card | |
| CN220514209U (en) | Centrifugal assembly | |
| CN219490025U (en) | Nucleic acid detection and analysis kit | |
| CN107389455B (en) | Cooling device and method for initial temperature of sample in magnetic drive oblique wave compression | |
| CN218596408U (en) | Micro-fluidic chip device based on recombinase amplification fluorescence probe method | |
| CN117511721B (en) | Real-time fluorescence quantitative PCR instrument based on high-flux microfluidic chip |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |