Detection chip and its operation method, detection device

Abstract

A detection chip, an operation method thereof, and a detection device, the detection chip comprising: a first substrate, comprising at least one storage cavity; and a first flexible layer, attached to a first side of the first substrate. The storage cavity has a first opening on the first side, and the first flexible layer is at the first opening of the at least one storage cavity and is configured to be in an open state or a closed state. The first side of the first substrate comprises a first bonding area and a second bonding area. When the first flexible layer is in a closed state, the first flexible layer is attached to the first side of the first substrate in the first bonding area and the second bonding area to at least close the first opening of the storage cavity. When the first flexible layer is in an open state, the first flexible layer is attached to the first side of the first substrate in the second bonding area and is separated from the first substrate in the first bonding area to open the first opening of the storage cavity.

Description

Technical Field

[0001] Embodiments of the present disclosure relate to a detection chip and a detection device that can be used with the detection chip, and an operating method of the detection chip. Background Art

[0002] Microfluidic chip technology integrates the basic operating units such as sample preparation, reaction, separation, and detection involved in the fields of biology, chemistry, and medicine onto a chip with micron-scale microchannels, automatically completing the entire process of reaction and analysis. The chip used in this process is called a microfluidic chip, also known as a chip laboratory (Lab-on-a-chip). Microfluidic chip technology has the advantages of small sample usage, fast analysis speed, easy to make portable instruments, and suitable for instant and on-site analysis. It has been widely used in many fields such as biology, chemistry, and medicine. Summary of the invention

[0003] According to at least one embodiment of the present disclosure, a detection chip is provided, which includes: a first substrate, including at least one storage cavity; and a first flexible layer, attached to a first side of the first substrate. The storage cavity has a first opening on the first side, and the first flexible layer is at the first opening of the at least one storage cavity and is configured to be in an open state or a closed state. The first side of the first substrate includes a first bonding area and a second bonding area. When the first flexible layer is in the closed state, the first flexible layer is attached to the first side of the first substrate in the first bonding area and the second bonding area to close the first opening of the storage cavity. When the first flexible layer is in the open state, the first flexible layer is attached to the first side of the first substrate in the second bonding area and is separated from the first substrate in the first bonding area to open the first opening of the storage cavity.

[0004] For example, in some embodiments, a first attachment strength of the first flexible layer to the first substrate at the first bonding area is less than a second attachment strength of the first flexible layer to the first substrate at the second bonding area.

[0005] For example, in some embodiments, the surface of the first substrate on the first side has a plurality of concave-convex structures at the first bonding area.

[0006] For example, in some embodiments, the maximum depth difference of the plurality of concave-convex structures in a direction perpendicular to the surface is in the range of 0.1 mm to 3 mm.

[0007] For example, in some embodiments, the first attachment strength is in the range of 1N-50N, and the second attachment strength is in the range of 50N-500N.

[0008] For example, in some embodiments, the surface of the first substrate on the first side is flush at the first bonding area and the second bonding area.

[0009] For example, in some embodiments, the first substrate is provided with a buffer groove and a substrate groove communicating with the buffer groove for forming at least one microfluidic channel on the surface of the first side. The buffer groove is at least partially arranged around the first opening of the storage cavity to define the first bonding area on the inner side of the buffer groove, and define the second bonding area on the outer side of the buffer groove.

[0010] For example, in some embodiments, the depth of the buffer groove in the vertical direction perpendicular to the surface is in the range of 0.01 mm-5 mm.

[0011] For example, in some embodiments, the width of the buffer groove in the horizontal direction parallel to the surface is in the range of 0.01 mm-5 mm.

[0012] For example, in some embodiments, the bottom surface of the buffer tank includes a hydrophobic surface.

[0013] For example, in some embodiments, the first substrate is an annular boss at the first bonding area, and a width of the annular boss is in the range of 0.01 mm-5 mm.

[0014] For example, in some embodiments, a cross-section of the annular boss decreases toward the first side.

[0015] For example, in some embodiments, the annular boss includes a plurality of sub-annular bosses.

[0016] For example, in some embodiments, the at least one storage cavity includes a plurality of storage cavities. Also, the surface of the first substrate on the first side includes a plurality of substrate grooves for forming a plurality of microfluidic channels.

[0017] For example, in some embodiments, at least some of the plurality of microfluidic channels are provided with switch valves for controlling the connection and disconnection of the corresponding at least some of the microfluidic channels.

[0018] For example, in some embodiments, the detection chip further includes an intermediate layer, and the first flexible layer is adhered to the first side of the first substrate through the intermediate layer. The intermediate layer includes a plurality of intermediate through grooves that penetrate the intermediate layer, and the positions of the plurality of intermediate through grooves correspond to the positions of the plurality of substrate grooves to form the plurality of microchannels. The switch valve includes a disconnection area that is arranged in the substrate groove to disconnect the substrate groove into two sub-substrate grooves and a valve action area that is arranged in the intermediate through groove, and the projection of the valve action area in the direction of the surface of the first substrate covers the disconnection area of ​​the substrate groove and a part of the two sub-substrate grooves, so that the two sub-substrate grooves can be connected through the valve action area.

[0019] For example, in some embodiments, the plurality of storage chambers include at least one liquid storage chamber, wherein the at least one liquid storage chamber is pre-stored with a reagent, and the reagent is sealed in the liquid storage chamber.

[0020] For example, in some embodiments, the plurality of storage cavities include at least one liquid storage cavity. The detection chip further includes a second flexible layer, the second flexible layer is attached to a second side of the first substrate opposite to the first side at the at least one liquid storage cavity, the liquid storage cavity has a second opening on the second side, and the second flexible layer at least closes the second opening of the liquid storage cavity.

[0021] For example, in some embodiments, the first substrate further includes a mixing chamber system, and the mixing chamber system includes a first mixing chamber and a second mixing chamber. The at least one liquid storage chamber includes an eluent chamber pre-stored with an eluent, a first cleaning chamber pre-stored with a first cleaning liquid, and a lysate chamber pre-stored with a lysate. The multiple microchannels include a first microchannel connecting the mixing chamber system and the lysate chamber, a second microchannel connecting the mixing chamber system and the first cleaning chamber, and a third microchannel connecting the mixing chamber system and the eluent chamber.

[0022] For example, in some embodiments, the diameter of the elution liquid chamber is smaller than the diameters of the lysis liquid chamber and the first cleaning liquid chamber.

[0023] For example, in some embodiments, the at least one liquid storage chamber further includes a second cleaning liquid chamber pre-stored with a second cleaning liquid and a third cleaning liquid chamber pre-stored with a third cleaning liquid. The plurality of microfluidics further include a fourth microfluidic channel connecting the mixing chamber system and the third cleaning liquid chamber, and a fifth microfluidic channel connecting the mixing chamber system and the second cleaning liquid chamber.

[0024] For example, in some embodiments, the first mixing chamber and the second mixing chamber are disposed in the middle of the first substrate, and the at least one liquid storage chamber is disposed on both sides of the middle.

[0025] For example, in some embodiments, the heights of the first mixing chamber and the second mixing chamber are higher than the heights of the lysis liquid chamber, the first cleaning liquid chamber, the second cleaning liquid chamber, and the third cleaning liquid chamber.

[0026] For example, in some embodiments, the first microfluidic channel connects the first mixing chamber and the lysis liquid chamber, the second microfluidic channel connects the first mixing chamber and the first cleaning liquid chamber, the third microfluidic channel connects the second mixing chamber and the elution liquid chamber, the fourth microfluidic channel connects the first mixing chamber and the third cleaning liquid chamber, and the fifth microfluidic channel connects the fourth microfluidic channel and the second cleaning liquid chamber.

[0027] For example, in some embodiments, the first substrate further includes a waste liquid chamber, the waste liquid chamber includes two waste liquid chamber openings on the first side and the second side, respectively, the two waste liquid chamber openings are covered by the first flexible layer and the second flexible layer, respectively, and the plurality of microchannels further include a sixth microchannel connecting the first mixing chamber and the waste liquid chamber. A waste liquid chamber through hole is provided in the second flexible layer. The detection chip further includes a first gas isolation membrane attached to the second flexible layer at the waste liquid chamber through hole, the first gas isolation membrane enables the waste liquid chamber to be connected and isolated from the external gas of the detection chip through the first gas isolation membrane and the waste liquid chamber through hole.

[0028] For example, in some embodiments, the detection chip further includes a second substrate, which includes an amplification chamber configured to store an amplification reagent. One end of the amplification chamber is closed, and the other end has an amplification chamber opening and is attached to the side of the first flexible layer opposite to the first substrate. The first substrate also includes a first mixing chamber and a gas-permeable chamber that is connected to the external gas of the detection chip but isolated from the liquid. The multiple microfluidic channels include a seventh flow channel and an eighth microfluidic channel, the amplification chamber is connected to the first mixing chamber through a first flow channel outlet formed in the first flexible layer and the seventh microfluidic channel, and the amplification chamber is connected to the gas-permeable chamber through a second flow channel outlet formed in the first flexible layer and the eighth microfluidic channel.

[0029] For example, in some embodiments, the second substrate is a one-piece piece.

[0030] For example, in some embodiments, the amplification chamber opening is in the shape of an elongated strip, and the width of the opening is in the range of 0.1 mm-3 mm.

[0031] For example, in some embodiments, the amplification chamber is in an elongated strip shape, and the first flow channel outlet and the second flow channel outlet are respectively disposed at two ends of an opening of the amplification chamber.

[0032] For example, in some embodiments, the second substrate is made of a material that is transparent to the detection light used for the detection chip, and the wall thickness of the amplification chamber is in the range of 0.1 mm-3 mm.

[0033] For example, in some embodiments, one end of the gas permeable cavity is covered by the first flexible layer. The detection chip further includes a second gas isolation membrane, which is attached to the second side of the first substrate to cover the other end of the gas permeable cavity, and the gas permeable cavity is connected and isolated from the external gas of the detection chip through the second gas isolation membrane.

[0034] For example, in some embodiments, both ends of the gas permeable cavity are covered by the first flexible layer and the second flexible layer, respectively. A gas permeable cavity through hole is provided in the second flexible layer. The detection chip further includes a second gas isolation membrane attached to the second flexible layer at the gas permeable cavity through hole, and the gas permeable cavity is connected and isolated from the external gas of the detection chip through the second gas isolation membrane and the gas permeable cavity through hole.

[0035] For example, in some embodiments, the first substrate includes a first positioning structure, the second substrate includes a second positioning structure, and the positions of the first positioning structure and the second positioning structure correspond to each other so as to position the second substrate relative to the first substrate.

[0036] For example, in some embodiments, the first substrate further includes a first mixing chamber and a second mixing chamber. The second mixing chamber includes two first mixing chamber openings respectively on the first side of the first substrate and the second side of the first substrate opposite to the first side, and the two first mixing chamber openings are respectively covered by the first flexible layer and the first cover. The first mixing chamber includes two second mixing chamber openings respectively on the first side of the first substrate and the second side of the first substrate, and the two second mixing chamber openings are respectively covered by the first flexible layer and the second cover. The plurality of microfluidics include a ninth microfluidic channel, and the ninth microfluidic channel connects the first mixing chamber and the second mixing chamber. The first substrate further includes a first air permeable groove surrounding the first mixing chamber and a second air permeable groove surrounding the second mixing chamber. The first air permeable groove includes a first outer wall and a first inner wall between the first mixing chamber and the first air permeable groove. The second air permeable groove includes a second outer wall and a second inner wall between the first mixing chamber and the second air permeable groove.

[0037] For example, in some embodiments, the first cover member includes a first cover flexible portion, a first sealing groove surrounding the first cover flexible portion, a first protrusion surrounding the first sealing groove, and a first supporting edge surrounding the first protrusion. The bottom surface of the first sealing groove abuts against the end surface of the first inner wall, the first protrusion is inserted into the first breathable groove, the first supporting edge is supported by the first outer wall, and the first protrusion is further provided with a first breathable channel facing the first inner wall and extending into the first sealing groove.

[0038] For example, in some embodiments, the detection chip further includes an adhesive ring. The adhesive ring is disposed between the end surface of the first supporting edge and the end surface of the first outer wall to bond the first supporting edge and the first outer wall. The adhesive ring is also disposed between the end surface of the second supporting edge and the end surface of the second outer wall to bond the second supporting edge and the second outer wall.

[0039] For example, in some embodiments, the first cover and the second cover form an integrated mixing cover, the first air permeable groove and the second air permeable groove are connected through a connecting air permeable groove, and the mixing cover is provided with a mixing cavity through hole at the connecting air permeable groove. The detection chip further includes a third gas isolation membrane attached to the mixing cover at the mixing cavity through hole, and the connecting air permeable groove is connected and isolated from the external gas of the detection chip through the third gas isolation membrane and the mixing cavity through hole.

[0040] For example, in some embodiments, the storage cavity includes a sample cavity. The sample cavity includes two sample cavity openings, one on the first side of the first substrate and one on the second side of the first substrate opposite to the first side, respectively, the two sample cavity openings are covered by the first flexible layer and the third cover, respectively, the plurality of microfluidic channels include a tenth microfluidic channel, the tenth microfluidic channel connects the first mixing cavity and the sample cavity. The first substrate also includes a third air permeable groove surrounding the sample cavity, the third air permeable groove includes a third outer wall and a third inner wall between the sample cavity and the third air permeable groove.

[0041] For example, in some embodiments, the third cover member includes a third cover flexible portion, a third sealing groove surrounding the third cover flexible portion, a third protrusion surrounding the third sealing groove, and a third supporting edge surrounding the third protrusion. The bottom surface of the third sealing groove abuts against the end surface of the third inner wall, the third protrusion is inserted into the third breathable groove, the third supporting edge is supported by the third outer wall, and the third protrusion is further provided with a third breathable channel facing the third inner wall and extending to the third sealing groove.

[0042] For example, in some embodiments, the detection chip further includes a first chip positioning structure configured to position and mount the detection chip to a detection device.

[0043] At least one embodiment of the present disclosure provides a detection device, comprising a second chip positioning structure and a first operating part. The second chip positioning structure is configured to install the detection chip as described above, and the first operating part is configured to work to change the volume of the storage cavity.

[0044] At least one embodiment of the present disclosure further provides a detection device, comprising a second chip positioning structure and a second operating part. The second chip positioning structure is configured to install the detection chip as described above. The second operating part is configured to be operable to press the second flexible layer to operate the switch valve.

[0045] At least one embodiment of the present disclosure also provides an operating method of a detection chip, comprising: adding a sample to the detection chip; mixing the sample with a lysis solution to form a mixed solution, wherein the lysis solution can lyse the sample to form a substance to be detected; driving the mixed solution to flow to a mixing chamber system, wherein the mixing chamber system includes a first mixing chamber and a second mixing chamber, and the mixing chamber system has magnetic beads, and the magnetic beads can adsorb the substance to be detected; driving the mixed solution to move back and forth between the first mixing chamber and the second mixing chamber to mix the mixed solution and the magnetic beads; separating and processing the magnetic beads to make the substance to be detected fall off the magnetic beads; and detecting the substance to be detected. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solutions of the embodiments of the present disclosure, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present disclosure and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0047] Figure 1 A perspective view of a detection chip according to at least one embodiment of the present disclosure is shown;

[0048] Figure 2 Shows Figure 1 An exploded perspective view of the detection chip;

[0049] Figure 3A Shows Figure 1 A top view of the first substrate of the detection chip;

[0050] Figure 3B Shows Figure 1 A bottom view of the first substrate of the detection chip;

[0051] Figure 4 Shows Figure 1 A top view of the middle layer of the detection chip;

[0052] Figure 5 Shows Figure 1 A plan view of the first flexible layer of the detection chip;

[0053] Fig. 6A Shows Figure 1 A plan view of a first portion of a second flexible layer of a detection chip;

[0054] Figure 6B Shows Figure 1 A plan view of a second portion of a second flexible layer of the detection chip;

[0055] Figure 7 Shows Figure 1 Bottom view of the detection chip;

[0056] Figure 8 shows a perspective view of a second substrate according to an embodiment of the present disclosure;

[0057] Fig. 9A A perspective view of a detection chip at a storage cavity according to an embodiment of the present disclosure is shown;

[0058] Fig. 9B Shows Fig. 9A A partial enlarged view of

[0059] Fig.10 A cross-sectional view of a detection chip at a storage cavity according to an embodiment of the present disclosure is shown, wherein the first flexible layer is in a closed state;

[0060] Fig.11 Another cross-sectional view of the detection chip at a storage cavity according to an embodiment of the present disclosure is shown, wherein the first flexible layer is in an open state;

[0061] Fig.12 A perspective view showing a first substrate at a storage cavity according to an embodiment of the present disclosure;

[0062] Fig.13 A longitudinal cross-sectional view of a detection chip at a storage cavity according to another embodiment of the present disclosure is shown, wherein the first flexible layer is in a closed state;

[0063] Fig.14 Another longitudinal cross-sectional view of the storage cavity in the figure is shown, wherein the first flexible layer is in an open state;

[0064] Fig.15 A transverse cross-sectional view at the storage cavity in the figure is shown;

[0065] Figures 16A-16D A schematic diagram showing a concave-convex structure according to an embodiment of the present disclosure is shown;

[0066] Fig.17 A schematic perspective view showing a portion of a switching valve according to an embodiment of the present disclosure;

[0067] Fig.18 Shows Fig.17 A perspective view of a base groove of a first base;

[0068] Figure 19-22 A cross-sectional view of a switch valve according to an embodiment of the present disclosure is shown, wherein: Fig.19 This is an exploded sectional view of the switch valve. Fig. 20 This is a cross-sectional view of the switch valve in a normally open state. Fig.21 This is a cross-sectional view of the switch valve in the fully open state. Fig. 22 is a cross-sectional view of the switch valve in a closed state;

[0069] Fig.23 A perspective view of a detection chip at a sample cavity according to an embodiment of the present disclosure is shown;

[0070] Fig.24 Shows Fig.23 Another perspective view of the detection chip at the sample chamber, wherein the third cover is removed;

[0071] Fig.25 Shows Fig.23 A cross-sectional perspective view of the detection chip at the sample cavity;

[0072] Fig.26 shows a perspective view of a third cover according to an embodiment of the present disclosure;

[0073] Fig. 27 A perspective view of a detection chip at a mixing system according to an embodiment of the present disclosure is shown;

[0074] Fig.28 Shows Fig. 27 Another perspective view of the detection chip at the mixing system, wherein the mixing cover is removed;

[0075] Fig.29 Shows Fig. 27 Another perspective view of the detection chip at the mixing system, wherein the third gas isolation membrane is removed;

[0076] Fig.30 A perspective view of a mixing cover according to an embodiment of the present disclosure is shown;

[0077] Fig.31shows a perspective view of a bonding ring according to an embodiment of the present disclosure;

[0078] Fig.32 A cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown;

[0079] Fig.33 A top view of a base groove at a mixing system according to an embodiment of the present disclosure is shown;

[0080] Figures 34A-34D Another cross-sectional view of the mixing system according to an embodiment of the present disclosure is shown, which shows the steps of re-dissolving the magnetic beads 930 and mixing with the reagents;

[0081] Figures 35A-35D Another cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, which shows the magnetic beads 930 collection and mixing steps;

[0082] Fig.36 and Fig.37 A partial top view and a partial bottom view of the detection chip are shown respectively;

[0083] Fig.38A A perspective view of a first substrate at a storage cavity according to another embodiment of the present disclosure is shown;

[0084] Fig.38B Shows Fig.38A A cross-sectional view of the first substrate at the storage cavity;

[0085] Fig.39 A perspective view of a first substrate at a storage cavity according to another embodiment of the present disclosure is shown;

[0086] Fig.40A A perspective view of a first substrate at a storage cavity according to another embodiment of the present disclosure is shown;

[0087] Fig.40B Shows Fig.40A A cross-sectional view of the first substrate at the storage cavity;

[0088] Fig.41 A perspective view of a first substrate at a storage cavity according to another embodiment of the present disclosure is shown. DETAILED DESCRIPTION

[0089] In order to make the purpose, technical solution and advantages of the embodiments of the present disclosure clearer, the technical solution of the embodiments of the present disclosure will be clearly and completely described below in conjunction with the drawings of the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, not all of the embodiments. Based on the described embodiments of the present disclosure, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present disclosure.

[0090] Unless otherwise defined, the technical terms or scientific terms used in the present disclosure should be understood by people with ordinary skills in the field to which the present disclosure belongs. The "first", "second" and similar words used in the present disclosure do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word cover the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0091] At least one embodiment of the present disclosure provides a detection chip, including a first substrate and a first flexible layer. The first substrate includes at least one storage cavity, and the first flexible layer is attached to the first side of the first substrate. The storage cavity has a first opening on the first side, and the first side of the first substrate includes a first bonding area and a second bonding area. The first flexible layer is at the first opening of at least one storage cavity and is configured to be in an open state or a closed state. When the first flexible layer is in a closed state, the first flexible layer is attached to the first side of the first substrate in the first bonding area and the second bonding area to at least close the first opening of the storage cavity. When the first flexible layer is in an open state, the first flexible layer is attached to the first side of the first substrate in the second bonding area and is separated from the first substrate in the first bonding area to open the first opening of the storage cavity. The storage cavity can be squeezed so that the first flexible layer is separated from the substrate at the first bonding area, thereby accurately and flexibly controlling the opening of the storage cavity.

[0092] At least one embodiment of the present disclosure provides a detection chip, which can include one or more of an elution liquid chamber, a first cleaning liquid chamber, a lysis liquid chamber, a second cleaning liquid chamber, a third cleaning liquid chamber, a sample chamber, a first mixing chamber, a second mixing chamber, a waste liquid chamber, a gas permeable chamber, and an amplification chamber as needed. The detection chip can integrate all or part of the steps of the detection process, thereby facilitating the design and operation of the detection, reducing the detection time, reducing the contamination during the detection process, etc.

[0093] At least one embodiment of the present disclosure provides a detection device, which is used to install the chip positioning structure of the detection chip as described above, so as to be used together with the detection chip as described above.

[0094] The detection chip of the embodiment of the present disclosure will be generally introduced below with reference to the accompanying drawings.

[0095] Figure 1 shows a perspective view of a detection chip according to at least one embodiment of the present disclosure, Figure 2 Shows Figure 1 Exploded perspective view of the detection chip. Figure 3A Shows Figure 1 A top view of the first substrate 100 of the detection chip, Figure 3B Shows Figure 1 A bottom view of the first substrate 100 of the detection chip, Figure 4 Shows Figure 1 A top view of the middle layer 500 of the detection chip, Figure 5 Shows Figure 1 A plan view of the first flexible layer 200 of the detection chip, Fig. 6A Shows Figure 1 A plan view of the first portion 310 of the second flexible layer 300 of the detection chip, Figure 6B Shows Figure 1 A plan view of the second portion 320 of the second flexible layer 300 of the detection chip, Figure 7 Shows Figure 1 FIG. 4 is a bottom view of the detection chip, wherein the second substrate 400 is not shown.

[0096] like Figure 1 and Figure 2 As shown, the detection chip according to at least one embodiment of the present disclosure includes a first substrate 100 , an intermediate layer 500 , a first flexible layer 200 , a second flexible layer 300 , a first cover 710 , a second cover 720 , a third cover 800 and a second substrate 400 .

[0097] like Figure 3A and Figure 3B As shown, the first substrate 100 is formed with a plurality of storage chambers, a plurality of mixing chambers, a waste liquid chamber 190, a venting chamber 1100 and a plurality of substrate grooves 100c. The first substrate 100 can be injection molded from polymer materials such as PMMA (polymethyl methacrylate), PC (polycarbonate), PP (polypropylene), PS (polystyrene) and the like.

[0098] The plurality of storage chambers include a sample chamber 130 and a plurality of liquid storage chambers. The plurality of liquid storage chambers include an eluent chamber 110 for storing an eluent, a first cleaning chamber 160 for storing a first cleaning chamber, a second cleaning chamber 150 for storing a second cleaning chamber, a third cleaning chamber 140 for storing a third cleaning chamber, and a lysing chamber 180 for storing a lysing chamber. In this embodiment, the detection chip includes three cleaning chambers, namely, a first cleaning chamber 160, a second cleaning chamber 150, and a third cleaning chamber 140, but the present disclosure is not limited thereto, and more or fewer cleaning chambers may be provided as required.

[0099] The plurality of mixing chambers include a first mixing chamber 170 and a second mixing chamber 120, which constitute a part of the mixing system of the detection chip. The setting of the mixing chamber allows the reagents to be mixed in the detection chip, thereby realizing the integration of various processing steps. The setting of the first mixing chamber 170 and the second mixing chamber 120 enables the reagents to be more fully mixed by repeatedly flowing between the two mixing chambers.

[0100] like Figure 3B As shown, a plurality of base grooves 100 c are formed at a first side 100 a of the first base 100 facing the first flexible layer 200 .

[0101] In some embodiments, the first substrate may be formed by integral injection molding, in which case the side of the storage cavity higher than the substrate is the second side, and the side opposite to the second side is the first side 100a.

[0102] The intermediate layer 500 is disposed between the first base 100 and the first flexible layer 200 to bond the first base 100 to the first flexible layer 200. Figure 4 As shown, the intermediate layer 500 includes a plurality of intermediate through grooves 510 , each of which is positioned to correspond to one of the plurality of base grooves 100 c and is combined with the first flexible layer 200 to form a plurality of micro-channels.

[0103] The middle layer 500 may be a double-sided adhesive layer, a glue layer, or a plastic film layer attached between the first substrate 100 and the first flexible layer 200 by heat pressing or the like.

[0104] Those skilled in the art will recognize that in other embodiments, the detection chip may not have the intermediate layer 500. For example, the first flexible layer 200 may be attached to the first substrate 100 by heat pressing, ultrasonic welding or laser welding, chemical solvent method, etc.

[0105] In this embodiment, the side of the first flexible layer 200 attached to the first substrate 100 is a flat surface. In addition, in some other embodiments, a groove may be provided in the surface of the side of the first flexible layer 200 attached to the first substrate 100 to constitute a part of the microchannel.

[0106] The first flexible layer 200 is attached to the first side 100a of the first base 100. The first flexible layer 200 has flexibility so that the first flexible layer 200 is in an open state or a closed state at at least one storage cavity of the first base 100. The description of the exemplary structure of the storage cavity according to the embodiment of the present disclosure can be referred to later in conjunction with, for example, Fig. 9A , 9B by Fig.10 and Fig.11Description. When the first flexible layer 200 is in a closed state, the storage cavity is closed, and the liquid in the storage cavity cannot flow out to the outside of the storage cavity. When the first flexible layer 200 is in an open state, the storage cavity is opened, and the liquid in the storage cavity can flow out to the microchannel connected to the storage cavity. The first flexible layer 200 can be made of polymer materials such as PE (polyethylene), PP, PS, PC, etc. For example, the surface properties of the first flexible layer 200 can be changed by surface modification, etc., so that the enzymes, nucleic acid molecules, etc. in the detection chip are not easy to adhere to the first flexible layer 200, so that the first flexible layer 200 has good biocompatibility.

[0107] The second flexible layer 300 is attached to the second side 100b of the first substrate 100 opposite to the first side 100a. Specifically, the second flexible layer 300 is attached to the second side 100b of the first substrate 100 at each of the liquid storage chamber, the waste liquid chamber 190 and the air permeable chamber 1100 to close the corresponding chambers. For example, the second flexible layer 300 may include a plurality of independent parts. For example, Fig. 6A and Figure 6B As shown, a portion of the second flexible layer 300 attached to the elution liquid chamber 110, the lysis liquid chamber 180, the waste liquid chamber 190 and the air permeable chamber 1100 forms an integrated first portion 310 of the second flexible layer 300, and a portion of the second flexible layer 300 attached to the first cleaning liquid chamber 160, the second cleaning liquid chamber 150 and the third cleaning liquid chamber 140 forms an integrated second portion 320 of the second flexible layer 300.

[0108] Those skilled in the art will appreciate that in other embodiments, the second flexible layer 300 may be divided into more or fewer independent parts. The second flexible layer 300 may be attached to the first substrate 100 by heat pressing, adhesive bonding, interlayer bonding, ultrasonic welding, laser welding, etc. The second flexible layer 300 may be made of polymer materials such as PE, PP, PS, etc.

[0109] The first cover 710, the second cover 720 and the third cover 800 are attached to the second side 100b of the first substrate 100 at the first mixing chamber 170, the second mixing chamber 120 and the sample chamber 130, respectively, to at least close the corresponding chambers. In the embodiment shown in the figure, the first cover 710 and the second cover 720 are formed as an integrated mixing cover 700; in other embodiments, the first cover 710 and the second cover 720 can be provided independently of each other. For example, in some embodiments, the first cover 710, the second cover 720 and the third cover 800 can be of the same structure, and of course, in other embodiments, the three can also be different.

[0110] Figure 8 A perspective view of a second base body 400 according to an embodiment of the present disclosure is shown.

[0111] like Figure 8 As shown, the second substrate 400 is formed with an amplification chamber 410 configured to store an amplification reagent. The amplification chamber 410 is closed at one end and has an opening at the other end, and is attached to the side of the first flexible layer 200 opposite to the first substrate 100 .

[0112] like Figure 4 and Figure 5 As shown, the first flexible layer 200 is provided with a first flow channel outlet 210 and a second flow channel outlet 220 in the form of through holes, and the intermediate layer 500 is provided with first and second intermediate layer through holes 520, 530 corresponding to the first flow channel outlet 210 and the second flow channel outlet 220, respectively. The opening of the amplification chamber 410 is connected to the first flow channel outlet 210 and the first intermediate layer through hole 520 in the intermediate layer 500, and is also connected to the second flow channel outlet 220 and the second intermediate layer through hole 530 in the intermediate layer 500. The second substrate 400 is injection molded by a polymer material such as PMMA, PC, PP, PS, etc., preferably made of PP and PC. In addition, in at least one example, the second substrate 400 can be surface-modified so that enzymes, nucleic acid molecules, etc. are not easily adhered to the surface of the second substrate 400.

[0113] The detection chip according to the embodiment of the present disclosure allows one or more or even all of the steps of sample addition, lysis, mixing, washing, amplification, detection, etc. to be integrated. Therefore, the operation of the detection chip is facilitated, the automation level of the detection chip is improved, and the time for performing related detection using the detection chip is reduced.

[0114] As described above, the first matrix 100 includes a sample chamber 130, an elution liquid chamber 110, a first cleaning liquid chamber 160, a second cleaning liquid chamber 150, a third cleaning liquid chamber 140, a lysis liquid chamber 180, a first mixing chamber 170, a second mixing chamber 120, a waste liquid chamber 190 and a gas permeable chamber 1100.

[0115] The second substrate 400 includes an amplification chamber 410. The first substrate 100, the intermediate layer 500 and the first flexible layer 200 together form a plurality of microfluidic channels, including a first microfluidic channel 1, a second microfluidic channel 2, a third microfluidic channel 3, a fourth microfluidic channel 4, a fifth microfluidic channel 5, a sixth microfluidic channel 6, a seventh microfluidic channel 7, an eighth microfluidic channel 8, a ninth microfluidic channel 9 and a tenth microfluidic channel 10.

[0116] The first mixing chamber 170 and the second mixing chamber 120 form a mixing chamber system. The sample chamber 130, the elution liquid chamber 110, the first cleaning liquid chamber 160, the second cleaning liquid chamber 150, the third cleaning liquid chamber 140, the lysis liquid chamber 180 and the waste liquid chamber 190 are all connected to the mixing chamber system to perform various processing steps on the sample.

[0117] Specifically, the first microfluidic channel 1 is configured to connect the first mixing chamber 170 and the lysis liquid chamber 180, the second microfluidic channel 2 is configured to connect the first mixing chamber 170 and the first cleaning liquid chamber 160, the third microfluidic channel 3 is configured to connect the second mixing chamber 120 and the elution liquid chamber 110, the fourth microfluidic channel 4 is configured to connect the first mixing chamber 170 and the third cleaning liquid chamber 140, the fifth microfluidic channel 5 is configured to connect the fourth microfluidic channel 4 and the second cleaning liquid chamber 150, the sixth microfluidic channel 6 is configured to connect the first mixing chamber 170 and the waste liquid chamber 190, the seventh microfluidic channel 7 is configured to connect the amplification chamber 410 and the first mixing chamber 170, the eighth microfluidic channel 8 is configured to connect the amplification chamber 410 and the air permeable chamber 1100, the ninth microfluidic channel 9 is configured to connect the first mixing chamber 170 and the second mixing chamber 120, and the tenth microfluidic channel 10 is configured to connect the first mixing chamber 170 and the sample chamber 130. In addition, the first microchannel 1, the second microchannel 2, the third microchannel 3, the fourth microchannel 4, the fifth microchannel 5, the sixth microchannel 6, the seventh microchannel 7, the eighth microchannel 8, the ninth microchannel 9 and the tenth microchannel 10 are respectively provided with the first switch valve 11, the second switch valve 21, the fourth switch valve 31, the fourth switch valve 41, the fifth switch valve 51, the sixth switch valve 61, the seventh switch valve 71, the eighth switch valve 81, the ninth switch valve 91 and the tenth switch valve 101 to control the opening and closing of the corresponding microchannels.

[0118] The mixing chamber system composed of the first mixing chamber 170 and the second mixing chamber 120 is arranged in the middle section of the detection chip, and other chambers (such as the sample chamber 130, the elution liquid chamber 110, the first cleaning liquid chamber 160, the second cleaning liquid chamber 150, the third cleaning liquid chamber 140, the lysis liquid chamber 180 and the waste liquid chamber 190) are arranged on both sides of the mixing chamber system to help make the length of each flow channel more uniform, reduce liquid residue or product instability caused by too long flow channels. Design the extension path and position of each flow channel so that the spacing between the flow channels is uniform to prevent the flow channels in some positions from being too dense, which increases the difficulty and cost of production.

[0119] The volume of each chamber can be selected as needed. For example, in one example, 100 μL of eluent, 500 μL of lysate and 500 μL of first cleaning solution are required. Therefore, the diameter of the eluent chamber 110 can be designed to be smaller than the lysate chamber 180 and the first cleaning solution chamber 160, and the diameters of the lysate chamber 180 and the first cleaning solution chamber 160 can be designed to be equal. If the eluent chamber 110 is designed to be larger, the eluent may not fill the bottom of the chamber, resulting in flow interruption. For example, the first mixing chamber 170 and the second mixing chamber 120 can be designed to have a higher height to accommodate more liquids to be mixed.

[0120] For example, at least one embodiment of the present disclosure also provides an operating method for a detection chip, which includes: adding a sample to the detection chip; mixing the sample with a lysis solution to form a mixed solution, the lysis solution can lyse the sample to form a substance to be detected; driving the mixed solution to flow to a mixing chamber system, the mixing chamber system includes a first mixing chamber and a second mixing chamber, the mixing chamber system has magnetic beads, and the magnetic beads can adsorb the substance to be detected; driving the mixed solution to move back and forth between the first mixing chamber and the second mixing chamber to mix the mixed solution and the magnetic beads; separating and processing the magnetic beads to make the substance to be detected fall off the magnetic beads; and detecting the substance to be detected.

[0121] For example, in at least one usage scenario, the detection chip may operate as follows.

[0122] S1, sample adding step: open the third cover 800, add the sample to be tested into the sample chamber 130, and then cover the third cover 800. For example, the volume of the sample to be tested can be 10 μL-1000 μL.

[0123] S2, sample release step: open the tenth switch valve 101, squeeze the sample chamber 130 to make the sample flow from the sample chamber 130 through the tenth microchannel 10 to the first mixing chamber 170, and then close the tenth switch valve 101.

[0124] S3, lysate release step: open the first switch valve 11, squeeze the lysate chamber 180 to make the lysate in the lysate chamber 180 flow from the lysate chamber 180 to the first mixing chamber 170 through the first microchannel 1, and then close the first switch valve 11. Therefore, the sample and the lysate are mixed in the sample chamber 130, so that biological substances such as cells in the sample are lysed to release the nucleic acid molecules therein.

[0125] S4, magnetic bead adsorption step: the ninth switch valve 91 is opened, and the sample and lysate mixture flows to the second mixing chamber 120 through the ninth microchannel 9 by squeezing the first mixing chamber 170 to re-dissolve the magnetic beads in the second mixing chamber 120. Therefore, in the second mixing chamber 120, the nucleic acid molecules in the mixture can be adsorbed by the magnetic beads.

[0126] S5, lysis and first mixing step: alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the mixed solution of the sample, the lysis solution and the magnetic beads moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally the mixed solution stays in the first mixing chamber 170, and then close the ninth switch valve 91.

[0127] The lysis and first mixing steps promote the full mixing of the sample, lysis solution and magnetic beads, so that the nucleic acid molecules are fully released and fully adsorbed by the magnetic beads. For example, the first mixing chamber 170 and the second mixing chamber 120 are squeezed alternately to make the mixed solution of the sample, lysis solution and magnetic beads move back and forth between the first mixing chamber 170 and the second mixing chamber 120 1-100 times to achieve the desired mixing effect.

[0128] S6, first magnetic bead collection and second mixing step: move the magnet from the outside of the detection chip close to the bottom of the second mixing chamber 120 (i.e., close to the side of the first flexible layer 200), open the ninth switch valve 91, squeeze the first mixing chamber 170 to make the mixed solution of the sample, lysate and magnetic beads flow from the first mixing chamber 170 to the second mixing chamber 120 through the ninth microfluidic channel 9, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the mixed solution of the sample, lysate and magnetic beads moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally make the mixed solution without magnetic beads stay in the first mixing chamber 170, while the magnetic beads adhered with nucleic acid molecules stay in the second mixing chamber 120 due to the adsorption of the magnet, and then remove the magnet and close the ninth switch valve 91.

[0129] After the first magnetic bead collection and the second mixing step, the magnetic beads adsorbed with nucleic acid molecules are separated from other components in the mixed solution. For example, the number of mixing times (ie, the number of round trips) can be set to 1-100 times.

[0130] S7, the first step of emptying the waste liquid: open the sixth switch valve 61, squeeze the first mixing chamber 170 to allow the mixed liquid without magnetic beads to flow from the first mixing chamber 170 to the waste liquid chamber 190 through the sixth microchannel 6 as waste liquid, and then close the sixth switch valve 61.

[0131] S8, first cleaning liquid releasing step: open the second switch valve 21, squeeze the first cleaning liquid chamber 160 to make the first cleaning liquid flow from the first cleaning liquid chamber 160 through the second microchannel 2 to the first mixing chamber 170, and then close the second switch valve 21.

[0132] S9, first cleaning and third mixing step: open the ninth switch valve 91, squeeze the first mixing chamber 170 to make the first cleaning solution in the first mixing chamber 170 flow to the second mixing chamber 120 through the ninth microfluidic channel 9 to redissolve the magnetic beads staying in the second mixing chamber 120, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, make the mixed solution of the first cleaning solution and the magnetic beads move back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, finally make the mixed solution stay in the first mixing chamber 170, and then close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0133] Impurities such as components in the sample and lysate will remain on the surface of the magnetic beads that have been adsorbed with nucleic acid molecules, and these impurities will affect subsequent amplification reactions and detection, etc. The first cleaning step and the subsequent second cleaning step and third cleaning step will remove these impurities. For example, the first cleaning solution in the first cleaning step can be used to clean protein molecules on the surface of nucleic acids.

[0134] S10, second magnetic bead collection and fourth mixing step: move the magnet from the outside of the detection chip close to the bottom of the second mixing chamber 120, open the ninth switch valve 91, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the mixed solution of the first cleaning solution and the magnetic beads moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally the mixed solution without magnetic beads stays in the first mixing chamber 170, while the magnetic beads adhered to the nucleic acid molecules stay in the second mixing chamber 120 due to the adsorption of the magnet, and then remove the magnet and close the ninth switch valve 91.

[0135] After the magnetic bead collection and the second mixing step, the magnetic beads adsorbed with nucleic acid molecules are separated from other components in the mixed solution. For example, the number of mixing times (ie, the number of round trips) can be set to 1-100 times.

[0136] S11, second step of emptying waste liquid: open the sixth switch valve 61, squeeze the first mixing chamber 170 to allow the mixed liquid without magnetic beads to flow from the first mixing chamber 170 to the waste liquid chamber 190 through the sixth microchannel 6 as waste liquid, and then close the sixth switch valve 61.

[0137] S12, second cleaning liquid release step: open the fourth switch valve 41 and the fifth switch valve 51, and squeeze the second cleaning liquid chamber 150 to make the second cleaning liquid flow from the second cleaning liquid chamber 150 through the fourth microchannel 4 and the fifth microchannel 5 to the first mixing chamber 170, and then close the fourth switch valve 41 and the fifth switch valve 51.

[0138] S13, second cleaning and fifth mixing step: open the ninth switch valve 91, squeeze the first mixing chamber 170 to make the second cleaning solution in the first mixing chamber 170 flow to the second mixing chamber 120 through the ninth microchannel 9 to redissolve the magnetic beads staying in the second mixing chamber 120, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, make the mixed solution of the second cleaning solution and the magnetic beads move back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microchannel 9, finally make the mixed solution stay in the first mixing chamber 170, and then close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0139] For example, the second cleaning solution in the second cleaning step can be used to clean small molecule impurities and salt ions on the surface of the magnetic beads.

[0140] S14, third magnetic bead collection and sixth mixing step: move the magnet from the outside of the detection chip close to the bottom of the second mixing chamber 120, open the ninth switch valve 91, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the mixed solution of the second cleaning solution and the magnetic beads moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microchannel 9, and finally the mixed solution without magnetic beads stays in the first mixing chamber 170, and the magnetic beads adhered to the nucleic acid molecules stay in the second mixing chamber 120 due to the adsorption of the magnet, and then remove the magnet and close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0141] S15, the third step of emptying the waste liquid: open the sixth switch valve 61, squeeze the first mixing chamber 170 to allow the mixed liquid without magnetic beads to flow from the first mixing chamber 170 to the waste liquid chamber 190 through the sixth microchannel 6 as waste liquid, and then close the sixth switch valve 61.

[0142] S16, third cleaning liquid releasing step: open the fourth switch valve 41, squeeze the third cleaning liquid chamber 140 to make the third cleaning liquid flow from the third cleaning liquid chamber 140 through the fourth microchannel 4 to the first mixing chamber 170, and then close the fourth switch valve 41.

[0143] S17, third cleaning and seventh mixing step: open the ninth switch valve 91, squeeze the first mixing chamber 170 to make the third cleaning solution in the first mixing chamber 170 flow to the second mixing chamber 120 through the ninth microfluidic channel 9 to redissolve the magnetic beads staying in the second mixing chamber 120, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, make the mixed solution of the first cleaning solution and the magnetic beads move back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, finally make the mixed solution stay in the first mixing chamber 170, and then close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0144] For example, the third cleaning liquid in the third cleaning step can be used to clean small molecules and salt ions remaining on the surface of the magnetic beads. For example, the third cleaning liquid is the same as the second cleaning liquid. Since the second cleaning liquid is the same as the third cleaning liquid, the second cleaning liquid chamber 150 and the third cleaning liquid chamber 140 are designed to share a portion of the fourth microfluidic channel 4 to reduce the volume of the detection chip and simplify the layout of the microfluidic channel of the detection chip.

[0145] In this embodiment, the magnetic beads are washed using three washing and mixing steps to remove impurities (such as small molecules, salt ions, protein molecules, etc.) on the surface of the magnetic beads. In other embodiments, a specific washing solution can be selected as needed to wash the magnetic beads in one, two, or more than three washing and mixing steps.

[0146] S18, fourth magnetic bead collection and eighth mixing step: move the magnet from the outside of the detection chip close to the bottom of the second mixing chamber 120, open the ninth switch valve 91, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the mixed solution of the second cleaning solution and the magnetic beads moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally the mixed solution without magnetic beads stays in the first mixing chamber 170, and the magnetic beads adhered to the nucleic acid molecules stay in the second mixing chamber 120 due to the adsorption of the magnet, and then remove the magnet and close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0147] S19, the fourth step of emptying the waste liquid: open the sixth switch valve 61, squeeze the first mixing chamber 170 to allow the mixed liquid without magnetic beads to flow from the first mixing chamber 170 to the waste liquid chamber 190 through the sixth microchannel 6 as waste liquid, and then close the sixth switch valve 61.

[0148] S20, magnetic bead drying step: heat the bottom of the second mixing chamber 120 and keep the temperature at a constant temperature, for example, in the range of 30-70° C. for a certain period of time. During the evaporation process of the drying water, the organic reagent (such as ethanol) remaining in the cleaning solution on the surface of the magnetic beads will be removed by volatilization.

[0149] S21, eluent release step: open the third switch valve 31, squeeze the eluent chamber 110 to make the eluent flow from the eluent chamber 110 through the third microchannel 3 to the second mixing chamber 120, and then close the third switch valve 31.

[0150] S22, elution and ninth mixing step: open the ninth switch valve 91, and squeeze the second mixing chamber 120 to make the eluent and magnetic bead mixture in the second mixing chamber 120 flow to the first mixing chamber 170 through the ninth microfluidic channel 9. While heating the first mixing chamber 170 and the second mixing chamber 120, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120 to make the eluent and magnetic bead mixture move back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally make the mixture stay in the first mixing chamber 170, and then close the ninth switch valve 91 and stop heating.

[0151] Under the combined action of heating and the eluent, the nucleic acid molecules will be detached from the surface of the magnetic beads. In addition, by fully mixing the eluent with the magnetic beads, the nucleic acid molecules can be more effectively detached from the surface of the magnetic beads. Ultimately, the liquid remaining in the first mixing chamber 170 will be the eluent containing the nucleic acid molecules, i.e., the sample to be amplified. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0152] S23, fifth magnetic bead collection and ninth mixing step: move the magnet from the outside of the detection chip close to the bottom of the second mixing chamber 120, open the ninth switch valve 91, alternately squeeze the first mixing chamber 170 and the second mixing chamber 120, so that the sample to be amplified moves back and forth between the first mixing chamber 170 and the second mixing chamber 120 through the ninth microfluidic channel 9, and finally the sample to be amplified without magnetic beads stays in the first mixing chamber 170, and the magnetic beads without nucleic acid molecules stay in the second mixing chamber 120 due to the adsorption of the magnet, and then remove the magnet and close the ninth switch valve 91. For example, the number of mixing times (i.e., the number of round trips) can be set to 1-100 times.

[0153] S24, the transfer step to the amplification chamber 410: open the seventh switch valve 71 and the eighth switch valve 81, squeeze the first mixing chamber 170 to make the sample to be amplified flow to the amplification chamber 410 through the seventh microfluidic channel 7 to redissolve the amplification reagent in the amplification chamber 410, and then flow to the gas permeable chamber 1100 through the eighth microfluidic channel 8 after filling the amplification chamber 410, and then close the seventh switch valve 71 and the eighth switch valve 81. Such a transfer step helps to fill the amplification chamber 410 with the sample to be amplified, and the excess eluent containing nucleic acid molecules will stay in the seventh microfluidic channel 7, the eighth microfluidic channel 8 or the gas permeable chamber 1100.

[0154] S25, amplification step: cyclically heating the amplification chamber to amplify the amplified sample therein, and then detecting the properties of the amplified sample, such as optical properties, to obtain a detection result.

[0155] The above operation process can be realized by operating the detection chip through the detection device. For example, after step S1, the detection chip can be placed in a proper position of the detection device, and then the detection chip can be operated by the relevant components of the detection device.

[0156] The detection chip according to the embodiment of the present disclosure can process the entire process from sample addition to detection of the original sample, and has a high degree of integration.

[0157] In addition, as described above, in the above embodiment, the first mixing chamber is used as an intermediate chamber to achieve effective integration and relative isolation of multiple processing steps.

[0158] Those skilled in the art should understand that the detection chip according to the embodiment of the present disclosure can be operated in a different order from the above, or one or more of the above steps can be omitted, or one or more of the above steps can be repeated. For example, when the added sample is a cell suspension that has been lysed, the steps related to lysis can be omitted.

[0159] In addition, those skilled in the art should understand that the detection chip can omit one or more of, for example, the sample chamber 130, the elution liquid chamber 110, the first cleaning liquid chamber 160, the second cleaning liquid chamber 150, the third cleaning liquid chamber 140, the lysis liquid chamber 180, the first mixing chamber 170, the second mixing chamber 120, the waste liquid chamber 190 and the air permeable chamber 1100 to meet the needs of different applications.

[0160] The open state and the closed state of the first flexible layer 200 at the storage cavity will be described below with reference to the accompanying drawings.

[0161] As described above, the first flexible layer 200 is in an open state or a closed state at at least one storage cavity of the first base 100 .

[0162] Refer to the following Figure 9A-Figure 12 The open state and the closed state of the first flexible layer 200 at the storage cavity are described in detail.

[0163] Fig. 9A A perspective view of a detection chip at a storage cavity according to an embodiment of the present disclosure is shown, wherein the first flexible layer 200 is in an open state. Fig. 9B Shows Fig. 9A A partial enlarged view of . Fig.10 FIG. 4 shows a cross-sectional view of a detection chip at a storage cavity according to an embodiment of the present disclosure, wherein the first flexible layer 200 is in a closed state. Fig.11 FIG. 4 shows another cross-sectional view of the detection chip at a storage cavity according to an embodiment of the present disclosure, wherein the first flexible layer 200 is in an open state. Fig.12 A perspective view of the first base 100 at the storage cavity according to an embodiment of the present disclosure is shown.

[0164] like Figure 9A-Figure 12 As shown, the storage cavity has a first opening at a first side 100a of the first substrate 100 and a second opening at a second side 100b opposite to the first side 100a. The first flexible layer 200 is attached to the first side 100a of the first substrate 100 at the storage cavity, and the second flexible layer 300 is attached to the second side 100b of the first substrate 100 opposite to the first side 100a at the storage cavity to close the second opening of the storage cavity at the second side 100b.

[0165] The first substrate 100 has a first bonding area 111a and a second bonding area 111b at the first side 100a. The first flexible layer 200 can be attached to the first side 100a of the first substrate 100 at the first bonding area 111a and the second bonding area 111b. The attachment strength is defined as the magnitude of the total tearing force required to completely separate the first flexible layer 200 from the first substrate 100. In this embodiment, the first attachment strength of the first flexible layer 200 to the first substrate 100 at the first bonding area 111a is less than the second attachment strength of the first flexible layer 200 to the first substrate 100 at the second bonding area 111b. The first bonding area 111a and the second bonding area 111b are respectively a weak bonding area and a strong bonding area. For example, the first attachment strength is in the range of 1N-50N, such as 35N, and the second attachment strength is in the range of 50N-500N, such as 100N. Here, the bonding area of ​​the first substrate 100 and the first flexible layer 200 at the first bonding area 111a is small, so the bonding force is weak. In addition, the attachment strength at the first bonding area 111a and the second bonding area 111b can also be controlled by differentiating the bonding process parameters of different areas, such as the temperature and pressure of heat press sealing, the bonding force of the glue, the surface microstructure of the substrate at different areas (e.g., concave-convex structure), etc.

[0166] The surface of the first side 100a of the first substrate 100 is provided with a buffer groove 111 and a substrate groove 100c for forming a microfluidic channel connected to the buffer groove 111. For example, the microfluidic channel can be a first microfluidic channel 1, a second microfluidic channel 2, a third microfluidic channel 3, etc. In this example, the buffer groove 111 is annular and is arranged around the storage cavity to define an annular first bonding area 111a on the inner side of the buffer groove 111 and a second bonding area 111b on the outer side of the buffer groove 111. When the first flexible layer 200 is in a closed state, it is attached to the first side 100a of the first substrate 100 at both the first bonding area 111a and the second bonding area 111b to at least close the first opening of the corresponding storage cavity, and when the first flexible layer 200 is in an open state, it is attached to the first side 100a of the first substrate 100 at the second bonding area 111b and is separated from the first substrate 100 at the first bonding area 111a to open the first opening of the corresponding storage cavity.

[0167] By providing the first and second adhesive regions 111a and 111b, the first flexible layer 200 can be precisely separated from the first substrate 100 at the first adhesive region 111a by pressing the second flexible layer 300 at the storage cavity, thereby allowing precise and flexible control over the opening and closing of the storage cavity.

[0168] For example, the storage chamber (e.g., the liquid storage chamber described above) may be pre-stored with reagents, which are sealed in the storage chamber and are allowed to be sold together with the detection chip. When using the detection chip, the user does not need to add reagents to the liquid storage chamber before starting to use the detection chip. Fig.10 The push rod 910 in the first flexible layer 200 is used to squeeze the second flexible layer 300, so that the first flexible layer 200 is converted from a closed state to an open state, thereby allowing the pre-stored reagent to be released. Therefore, the use of the detection chip is facilitated. The storage cavity according to an embodiment of the present disclosure allows the long-term storage of the reagent, for example, up to one year. In addition, the first substrate 100, the first flexible layer 200 and the second flexible layer 300 can be designed to isolate air and liquid, so that the reagent stored in the storage cavity is isolated from air and external substances and remains stable during the transportation of the detection chip. In addition, by controlling the stroke of the push rod 910, the amount of the reagent in the extruded liquid storage cavity can be quantitatively controlled, thereby achieving the effect of quantitative release.

[0169] Due to the existence of buffer tank 111, bonding area is separated into two bonding areas, first bonding area 111a and second bonding area 111b. Therefore, when reagent is released from liquid storage chamber due to the separation of first substrate 100 and first flexible layer 200 at first bonding area 111a, it will first enter buffer tank 111 to slow down the instantaneous flow rate of reagent, eliminate the impact of reagent on second bonding area 111b, avoid separating second bonding area 111b from first substrate 100, eliminate the violent rush phenomenon. Then, reagent will enter microfluidic channel from buffer tank 111, thereby entering other chambers via microfluidic channel. In addition, because the bonding area of ​​first substrate 100 and first flexible layer 200 at first bonding area 111a is small and bonding force is weak, when reagent in storage chamber is squeezed, it only needs to separate any part of first bonding area 111a from first substrate 100 at first, which can cause the whole first bonding area 111a to separate from first substrate 100, thus ensuring the success rate of reagent release. Therefore, the buffer groove 111 makes it easier to destroy the attachment of the first flexible layer 200 to the first substrate 100 at the first bonding area 111a, while preventing the attachment of the first flexible layer 200 to the first substrate 100 at the second bonding area 111b from being destroyed, and accurately controls the position and size of the portion where the storage cavity is connected to the microchannel when the first flexible layer 200 is in an open state.

[0170] The depth of buffer tank 111 can be in the range of 0.01mm-5mm, for example 0.5mm, and the width (when buffer tank 111 is annular, the difference between its outer diameter and inner diameter) can be in the range of 0.01mm-5mm, for example 1mm. The increase of depth and width can cause reagent to remain, causing the quantitative release of reagent to be inaccurate. The reduction of depth and width can cause fluid resistance to increase, which is unfavorable for the release of reagent. When buffer tank 111 is annular, its inner diameter is determined by the size of the storage cavity, for example, in the range of 3-20mm. The bottom surface of buffer tank 111 can be a hydrophobic surface, which is conducive to reducing reagent residue.

[0171] In this embodiment, the first substrate 100 is an annular boss 113 at the first bonding area 111a, and the width of the annular boss 113 is, for example, in the range of 0.01mm-5mm, and its inner diameter is determined by the size of the storage cavity, for example, in the range of 3mm-20mm. In this embodiment, the surface of the first substrate 100 at the bonding area of ​​the first bonding area 111a is substantially flat. For example, the first flexible layer 200 can be attached to the first side 100a of the first substrate 100 through the intermediate layer 500 as described above.

[0172] Fig.10 and Fig.11The process of the push rod 910 squeezing the second flexible layer 300 is shown. The push rod 910 is used to act on the second flexible layer 300 to deform the second flexible layer 300, and the closed volume formed by the storage cavity is compressed. When the push rod 910 causes the deformation of the second flexible layer 300 to be large enough, the pressure inside the storage cavity is large enough, and the first flexible layer 200 is deformed due to the squeezing, so that the first flexible layer 200 is separated from the first substrate 100 at the first bonding area. When the first flexible layer 200 is separated from the first substrate 100 at the first bonding area 111a, the closed volume in the storage cavity is destroyed, and the storage cavity is connected to the buffer tank 111. The second flexible layer 300 is continuously squeezed to push the reagent into the buffer tank 111 and then enter the microchannel via the buffer tank 111, thereby releasing the reagent to other working areas of the detection chip. During the squeezing process of the second flexible layer 300, the second flexible layer 300 and the first flexible layer 200 are subjected to the squeezing force F1, and the tearing force of the first flexible layer 200 separating from the first substrate 100 at the first bonding area 111a is F2, and F2≈F1. Due to the existence of the buffer groove 111, the tearing force of the first flexible layer 200 separating from the first substrate 100 at the second bonding area 111b is F3, and F3≈0. Therefore, the bonding area is separated into two force-bearing areas, the first bonding area 111a and the second bonding area 111b. In addition, when the first flexible layer 200 is separated from the first substrate 100 at the first bonding area 111a and the reagent enters the buffer groove 111 and the microfluidic channel, the tearing force of the first flexible layer 200 separating from the first substrate 100 at the second bonding area 111b increases from F3 to F4. Since the reagent flows slowly in the microfluidic channel, F4 is much smaller than F2. The bonding of the first flexible layer 200 to the first substrate 100 in the second bonding area 111 b will not be damaged.

[0173] Those skilled in the art will appreciate that the buffer groove 111 may also be in other shapes and may only be disposed partially around the storage cavity. The buffer groove 111 disposed substantially completely around the storage cavity is conducive to better defining the first bonding area 111a and the second bonding area 111b.

[0174] Fig.38A FIG. 1 shows a perspective view of a first substrate 100 at a storage cavity according to another embodiment of the present disclosure. Fig.38B Shows Fig.38A A cross-sectional view of the first substrate 100 at the storage cavity.

[0175] like Figures 38A-38BAs shown, the annular boss 113 may include a plurality of sub-annular bosses surrounding each other, the plurality of sub-annular bosses jointly forming a concave-convex structure at the first bonding area 111a, and the top surfaces of the plurality of sub-annular bosses jointly forming a first bonding area 111a as a weak bonding area. Since the first bonding area 111a is divided into a plurality of sub-bonding areas corresponding to the plurality of sub-annular bosses, the first bonding strength at the first bonding area 111a is further reduced. Here, the depth of the groove for forming the plurality of sub-annular bosses is equal to the depth of the buffer groove 111. In other embodiments, the depth of the groove for forming the plurality of sub-annular bosses may be less than the depth of the buffer groove 111.

[0176] exist Figures 38A-38B In the illustrated embodiment, there are two sub-annular bosses. Fig.39 A perspective view of a first substrate 100 at a storage cavity according to another embodiment of the present disclosure is shown, wherein there are three sub-annular bosses, but the present disclosure is not limited thereto. The width of each sub-annular boss may be in the range of 0.001 mm-100 mm, and the interval between them may be in the range of 0.001 mm-100 mm.

[0177] Fig.40A FIG. 1 shows a perspective view of a first substrate 100 at a storage cavity according to another embodiment of the present disclosure. Fig.40B Shows Fig.40A A cross-sectional view of the first substrate 100 at the storage cavity.

[0178] The cross section of the annular boss 113 may decrease toward the first side 100a, for example, Fig.40A and 40B The arc-shaped cross section shown may alternatively be a trapezoidal cross section, etc. Since the cross section of the annular boss 113 decreases toward the first side 100a, the first bonding strength at the first bonding area 111a, which is a weak bonding area, is further reduced.

[0179] Fig.41 FIG. 2 shows a perspective view of a first substrate 100 at a storage cavity according to another embodiment of the present disclosure. Fig.41 As shown, the annular boss 113 may include a plurality of sub-annular bosses (two in this case), and the cross sections of the plurality of sub-annular bosses are all arc-shaped cross sections. The width (i.e., bottom width) of each sub-annular boss may be in the range of 0.001 mm-100 mm, and the intervals thereof may be in the range of 0.001 mm-100 mm. Fig.13 A longitudinal cross-sectional view of a detection chip at a storage cavity according to another embodiment of the present disclosure is shown, wherein the first flexible layer 200 is in a closed state. Fig.14 Another longitudinal cross-sectional view of the storage cavity is shown in the figure, wherein the first flexible layer 200 is in an open state. Fig.15 A transverse cross-sectional view at the storage cavity in the figure is shown.

[0180] like Figure 13-15 As shown, the first flexible layer 200 is attached to the first side 100a of the first substrate 100 at the storage cavity, and the second flexible layer 300 is attached to the second side 100b of the first substrate 100 opposite to the first side 100a at the storage cavity to close the storage cavity at the second side 100b. The surface of the first substrate 100 on the first side 100a is divided into a first bonding area 111c and a second bonding area 111d. The first bonding area 111c is arranged between the storage cavity of the first substrate 100 and the substrate groove 100c for forming a microfluidic channel. For example, the microfluidic channel can be a first microfluidic channel 1, a second microfluidic channel 2, a third microfluidic channel 3, etc. The first flexible layer 200 can be attached to the first side 100a of the first substrate 100 at the first bonding area 111c and the second bonding area 111d. When the first flexible layer 200 is in a closed state, it is attached to the first side 100a of the first substrate 100 at both the first bonding area 111c and the second bonding area 111d to at least close the corresponding storage cavity, and when the first flexible layer 200 is in an open state, it is attached to the first side 100a of the first substrate 100 at the second bonding area 111d and is separated from the first substrate 100 at the first bonding area 111c to liquid-open the corresponding storage cavity.

[0181] By providing the first adhesive region 111c and the second adhesive region 111d, the first flexible layer 200 can be separated from the first substrate 100 at the first adhesive region 111c by pressing the second flexible layer 300 at the storage cavity, thereby allowing precise and flexible control of opening and closing of the storage cavity.

[0182] Furthermore, the reagents may be pre-stored in the storage chamber and allowed to be sold together with the detection chip.

[0183] In this embodiment, the first attachment strength of the first flexible layer 200 to the first substrate 100 at the first bonding area 111c is less than the second attachment strength to the first substrate 100 at the second bonding area 111d. The first bonding area 111c is used as a weak bonding area, and the second bonding area 111d is used as a strong bonding area. The weak bonding area is realized by a plurality of concave-convex structures 112 at the first bonding area 111c on the surface of the first substrate 100 facing the first flexible layer 200.

[0184] Figures 16A-16D Schematic diagrams of a concave-convex structure 112 according to an embodiment of the present disclosure are shown, which respectively show a perspective schematic diagram and a cross-sectional schematic diagram of the concave-convex structure.

[0185] like Figures 16A-16DAs shown, the concave-convex structure 112 can be in the shape of an elongated strip, and a plurality of the concave-convex structures 112 in the shape of an elongated strip are arranged at a distance from each other. For example, the concave-convex structure 112 can be arranged at a period in the range of 0.1 mm to 5 mm. For example, the maximum depth difference of the concave-convex structure 112 in the vertical direction perpendicular to the surface of the first substrate 100 is in the range of 0.1 mm to 3 mm. For example, the number of the concave-convex structures 112 is 2 to 5. For example, the concave-convex structure 112 can include a plurality of wavy concave-convex structures 112 (or arc-shaped cross-section concave-convex structures 112, such as Fig.16A ), which are arranged at intervals of 1 mm (i.e., the distance between the vertices of adjacent concave-convex structures 112), the wave convex height is 0.2 mm, and the wave bottom width is 0.5 mm. For example, the concave-convex structure 112 may include a plurality of rectangular cross-section concave-convex structures 112 (such as Fig. 16B ), which are arranged at intervals of 1 mm (i.e., the distance between the vertices of adjacent concave-convex structures 112), the rectangular protrusion height is 0.2 mm, and the rectangle width is 0.1 mm. For example, the concave-convex structure 112 may include a plurality of trapezoidal cross-section concave-convex structures 112 (such as Fig. 16C ), which are arranged at intervals of 1 mm (i.e., the distance between the vertices of adjacent concavo-convex structures 112), the height of the trapezoidal protrusion is 0.2 mm, and the width of the trapezoidal bottom is 0.5 mm. In addition or alternatively, a plurality of concavo-convex structures 112 can be arranged in an array. For example, the concavo-convex structure 112 can be a circular groove (such as Fig.16D ), the number of which is 2-20, the spacing between the circular grooves is 0.1mm, and the diameter of the circular hole is 0.45mm. In addition, for example, the concave-convex structure 112 can also be a rectangular groove, a triangular groove, etc. Each concave-convex structure 112 can be the same or different. The bonding area between the concave-convex structure 112 of the first substrate 100 and the first flexible layer 200 is small, so the bonding force is reduced. At the same time, the arrangement of multiple concave-convex structures 112 improves the structural stability of the first substrate 100 at the first bonding area 111c. When one of the concave-convex structures 112 fails, the other concave-convex structures 112 still play a sealing role. The concave-convex structure 112 can be integrally formed with the first substrate 100 by methods such as injection molding or machining, so the processing is simple. In addition, there is no need to adopt different bonding methods or bonding parameters at the first bonding area 111c and the second bonding area 111d to form bonding areas with different bonding strengths. The division of the first bonding area 111c and the second bonding area 111d is achieved by using a plurality of concave-convex structures 112 instead of controlling process parameters such as temperature, pressure, and materials, thereby simplifying the preparation process of the detection chip, facilitating large-scale manufacturing, and improving the yield of the detection chip.

[0186] The switch valve in the microfluidic channel of the embodiment will be described below with reference to the accompanying drawings.

[0187] As described above, the first microchannel 1, the second microchannel 2, the third microchannel 3, the fourth microchannel 4, the fifth microchannel 5, the sixth microchannel 6, the seventh microchannel 7, the eighth microchannel 8, the ninth microchannel 9 and the tenth microchannel 10 are respectively provided with the first switch valve 11, the second switch valve 21, the third switch valve 31, the fourth switch valve 41, the fifth switch valve 51, the sixth switch valve 61, the seventh switch valve 71, the eighth switch valve 81, the ninth switch valve 91 and the tenth switch valve 101 to control the opening and closing of the corresponding microchannels.

[0188] The following describes a switch valve according to at least one embodiment of the present disclosure. The first to tenth switch valves 101 may be designed as such a switch valve.

[0189] Fig.17 A schematic perspective view showing a portion of a switching valve according to an embodiment of the present disclosure, Fig.18 Shows Fig.17 A perspective view of a base groove 100c of the first base 100, Figure 19-22 A cross-sectional view of a switch valve according to an embodiment of the present disclosure is shown. The cross-sectional view may be taken along Fig.17 The midline LL is intercepted. Fig.19 This is an exploded sectional view of the switch valve. Fig. 20 This is a cross-sectional view of the switch valve in a normally open state. Fig.21 This is a cross-sectional view of the switch valve in the fully open state. Fig. 22 It is a cross-sectional view of the switch valve in the closed state.

[0190] like Figure 17-22 As shown, in at least one example, the switch valve is formed by the base groove 100c of the first base 100, the middle layer through groove 510 in the middle layer 500, and the first flexible layer 200. The base groove 100c has a disconnection area that disconnects the base groove 100c into two sub-base grooves, and the middle through groove 510 includes a valve action area 511. The projection of the valve action area 511 in the direction of the surface of the first base 100 overlaps with the disconnection area of ​​the base groove 100c and the projection of a part of the two sub-base grooves, so that the two sub-base grooves 100c can be connected through the valve action area 511.

[0191] In the absence of external force, the switch valve is in the open state (see Fig. 20 When the fluid flows to the switch valve, the first flexible layer 200 will be deformed by the fluid pressure, so that the minimum gap between the first flexible layer 200 and the first substrate 100 at the pressing valve action area 511 is enlarged, so that the fluid can fully flow through the switch valve in the fully open state (see Fig.21When the switch valve is in an open state, the minimum gap is, for example, in the range of 0.001 mm to 1 mm. When the valve action area 511 is pressed (for example, by pressing the rod), the first flexible layer 200 will deform to fit with the first substrate 100, so that the minimum gap is 0, thereby closing the switch valve (see Fig. 22 ).

[0192] For example, in other embodiments, the opening and closing of the switch valve may also be controlled by other suitable means, such as by an air pump. In this case, the air pump may apply air pressure (positive pressure or negative pressure) to the first flexible layer 200 to deform the first flexible layer 200, thereby controlling the opening and closing of the switch valve. For example, the air pump may also control the degree of opening of the switch valve by the intensity of the air pressure applied to the first flexible layer 200.

[0193] For example, the width of the substrate groove 100c can be in the range of 0.1mm-3mm, and its depth can be in the range of 0.2mm-2mm. For example, the valve action area 511 can be circular, and its diameter is in the range of 0.2mm-6mm. The projection of the valve action area 511 in the direction of the surface of the first substrate 100 overlaps with the projection of the disconnected area of ​​the substrate groove 100c and a part of the two sub-substrate grooves. For example, the area of ​​the overlapping area can be, for example, 5%-50%, for example, 10%-30% of the area of ​​the valve action area 511. Such a range of the area of ​​the overlapping area helps the valve to be completely closed and fully opened.

[0194] The sample chamber will be described below with reference to the accompanying drawings.

[0195] As described above, the first substrate 100 includes the sample cavity 130, which has a sample cavity opening at the first side 100a of the first substrate 100 and a sample cavity opening at the second side 100b of the first substrate 100. The sample cavity opening at the first side 100a of the first substrate 100 is covered by the first flexible layer 200, and the sample cavity opening at the second side 100b of the first substrate 100 is covered by the third cover member 800.

[0196] As one of the storage chambers, the first flexible layer 200 attached to the sample chamber 130 may be in a closed state first. When adding a sample to the sample chamber 130 , the sample added to the sample chamber 130 does not flow to other working areas but remains in the sample chamber 130 .

[0197] Fig.23 A perspective view of a detection chip at a sample cavity 130 according to an embodiment of the present disclosure is shown. Fig.24 Shows Fig.23 Another perspective view of the detection chip at the sample chamber 130, wherein the third cover 800 is removed. Fig.25 Shows Fig.23 A cross-sectional perspective view of the detection chip at the sample cavity 130. Fig.26 A perspective view of a third cover 800 according to an embodiment of the present disclosure is shown.

[0198] like Figure 23-26 As shown, the first substrate 100 includes a third air permeable groove 131 surrounding the sample cavity 130, which is used to selectively connect the sample cavity 130 with the external gas, thereby eliminating the negative pressure state in the sample cavity 130. The third air permeable groove 131 is formed by a third outer wall 132 and a third inner wall 133 between the sample cavity 130 and the third air permeable groove 131. A reinforcing rib connecting the third outer wall 132 and the third inner wall 133 may be provided in the third air permeable groove 131, which is used to enhance the structural strength of the third outer wall 132 and the third inner wall 133, for example, to prevent the third outer wall 132, the third inner wall 133, etc. from being damaged during the process of covering the third cover 800. For example, the reinforcing ribs may be arranged at equal intervals in the circumferential direction.

[0199] like Fig.23 and 24 As shown, the third cover 800 can be opened and closed. The third cover 800 can be opened to add a sample to the sample chamber 130 and then the third cover 800 can be closed.

[0200] The third cover 800 includes a third cover flexible portion 811, a third sealing groove 812 surrounding the third cover flexible portion 811, a third protrusion 813 surrounding the sealing surface, and a third supporting edge 814 surrounding the third protrusion 813. The bottom surface of the third sealing groove 812 abuts against the end surface of the third inner wall 133, the third protrusion 813 is inserted into the third breathable groove 131, and the third supporting edge 814 is supported by the third outer wall 132. The third protrusion 813 is also provided with a third breathable channel 816 facing the third inner wall 133 and extending into the third sealing groove 812. The third breathable channel 816 can be multiple, for example, four, and is arranged at intervals along the circumference of the third cover 800. The third air permeable channel 816 is configured to connect the sample chamber 130 and the third air permeable groove 131 when the third cover flexible portion 811 is not pressed, so that gas can be discharged from the sample chamber 130 into the third air permeable groove 131 and enter the sample chamber 130 from the third air permeable groove 131. By pressing the third cover flexible portion 811, the bottom surface of the third sealing groove 812 will be tightly against the third inner wall 133 so that the sample chamber 130 is sealed. Therefore, when the third cover flexible portion 811 is pressed, the sample chamber 130 cannot be connected to the third air permeable groove 131 through the third air permeable channel 816. In this way, when the third cover flexible portion 811 is deformed by being squeezed (for example, by the push rod 910), the first flexible layer 200 at the sample chamber 130 will be converted from a closed state to an open state, so that the sample chamber 130 is opened and the sample in the sample chamber 130 can flow out. When the external force is removed and the third cover flexible portion 811 is not squeezed, the air outside the detection chip will enter the sample chamber 130 through the third ventilation groove 131 and the third ventilation channel 816 , thereby eliminating the negative pressure state of the sample chamber 130 .

[0201] The thickness of the third inner wall 133 may be in the range of 0.1 mm-5 mm, and the thickness of the third outer wall 132 may be in the range of 0.1 mm-5 mm. The width of the third supporting edge 814 may be in the range of 1 mm-5 mm, so that the third supporting edge 814 has a certain strength.

[0202] In addition, the third cover 800 may include a flip protrusion 815 disposed in the third supporting edge 814 , the position of which corresponds to the flip groove 135 disposed in the third outer wall 132 , so as to facilitate the user to open the third cover 800 .

[0203] The mixing system of the embodiment will be described below with reference to the accompanying drawings.

[0204] As described above, the mixing system includes a first mixing chamber 170 and a second mixing chamber 120, which are connected to each other through the ninth microfluidic channel 9. The first mixing chamber 170 has two first mixing chamber openings at the first side 100a of the first substrate 100 and the second side 100b of the first substrate 100, respectively. The second mixing chamber 120 has two second mixing chamber openings at the first side 100a of the first substrate 100 and the second side 100b of the first substrate 100, respectively. The first mixing chamber opening located at the first side 100a and the second mixing chamber opening located at the first side 100a are covered by the first flexible layer 200, the first mixing chamber opening located at the second side 100b is covered by the first cover 710, and the second mixing chamber opening located at the second side 100b is covered by the second cover 720. In this embodiment, the first cover 710 and the second cover 720 are formed as an integrated mixing cover 700, which is convenient for operation and allows sharing of structures such as the mixing chamber through hole 730 described below, thereby reducing the volume and manufacturing cost of the mixing system.

[0205] Fig. 27 A perspective view of a detection chip at a mixing system according to an embodiment of the present disclosure is shown. Fig.28 Shows Fig. 27 Another perspective view of the detection chip at the mixing system, with the mixing cover 700 removed. Fig.29 Shows Fig. 27 Another perspective view of the detection chip at the mixing system, where the third gas isolation membrane 630 is removed. Fig.30 A perspective view of a mixing cap 700 according to an embodiment of the present disclosure is shown. Fig.31 A perspective view of a bond ring 740 is shown in accordance with an embodiment of the present disclosure.

[0206] Fig.32 A cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown. Fig.33 A top view of a base groove 100c at a mixing system according to an embodiment of the present disclosure is shown.

[0207] like Figure 27-33As shown, the first substrate 100 includes a first air-permeable groove 171 surrounding the first mixing cavity 170 and a second air-permeable groove 121 surrounding the second mixing cavity 120, the first air-permeable groove 171 is formed by a first outer wall 172 and a first inner wall 173 between the first mixing cavity 170 and the first air-permeable groove 171, and the second air-permeable groove 121 is formed by a second outer wall 122 and a second inner wall 123 between the first mixing cavity 170 and the second air-permeable groove 121. A reinforcing rib connecting the first outer wall 172 and the first inner wall 173 may be provided in the first air-permeable groove 171 to enhance the structural strength of the first mixing cavity 170. A reinforcing rib connecting the second outer wall 122 and the second inner wall 123 may be provided in the second air-permeable groove 121 to enhance the structural strength of the second mixing cavity 120.

[0208] The first cover 710 includes a first cover flexible portion 711, a first sealing groove 712 surrounding the first cover flexible portion 711, a first protrusion 713 surrounding the first sealing groove 712, and a first supporting edge 714 surrounding the first protrusion 713. The bottom surface of the first sealing groove 712 abuts against the end surface of the first inner wall 173, the first protrusion 713 is inserted into the first venting groove 171, and the first supporting edge 714 is supported by the first outer wall 172. The first protrusion 713 is also provided with a first venting channel 716 facing the first inner wall 173 and extending into the first sealing groove 712. The first venting channel 716 is configured to connect the first mixing chamber 170 and the first venting groove 171 when the first cover flexible portion 711 is not pressed, so that gas can be discharged from the first mixing chamber 170 to the first venting groove 171 through it and enter the first mixing chamber 170 from the first venting groove 171. By pressing the first cover flexible portion 711, the bottom surface of the first sealing groove 712 will be tightly against the first inner wall 173 so that the first mixing chamber 170 is sealed. Therefore, when the first cover flexible portion 711 is pressed, the first mixing chamber 170 cannot be communicated with the first breathable groove 171 through the first breathable channel 716. In this way, when the first cover flexible portion 711 is deformed due to being squeezed (for example, by the ejector 910), the first flexible layer 200 at the first mixing chamber 170 will be converted from a closed state to an open state, so that the first mixing chamber 170 is opened and the reagent in the first mixing chamber 170 can flow out. When the external force is removed and the first cover flexible portion 711 is not squeezed, the air outside the detection chip will enter the first mixing chamber 170 through the first breathable groove 171 and the first breathable channel 716, thereby eliminating the negative pressure state of the first mixing chamber 170.

[0209] The second cover 720 includes a second cover flexible portion 721, a second sealing groove 722 surrounding the second cover flexible portion 721, a second protrusion 723 surrounding the second sealing groove 722, and a second supporting edge 724 surrounding the second protrusion 723. The bottom surface of the second sealing groove 722 abuts against the end surface of the second inner wall 123, the second protrusion 723 is inserted into the second venting groove 121, and the second supporting edge 724 is supported by the second outer wall 122. The second protrusion 723 is also provided with a second venting channel 726 facing the second inner wall 123 and extending into the second sealing groove 722. The second venting channel 726 is configured to connect the second mixing chamber 120 and the second venting groove 121 when the second cover flexible portion 721 is not pressed, so that gas can be discharged from the second mixing chamber 120 into the second venting groove 121 and enter the second mixing chamber 120 from the second venting groove 121. By pressing the second cover flexible part 721, the bottom surface of the second sealing groove 722 will be tightly against the second inner wall 123 so that the second mixing chamber 120 is sealed. Therefore, when the second cover flexible part 721 is pressed, the second mixing chamber 120 cannot be communicated with the second breathable groove 121 through the second breathable channel 726. In this way, when the second cover flexible part 721 is deformed by being squeezed (for example, by the ejector 910), the second flexible layer 300 at the second mixing chamber 120 will be converted from a closed state to an open state, so that the second mixing chamber 120 is opened and the reagent in the second mixing chamber 120 can flow out. When the external force is removed and the second cover flexible part 721 is not squeezed, the air outside the detection chip will enter the second mixing chamber 120 through the second breathable groove 121 and the second breathable channel 726, thereby eliminating the negative pressure state of the second mixing chamber 120.

[0210] In this embodiment, the first ventilation groove 171 and the second ventilation groove 121 are connected to each other through the connecting ventilation groove 174. The first cover 710 and the second cover 720 are formed as an integrated mixing cover 700.

[0211] The mixing cover 700 is provided with a mixing cavity through hole 730 at the connection air-permeable groove 174. The detection chip also includes a third gas isolation membrane 630 attached to the mixing cover 700 at the mixing cavity through hole 730, so that the connection air-permeable groove 174 is connected to the external gas of the detection chip through the third gas isolation membrane 630 and the mixing cavity through hole 730 and is liquid-isolated. When the first cover 710 and the second cover 720 are separately arranged, the through hole in the first cover 710, the through hole in the second cover 720 and the corresponding gas isolation membrane can be respectively arranged at the first air-permeable groove 171 and the second air-permeable groove 121. The third gas isolation membrane 630 can be a composite membrane, which includes a polyester substrate and a hydrophobic ePTFE membrane. In addition, in the illustrated embodiment, the third gas isolation membrane 630 is attached to the mixing cover 700 from the side of the mixing cover 700 away from the first substrate 100. However, those skilled in the art will appreciate that the third gas barrier film 630 may also be attached to the mixing cover 700 from the other side of the mixing cover 700 .

[0212] In addition, the mixing chamber further includes a bonding ring 740, which is disposed between the first supporting edge 714 and the first outer wall 172 and between the second supporting edge 724 and the second outer wall 122, so as to bond the first supporting edge 714 to the first outer wall 172 and the second supporting edge 724 to the second outer wall 122. Those skilled in the art should understand that when the first cover 710 and the second cover 720 are separately disposed, the bonding ring 740 will also be separated into two separate bonding rings.

[0213] When performing a mixing step, for example, transferring the fluid in the second mixing chamber 120 to the first mixing chamber 170, the ninth switch valve 91 can be opened to squeeze the second cover flexible portion 721 of the second cover member 720 of the second mixing chamber 120 to deform it. When the second cover flexible portion 721 is squeezed, the second mixing chamber 120 is isolated from the external air, and the reagent in the second mixing chamber 120 will flow into the first mixing chamber 170 via the ninth microfluidic channel 9. At this time, the air in the first mixing chamber 170 will be discharged to the outside of the first mixing chamber 170 through the first air permeable channel 716, the first air permeable groove 171, the mixing chamber through hole 730, and the third gas isolation membrane 630, so that the reagent can flow smoothly from the second mixing chamber 120 to the first mixing chamber 170. When the second mixing chamber 120 is no longer squeezed, external air will enter the second mixing chamber 120 through the second air permeable channel 726, the second air permeable groove 121, the mixing chamber through hole 730, and the third gas isolation membrane 630 to eliminate the negative pressure state in the second mixing chamber 120 caused by the second cover flexible portion 721 no longer deforming. The process of transferring the fluid in the first mixing chamber 170 to the second mixing chamber 120 is similar. Fig.32The arrows in FIG. 1 show the flow of air and reagent during the process of transferring the fluid in the second mixing chamber 120 to the first mixing chamber 170 .

[0214] When the step of releasing the reagent into the first mixing chamber 170 is performed, the air in the first mixing chamber 170 will be discharged to the outside of the first mixing chamber 170 through the first air permeable channel 716, the first air permeable groove 171, the mixing chamber through hole 730, and the third gas isolation membrane 630, so that the reagent can flow smoothly into the first mixing chamber 170.

[0215] Figures 34A-34D Another cross-sectional view of the mixing system according to an embodiment of the present disclosure is shown, which illustrates the steps of re-dissolving the magnetic beads 930 and mixing them with the reagents, and the arrows in the figure indicate the flow of air.

[0216] like Fig.34A As shown, the magnet 920 is moved away from the second mixing chamber 120, and the reagent is placed in the first mixing chamber 170. The ninth switch valve 91 is opened, and the reagent flows from the first mixing chamber 170 to the second mixing chamber 120 via the ninth microchannel 9 by squeezing the first cover flexible portion 711 of the first cover member 710 to redissolve the magnetic beads 930 in the second mixing chamber 120. At this time, the air is discharged to the outside of the second mixing chamber 120 via the second air permeable channel 726, the second air permeable groove 121, the mixing chamber through hole 730, and the third gas isolation membrane 630. Fig.34B As shown, the squeezing of the first cover flexible portion 711 of the first cover member 710 is released. At this time, air enters the first mixing chamber 170 through the first air permeable channel 716, the first air permeable groove 171, the mixing chamber through hole 730, and the third gas isolation membrane 630. Fig.34C As shown, by squeezing the second cover flexible portion 721 of the second cover member 720, the reagent mixed with the magnetic beads 930 flows from the second mixing chamber 120 to the first mixing chamber 170 via the ninth microfluidic channel 9. Fig.34D As shown, the squeezing of the second cover flexible portion 721 of the second cover member 720 is released. At this time, air enters the second mixing chamber 120 through the second air permeable channel 726, the second air permeable groove 121, the mixing chamber through hole 730, and the third gas isolation membrane 630.

[0217] Figures 35A-35D Another cross-sectional view of the mixing system according to an embodiment of the present disclosure is shown, which illustrates the magnetic beads 930 collection and mixing steps, and the arrows in the figure indicate the flow of air.

[0218] like Fig.35AAs shown, the magnet 920 is away from the second mixing chamber 120, and the reagent mixed with the magnetic beads 930 is placed in the first mixing chamber 170. The ninth switch valve 91 is opened, and the reagent mixed with the magnetic beads 930 flows from the first mixing chamber 170 to the second mixing chamber 120 via the ninth microchannel 9 by squeezing the first cover flexible portion 711 of the first cover member 710. At this time, the air is discharged to the outside of the second mixing chamber 120 via the second air permeable channel 726, the second air permeable groove 121, the mixing chamber through hole 730, and the third gas isolation membrane 630. Fig.35B As shown, the squeezing of the first cover flexible portion 711 of the first cover member 710 is released, and the magnet 920 is moved close to the second mixing chamber 120. At this time, air enters the first mixing chamber 170 through the first air permeable channel 716, the first air permeable groove 171, the mixing chamber through hole 730, and the third gas isolation membrane 630. Fig.35C As shown, by squeezing the second cover flexible portion 721 of the second cover member 720, the reagent flows from the second mixing chamber 120 to the first mixing chamber 170 via the ninth microfluidic channel 9. At this time, due to the attraction of the magnet 920 to the magnetic beads 930, the magnetic beads 930 remain in the second mixing chamber 120. Fig.35D As shown, the squeezing of the second cover flexible portion 721 of the second cover member 720 is released. At this time, air enters the second mixing chamber 120 through the second air permeable channel 726, the second air permeable groove 121, the mixing chamber through hole 730, and the third gas isolation membrane 630.

[0219] The waste liquid chamber of the embodiment of the present disclosure will be introduced below with reference to the accompanying drawings.

[0220] like Figure 1-7As shown, the first substrate 100 includes a waste liquid chamber 190, which has a waste liquid chamber opening on the first side 100a of the first substrate 100 and a waste liquid chamber opening on the second side 100b of the first substrate 100. The waste liquid chamber opening on the first side 100a of the first substrate 100 is covered by the first flexible layer 200, and the waste liquid chamber opening on the second side 100b of the first substrate 100 is covered by the second flexible layer 300 for collecting and storing waste liquid. A waste liquid chamber through hole 311 may be provided in the first portion 310 of the second flexible layer 300, and the waste liquid chamber through hole 311 is aligned with the waste liquid chamber 190 in the first substrate 100. In addition, the detection chip also includes a first gas isolation membrane 610, which is attached to the waste liquid chamber through hole 311 of the second flexible layer 300 and can be a waterproof and breathable membrane that prevents liquid from penetrating and allows gas to penetrate. Therefore, the waste liquid chamber 190 is gaseously connected and liquid-isolated with the outside of the detection chip through the first gas isolation membrane 610 and the waste liquid chamber through hole 311. Therefore, the waste liquid can flow into the waste liquid chamber 190 while the gas in the waste liquid chamber 190 flows out of the waste liquid chamber 190, while the waste liquid in the waste liquid chamber 190 will not flow out of the waste liquid chamber 190 and pollute the detection environment. The first gas isolation membrane 610 can be a composite membrane, which includes a polyester substrate and a hydrophobic ePTFE membrane. In addition, in the illustrated embodiment, the first gas isolation membrane 610 is attached to the second flexible layer 300 from the side of the second flexible layer 300 away from the first substrate 100. However, it should be understood by those skilled in the art that the first gas isolation membrane 610 can also be attached to the second flexible layer 300 from the other side of the second flexible layer 300.

[0221] The amplification chamber of the embodiment of the present disclosure will be introduced below with reference to the accompanying drawings.

[0222] Fig.36 and Fig.37 A partial top view and a partial bottom view of the detection chip are shown respectively.

[0223] As described above, the second base 400 is attached to the side of the first flexible layer 200 opposite to the first base 100. Fig.36 and Fig.37 As shown, the first substrate 100 includes a first positioning structure 1140, and the second substrate 400 includes a second positioning structure 420. The first positioning structure 1140 corresponds to the position of the second positioning structure 420 to position the second substrate 400 relative to the first substrate 100. The second substrate 400 is attached to the first flexible layer 200 by attaching the bonding surface of the second substrate 400 to the first flexible layer 200.

[0224] The second substrate 400 is formed with an amplification chamber 410 configured to store amplification reagents. For example, the amplification reagents may be freeze-dried molecules pre-stored in the amplification chamber 410, which include enzymes, probes, primers and other reagents required for amplification.

[0225] like Figure 8 , Fig.36 , Fig.37 As shown, one end of the amplification chamber 410 is closed by itself, and the other end has an opening. The opening can be in the shape of an elongated strip. The second substrate 400 is positioned relative to the first substrate 100 so that one end of the opening is aligned with the first flow channel outlet 210 in the first flexible layer 200 and the first intermediate layer through hole 520 in the intermediate layer 500, and the other end of the opening is aligned with the second flow channel outlet 220 in the first flexible layer 200 and the second intermediate layer through hole 530 in the intermediate layer 500. Therefore, one end of the opening of the amplification chamber 410 is connected to the seventh microfluidic channel 7 through the first flow channel outlet 210 and the first intermediate layer through hole 520 in the intermediate layer 500, and the other end of the opening is connected to the eighth microfluidic channel 8 through the second flow channel outlet 220 and the second intermediate layer through hole 530 in the intermediate layer 500. The seventh microfluidic channel 7 and the eighth microfluidic channel 8 are respectively connected to the two ends of the long strip opening, which helps to fill the amplification chamber 410 with the reagent to be amplified (for example, an eluent containing nucleic acid molecules) and prevent the generation of bubbles, thereby achieving accurate quantification of the volume of the reagent to be detected entering the amplification chamber 410. Therefore, the reagent to be amplified can flow from the first mixing chamber 170 to the amplification chamber 410 via the seventh microfluidic channel 7 and one end of the opening, and after filling the amplification chamber 410, it flows to the air permeable chamber 1100 via the other end of the opening and the eighth microfluidic channel 8, and the air permeable chamber 1100 can discharge air to the outside of the detection chip.

[0226] The size of the first flow channel outlet 210 in the first flexible layer 200 and the first intermediate layer through hole 520 in the intermediate layer 500 is designed to be larger than the width of the seventh microfluidic channel 7, and the size of the second flow channel outlet 220 in the first flexible layer 200 and the second intermediate layer through hole 530 in the intermediate layer 500 is designed to be larger than the width of the eighth microfluidic channel 8, so as to avoid the blockage of the seventh microfluidic channel 7 and the eighth microfluidic channel 8 due to the alignment error during the manufacture of the detection chip. For example, the seventh microfluidic channel 7 and the eighth microfluidic channel 8 can be 1 mm, and the diameters of the first flow channel outlet 210 and the second flow channel outlet 220 in the first flexible layer 200 and the first intermediate layer through hole 520 and the second intermediate layer through hole 530 in the intermediate layer 500 can be 1.2 mm.

[0227] In an embodiment according to the present disclosure, by reasonably designing the shape and size of the amplification chamber 410, the detected reagent entering the amplification chamber 410 can actively wet the volume of the amplification chamber 410 under capillary action, reducing the generation of bubbles in the amplification chamber 410. For example, the volume of the amplification chamber 410 can be in the range of 10-100 μL, such as 50 μL. At the same nucleic acid fragment concentration, the larger the volume of the amplification system, the lower the detection limit, the higher the sensitivity, and the better the chip performance; on the other hand, the larger the volume of the amplification system, the slower the heating and cooling rate of the amplification system at the same power, the longer the amplification time, and the worse the chip performance. The heating efficiency and the detection limit can be balanced at a reagent volume of 50 μL. By filling the reagent with a rigid amplification chamber 410 with a fixed volume, the volume of the detected reagent entering the amplification chamber 410 can be quantified. For example, the width of the opening can be in the range of 0.1 mm-3 mm, such as 1.5 mm. A smaller opening width is conducive to the detection reagent flowing to the amplification chamber 410 by means of capillary force.

[0228] The second substrate 400 is injection molded from a polymer material such as PMMA, PC, PP, PS, etc., preferably PP and PC. In addition, the second substrate 400 may be surface-modified so that enzymes, nucleic acid molecules, etc. are not easily adhered to the surface of the second substrate 400.

[0229] The second substrate 400 can be an integrated part, so it has a simple structure and is easy to process, thereby reducing the cost of the detection chip.

[0230] For example, the second substrate 400 can be made of a material that is transparent to the detection light. The detection light includes light that is incident into the amplification chamber 410 to excite the reagent in the amplification chamber 410 to emit a fluorescent signal, and also includes a fluorescent signal emitted by the reagent in the amplification chamber 410. The second substrate 400 made of a transparent material helps to optically detect the amplification reaction occurring in the amplification chamber 410. For example, the wall thickness of the amplification chamber 410 can be in the range of 0.1 mm-3 mm, such as 0.5 mm. The thin wall thickness can improve the heating efficiency of the detection device for the reagent in the amplification chamber 410.

[0231] A gas permeable cavity through hole 312 may be provided in the first part 310 of the second flexible layer 300, and the gas permeable cavity through hole 312 is aligned with the gas permeable cavity 1100 in the first substrate 100. In addition, the detection chip also includes a second gas isolation membrane 620, which is attached to the gas permeable cavity through hole 312 of the second flexible layer 300 and can be a waterproof gas permeable membrane that prevents liquid from penetrating and allows gas to penetrate. Therefore, the gas permeable cavity 1100 is connected to the external gas of the detection chip and liquid-isolated through the second gas isolation membrane 620 and the gas permeable cavity through hole 312. The waterproof performance of the gas permeable cavity 1100 prevents aerosols and volatile steam from being discharged from the gas permeable cavity 1100 during the amplification process, and the gas permeability of the gas permeable cavity 1100 enables the air inside the detection chip to be discharged when the detected reagent enters the amplification cavity 410. The second gas isolation membrane 620 can be a composite membrane, which includes a polyester substrate and a hydrophobic ePTFE membrane. Furthermore, in the illustrated embodiment, the second gas barrier film 620 is attached to the second flexible layer 300 from a side of the second flexible layer 300 that is away from the first substrate 100. However, those skilled in the art will appreciate that the second gas barrier film 620 may also be attached to the second flexible layer 300 from the other side of the second flexible layer 300.

[0232] The detection chip of the embodiment of the present disclosure can be installed in a proper position of the detection device so as to cooperate with the detection device to complete the detection process.

[0233] The detection chip includes a first chip positioning structure for positioning the chip to a suitable position of the detection device. The first chip positioning structure may include a chip positioning hole 1120 provided in the first substrate 100, which may be a circular through hole with a diameter in the range of 0.1 mm-10 mm, and is used to cooperate with a positioning pin as a second chip positioning structure on the detection device to position the chip on the detection device. In addition, the first chip positioning structure may include a long chip positioning groove 1130 located on both sides of the detection chip, which is used to match with a fixing device on the detection device to fix the chip to the detection device.

[0234] like Figure 4 , Figure 5 and Figure 7 As shown, the first flexible layer 200 and the intermediate layer 500 are both provided with through holes which are aligned with the chip positioning holes 1120 on the first substrate 100 and have the same size as each other, and are used to cooperate with the positioning pins on the detection device.

[0235] The detection device may include the push rod 910 as described above as the first operating part for changing the volume of the storage chamber. There may be multiple push rods 910, and the multiple push rods 910 are respectively arranged above different storage chambers to change the volume of the storage chamber. Alternatively, the detection device may include a push rod 910 that can move above different storage chambers.

[0236] The detection device may include a second operating part, such as a pressing rod, which presses the second flexible layer 300 to operate the switch valve. The pressing rod may include a plurality of pressing rods or be configured to be movable.

[0237] In addition, the detection device may also include a heating device for heating the amplification chamber 410, a detection device for detecting the amplification reaction in the amplification chamber 410, a heating device for heating the first mixing chamber 170 and the second mixing chamber 120, a magnet 920 for fixing magnetic beads 930, etc.

[0238] The scope of the present disclosure is not limited by the above-described embodiments but by the appended claims and their equivalents.

Claims

1. A detection chip, comprising: A first substrate including at least one storage cavity; as well as a first flexible layer attached to a first side of the first substrate, The storage cavity has a first opening on the first side, and the first flexible layer is located at the first opening of the at least one storage cavity and is configured to be in an open state or a closed state. The first side of the first substrate comprises a first bonding area and a second bonding area, When the first flexible layer is in the closed state, the first flexible layer is attached to the first side of the first substrate at the first bonding area and the second bonding area to close the first opening of the storage cavity, When the first flexible layer is in the open state, the first flexible layer is attached to the first side of the first substrate at the second bonding area and is separated from the first substrate at the first bonding area to open the first opening of the storage cavity, The at least one storage cavity includes a plurality of storage cavities, and the surface of the first substrate on the first side includes a plurality of substrate grooves for forming a plurality of microchannels, the plurality of storage cavities include at least one liquid storage cavity, and the at least one liquid storage cavity includes an elution liquid cavity pre-stored with an elution liquid, a first cleaning liquid cavity pre-stored with a first cleaning liquid, and a lysis liquid cavity pre-stored with a lysis liquid.

2. The detection chip according to claim 1, wherein: A first attachment strength of the first flexible layer to the first substrate at the first bonding area is less than a second attachment strength of the first flexible layer to the first substrate at the second bonding area.

3. The detection chip according to claim 2, wherein: The surface of the first substrate on the first side has a plurality of concavo-convex structures at the first bonding area.

4. The detection chip according to claim 3, wherein: The maximum depth difference of the plurality of concavo-convex structures in a direction perpendicular to the surface is in the range of 0.1 mm to 3 mm.

5. The detection chip according to any one of claims 2 to 4, wherein: The first attachment strength is in the range of 1N-50N, and the second attachment strength is in the range of 50N-500N.

6. The detection chip according to claim 1, wherein: The surface of the first substrate on the first side is flush at the first bonding area and the second bonding area.

7. The detection chip according to claim 1, wherein: The first substrate is provided with a buffer groove and a substrate groove communicating with the buffer groove and used to form at least one microchannel on the surface of the first side. The buffer groove is disposed at least partially around the first opening of the storage cavity to define the first bonding area at an inner side of the buffer groove and the second bonding area at an outer side of the buffer groove.

8. The detection chip according to claim 7, wherein: The depth of the buffer groove in a vertical direction perpendicular to the surface is in the range of 0.01-5 mm.

9. The detection chip according to claim 7, wherein: The width of the buffer groove in a horizontal direction parallel to the surface is in the range of 0.01 mm to 5 mm.

10. The detection chip according to claim 7, wherein: The bottom surface of the buffer tank includes a hydrophobic surface.

11. The detection chip according to claim 7, wherein: The first substrate is an annular boss at the first bonding area, and the width of the annular boss is in the range of 0.01 mm-5 mm.

12. The detection chip according to claim 11, wherein: The cross section of the annular boss decreases toward the first side.

13. The detection chip according to claim 11, wherein: The annular boss includes a plurality of sub-annular bosses.

14. The detection chip according to claim 1, wherein: At least some of the plurality of microfluidic channels are provided with switch valves for controlling the connection and disconnection of the corresponding at least some of the microfluidic channels.

15. The detection chip according to claim 14, further comprising: an intermediate layer, the first flexible layer being adhered to the first side of the first substrate by the intermediate layer, The intermediate layer includes a plurality of intermediate through grooves penetrating the intermediate layer, and the positions of the plurality of intermediate through grooves correspond to the positions of the plurality of base grooves to form the plurality of microchannels. The switch valve includes a disconnection area arranged in the base groove to disconnect the base groove into two sub-base grooves, and a valve action area arranged in the middle through groove. The projection of the valve action area in the direction of the surface of the first base covers the disconnection area of ​​the base groove and a part of the two sub-base grooves, so that the two sub-base grooves can be connected through the valve action area.

16. The detection chip according to claim 1, wherein: The plurality of storage chambers include at least one liquid storage chamber, wherein the at least one liquid storage chamber stores a reagent in advance, and the reagent is sealed in the liquid storage chamber.

17. The detection chip according to claim 1, wherein: The detection chip also includes a second flexible layer, which is attached to a second side of the first substrate opposite to the first side at the at least one liquid storage cavity, the liquid storage cavity has a second opening on the second side, and the second flexible layer at least closes the second opening of the liquid storage cavity.

18. The detection chip according to claim 17, wherein: The first substrate further includes a mixing chamber system, wherein the mixing chamber system includes a first mixing chamber and a second mixing chamber. The multiple microfluidic channels include a first microfluidic channel connecting the mixing chamber system and the lysis liquid chamber, a second microfluidic channel connecting the mixing chamber system and the first cleaning liquid chamber, a third microfluidic channel connecting the mixing chamber system and the elution liquid chamber, and a ninth microfluidic channel connecting the first mixing chamber and the second mixing chamber.

19. The detection chip according to claim 18, wherein: The diameter of the elution liquid chamber is smaller than the diameters of the lysis liquid chamber and the first cleaning liquid chamber.

20. The detection chip according to claim 18, wherein: The at least one liquid storage chamber further comprises a second cleaning liquid chamber pre-stored with a second cleaning liquid and a third cleaning liquid chamber pre-stored with a third cleaning liquid, The plurality of microchannels further include a fourth microchannel connecting the mixing chamber system and the third cleaning liquid chamber, and a fifth microchannel connecting the mixing chamber system and the second cleaning liquid chamber.

21. The detection chip according to claim 20, wherein: The first mixing chamber and the second mixing chamber are arranged in the middle of the first substrate, and the at least one liquid storage chamber is arranged on both sides of the middle.

22. The detection chip according to claim 21, wherein: The heights of the first mixing chamber and the second mixing chamber are higher than the heights of the lysing liquid chamber, the first cleaning liquid chamber, the second cleaning liquid chamber, and the third cleaning liquid chamber.

23. The detection chip according to claim 20, wherein: The first microfluidic channel connects the first mixing chamber and the lysis liquid chamber, the second microfluidic channel connects the first mixing chamber and the first cleaning liquid chamber, the third microfluidic channel connects the second mixing chamber and the elution liquid chamber, the fourth microfluidic channel connects the first mixing chamber and the third cleaning liquid chamber, and the fifth microfluidic channel connects the fourth microfluidic channel and the second cleaning liquid chamber.

24. The detection chip according to claim 18, wherein: The first substrate further includes a waste liquid chamber, the waste liquid chamber includes two waste liquid chamber openings respectively on the first side and the second side, the two waste liquid chamber openings are respectively covered by the first flexible layer and the second flexible layer, the plurality of microchannels further includes a sixth microchannel connecting the first mixing chamber and the waste liquid chamber, The second flexible layer is provided with a waste liquid cavity through hole, The detection chip also includes a first gas isolation membrane attached to the second flexible layer at the waste liquid chamber through hole, wherein the first gas isolation membrane enables the waste liquid chamber to be gaseously connected and liquid-isolated from the outside of the detection chip through the first gas isolation membrane and the waste liquid chamber through hole.

25. The detection chip according to claim 17, further comprising: The second substrate comprises an amplification chamber configured to store an amplification reagent, wherein: One end of the amplification chamber is closed, the other end has an amplification chamber opening and is attached to a side of the first flexible layer opposite to the first substrate, The first substrate further includes a first mixing cavity and a gas permeable cavity which is connected to the external gas of the detection chip but isolated from the liquid. The multiple microfluidic channels include a seventh microfluidic channel and an eighth microfluidic channel. The amplification chamber is connected to the first mixing chamber through a first channel outlet formed in the first flexible layer and the seventh microfluidic channel. The amplification chamber is connected to the air permeable chamber through a second channel outlet formed in the first flexible layer and the eighth microfluidic channel.

26. The detection chip according to claim 25, wherein: The second substrate is a one-piece piece.

27. The detection chip according to claim 25, wherein The amplification chamber opening is in the shape of an elongated strip, and the width of the opening is in the range of 0.1 mm to 3 mm.

28. The detection chip according to claim 25, wherein The amplification chamber is in an elongated strip shape, and the first flow channel outlet and the second flow channel outlet are respectively arranged at two ends of the amplification chamber opening.

29. The detection chip according to claim 25, wherein The second substrate is made of a material that is transparent to the detection light used for the detection chip, and the wall thickness of the amplification chamber is in the range of 0.1 mm to 3 mm.

30. The detection chip according to claim 25, wherein One end of the air permeable cavity is covered by the first flexible layer, The detection chip further includes a second gas isolation membrane attached to the second side of the first substrate to cover the other end of the gas permeable cavity, and the gas permeable cavity is gaseously connected and liquid-isolated from the outside of the detection chip through the second gas isolation membrane.

31. The detection chip according to claim 25, wherein Both ends of the air permeable cavity are covered by the first flexible layer and the second flexible layer respectively. The second flexible layer is provided with a vent hole. The detection chip further includes a second gas isolation membrane attached to the second flexible layer at the gas permeable cavity through hole, and the gas permeable cavity is connected to and isolated from external gas of the detection chip through the second gas isolation membrane and the gas permeable cavity through hole.

32. The detection chip according to claim 25, wherein The first substrate includes a first positioning structure, The second substrate includes a second positioning structure, The positions of the first positioning structure and the second positioning structure correspond to each other so as to position the second substrate relative to the first substrate.

33. The detection chip according to claim 11, wherein: The first matrix further includes a first mixing chamber and a second mixing chamber. The first mixing chamber comprises two first mixing chamber openings respectively located at the first side of the first substrate and at a second side of the first substrate opposite to the first side, and the two first mixing chamber openings are respectively covered by the first flexible layer and the first cover. The second mixing chamber comprises two second mixing chamber openings respectively located on the first side of the first substrate and the second side of the first substrate, and the two second mixing chamber openings are respectively covered by the first flexible layer and the second cover. The plurality of microchannels include a ninth microchannel, and the ninth microchannel connects the first mixing chamber and the second mixing chamber. The first substrate further includes a first air permeable groove surrounding the first mixing cavity and a second air permeable groove surrounding the second mixing cavity. The first air permeable groove comprises a first outer wall and a first inner wall between the first mixing cavity and the first air permeable groove. The second air permeable groove includes a second outer wall and a second inner wall between the first mixing cavity and the second air permeable groove.

34. The detection chip according to claim 33, wherein: The first cover member comprises a first cover flexible portion, a first sealing groove surrounding the first cover flexible portion, a first protrusion surrounding the first sealing groove, and a first supporting edge surrounding the first protrusion. The bottom surface of the first sealing groove abuts against the end surface of the first inner wall, the first protrusion is inserted into the first breathable groove, the first supporting edge is supported by the first outer wall, and the first protrusion is also provided with a first breathable channel facing the first inner wall and extending into the first sealing groove.

35. The detection chip according to claim 34, further comprising an adhesive ring, in, The bonding ring is arranged between the end surface of the first supporting edge and the end surface of the first outer wall to bond the first supporting edge and the first outer wall. The second cover member includes a second cover flexible portion, a second sealing groove surrounding the second cover flexible portion, a second protrusion surrounding the second sealing groove, and a second supporting edge surrounding the second protrusion. The bonding ring is also arranged between the end surface of the second supporting edge and the end surface of the second outer wall to bond the second supporting edge and the second outer wall.

36. The detection chip according to claim 34, wherein The first cover and the second cover form an integrated mixing cover, the first ventilation groove and the second ventilation groove are connected through a connecting ventilation groove, and the mixing cover is provided with a mixing cavity through hole at the connecting ventilation groove. The detection chip further includes a third gas isolation membrane attached to the mixing cover at the mixing cavity through hole, and the connecting gas permeable groove is connected and isolated from the external gas of the detection chip through the third gas isolation membrane and the mixing cavity through hole.

37. The detection chip according to claim 1, wherein: The storage chamber includes a sample chamber, and the first matrix also includes a first mixing chamber, The sample chamber comprises two sample chamber openings respectively on the first side of the first substrate and on the second side of the first substrate opposite to the first side, the two sample chamber openings are respectively covered by the first flexible layer and the third cover, the plurality of microfluidic channels comprises a tenth microfluidic channel, the tenth microfluidic channel connects the first mixing chamber and the sample chamber, The first substrate further includes a third gas permeable groove surrounding the sample cavity, and the third gas permeable groove includes a third outer wall and a third inner wall between the sample cavity and the third gas permeable groove.

38. The detection chip according to claim 1, further comprising: The first chip positioning structure is configured to position and install the detection chip to the detection device.

39. A detection device comprising A second chip positioning structure is configured to install the detection chip according to any one of claims 1 to 38, and The first operating portion is configured to be operable to change the volume of the storage chamber.

40. A detection device comprising The second chip positioning structure is configured to install the detection chip as claimed in claim 14, wherein: The detection chip further includes a second flexible layer attached to a second side of the first substrate opposite to the first side at the at least one liquid storage cavity, and The second operating portion is configured to be operable to press the second flexible layer to operate the switch valve.

41. A method for operating the detection chip according to claim 1, comprising: adding a sample to the detection chip; Mixing the sample with a lysing solution to form a mixed solution, wherein the lysing solution can lyse the sample to form a substance to be detected; Driving the mixed liquid to flow to a mixing chamber system, wherein the mixing chamber system comprises a first mixing chamber and a second mixing chamber, and the mixing chamber system has magnetic beads, and the magnetic beads can adsorb the substance to be detected; Driving the mixed solution to move back and forth between the first mixing chamber and the second mixing chamber to mix the mixed solution and the magnetic beads; Separating and treating the magnetic beads to make the substance to be detected fall off the magnetic beads; and The substance to be detected is detected.

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