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 which are recessed towards the interior of the chip main body along the extension direction of the chip main body, the accommodating cavities are mutually independent, and the sliding cover is provided with at least two first through holes which penetrate through the surface of the sliding cover. A cylindrical channel protruding out of the surface of the sliding cover is arranged on the surface of the side, away from the chip body, of the sliding cover, the cylindrical channel is located above the first through hole, and an elastic sealing ring is arranged in the cylindrical channel.

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 moves the sliding cover to connect the first through hole with different accommodating cavities so as to carry out experiments (such as DNA extraction, methylation purification and PCR detection). Since an elastic sealing ring is provided in the cylindrical channel, the accommodating cavity is not connected to the outside world during the experiment, thereby ensuring the sealing of the chip.

[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, and each accommodating cavity is independent of each other.

[0011] 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, and an elastic sealing ring is provided in the cylindrical channel.

[0012] 2. The chip according to item 1, wherein at least two of the first through holes are arranged in sequence along the extension direction of the sliding cover.

[0013] 3. The chip according to item 1, 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.

[0014] 4. The chip according to item 3, wherein a channel cover for closing the open channel is provided at the top of the open channel.

[0015] 5. The chip according to item 3, wherein the opening channel and the cylindrical channel are located on the center line of a surface of the sliding cover facing away from the chip body;

[0016] The height of the open channel is smaller than the height of the cylindrical channel.

[0017] 6. A chip according to item 3, wherein a mounting plate perpendicular to the sliding cover surface is provided on the side surface of the sliding cover away from the chip body, the mounting plate is located between the opening channel and the cylindrical channel, and one end of the mounting plate is connected to the outer wall of the cylindrical channel.

[0018] 7. The chip according to item 3, 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,

[0019] The slide is provided with an opening matched with the accommodating cavity.

[0020] 8. The chip according to item 3, wherein the plurality of accommodating cavities are divided into a first accommodating cavity, a second accommodating cavity and a third accommodating cavity,

[0021] The first accommodating cavity is close to the end of the chip body, the first accommodating cavity comprises a first accommodating cavity body and a detection tube detachably connected to the bottom thereof, and the first accommodating cavity body is in communication with the detection tube;

[0022] The opening size of the second accommodating cavity is smaller than the opening size of the third accommodating cavity.

[0023] 9. The chip according to item 8, wherein the second accommodating cavity comprises a second accommodating cavity A and a second accommodating cavity B, the second accommodating cavity A is used to store reagents, and the second accommodating cavity B is used to provide an experimental space for experiments;

[0024] The third accommodating chamber includes a third accommodating chamber A, a third accommodating chamber B and a third accommodating chamber C. The third accommodating chamber A is used to store reagents and also provides an experimental space for experiments; the third accommodating chamber B is used to provide an experimental space for experiments; and the third accommodating chamber C is used to store reagents or instruments.

[0025] 10. The chip according to item 8, wherein the number of the first accommodating cavities is at least 1, the number of the second accommodating cavities is at least 2, and the number of the third accommodating cavities is at least 3.

[0026] 11. The chip according to item 8, wherein the openings of the first accommodating cavity, the second accommodating cavity and the third accommodating cavity are respectively circular, elliptical or n-gonal, where n is greater than or equal to 3.

[0027] The integrated microfluidic chip provided in the present application, when in use, is constructed by moving the sliding cover so that the first through hole is connected to different accommodating chambers, thereby conducting experiments (such as DNA extraction, methylation purification, and PCR detection). Since an elastic sealing ring is provided in the cylindrical channel, the accommodating chamber is not connected to the outside world during the experiment, thereby ensuring the sealing of the chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Schematic diagram of the integrated microfluidic chip structure provided in this application.

[0029] Figure 2 This is a schematic diagram of the structure of the sliding cover provided in this application.

[0030] Description of Reference Numerals

[0031] 1-slide cover, 2-chip body, 3-cylindrical channel, 4-opening channel, 5-channel cover, 6-mounting plate, 7-first accommodating chamber, 8-slide groove, 9-slide sheet, 10-detection tube, 11-third accommodating chamber, 12-second accommodating chamber, 13-syringe, 14-magnetic rod sleeve. DETAILED DESCRIPTION

[0032] 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.

[0033] 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 2 and a slide cover 1, wherein the slide cover 1 is movably arranged on the top of the chip body 2, and when in use, the slide cover 1 moves along the extension direction of the chip body 2,

[0034] The chip body 2 is provided with at least two accommodating cavities recessed into the chip body 2 along its extension direction, and each accommodating cavity is independent of each other and not connected to each other.

[0035] At least two first through holes penetrating the surface of the slide cover 1 are arranged on the slide cover 1, and a cylindrical channel 3 protruding from the surface of the slide cover 1 is arranged on the side surface of the slide cover 1 away from the chip body 2, and the cylindrical channel 3 is located above the first through holes, and an elastic sealing ring is arranged in the cylindrical channel 3. When in use, the first through hole is connected to the accommodating cavity.

[0036] Herein, the extension direction of the chip body 2 is the long side direction of the chip, and the extension direction of the chip body 2 is perpendicular to the recessed direction of the accommodating cavity. The extension direction of the chip body 2 is consistent with the extension direction of the sliding cover 1.

[0037] 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.

[0038] 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 a disposable syringe or a magnetic rod sleeve to seal or experiment the chip body. The inner diameters of the multiple 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 is smaller than the inner diameter of the first through hole used to match the magnetic rod sleeve.

[0039] 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.

[0040] 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.

[0041] The inner diameter of the cylindrical channel 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 disposable syringes and magnetic rod sleeves.

[0042] In some embodiments, the slide cover 1 is provided with three first through holes penetrating the surface of the slide cover 1, and the three 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.

[0043] 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.

[0044] 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 2, 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 chip body 2 is not connected to the external environment.

[0045] When the chip is used for DNA extraction and amplification, among the multiple accommodating cavities, some of the accommodating cavities store experimental reagents, some of the accommodating cavities provide operating space for DNA extraction, and some of the accommodating cavities provide space for DNA amplification.

[0046] 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 2, and the opening channel 4 is located directly above the second through hole.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] Furthermore, a channel cover 5 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 5 are connected in a hinged manner.

[0051] 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 2 .

[0052] The height of the opening channel 4 is smaller than the height of the cylindrical channel 3 .

[0053] In the present application, a mounting plate 6 perpendicular to the surface of the sliding cover 1 is provided on the side surface of the sliding cover 1 away from the chip body 2, 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.

[0054] 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.

[0055] 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 .

[0056] In the present application, the slide cover 1 has a slide groove 8 on one side close to the chip body 2, and the chip body 2 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.

[0057] 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.

[0058] The chip body 2 is composed of a slide 9 and a housing cavity body, wherein the slide 9 is located at the top of the housing cavity body, and the slide 9 and the housing cavity body are integrally formed, the width of the slide 9 is greater than the width of the housing cavity body, and the opening on the slide 9 is connected to the housing cavity body.

[0059] 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.

[0060] In the present application, the plurality of accommodating chambers are divided into a first accommodating chamber 7, a second accommodating chamber 12 and a third accommodating chamber 11, wherein the first accommodating chamber 7 is close to the end of the chip body 2, and the first accommodating chamber 7 includes a first accommodating chamber body and a detection tube 10 detachably connected to the bottom thereof, and the first accommodating chamber body is in communication with the detection tube 10. The purpose of designing the first accommodating chamber 7 in this way is to ensure that the tip of the disposable pipette and the internal reagent are in a sealed state during the process of transferring the liquid to the detection tube 10.

[0061] The opening size of the second accommodating cavity 12 is smaller than the opening size of the third accommodating cavity 11 .

[0062] Opening size in this context refers to the diameter or equivalent diameter.

[0063] The capacity of the first accommodating chamber 7 is 1000ml-5000ml, for example, it can be 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, 3500ml, 4000ml, 4500ml, 5000ml, etc.

[0064] The capacity of the second containing chamber 12 is 100ml-3000ml, for example, it can be 100ml, 300ml, 500ml, 800ml, 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, etc.

[0065] The capacity of the third containing chamber 11 is 100ml-15000ml, for example, it can be 100ml, 300ml, 500ml, 800ml, 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, 4000ml, 5000ml, 8000ml, 10000ml, 12000ml, 15000ml, etc.

[0066] The number of the first accommodating cavities 7 is at least one, the number of the second accommodating cavities 12 is at least two, and the number of the third accommodating cavities 11 is at least three.

[0067] The number of the first accommodating cavities 7 can be 1, 2, 3, 4, 5, 6, 7, 8, etc., the number of the second accommodating cavities 12 can be 2, 3, 4, 5, 6, 7, 8, etc., and the number of the third accommodating cavities 11 can be 3, 4, 5, 6, 7, 8, etc. The specific data can be determined according to actual needs.

[0068] Furthermore, the second accommodating chamber includes a second accommodating chamber A and a second accommodating chamber B, the second accommodating chamber A is used to store reagents, and the second accommodating chamber B is used to provide an experimental space for experiments;

[0069] The third accommodating chamber includes a third accommodating chamber A, a third accommodating chamber B and a third accommodating chamber C. The third accommodating chamber A is used to store reagents and also provides an experimental space for experiments; the third accommodating chamber B is used to provide an experimental space for experiments; and the third accommodating chamber C is used to store reagents or instruments.

[0070] In some embodiments, when the chip is used for DNA extraction, methylation purification and PCR detection, the second accommodating chamber includes a second accommodating chamber A and a second accommodating chamber B, wherein the second accommodating chamber A is used to store reagents required in the DNA extraction process or amplification process, and the second accommodating chamber B is used to provide an experimental space for the DNA extraction or amplification process. The third accommodating chamber includes a third accommodating chamber A, a third accommodating chamber B and a third accommodating chamber C, wherein the third accommodating chamber A is used to store reagents required in the DNA extraction process and also provides an experimental space for the DNA extraction or amplification; the third accommodating chamber B is used for sulfite conversion of DNA; and the third accommodating chamber C is used to store reagents or magnetic beads in the DNA extraction process.

[0071] In some embodiments, the number of the first accommodating chamber 7 is 1, the number of the second accommodating chamber 12 can be 5, which are respectively the second accommodating chamber A1, the second accommodating chamber B2, the second accommodating chamber A3, the second accommodating chamber A4, and the second accommodating chamber B5, and the number of the third accommodating chamber 11 can be 8, which are respectively the third accommodating chamber A1, the third accommodating chamber A2, the third accommodating chamber B3, the third accommodating chamber A4, the third accommodating chamber A5, the third accommodating chamber C6, the third accommodating chamber C7, and the third accommodating chamber C8. From one end of the chip body 2, the first accommodating chamber 7, the second accommodating chamber A1, the second accommodating chamber B2, the second accommodating chamber A3, the third accommodating chamber A1, the second accommodating chamber A4, the second accommodating chamber B5, the third accommodating chamber A2, the third accommodating chamber B3, the third accommodating chamber A4, the third accommodating chamber A5, the third accommodating chamber C6, the third accommodating chamber C7, and the third accommodating chamber C8 are arranged in order.

[0072] The first containing chamber 7 is used to store PCR reaction reagents and perform PCR reactions.

[0073] The second accommodating chambers A1, A3, A4 are used to store reagents required in the DNA extraction process or amplification process, such as paraffin oil, sulfiding reagents and eluents.

[0074] The second accommodating chambers B2 and B5 are used to provide space for the DNA extraction or amplification process, and can also be used to place the syringe 13 used for injection.

[0075] The third accommodating chambers A1, A2, A4, and A5 are used to store reagents required in the DNA extraction process, such as elution solution, rinse solution, and lysis solution + magnetic beads, and provide experimental space for DNA extraction or amplification. At the same time, the third accommodating chamber A1 can also be used to place the magnetic rod sleeve 14.

[0076] The third containing chamber B3 is used for sulfite conversion and elution of DNA.

[0077] The third accommodating chambers C6, C7 and C8 are used to store lysis solution + magnetic beads, binding solution and washing solution in the DNA extraction process.

[0078] The capacity of the first accommodating cavity body is 500ml-4000ml, and the detection tube is a conical tube with a capacity of 500ml-1000ml.

[0079] The capacity of the second accommodating chamber A1 is 500ml-1500ml.

[0080] The capacity of the second accommodating chamber B2 is 800ml-2000ml.

[0081] The capacity of the second accommodating chamber A3 is 600ml-1800ml.

[0082] The capacity of the second accommodating chamber A4 is 100ml-500ml.

[0083] The capacity of the second accommodating chamber B5 is 800ml-2000ml.

[0084] The capacity of the third containing chamber A1 is 3000ml-8000ml.

[0085] The capacity of the third accommodating chamber A2 is 3000ml-8000ml.

[0086] The capacity of the third accommodating chamber B3 is 2000ml-6000ml.

[0087] The capacity of the third accommodating chamber A4 is 3000ml-8000ml.

[0088] The capacity of the third accommodating chamber A5 is 5000ml-15000ml.

[0089] The capacity of the third containing chamber C6 is 1000ml-5000ml.

[0090] The capacity of the third containing chamber C7 is 3000ml-8000ml.

[0091] The capacity of the third containing chamber C8 is 100ml-500ml.

[0092] The openings of the first accommodating cavity 7, the second accommodating cavity and the third accommodating cavity 11 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.

[0093] In some embodiments, the opening of the first accommodating cavity 7 is circular, the opening of the second accommodating cavity 12 is circular, and the opening of the third accommodating cavity 11 is circular or rectangular.

[0094] Example

[0095] 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.

[0096] Example 1

[0097] The integrated microfluidic chip of this embodiment is used for DNA extraction, methylation purification and PCR detection.

[0098] The integrated microfluidic chip comprises a chip body 2 and a slide cover 1, wherein the slide cover 1 is movably arranged on the top of the chip body 2.

[0099] The chip body 2 is provided with a first accommodating cavity 7, a second accommodating cavity 12 and a third accommodating cavity 11 recessed into the interior of the chip body 2 along its extension direction, and each accommodating cavity is independent of each other and not connected with each other. The number of the first accommodating cavity 7 is 1, the number of the second accommodating cavity 12 can be 5, which are respectively the second accommodating cavity A1, the second accommodating cavity B2, the second accommodating cavity A3, the second accommodating cavity A4, and the second accommodating cavity B5; the number of the third accommodating cavity 11 can be 8, which are respectively the third accommodating cavity A1, the third accommodating cavity A2, the third accommodating cavity B3, the third accommodating cavity A4, the third accommodating cavity A5, the third accommodating cavity C6, the third accommodating cavity C7, and the third accommodating cavity C8. From one end of the chip body 2, there are the first accommodating cavity 7, the second accommodating cavity A1, the second accommodating cavity B2, the second accommodating cavity A3, the third accommodating cavity A1, the second accommodating cavity A4, the second accommodating cavity B5, the third accommodating cavity A2, the third accommodating cavity B3, the third accommodating cavity A4, the third accommodating cavity A5, the third accommodating cavity C6, the third accommodating cavity C7, and the third accommodating cavity C8. The opening of the first accommodating cavity 7 is circular, the opening of the second accommodating cavity is circular, the opening of the third accommodating cavity A1 and the third accommodating cavity C8 are circular, the opening of the third accommodating cavity A2, the third accommodating cavity B3, the third accommodating cavity A4, the third accommodating cavity A5, the third accommodating cavity C6, and the third accommodating cavity C7 are rectangular, and the first accommodating cavity 7 is composed of the main body of the first accommodating cavity 7 and the detection tube 10 detachably connected to the bottom thereof, and the main body of the first accommodating cavity 7 is connected to the detection tube 10.

[0100] The capacity of the first accommodating chamber body is 2000 ml, and the detection tube is a conical tube with a capacity of 900 ml.

[0101] The capacity of the second accommodating chamber A1 is 950 ml.

[0102] The capacity of the second accommodating chamber B2 is 1500 ml.

[0103] The capacity of the second accommodating chamber A3 is 950 ml.

[0104] The capacity of the second accommodating chamber A4 is 400 ml.

[0105] The capacity of the second accommodating chamber B5 is 1500 ml.

[0106] The capacity of the third containing chamber A1 is 5000 ml.

[0107] The capacity of the third containing chamber A2 is 5000 ml.

[0108] The capacity of the third accommodating chamber B3 is 3500 ml.

[0109] The capacity of the third accommodating chamber A4 is 5000 ml.

[0110] The capacity of the third accommodating chamber A5 is 9000 ml.

[0111] The capacity of the third containing chamber C6 is 2000 ml.

[0112] The capacity of the third containing chamber C7 is 5000 ml.

[0113] The capacity of the third containing chamber C8 is 300 ml.

[0114] The slide cover 1 is provided with three first through holes and one second through hole that penetrate the surface of the slide cover 1. The three first through holes and the second through hole are arranged in sequence along the extension direction of the slide cover 1. The three 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 5 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.

[0115] An arc-shaped mounting plate 6 having a right-angled side perpendicular to the surface of the slide cover 1 is arranged on the surface of the slide cover 1 facing away from the chip body 2. 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 slide groove 8 is arranged on the side of the slide cover 1 close to the chip body 2, and a slide sheet 9 matching with the slide groove 8 is arranged on the side of the chip body 2 close to the slide cover 1, and an opening matching with the accommodating cavity is arranged on the slide sheet 9.

[0116] When the chip of this embodiment is used, the open channel 4 is first moved to the top of the third accommodating chamber b5 by moving the sliding cover 1, and the plasma sample is added into the third accommodating chamber b5 through the opening channel 4; then the sliding cover 1 is moved so that the accommodating chamber in which the syringe 13 is placed is moved to the top of the accommodating chamber in which the reagents required for the DNA lysis process are stored, the reagents are aspirated, and then the syringe 13 is moved to the top of the third accommodating chamber b5 through a certain sliding cover 1, so that the reagents required for the DNA lysis process are transferred into the third accommodating chamber b5; then the accommodating chamber in which the magnetic rod sleeve 14 is placed is moved to the top of the third accommodating chamber b5, and the magnetic rod sleeve 14 is used to perform mixing and shaking actions in the third accommodating chamber b5, so that the DNA is lysed from the plasma; after the lysis process is completed, the opening channel 4 is added into the third accommodating chamber b5 through the opening channel 4; then the opening channel 4 is added into the third accommodating chamber b5 through the sliding cover 1, and the opening channel 4 is added into the third accommodating chamber b5 through the opening channel 4; then the opening channel 4 is added into the third accommodating chamber b5 through the opening channel 4; The movable slide 1 transfers the magnetic beads to the third accommodating chamber b4 and performs elution by shaking; after completion, the magnetic beads are transferred to the third accommodating chamber b3 by moving the slide 1, and the reagents required for sulfite conversion are transferred to the third accommodating chamber b3, and sulfite conversion is performed in the third accommodating chamber b3, and elution by shaking is performed. After completion, the magnetic beads are transferred to the third accommodating chamber b2 by moving the slide 1 and through the magnetic rod sleeve 14 for washing. After washing is completed, the magnetic beads are transferred to the third accommodating chamber b1 by moving the slide 1 and perform elution by shaking. After elution is completed, the magnetic beads are transferred to the third accommodating chamber b2 by moving the slide 1, and the eluent is transferred to the first accommodating chamber 7, and then the paraffin oil in the second accommodating chamber a1 is transferred to the first accommodating chamber 7 by moving the slide 1 for PCR reaction. Therefore, the chip of the present application uses two disposable syringes 13 to transfer liquid; uses a magnetic rod sleeve to shake and mix the liquid and elute the magnetic beads; all reagents required for the experiment are pre-packaged in the detection box; the same reagents are stored in the same hole and transferred through the disposable syringe 13 when used; the detection tube 10 for PCR amplification is independently designed and can also be used in a conventional PCR instrument, with a simple structure and easy operation, which greatly saves equipment costs.

[0117] 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, and each accommodating cavity is independent of each other. 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, and an elastic sealing ring is provided in the cylindrical channel.

2. The chip according to claim 1, wherein: At least two of the first through holes are arranged in sequence along the extending direction of the sliding cover.

3. The chip according to claim 1, wherein: A second through hole is arranged on the slide cover, and an opening channel protruding from the surface of the slide cover is arranged on a side surface of the slide cover away from the chip body, and the opening channel is located above the second through hole.

4. The chip according to claim 3, wherein: A channel cover for closing the open channel is arranged on the top of the open channel.

5. The chip according to claim 3, wherein: The opening channel and the cylindrical channel are located on the midline of a surface of the sliding cover on a side away from the chip body; The height of the open channel is smaller than the height of the cylindrical channel.

6. The chip according to claim 3, wherein: A mounting plate perpendicular to the surface of the slide cover is arranged on a side surface of the slide cover away from the chip body. The mounting plate is located between the opening channel and the cylindrical channel, and one end of the mounting plate is connected to the outer wall of the cylindrical channel.

7. The chip according to claim 3, wherein: The sliding cover has a sliding groove on one side close to the chip body, and the chip body has a sliding sheet matched with the sliding groove on one side close to the sliding cover. The slide is provided with an opening matched with the accommodating cavity.

8. The chip according to claim 3, wherein: The plurality of accommodating chambers are divided into a first accommodating chamber, a second accommodating chamber and a third accommodating chamber, The first accommodating cavity is close to the end of the chip body, the first accommodating cavity comprises a first accommodating cavity body and a detection tube detachably connected to the bottom thereof, and the first accommodating cavity body is in communication with the detection tube; The opening size of the second accommodating cavity is smaller than the opening size of the third accommodating cavity.

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

10. The chip according to claim 8, wherein: The number of the first accommodating cavities is at least 1, the number of the second accommodating cavities is at least 2, and the number of the third accommodating cavities is at least 3; or The openings of the first accommodating cavity, the second accommodating cavity and the third accommodating cavity are respectively circular, elliptical or n-sided, where n is greater than or equal to 3.

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