Integrated micro-fluidic chip

Abstract

The invention discloses an integrated micro-fluidic chip, the chip comprises a chip main body and a sliding cover, the sliding cover is movably arranged at the top of the chip main body, and the sliding cover can move along the extension direction of the chip main body. The chip main body is provided with at least two accommodating cavities recessed towards the interior of the chip main body along the extension direction of the chip main body, and the plurality of accommodating cavities comprise a first accommodating cavity and a second accommodating cavity; the first containing cavity comprises a first containing cavity main body, a reversing column and a detection tube, the first containing cavity main body and the chip main body are integrally formed, the reversing column is arranged in the first containing cavity main body in a sleeved mode, and the reversing column can rotate relative to the first containing cavity main body; the reversing column is arranged in the second accommodating cavity, so that the reversing column is communicated with the second accommodating cavity, the detection tube is arranged at one end, far away from the chip main body, of the reversing column, the detection tube is communicated with the reversing column, and the second accommodating cavity is used for storing a reagent and / or providing an experiment space.

Description

Technical Field

[0001] The present application relates to the field of biomedical engineering, and in particular to an integrated microfluidic chip. Background Art

[0002] Nucleic acid is the general term for deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). It is a biological macromolecular compound formed by the polymerization of many nucleotide monomers and is one of the most basic substances of life. Nucleic acid is a class of biological polymers, an indispensable component of all known life forms, the most important substance among all biological molecules, and widely present in all animal and plant cells and microorganisms. Nucleic acid is composed of nucleotides, and nucleotide monomers are composed of pentose, phosphate group and nitrogenous base. If the pentose is ribose, the polymer formed is RNA; if the pentose is deoxyribose, the polymer formed is DNA.

[0003] Methylation refers to the process of catalytically transferring methyl groups from active methyl compounds to other compounds, which can form various methyl compounds, or chemically modify certain proteins or nucleic acids to form methylated products. In biological systems, methylation is catalyzed by enzymes, which involves heavy metal modification, regulation of gene expression, regulation of protein function, and RNA processing.

[0004] DNA methylation in vertebrates generally occurs at CpG sites (cytosine-phosphate-guanine sites, i.e., sites in the DNA sequence where guanine is immediately following cytosine). Cytosine is converted to 5-methylcytosine by DNA methyltransferase. About 80%-90% of CpG sites in human genes are methylated, but in certain specific areas, such as CpG islands rich in cytosine and guanine, they are not methylated. This is related to promoters in 56% of mammalian genes, including all widely expressed genes. 1%-2% of the human genome is CpG clusters, and CpG methylation is inversely proportional to transcriptional activity.

[0005] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be regarded as a special DNA replication outside the body. The biggest feature of PCR is that it can greatly increase trace amounts of DNA. Therefore, whether it is ancient organisms in fossils, the remains of historical figures, or the hair, skin or blood left by the murderer in a homicide case decades ago, as long as a little bit of DNA can be separated, it can be amplified and compared using PCR. This is also the power of "trace evidence". PCR uses the fact that DNA denatures at a high temperature of 95°C in vitro to become a single strand. At low temperatures (usually around 60°C), primers and single strands combine according to the principle of complementary base pairing. The temperature is then adjusted to the optimal reaction temperature of DNA polymerase (around 72°C), and DNA polymerase synthesizes complementary chains along the direction from phosphate to pentose (5'-3'). The PCR instrument manufactured based on polymerase is actually a temperature control device that can well control the denaturation temperature, renaturation temperature, and extension temperature.

[0006] At present, there are three main methods for nucleic acid extraction and amplification: 1) Manual experimental scheme, in which the experimenter uses pipettes, constant temperature oscillators, mixers and other equipment in a specific experimental environment, and extracts DNA from the sample according to the use requirements of the reagent instructions, methylates and purifies it, and finally performs PCR amplification; however, the experimental process of this scheme is complicated, the experimental time is long, and the experimental efficiency is low; there are many types of small equipment used in the experiment; and there are extremely high requirements for the laboratory environment; 2) Semi-automatic equipment experimental scheme, in which the experimenter uses a pipette to add reagents to the equipment consumables according to the equipment use requirements in a specific experimental environment, and then runs the semi-automatic equipment to perform nucleic acid extraction and methylation conversion experiments. After the experiment is completed, the experimenter manually transfers the DNA and performs PCR amplification on the PCR instrument. However, this scheme requires manual supervision and manual sample addition during the experiment; it has extremely high requirements for the laboratory environment; 3) Fully automatic equipment experimental scheme, in which the experimenter directly operates the automated equipment, and the equipment automatically completes nucleic acid extraction, methylation conversion and PCR amplification. The equipment of this scheme occupies a large area and is expensive; the equipment development is difficult. Summary of the invention

[0007] Based on the above-mentioned defects, the present application provides an integrated microfluidic chip, which can perform experiments (such as DNA extraction, methylation purification and PCR detection) by moving the sliding cover to connect the first through hole with different accommodating cavities.

[0008] This application provides the following technical solutions.

[0009] 1. An integrated microfluidic chip, wherein the chip comprises a chip body and a slide cover, wherein the slide cover is movably arranged on the top of the chip body, and the slide cover can move along the extension direction of the chip body,

[0010] The chip body is provided with at least two accommodating cavities recessed into the chip body along its extension direction.

[0011] The plurality of accommodating chambers include a first accommodating chamber and a second accommodating chamber;

[0012] The first accommodating cavity comprises a first accommodating cavity body, a commutating column and a detection tube. The first accommodating cavity body and the chip body are integrally formed. The commutating column is sleeved in the first accommodating cavity body, and the commutating column can rotate relative to the first accommodating cavity body, so that the commutating column is connected with the second accommodating cavity. The detection tube is arranged at one end of the commutating column away from the chip body, and the detection tube is connected with the commutating column.

[0013] The second containing cavity is used for storing reagents and / or for providing experimental space.

[0014] 2. The chip according to item 1, wherein the reversing column is provided with a plurality of ventilation channels and a plurality of microchannels,

[0015] The ventilation air channel penetrates the reversing column along the radial direction of the reversing column;

[0016] The microfluidic channel is connected to the detection tube;

[0017] The reversing column can be rotated to make the microchannel communicate with the second accommodating chamber, thereby making the second accommodating chamber communicate with the detection tube.

[0018] 3. The chip according to item 2, wherein the commutation column comprises a commutation column body and a connecting column, the connecting column is arranged at one end of the commutation column body away from the chip body, and the axis of the connecting column coincides with the axis of the commutation column body;

[0019] The connecting column is sleeved in the detection tube.

[0020] 4. The chip according to item 3, wherein the microfluidic channel comprises a connecting channel and a guiding channel, the connecting channel is used to introduce the reagent or sample in the second accommodating chamber into the reversing column body; one end of the connecting channel is used to communicate with the second accommodating chamber, and the other end thereof is connected to the guiding channel;

[0021] The guide channel is used to communicate with the detection tube, and the guide channel is used to introduce the reagent or sample introduced through the connecting channel into the detection tube.

[0022] 5. The chip according to item 4, wherein the flow guide channel includes a first flow channel and a second flow channel; the connecting channel includes a third flow channel, a fourth flow channel, a fifth flow channel and a sixth flow channel,

[0023] The first flow channel is arranged along the axis of the reversing column body, and the second flow channel is parallel to the first flow channel;

[0024] The third flow channel and the fourth flow channel are both arranged along the radial direction of the reversing column body, and are communicated with the first flow channel;

[0025] The fifth flow channel and the sixth flow channel are both arranged along the radial direction of the reversing column body, and are communicated with the second flow channel;

[0026] Along the axial direction of the reversing column body, the third flow channel, the fourth flow channel and the fifth flow channel are arranged at different heights;

[0027] Along the axial direction of the reversing column body, the fifth flow channel and the sixth flow channel are arranged at the same height.

[0028] 6. The chip according to item 5, wherein along the axial direction of the reversing column body, from one end close to the chip body to one end close to the detection tube, the third flow channel, the fourth flow channel, and the fifth flow channel are arranged in sequence;

[0029] The plane where the third flow channel is located is plane A,

[0030] The projection of the fourth flow channel on the plane A forms an angle β with the third flow channel;

[0031] The projection of the fourth flow channel on the plane A and the projection of the fifth flow channel on the plane A are on the same straight line;

[0032] The fifth flow channel forms an angle α with the sixth flow channel.

[0033] 7. The chip according to item 5, wherein the first flow channel takes one end of the third flow channel as a starting point and extends along the axis of the reversing column body until it passes through the connecting column;

[0034] The second flow channel extends from one end of the fourth flow channel and the fifth flow channel to the side wall of the connecting column and penetrates the side wall of the connecting column.

[0035] 8. The chip according to item 6, wherein the ventilation airway comprises a first ventilation airway, a second ventilation airway and a third ventilation airway, and along the axial direction of the reversing column body, the first ventilation airway, the second ventilation airway and the third ventilation airway are arranged at different heights;

[0036] In the reversing column body, along the axial direction of the reversing column body, the first ventilation channel and the third flow channel are located at the same height;

[0037] In the reversing column body, along the axial direction of the reversing column body, the second ventilation channel and the fourth flow channel are located at the same height;

[0038] In the reversing column body, along the axial direction of the reversing column body, the third ventilation channel and the fifth flow channel are located at the same height.

[0039] 9. The chip according to item 8, wherein a projection of the first ventilation channel on the plane A coincides with a projection of the second ventilation channel on the plane A;

[0040] The projection of the first ventilation channel on the plane A and the projection of the third ventilation channel on the plane A form an angle θ.

[0041] 10. The chip according to item 8, wherein the second accommodating cavity comprises a second accommodating cavity A, a second accommodating cavity B, and a second accommodating cavity C,

[0042] The second receiving chamber A is used to store reagents.

[0043] The second accommodating chamber B is used to provide an experimental space for experiments;

[0044] The second containing chamber C is used to store reagents and also provides experimental space for experiments.

[0045] 11. The chip according to item 10, wherein a second accommodating cavity A is provided on one side of the first accommodating cavity, and a second accommodating cavity C is provided on the other side.

[0046] The second accommodating chamber A comprises a second accommodating chamber A1 and a second accommodating chamber A2 arranged side by side;

[0047] The second accommodating chamber C is in communication with the fourth flow channel or the second ventilation channel;

[0048] The second accommodating chamber A1 is in communication with the third flow channel or the first ventilation channel;

[0049] The second accommodating chamber A2 is communicated with the fifth flow channel, the sixth flow channel or the third ventilation channel.

[0050] 12. The chip according to item 10, wherein the number of the first accommodating cavities is at least 1, the number of the second accommodating cavities A is at least 2, the number of the second accommodating cavities B is at least 1, and the number of the second accommodating cavities C is at least 2; or

[0051] The openings of the first accommodating chamber, the second accommodating container A, the second accommodating chamber B and the second accommodating chamber C are respectively circular, elliptical or n-sided, where n is greater than or equal to 3.

[0052] 13. The chip according to any one of items 1 to 12, wherein the slide cover is provided with at least two first through holes penetrating the slide cover surface, and a cylindrical channel protruding from the slide cover surface is provided on a side surface of the slide cover away from the chip body, and the cylindrical channel is located above the first through holes;

[0053] An elastic sealing ring is arranged in the cylindrical passage.

[0054] 14. The chip according to item 13, wherein at least two of the first through holes are arranged in sequence along the extension direction of the sliding cover.

[0055] 15. The chip according to item 13, wherein a second through hole is provided on the slide cover, and an opening channel protruding from the surface of the slide cover is provided on a side surface of the slide cover away from the chip body, and the opening channel is located above the second through hole.

[0056] 16. The chip according to item 15, wherein a channel cover for closing the open channel is provided at the top of the open channel.

[0057] 17. The chip according to item 15, wherein the opening channel and the cylindrical channel are located on a center line of a surface of the sliding cover facing away from the chip body;

[0058] The height of the open channel is smaller than the height of the cylindrical channel.

[0059] 18. The chip according to item 15, wherein the side of the slide cover close to the chip body has a slide groove, and the side of the chip body close to the slide cover has a slide sheet matched with the slide groove,

[0060] The slide is provided with an opening matched with the accommodating cavity.

[0061] The integrated microfluidic chip provided by the present application, when in use, by moving the slide cover, so that the first through hole is connected to different accommodating chambers, so as to conduct experiments, such as extracting and methylating DNA in the second accommodating chamber. After methylating purification, the reversing column of the first accommodating chamber rotates relative to the main body of the first accommodating chamber, so that the reversing column is connected to the second accommodating chamber, and the purified DNA and PCR detection reagents in the second accommodating chamber enter the first accommodating chamber for PCR detection, thereby simplifying the operation process, reducing the time for preparation and operation, and improving work efficiency. Since the reagents can be pre-packaged in the second accommodating chamber in the present application, it can not only prevent external contamination, protect the experimenters and the environment, but also maintain the stability of the experimental conditions. At the same time, the pre-packaged reagents also ensure the consistency and repeatability of the experiment, reducing the risk of operational errors and cross contamination. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 Schematic diagram of the integrated microfluidic chip structure provided in this application.

[0063] Figure 2 This is a schematic diagram of the structure of the reversing column provided in this application.

[0064] Description of Reference Numerals

[0065] 1-sliding cover, 2-reversing column, 3-cylindrical channel, 4-opening channel, 5-first accommodating chamber, 6-mounting plate, 7-first accommodating chamber body, 8-sliding groove, 9-sliding sheet, 10-detection tube, 11-second flow channel, 12-card slot, 13-syringe, 14-magnetic rod sleeve, 15-connecting column, 16-first ventilation channel, 17-second ventilation channel, 18-third ventilation channel, 19-third flow channel, 20-fourth flow channel, 21-fifth flow channel, 22-sixth flow channel, 23-first flow channel. DETAILED DESCRIPTION

[0066] The following is a description of the exemplary embodiments of the present application, including various details of the embodiments of the present application to facilitate understanding, which should be considered as merely exemplary. Therefore, it should be recognized by those of ordinary skill in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for the sake of clarity and conciseness, the description of well-known functions and structures is omitted in the following description.

[0067] like Figure 1 as well as Figure 2 As shown, the present application provides an integrated microfluidic chip, wherein the chip comprises a chip body and a slide cover 1, wherein the slide cover 1 is movably arranged on the top of the chip body, and when in use, the slide cover 1 moves along the extension direction of the chip body.

[0068] The chip body is provided with at least two accommodating cavities recessed into the chip body along its extension direction.

[0069] The number of the accommodating cavities may be 2, 3, 4, 5, 6, 7, 8, 9, 10, etc., and the number of the accommodating cavities may be determined according to actual needs.

[0070] The plurality of accommodating chambers include a first accommodating chamber 5 and a second accommodating chamber;

[0071] The first accommodating chamber 5 includes a first accommodating chamber body 7, a reversing column 2 and a detection tube 10. The first accommodating chamber body 7 is integrally formed with the chip body. The reversing column 2 is sleeved in the first accommodating chamber body 7, and the reversing column 2 can rotate relative to the first accommodating chamber body 7, so that the reversing column 2 is connected to the second accommodating chamber. The detection tube 10 is arranged at one end of the reversing column 2 away from the chip body, and the detection tube 10 is connected to the reversing column 2; for example, when the chip is used for nucleic acid extraction, methylation purification and PCR detection, DNA extraction and methylation purification are carried out in the second accommodating chamber. After methylation purification, the reversing column 2 of the first accommodating chamber 5 is rotated relative to the first accommodating chamber body 7, so that the reversing column 2 is connected to the second accommodating chamber, and the purified DNA and PCR detection reagent in the second accommodating chamber enter the first accommodating chamber 5 for PCR detection.

[0072] The second accommodating cavity is integrally formed with the chip body, and is used for storing reagents and / or for providing an experimental space.

[0073] The number of the second accommodating cavities may be 2, 3, 4, 5, 6, 7, 8, 9, etc., and the number of the second accommodating cavities may be determined according to actual needs.

[0074] In the present application, a plurality of ventilation channels and a plurality of microchannels are provided in the reversing column 2, and the ventilation channels penetrate the reversing column 2 in the radial direction of the reversing column 2; the microchannels are connected to the detection tube 10;

[0075] The reversing column 2 can be rotated to connect the microchannel with the second accommodating chamber, thereby connecting the second accommodating chamber with the detection tube 10 .

[0076] When the second accommodating chamber needs to be in a negative pressure state, the reversing column 2 can be rotated to connect the ventilation channel with the second accommodating chamber, and the second accommodating chamber can be connected with the external environment, and then the piston is pulled to make the second accommodating chamber in a negative pressure state. When the reagent or sample in the second accommodating chamber needs to be injected into the detection tube, the reversing column 2 can be rotated to connect the microchannel with the second accommodating chamber, so that under the action of negative pressure, the reagent or sample in the second accommodating chamber is pressed into the microchannel and then enters the detection tube.

[0077] Furthermore, the commutation column 2 includes a commutation column 2 body and a connecting column 15, wherein the connecting column 15 is arranged at one end of the commutation column 2 body away from the chip body, and the connecting column 15 coincides with the axis of the commutation column 2 body; the connecting column 15 is sleeved in the detection tube 10.

[0078] A clamping slot 12 is further provided at one end of the main body of the reversing column 2 away from the connecting column 15 , and the reversing column 2 is rotated through the clamping slot 12 .

[0079] The outer contour of the body of the reversing column 2 can be cylindrical, truncated cone, prism or prism, for example, cylindrical.

[0080] The profiles of the ventilation channel and the microchannel may be cylindrical, truncated cone, prism-shaped or prismatic channels.

[0081] The length of the ventilation passage is consistent with the equivalent diameter of the main body of the reversing column 2 . For example, if the main body of the reversing column 2 is a cylindrical structure, the length of the ventilation passage is equal to the diameter of the main body of the reversing column 2 .

[0082] The outer contour of the connecting column 15 is conical or cylindrical, for example, it can be a cone, a pyramid, a cylinder or a prism.

[0083] The equivalent diameter of one end of the connecting post 15 close to the body of the reversing post 2 is smaller than the equivalent diameter of one end of the body of the reversing post 2 close to the connecting post 15 .

[0084] Furthermore, the microfluidic channel includes a connecting channel and a guiding channel, wherein the connecting channel is used to introduce the reagent or sample in the second accommodating chamber into the body of the reversing column 2; one end of the connecting channel is used to communicate with the second accommodating chamber, and the other end thereof is connected with the guiding channel;

[0085] The guide channel is used to communicate with the detection tube 10 ; the guide channel is used to introduce the reagent or sample introduced through the connecting channel into the detection tube 10 .

[0086] The connecting channel is arranged along the radial direction of the body of the reversing column 2 , the equivalent diameter of the radial section where the connecting channel is located is R, and the length of the connecting channel is less than or equal to R.

[0087] The length of the guide channel is smaller than the height of the body of the reversing column 2 .

[0088] Furthermore, the flow guiding channel includes a first flow channel 23 and a second flow channel 11 ; the connecting channel includes a third flow channel 19 , a fourth flow channel 20 , a fifth flow channel 21 and a sixth flow channel 22 .

[0089] The first flow channel 23 is arranged along the axis of the body of the reversing column 2, and the second flow channel 11 is parallel to the first flow channel 23; the length of the first flow channel 23 is greater than the length of the second flow channel 11. The equivalent diameter of the first flow channel 23 may be equal to or different from the equivalent diameter of the second flow channel 11.

[0090] The third flow channel 19 and the fourth flow channel 20 are both arranged along the radial direction of the body of the reversing column 2 and are connected to the first flow channel 23 ; the third flow channel 19 and the fourth flow channel 20 are equal in length.

[0091] The fifth flow channel 21 and the sixth flow channel 22 are both arranged along the radial direction of the body of the reversing column 2 and are communicated with the second flow channel 11 ; the fifth flow channel 21 and the sixth flow channel 22 are equal in length.

[0092] Along the axial direction of the body of the reversing column 2, the third flow channel 19, the fourth flow channel 20 and the fifth flow channel 21 are arranged at different heights;

[0093] Along the axial direction of the main body of the reversing column 2 , the fifth flow channel 21 and the sixth flow channel 22 are arranged at the same height.

[0094] Furthermore, along the axial direction of the body of the reversing column 2, from one end close to the chip body to one end close to the detection tube 10, the third flow channel 19, the fourth flow channel 20, and the fifth flow channel 21 are sequentially arranged;

[0095] The plane where the third flow channel 19 is located is plane A.

[0096] The projection of the fourth flow channel 20 on the plane A forms an angle β with the third flow channel 19, and β is 0-180°, for example, it can be 0°, 10°, 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, 170°, 180°, etc.

[0097] The projection of the fourth flow channel 20 on the plane A and the projection of the fifth flow channel 21 on the plane A are on the same straight line;

[0098] The fifth flow channel 21 forms an angle α with the sixth flow channel 22, and α is 0-180° and does not include 0°, for example, it can be 10°, 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, 170°, 180°, etc.

[0099] Furthermore, the first flow channel 23 takes one end of the third flow channel 19 as a starting point and extends along the axis of the body of the reversing column 2 until it passes through the connecting column 15;

[0100] The second flow channel 11 extends from one end of the fourth flow channel 20 and the fifth flow channel 21 to the side wall of the connecting column 15 and penetrates the side wall of the connecting column 15 .

[0101] Furthermore, the ventilation passage comprises a first ventilation passage 16, a second ventilation passage 17 and a third ventilation passage 18, and along the axial direction of the body of the reversing column 2, the first ventilation passage 16, the second ventilation passage 17 and the third ventilation passage 18 are arranged at different heights;

[0102] In the body of the reversing column 2, along the axial direction of the body of the reversing column 2, the first ventilation channel 16 and the third flow channel 19 are located at the same height;

[0103] In the body of the reversing column 2, along the axial direction of the body of the reversing column 2, the second ventilation channel 17 and the fourth flow channel 20 are located at the same height;

[0104] In the body of the reversing column 2 , along the axial direction of the body of the reversing column 2 , the third ventilation passage 18 and the fifth flow passage 21 are located at the same height.

[0105] Further, the projection of the first ventilation airway 16 on the plane A coincides with the projection of the second ventilation airway 17 on the plane A;

[0106] The projection of the first ventilation duct 16 on the plane A and the projection of the third ventilation duct 18 on the plane A form an angle θ, where θ is 0-180°, for example, 10°, 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, 170°, 180°, etc.

[0107] In the present application, the second accommodating chamber includes a second accommodating chamber A, a second accommodating chamber B, and a second accommodating chamber C.

[0108] The second receiving chamber A is used to store reagents.

[0109] The second accommodating chamber B is used to provide an experimental space for experiments;

[0110] The second containing chamber C is used to store reagents and also provides experimental space for experiments.

[0111] Furthermore, a second accommodating cavity A is disposed on one side of the first accommodating cavity 5, and a second accommodating cavity C is disposed on the other side.

[0112] The second accommodating chamber A comprises a second accommodating chamber A1 and a second accommodating chamber A2 arranged side by side;

[0113] The second accommodating chamber C is connected to the fourth flow channel 20 or the second ventilation channel 17; when the second accommodating chamber C is connected to the fourth flow channel 20, the sample or reagent in the second accommodating chamber C can flow into the detection tube 10 through the fourth flow channel 20 and the first flow channel 23. When the second accommodating chamber C is connected to the second ventilation channel 17, the second accommodating chamber C is connected to the outside atmosphere.

[0114] The second accommodating chamber A1 is connected to the third flow channel 19 or the first ventilation channel 16; when the second accommodating chamber A1 is connected to the third flow channel 19, the sample or reagent in the second accommodating chamber A1 can flow into the detection tube 10 through the third flow channel 19 and the first flow channel 23. When the second accommodating chamber A1 is connected to the first ventilation channel 16, the second accommodating chamber A1 is connected to the outside atmosphere.

[0115] The second accommodating chamber A2 is in communication with the fifth flow channel 21, the sixth flow channel 22 or the third ventilation channel 18. When the second accommodating chamber A2 is in communication with the fifth flow channel 21, the sample or reagent in the second accommodating chamber A2 can flow into the detection tube 10 through the fifth flow channel 21 and the second flow channel 11, and the sixth flow channel 22 is in communication with the atmosphere, so that the detection tube 10 is in communication with the atmosphere so that the sample or reagent in the second accommodating chamber A2 can flow smoothly into the detection tube 10. When the second accommodating chamber A2 is in communication with the third ventilation channel 18, the second accommodating chamber A2 is in communication with the outside atmosphere.

[0116] Further, the number of the first accommodating cavities 5 is at least 1, the number of the second accommodating cavities A is at least 2, the number of the second accommodating cavities B is at least 1, and the number of the second accommodating cavities C is at least 2.

[0117] The openings of the first accommodating cavity 5, the second accommodating container A, the second accommodating cavity B and the second accommodating cavity C are respectively circular, elliptical or n-gonal, where n is greater than or equal to 3. The n-gonal can be a rectangle, a regular hexagon, a regular octagon, a regular dodecagon, a regular tetragon and the like.

[0118] The number of the first accommodating cavities 5 can be 2, 3, 4, 5, 6, 7, 8, etc., and the number of the second accommodating cavities A can be 2, 3, 4, 5, 6, 7, 8, etc., and the specific data can be determined according to actual needs. The number of the second accommodating cavities B can be 1, 2, 3, 4, 5, 6, 7, 8, etc. The specific data can be determined according to actual needs. The number of the second accommodating cavities C can be 2, 3, 4, 5, 6, 7, 8, etc. The specific data can be determined according to actual needs.

[0119] In some embodiments, the opening of the first accommodating cavity 5 is square, the opening of the second accommodating cavity A is circular or rectangular, the opening of the second accommodating cavity B is circular, and the opening of the second accommodating cavity C is rectangular or circular.

[0120] In some embodiments, when the chip is used for DNA extraction, methylation purification and PCR detection, the number of the first accommodating chamber 5 is 1, the number of the second accommodating chambers A is 8, which are respectively second accommodating chamber A1, second accommodating chamber A2, second accommodating chamber A3, second accommodating chamber A4, second accommodating chamber A5, second accommodating chamber A6, second accommodating chamber A7, and second accommodating chamber A8, the number of the second accommodating chamber B is 1, and the number of the second accommodating chambers C is 4, which are respectively second accommodating chamber C1, second accommodating chamber C2, second accommodating chamber C3, second accommodating chamber C4, second accommodating chamber C5, second accommodating chamber C6, second accommodating chamber C7, second accommodating chamber C8, second accommodating chamber C9, second accommodating chamber C10, second accommodating chamber C11, second accommodating chamber C2, second accommodating chamber C3, second accommodating chamber C4, second accommodating chamber C5, second accommodating chamber C6, second accommodating chamber C7, second accommodating chamber C8, second accommodating chamber C9, second accommodating chamber C12, second accommodating chamber C3, second accommodating chamber C5, second accommodating chamber C6, second accommodating chamber C7, second accommodating chamber C8, second accommodating chamber The accommodating chamber C4, wherein the second accommodating chamber A1 and the second accommodating chamber A2 are located on the same side of the first accommodating chamber 5, and the second accommodating chamber A1 and the second accommodating chamber A2 extend along the width direction of the chip body, and the other side of the first accommodating chamber 5 is sequentially arranged with the second accommodating chamber C1, the second accommodating chamber A3, the second accommodating chamber C2, the second accommodating chamber A4, the second accommodating chamber B, the second accommodating chamber A5, the second accommodating chamber A6, the second accommodating chamber C3, the second accommodating chamber A7, the second accommodating chamber A8, and the second accommodating chamber C4 along the extension direction of the chip body. The second accommodating chamber A1 and the second accommodating chamber A2 are independent of each other and do not communicate with each other, and the second accommodating chamber C1, the second accommodating chamber A3, the second accommodating chamber C2, the second accommodating chamber A4, the second accommodating chamber B, the second accommodating chamber A5, the second accommodating chamber A6, the second accommodating chamber C3, the second accommodating chamber A7, the second accommodating chamber A8, and the second accommodating chamber C4 are independent of each other and do not communicate with each other. The purpose of this design is to ensure that during the process of transferring the liquid to the detection tube 10, the gun tip and the internal reagent of the disposable pipette are in a sealed state.

[0121] The bottom of the second accommodating chamber A1 is provided with a first piston connected to the connecting channel or the venting airway. The first piston is used to provide negative pressure to the sample or reagent in the second accommodating chamber A1 so that the sample or reagent can enter the microchannel for circulation.

[0122] The bottom of the second accommodating chamber A2 is provided with a second piston connected to the connecting channel or the venting airway. The second piston is used to provide negative pressure to the sample or reagent in the second accommodating chamber A2 so that the sample or reagent can enter the microchannel for circulation.

[0123] The bottom of the second accommodating chamber C1 is provided with a third piston connected to the connecting channel or the venting channel. The function of the third piston is to provide negative pressure to the sample or reagent in the second accommodating chamber C1 so that the sample or reagent can enter the microchannel for circulation.

[0124] The first containing chamber 5 is used for performing PCR reaction.

[0125] The second accommodating chambers A1, A2, A5, A6, A7, and A8 are used to store reagents required for the DNA extraction process or amplification process, such as paraffin oil, PCR reaction reagents, lysis solution + magnetic beads, sulfidation reagents, anhydrous ethanol, proteinase K, etc.

[0126] The second accommodating chamber A3 is used to store a syringe 13 used for injection; the second accommodating chamber A4 is used to store a magnetic rod sleeve 14.

[0127] The second containing chamber B is used for sulfite conversion and elution of DNA.

[0128] The second accommodating chamber C4 is used to store lysis solution+magnetic beads in the DNA extraction process, and DNA extraction is performed in the second accommodating chamber C4.

[0129] The second containing chamber C3 is used to store the lysate during the DNA extraction process, and the DNA extraction is continued in the second containing chamber C3.

[0130] The second containing chamber C2 is used to store alcohol-free rinsing liquid, and the DNA is rinsed in the second containing chamber C2.

[0131] The second containing chamber C1 is used to store an elution liquid, and DNA is eluted in the second containing chamber C1.

[0132] The detection tube 10 is a conical tube with a capacity of 500ml-4000ml.

[0133] The capacity of the second accommodating chamber A1 is 300ml-1500ml.

[0134] The capacity of the second accommodating chamber A2 is 300ml-1500ml.

[0135] The capacity of the second accommodating chamber A3 is 1000ml-5000ml.

[0136] The capacity of the second accommodating chamber A4 is 500ml-3000ml.

[0137] The capacity of the second accommodating chamber A5 is 1000ml-5000ml.

[0138] The capacity of the second accommodating chamber A6 is 300ml-1500ml.

[0139] The capacity of the second accommodating chamber A7 is 2000ml-6000ml.

[0140] The capacity of the second accommodating chamber A8 is 300ml-1500ml.

[0141] The capacity of the second accommodating chamber B is 2000ml-6000ml.

[0142] The capacity of the second containing chamber C1 is 300ml-1500ml.

[0143] The capacity of the second containing chamber C2 is 300ml-1500ml.

[0144] The capacity of the second containing chamber C3 is 2000ml-8000ml.

[0145] The capacity of the second containing chamber C4 is 5000ml-15000ml.

[0146] In the present application, at least two first through holes penetrating the surface of the slide cover 1 are provided on the slide cover 1, and a cylindrical channel 3 protruding from the surface of the slide cover 1 is provided on the side surface of the slide cover 1 away from the chip body, and the cylindrical channel 3 is located above the first through holes, and an elastic sealing ring is provided in the cylindrical channel 3. When in use, the first through hole is connected to the accommodating cavity.

[0147] In this article, the extension direction of the chip body is the long side direction of the chip, and the extension direction of the chip body is perpendicular to the recessed direction of the accommodating cavity. The extension direction of the chip body is consistent with the extension direction of the sliding cover 1.

[0148] The number of the first through holes on the sliding cover 1 may be 2, 3, 4, 5, 6, 7 or the like, and the number of the first through holes may be determined according to actual needs.

[0149] The inner diameter of the first through hole is 1mm-100mm, for example, it can be 1mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc., and the spacing is 10mm-30mm, for example, it can be 10mm, 15mm, 20mm, 25mm, 30mm, etc. The first through hole can be used to cooperate with the disposable syringe 13 or the magnetic rod sleeve 14 to seal or experiment the chip body. The inner diameters of the plurality of first through holes can be the same or different, for example, the inner diameter of the first through hole used to match the disposable syringe 13 is smaller than the inner diameter of the first through hole used to match the magnetic rod sleeve 14.

[0150] Further, at least two of the first through holes are arranged in sequence along the extension direction of the slide cover 1. In some embodiments, the first through holes are arranged in sequence along the extension direction of the slide cover 1, and adjacent first through holes are arranged at equal intervals. The interval can match the accommodating cavity of the chip body, and the interval is consistent with the distance between the accommodating cavities, so that the chip is easy to adapt to the device.

[0151] Furthermore, the number of the cylindrical channels 3 is equal to the number of the first through holes, and the inner diameter of the cylindrical channel 3 is greater than or equal to the inner diameter of the first through hole. The bottom edge of the cylindrical channel 3 is arranged around the edge of the first through hole.

[0152] The inner diameter of the cylindrical passage 3 is 2mm-120mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc., and its height is 5mm-120mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc. This design is convenient for the device to take and put the disposable syringe 13 and the magnetic rod sleeve 14.

[0153] In some embodiments, the slide cover 1 is provided with two first through holes penetrating the surface of the slide cover 1, and the two first through holes are arranged in sequence along the extension direction of the slide cover 1. A cylindrical channel 3 is provided on the surface of the slide cover 1 around the edge of the first through holes, and the cylindrical channel 3 corresponds to the first through holes one by one.

[0154] Furthermore, the outer periphery of the sealing ring is connected to the inner wall of the cylindrical channel 3, and the middle of the sealing ring has an opening, and the sealing ring is an elastic sealing ring. When the device is in use, the syringe 13 or the magnetic rod sleeve 14 passes through the opening and enters the first through hole to enter the accommodating cavity. The inner diameter of the opening of the sealing ring is smaller than the outer diameter of the syringe 13 and the magnetic rod sleeve 14. This design isolates the accommodating cavity from the external environment, thereby ensuring the sealing of the chip.

[0155] When methylation treatment is performed through the chip, the syringe 13 or the magnetic rod sleeve 14 penetrates the cylindrical channel 3 and extends into the accommodating cavity. The sealing ring in the cylindrical channel 3 is in tight and sealed contact with the side wall of the syringe 13 or the magnetic rod sleeve 14 to prevent the outside from communicating with the accommodating cavity; when the sliding cover 1 moves along the extension direction of the chip body, the syringe 13 or the magnetic rod sleeve 14 moves out of the accommodating cavity, but the syringe 13 or the magnetic rod sleeve 14 does not extend out of the cylindrical channel 3, that is, the end of the syringe 13 or the magnetic rod sleeve 14 close to the accommodating cavity is still between the sealing ring and the accommodating cavity to ensure that the inside of the chip body is connected with the external environment.

[0156] In the present application, a second through hole is provided on the slide cover 1, and an opening channel 4 protruding from the surface of the slide cover 1 is provided on a side surface of the slide cover 1 away from the chip body, and the opening channel 4 is located directly above the second through hole.

[0157] The number of the second through holes may be 1, 2, 3, 4, 5, 6, 7, 8, 9, etc., and the number of the second through holes may be determined according to actual needs.

[0158] The inner diameter of the second through hole is 2mm-100mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc. The inner diameter of the opening channel 4 is 5mm-120mm, for example, it can be 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc., and the height is 2mm-100mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc. The experimental sample is added into the corresponding accommodating cavity through the second through hole and the opening channel 4.

[0159] Furthermore, the number of the second through holes is the same as the number of the open channels 4. On the surface of the sliding cover 1, the bottom of the open channels 4 is arranged around the second through holes, and the inner diameter of the open channels 4 is greater than or equal to the inner diameter of the second through holes. The center of the second through hole is located on the center line of all the first through holes.

[0160] Furthermore, a channel cover for closing the open channel 4 is provided on the top of the open channel 4. In some embodiments, the open channel 4 and the channel cover are connected in a hinged manner.

[0161] The opening channel 4 and the cylindrical channel 3 are located on the center line of the surface of the sliding cover 1 on one side away from the chip body.

[0162] The height of the opening channel 4 is smaller than the height of the cylindrical channel 3 .

[0163] In the present application, a mounting plate 6 perpendicular to the surface of the slide cover 1 is provided on the side surface of the slide cover 1 away from the chip body, the mounting plate 6 is located between the opening channel 4 and the cylindrical channel 3, and one end of the mounting plate 6 is connected to the outer wall of the cylindrical channel 3.

[0164] The mounting plate 6 is in the shape of an arc-shaped plate with right-angled sides and is used to cooperate with the device to move the slide cover 1 so that the experimenter can grab the chip.

[0165] The lengths of the right-angled sides of the mounting plate 6 are L1 and L2 respectively, L1 is the length of the side in contact with the sliding cover 1 , and L2 is the length of the side in contact with the cylindrical channel 3 .

[0166] In the present application, the slide cover 1 has a slide groove 8 on one side close to the chip body, and the chip body has a slide sheet 9 matched with the slide groove 8 on one side close to the slide cover 1, and an opening matched with the accommodating cavity is provided on the slide sheet 9.

[0167] The extending direction of the slide groove 8, the extending direction of the slide cover 1 and the extending direction of the slide sheet 9 are consistent.

[0168] The chip body is composed of a slide 9 and a receiving cavity. The slide 9 is located at the top of the receiving cavity, and the slide 9 and the receiving cavity are integrally formed.

[0169] When the sliding cover 1 needs to move, the sliding groove 8 moves along the extending direction of the sliding sheet 9, so that the first through hole is connected with the corresponding accommodating cavity.

[0170] Example

[0171] The materials and test methods used in the examples of this application are generally and / or specifically described. In the following examples, unless otherwise specified, % means wt%, i.e., weight percentage. The reagents or instruments used without indicating the manufacturer are all conventional reagent products that can be obtained commercially.

[0172] Example 1

[0173] The integrated microfluidic chip of this embodiment is used for DNA extraction, methylation purification and PCR detection.

[0174] The integrated microfluidic chip comprises a chip body and a slide cover 1, wherein the slide cover 1 is movably arranged on the top of the chip body.

[0175] The chip body is provided with a first accommodating cavity 5 and a second accommodating cavity recessed into the chip body along its extension direction. The number of the first accommodating cavity 5 is 1, the number of the second accommodating cavity A is 8, which are respectively second accommodating cavity A1, second accommodating cavity A2, second accommodating cavity A3, second accommodating cavity A4, second accommodating cavity A5, second accommodating cavity A6, second accommodating cavity A7, and second accommodating cavity A8. The number of the second accommodating cavity B is 1, and the number of the second accommodating cavity C is 4, which are respectively second accommodating cavity C1, second accommodating cavity C2, second accommodating cavity C3, The second accommodating cavity C4, wherein the second accommodating cavity A1 and the second accommodating cavity A2 are located on the same side of the first accommodating cavity 5, and the second accommodating cavity A1 and the second accommodating cavity A2 extend along the width direction of the chip body, and the other side of the first accommodating cavity 5 is sequentially arranged with the second accommodating cavity C1, the second accommodating cavity A3, the second accommodating cavity C2, the second accommodating cavity A4, the second accommodating cavity B, the second accommodating cavity A5, the second accommodating cavity A6, the second accommodating cavity C3, the second accommodating cavity A7, the second accommodating cavity A8, and the second accommodating cavity C4 along the extension direction of the chip body. The second accommodating cavity A1 and the second accommodating cavity A2 are independent of each other and do not communicate with each other, and the second accommodating cavity C1, the second accommodating cavity A3, the second accommodating cavity C2, the second accommodating cavity A4, the second accommodating cavity B, the second accommodating cavity A5, the second accommodating cavity A6, the second accommodating cavity C3, the second accommodating cavity A7, the second accommodating cavity A8, and the second accommodating cavity C4 are independent of each other and do not communicate with each other.

[0176] The first accommodating cavity 5 includes a first accommodating cavity body 7, a commutating column 2 and a detection tube 10. The first accommodating cavity body 7 is integrally formed with the chip body. The commutating column 2 is sleeved in the first accommodating cavity body 7, and the commutating column 2 can rotate relative to the first accommodating cavity body 7, so that the commutating column 2 is connected with the second accommodating cavity. The commutating column 2 includes a cylindrical commutating column 2 body and a conical connecting column 15. The connecting column 15 is arranged at one end of the commutating column 2 body away from the chip body, and the axis of the connecting column 15 coincides with the axis of the commutating column 2 body; the connecting column 15 is sleeved in the detection tube 10.

[0177] The reversing column 2 is provided with a first flow channel 23, a second flow channel 11, a third flow channel 19, a fourth flow channel 20, a fifth flow channel 21, a sixth flow channel 22, a first ventilation channel 16, a second ventilation channel 17, and a third ventilation channel 18. The first flow channel 23 is arranged along the axis of the body of the reversing column 2, and the second flow channel 11 is parallel to the first flow channel 23; the length of the first flow channel 23 is greater than the length of the second flow channel 11. The third flow channel 19 and the fourth flow channel 20 are both arranged along the radial direction of the body of the reversing column 2, and are connected to the first flow channel 23; the length of the third flow channel 19 is equal to that of the fourth flow channel 20. The fifth flow channel 21 and the sixth flow channel 22 are both arranged along the radial direction of the body of the reversing column 2, and are connected to the second flow channel 11; the length of the fifth flow channel 21 is equal to that of the sixth flow channel 22. Along the axial direction of the body of the reversing column 2, the third flow channel 19, the fourth flow channel 20 and the fifth flow channel 21 are arranged at different heights; along the axial direction of the body of the reversing column 2, the fifth flow channel 21 and the sixth flow channel 22 are arranged at the same height. In the body of the reversing column 2, along the axial direction of the body of the reversing column 2, the first ventilation channel 16 and the third flow channel 19 are located at the same height; in the body of the reversing column 2, along the axial direction of the body of the reversing column 2, the second ventilation channel 17 and the fourth flow channel 20 are located at the same height; in the body of the reversing column 2, along the axial direction of the body of the reversing column 2, the third ventilation channel 18 and the fifth flow channel 21 are located at the same height. The plane where the third flow channel 19 is located is plane A, the projection of the fourth flow channel 20 on the plane A forms an angle of 90° with the third flow channel 19, the fifth flow channel 21 forms an angle of 90° with the sixth flow channel 22, and the projection of the first ventilation channel 16 on the plane A forms an angle of 60° with the projection of the third ventilation channel 18 on the plane A. By rotating the reversing post 2, the second accommodating chamber C1 is connected to the fourth flow channel 20 or the second ventilation channel 17; by rotating the reversing post 2, the second accommodating chamber A1 is connected to the third flow channel 19 or the first ventilation channel 16; by rotating the reversing post 2, the second accommodating chamber A2 is connected to the fifth flow channel 21, the sixth flow channel 22 or the third ventilation channel 18. A card slot 12 is also provided at one end of the reversing post 2 body away from the connecting post 15, and the reversing post 2 is rotated by the card slot 12.

[0178] The detection tube 10 is a conical tube with a capacity of 900 ml.

[0179] The capacity of the second accommodating chamber A1 is 600 ml.

[0180] The capacity of the second accommodating chamber A2 is 600 ml.

[0181] The capacity of the second accommodating chamber A3 is 3500 ml.

[0182] The capacity of the second accommodating chamber A4 is 1500 ml.

[0183] The capacity of the second accommodating chamber A5 is 3500 ml.

[0184] The capacity of the second accommodating chamber A6 is 600 ml.

[0185] The capacity of the second accommodating chamber A7 is 4000 ml.

[0186] The capacity of the second accommodating chamber A8 is 600 ml.

[0187] The capacity of the second accommodating chamber B is 4500 ml.

[0188] The capacity of the second containing chamber C1 is 600 ml.

[0189] The capacity of the second containing chamber C2 is 600 ml.

[0190] The capacity of the second containing chamber C3 is 5000 ml.

[0191] The capacity of the second accommodating chamber C4 is 9500 ml.

[0192] The slide cover 1 is provided with two first through holes and one second through hole that penetrate the surface of the slide cover 1. The two first through holes and the second through hole are arranged in sequence along the extension direction of the slide cover 1. The two first through holes are arranged at equal intervals, and the interval is 15.5 mm. On the surface of the slide cover 1, a cylindrical channel 3 is arranged around the edge of the first through hole, and an open channel 4 is arranged around the edge of the second through hole. An elastic sealing ring is arranged around the inner wall of the cylindrical channel 3, and an opening is provided in the middle of the sealing ring. A channel cover for closing the open channel 4 is hinged at the top of the open channel 4. The height of the open channel 4 is 4.5 mm. The inner diameter is 12 mm. The height of the cylindrical channel 3 is 5.5 mm, and the inner diameter is 11 mm. The inner diameter of the first through hole is 8 mm, and the inner diameter of the second through hole is 11 mm.

[0193] An arc-shaped mounting plate 6 having a right-angled side perpendicular to the surface of the sliding cover 1 is arranged on the surface of the sliding cover 1 facing away from the chip body, the mounting plate 6 is located between the opening channel 4 and the cylindrical channel 3, and one end of the mounting plate 6 is connected to the outer wall of the cylindrical channel 3. A sliding groove 8 is arranged on the side of the sliding cover 1 close to the chip body, and a sliding sheet 9 matching with the sliding groove 8 is arranged on the side of the chip body close to the sliding cover 1, and an opening matching with the accommodating cavity is arranged on the sliding sheet 9.

[0194] When the chip of this embodiment is used, the open channel 4 is first moved to the top of the second accommodating chamber C4 by moving the slide cover 1, and the plasma sample is added to the second accommodating chamber C4 through the open channel 4; then the disposable syringe is moved to the top of the second accommodating chamber A8 by moving the slide cover 1, and the proteinase K in the syringe is transferred to the second accommodating chamber C4; the disposable syringe is moved to the top of the second accommodating chamber A7 by moving the slide cover 1, and the anhydrous ethanol in the syringe is transferred to the second accommodating chamber C4; the magnetic rod sleeve is made to perform mixing and shaking in the second accommodating chamber C4 by moving the slide cover 1, and after the mixing and shaking is completed, the magnetic rod sleeve is made to perform mixing and shaking in the second accommodating chamber C4 by moving the slide cover 1. The slide cover 1 transfers the disposable syringe to the top of the second accommodating chamber A7, and transfers the anhydrous ethanol to the second accommodating chamber C4; the slide cover 1 is moved to transfer the magnetic rod of the magnetic rod sleeve to the top of the second accommodating chamber C4, and the magnetic beads therein are transferred to the second accommodating chamber C3, and vibrates; the slide cover 1 is moved to transfer the disposable syringe to the top of the second accommodating chamber C1, and the eluent therein is transferred to the second accommodating chamber B; the slide cover 1 is moved to transfer the magnetic rod of the magnetic rod sleeve to the top of the second accommodating chamber C3, and the magnetic beads therein are transferred to the second accommodating chamber B, and vibrates; the slide cover 1 is moved to transfer the magnetic rod The magnetic rod of the sleeve is transferred to the top of the second accommodating chamber B, and the magnetic beads therein are transferred to the second accommodating chamber C3 and shaken; the disposable syringe is transferred to the top of the second accommodating chamber A6 by moving the sliding cover 1, and the sulfidation reagent therein is transferred to the second accommodating chamber B; the disposable syringe is transferred to the top of the second accommodating chamber A5 by moving the sliding cover 1, and the lysate and magnetic beads therein are shaken and mixed, and then transferred to the second accommodating chamber B; the disposable syringe is transferred to the top of the second accommodating chamber A7 by moving the sliding cover 1, and the anhydrous ethanol therein is transferred to the second accommodating chamber B; The cover 1 transfers the magnetic rod sleeve to the top of the second accommodating chamber B and oscillates therein; the magnetic rod of the magnetic rod sleeve is transferred to the top of the second accommodating chamber B by moving the sliding cover 1, and the magnetic beads therein are transferred to the second accommodating chamber C2 and oscillate; the magnetic rod of the magnetic rod sleeve is transferred to the top of the second accommodating chamber C2 by moving the sliding cover 1, and the magnetic beads therein are transferred to the second accommodating chamber C1 and oscillate; the magnetic rod of the magnetic rod sleeve is transferred to the top of the second accommodating chamber C1 by moving the sliding cover 1, and the magnetic beads therein are transferred to the second accommodating chamber C2, and the purified DNA sample remains in the second accommodating chamber C1;The second accommodating chamber A1 is connected to the third flow channel 19 by rotating the reversing column 2 of the first accommodating chamber 5, so that the paraffin oil in the second accommodating chamber A1 is transferred to the detection tube 10, and then the reversing column 2 of the first accommodating chamber 5 is rotated to connect the second accommodating chamber A2 with the fifth flow channel 21, so that the PCR reaction reagent in the second accommodating chamber A2 is transferred to the detection tube 10, and then the reversing column 2 of the first accommodating chamber 5 is rotated so that the second accommodating chamber C1 is connected to the fourth flow channel 20 of the reversing column 2, so that the purified DNA sample is transferred to the detection tube 10, and the purified DNA sample reacts with the paraffin oil and the PCR reaction reagent. The sliding cover can cooperate with the chip body to achieve sealing, and can slide slowly on the chip body. All reagents required for the experiment are pre-packaged in the chip; the same reagents are stored in the same hole and transferred through a disposable syringe 13 when used; the detection tube 10 for PCR amplification is independently designed and can be used in a conventional PCR instrument. It has a simple structure and is easy to operate, which greatly saves equipment costs. ;

[0195] Although the embodiments of the present application are described above in conjunction with the accompanying drawings, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can also make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the protection of the present application.

Claims

1. An integrated microfluidic chip, wherein: The chip comprises a chip body and a slide cover, wherein the slide cover is movably arranged on the top of the chip body and can move along the extension direction of the chip body. The chip body is provided with at least two accommodating cavities recessed into the chip body along its extension direction. The plurality of accommodating chambers include a first accommodating chamber and a second accommodating chamber; The first accommodating cavity comprises a first accommodating cavity body, a commutating column and a detection tube. The first accommodating cavity body and the chip body are integrally formed. The commutating column is sleeved in the first accommodating cavity body, and the commutating column can rotate relative to the first accommodating cavity body, so that the commutating column is connected with the second accommodating cavity. The detection tube is arranged at one end of the commutating column away from the chip body, and the detection tube is connected with the commutating column. The second containing cavity is used for storing reagents and / or for providing experimental space.

2. The chip according to claim 1, wherein: The reversing column is provided with a plurality of ventilation channels and a plurality of micro-channels. The ventilation air channel penetrates the reversing column along the radial direction of the reversing column; The microfluidic channel is in communication with the detection tube; The reversing column can be rotated to make the microchannel communicate with the second accommodating chamber, thereby making the second accommodating chamber communicate with the detection tube.

3. The chip according to claim 2, wherein: The commutation column comprises a commutation column body and a connecting column, wherein the connecting column is arranged at one end of the commutation column body away from the chip body, and the axis of the connecting column coincides with the axis of the commutation column body; The connecting column is sleeved in the detection tube.

4. The chip according to claim 3, wherein: The microfluidic channel includes a connecting channel and a guiding channel, wherein the connecting channel is used to introduce the reagent or sample in the second accommodating chamber into the reversing column body; one end of the connecting channel is used to communicate with the second accommodating chamber, and the other end thereof is connected with the guiding channel; The guide channel is used to communicate with the detection tube, and the guide channel is used to introduce the reagent or sample introduced through the connecting channel into the detection tube.

5. The chip according to claim 4, wherein: The flow guide channel includes a first flow channel and a second flow channel; the connecting channel includes a third flow channel, a fourth flow channel, a fifth flow channel and a sixth flow channel, The first flow channel is arranged along the axis of the reversing column body, and the second flow channel is parallel to the first flow channel; The third flow channel and the fourth flow channel are both arranged along the radial direction of the reversing column body, and are communicated with the first flow channel; The fifth flow channel and the sixth flow channel are both arranged along the radial direction of the reversing column body, and are communicated with the second flow channel; Along the axial direction of the reversing column body, the third flow channel, the fourth flow channel and the fifth flow channel are arranged at different heights; Along the axial direction of the reversing column body, the fifth flow channel and the sixth flow channel are arranged at the same height.

6. The chip according to claim 5, wherein: Along the axial direction of the reversing column body, from one end close to the chip body to one end close to the detection tube, the third flow channel, the fourth flow channel, and the fifth flow channel are arranged in sequence; The plane where the third flow channel is located is plane A, The projection of the fourth flow channel on the plane A forms an angle β with the third flow channel; The projection of the fourth flow channel on the plane A and the projection of the fifth flow channel on the plane A are on the same straight line; The fifth flow channel forms an angle α with the sixth flow channel.

7. The chip according to claim 5, wherein: The first flow channel takes one end of the third flow channel as a starting point and extends along the axis of the reversing column body until it passes through the connecting column; The second flow channel extends from one end of the fourth flow channel and the fifth flow channel to the side wall of the connecting column and penetrates the side wall of the connecting column.

8. The chip according to claim 6, wherein: The ventilation airway comprises a first ventilation airway, a second ventilation airway and a third ventilation airway, and along the axial direction of the reversing column body, the first ventilation airway, the second ventilation airway and the third ventilation airway are arranged at different heights; In the reversing column body, along the axial direction of the reversing column body, the first ventilation channel and the third flow channel are located at the same height; In the reversing column body, along the axial direction of the reversing column body, the second ventilation channel and the fourth flow channel are located at the same height; In the reversing column body, along the axial direction of the reversing column body, the third ventilation channel and the fifth flow channel are located at the same height.

9. The chip according to claim 8, wherein: The projection of the first ventilation airway on the plane A coincides with the projection of the second ventilation airway on the plane A; The projection of the first ventilation channel on the plane A and the projection of the third ventilation channel on the plane A form an angle θ.

10. The chip according to claim 8, wherein: The second accommodating chamber includes a second accommodating chamber A, a second accommodating chamber B, and a second accommodating chamber C. The second receiving chamber A is used to store reagents. The second accommodating chamber B is used to provide an experimental space for experiments; The second containing chamber C is used to store reagents and also provides experimental space for experiments.

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