Detection chip and detection device

Abstract

A detection chip and a detection device are provided. The detection chip comprises: a substrate having a first surface and comprising at least one cavity having a first opening in the first surface; at least one storage container movably accommodated in the at least one cavity through the first opening, the at least one storage container comprising a second opening and a first sealing film sealing the second opening; wherein the at least one cavity comprises a puncture structure arranged in the cavity for puncturing the first sealing film when the storage container moves to make the first sealing film contact the puncture structure, so that a reagent in the storage container enters the cavity.

Description

Technical Field

[0001] Embodiments of this disclosure relate to a detection chip and a detection device that can be used with the detection chip. Background Technology

[0002] Microfluidic chip technology integrates basic operational units involved in sample preparation, reaction, separation, and detection in fields such as biology, chemistry, and medicine onto a chip with micron-scale microchannels, automating the entire process of reaction and analysis. The chip used in this process is called a microfluidic chip, also known as a lab-on-a-chip. Microfluidic chip technology offers advantages such as small sample volume, fast analysis speed, ease of fabrication into portable instruments, and suitability for point-of-care and on-site analysis, and has been widely applied in many fields including biology, chemistry, and medicine. Summary of the Invention

[0003] According to at least one embodiment of the present disclosure, a detection chip is provided, the detection chip including a substrate and at least one storage container; the substrate has a first surface and includes at least one cavity and at least one fluid channel leading to the at least one cavity, the cavity having a first opening in the first surface; the at least one storage container is movably accommodated in the at least one cavity through the first opening, the at least one storage container including a second opening and a first sealing membrane sealing the second opening; wherein, the at least one cavity includes a puncture structure disposed in the cavity for puncturing the first sealing membrane to allow reagents in the storage container to enter the fluid channel.

[0004] For example, in some embodiments, the at least one storage container further includes a third opening and a second sealing membrane sealing the third opening, with the first sealing membrane and the second sealing membrane defining a liquid storage space for containing reagents.

[0005] For example, in some embodiments, the storage container further includes a removable lid that is installed at the third opening and covers the second sealing film.

[0006] For example, in some embodiments, the detection chip further includes a sealing cap that is connected to the substrate and seals a first opening in the substrate.

[0007] For example, in some embodiments, the sealing cap snaps onto the substrate.

[0008] For example, in some embodiments, the sealing cap includes a cap body and a snap-fit ​​assembly, the snap-fit ​​assembly including at least one first snap disposed on the outer peripheral portion of the cap body and a second snap disposed on the central portion of the cap body.

[0009] For example, in some embodiments, the sealing cap further includes a sealing ring disposed inside the outer peripheral portion of the cap body, wherein the cap body and the snap-fit ​​assembly are formed of a rigid material, and the sealing ring is formed of a flexible material.

[0010] For example, in some embodiments, the cover body, the snap-fit ​​assembly, and the sealing ring are integrally formed by two-color injection molding or assembled by overmolding.

[0011] For example, in some embodiments, the sealing cap is provided with a covering film at the first opening, the covering film being an elastic film.

[0012] For example, in some embodiments, the surface of the covering film facing the first opening is provided with a protruding structure.

[0013] For example, in some embodiments, the protrusion structure includes a central protrusion and a plurality of peripheral protrusions distributed circumferentially around the central protrusion, wherein the central protrusion is in the shape of a line, a cross, or a circle, and the plurality of peripheral protrusions are in the shape of a line or a circle.

[0014] For example, in some embodiments, the detection chip further includes at least one first seal disposed between the storage container and the cavity to seal the gap between the storage container and the cavity.

[0015] For example, in some embodiments, the outer surface of the storage container is provided with at least one groove, or the inner surface of the cavity is provided with at least one groove, and the at least one first seal is installed in the at least one groove, wherein the at least one first seal is an elastic ring.

[0016] For example, in some embodiments, the storage container includes a drainage channel and an inner wall having a first angle at the end facing the second opening, the drainage channel connecting the inner wall and the second opening.

[0017] For example, in some embodiments, the first angle ranges from 5 degrees to 60 degrees.

[0018] For example, in some embodiments, the cavity further includes a liquid channel, and the matrix further includes a fluid channel, the liquid channel being disposed at the puncture structure and communicating with the cavity and the fluid channel.

[0019] For example, in some embodiments, the puncture structure has a needle-shaped protrusion at one end facing the storage container, the center point of the needle-shaped protrusion coincides with the orthographic projection of the center of the bottom surface of the cavity, and the size of the needle-shaped protrusion is smaller than the size of the second opening.

[0020] For example, in some embodiments, the needle-like protrusion is pyramidal in shape with multiple side edges, and the opening of the fluid channel in the cavity is located between two of the multiple side edges and connected to the base and one side of the pyramid.

[0021] For example, in some embodiments, the needle-like protrusion is a cone, and the opening of the fluid channel in the cavity is connected to the bottom surface of the cone.

[0022] For example, in some embodiments, the area of ​​the opening of the fluid channel in the cavity is smaller than the area of ​​the bottom surface of the needle-shaped protrusion.

[0023] For example, in some embodiments, the bottom of the cavity is provided with a flange configured to contact the storage container and confine the storage container to an extreme position, whereby the puncture structure punctures the first sealing film.

[0024] For example, in some embodiments, the detection chip further includes a plurality of fluid channels and a mixing structure, wherein at least one of the plurality of cavities is in communication with the mixing structure through at least one of the plurality of fluid channels. The mixing structure includes a push rod, a first gas isolation membrane, and a first mixing chamber and a second mixing chamber that are in communication with each other. The first gas isolation membrane seals the opening of the first mixing chamber. The push rod is slidably and sealingly installed in the second mixing chamber or disposed separately on the opening side of the second mixing chamber relative to the detection chip. The reciprocating motion of the push rod in the second mixing chamber causes the reagent to be transported between the first mixing chamber and the second mixing chamber.

[0025] For example, in some embodiments, the push rod includes an operating portion disposed at one end of the push rod and protruding from the surface of the push rod.

[0026] For example, in some embodiments, the push rod further includes at least one groove and a sealing ring, the at least one groove being disposed on the outer surface of the push rod, and the sealing ring being installed in the at least one groove.

[0027] For example, in some embodiments, the mixing structure includes an air vent, a first air vent membrane, a second air vent membrane, a power chamber, and a mixing chamber communicating with the power chamber, wherein the power chamber and the plurality of cavities are respectively connected through multiple fluid channels.

[0028] For example, in some embodiments, the vent is located on the sealing cap.

[0029] For example, in some embodiments, the first breathable membrane covers the vent, and the substrate communicates with the outside atmosphere through the vent and the first breathable membrane.

[0030] For example, in some embodiments, the second breathable membrane covers the power chamber, which is connected to an external air source system through the second breathable membrane to receive positive or negative pressure applied by the external air source system.

[0031] For example, in some embodiments, the detection chip further includes a magnetic bead capture cavity, which is disposed between the power cavity and the mixing cavity and communicates with the power cavity and the mixing cavity respectively, wherein the area or size of the middle region of the magnetic bead capture cavity is larger than the area or size of the two end regions.

[0032] For example, in some embodiments, a flow channel converging structure is provided between multiple cavities.

[0033] For example, in some embodiments, a flow channel converging structure is provided between the power cavity and the plurality of cavities.

[0034] For example, in some embodiments, the central portion of the flow channel converging structure is circular, and multiple trapezoidal structures are evenly distributed around the central portion.

[0035] For example, in some embodiments, the substrate has a second surface opposite to the first surface, and the plurality of fluid channels are disposed in the second surface.

[0036] For example, in some embodiments, the plurality of fluid channels include a plurality of substrate grooves formed in the second surface, and the detection chip further includes a sealing layer on the second surface to at least cover the plurality of substrate grooves to form fluid channels.

[0037] For example, in some embodiments, the detection chip further includes an adhesive layer on the second surface, wherein the adhesive layer is disposed between the substrate and the sealing layer and configured to bond the substrate and the sealing layer to each other, the adhesive layer exposing at least the plurality of substrate recesses in the substrate and separating the plurality of fluid channels from each other.

[0038] For example, in some embodiments, the fluid channel further includes multiple flow paths and multiple switching valves, each of which is located in one of the multiple flow paths and is configured to control the connection and disconnection of at least a portion of the corresponding fluid channel. The substrate has a substrate groove on the second surface for forming at least one of the multiple flow paths.

[0039] For example, in some embodiments, the second surface has a partition portion in the membrane valve portion, the partition portion having a platform portion that separates substrate grooves belonging to the same flow path.

[0040] For example, in some embodiments, the platform portion may be flush with the rest of the second surface or have a smaller recess depth compared to the substrate groove.

[0041] For example, in some embodiments, the detection chip further includes a sample chamber sealed with an isolation membrane, the isolation membrane being an adhesive single-sided adhesive.

[0042] For example, in some embodiments, the detection chip further includes an amplification chamber, which is a quantitative chamber.

[0043] For example, in some embodiments, the amplification cavity has an interconnected reaction zone and a debubbling zone.

[0044] For example, in some embodiments, the defoaming zone is located upstream and / or downstream of the reaction zone.

[0045] For example, in some embodiments, the reaction zone is formed in a spindle shape, and the defoaming zone is formed in a circle.

[0046] For example, in some embodiments, the detection chip further includes a driving cavity that is in fluid communication with the amplification cavity and the mixing structure.

[0047] For example, in some embodiments, the detection chip further includes a driving assembly movably housed in the driving cavity, the driving assembly including a driving cover and an elastic member disposed between the driving cover and the driving cavity, and biasing the driving cover.

[0048] For example, in some embodiments, the drive assembly further includes at least one second seal disposed between the drive cover and the drive cavity to seal the gap between the drive cover and the drive cavity. The outer surface of the drive cover is provided with at least one groove, or the inner surface of the drive cavity is provided with at least one groove, and the at least one second seal is correspondingly installed in the at least one groove.

[0049] For example, in some embodiments, a sliding structure is provided between the drive cavity and the drive cover.

[0050] For example, in some embodiments, the detection chip further includes a first chip positioning structure configured to position and mount the detection chip onto the detection device.

[0051] At least one embodiment of this disclosure provides a detection device, including a second chip positioning structure and a first operating portion. The second chip positioning structure is configured to mount a detection chip as described above, and the first operating portion is configured to operate the storage container.

[0052] At least one embodiment of this disclosure provides a detection device, including a second chip positioning structure and a second operating portion. The second chip positioning structure is configured to mount a detection chip as described above, and the second operating portion is configured to operate the push rod.

[0053] At least one embodiment of this disclosure provides a detection device, including a second chip positioning structure and a third operating portion. The second chip positioning structure is configured to mount a detection chip as described above, and the third operating portion is configured to operate the switching valve. Attached Figure Description

[0054] To more clearly illustrate the technical solutions of the embodiments of this disclosure, the accompanying drawings used in the embodiments will be briefly described below. It should be understood that the following drawings only show some embodiments of this disclosure and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0055] Figure 1 A perspective view of a detection chip according to at least one embodiment of the present disclosure is shown;

[0056] Figure 2 It shows Figure 1 Exploded perspective view of the detection chip;

[0057] Figure 3A It shows Figure 1 A top view of the substrate of the detection chip;

[0058] Figure 3B It shows Figure 1 A bottom view of the substrate of the detection chip;

[0059] Figure 4A An exploded perspective view of a storage container according to at least one embodiment of the present disclosure is shown;

[0060] Figure 4B It shows Figure 4B A cross-sectional perspective view of the storage container;

[0061] Figure 5A A perspective view of a cavity illustrating a puncture structure according to at least one embodiment of the present disclosure is shown;

[0062] Figure 5B It shows Figure 5A A magnified top view of a portion of the punctured structure;

[0063] Figure 5C A partially enlarged top view of another embodiment of the puncture structure is shown;

[0064] Figures 6A-6CA schematic diagram of the reagent release process from the storage container is shown;

[0065] Figure 7A An exploded perspective view of a sample container according to at least one embodiment of the present disclosure is shown;

[0066] Figure 7B It shows Figure 7A A perspective view of the lid of the sample container;

[0067] Figure 7C It shows Figure 7A A perspective view of the sample container, with the lid removed;

[0068] Figure 7D A cross-sectional view of the assembled sample container is shown;

[0069] Figure 7E A perspective view shows the sample container installed in the sample chamber of the detection chip;

[0070] Figure 8 A schematic perspective view of a portion of a switching valve according to an embodiment of the present disclosure is shown;

[0071] Figure 9 It shows Figure 8 Perspective view of the matrix groove in the middle matrix;

[0072] Figures 10-13 A cross-sectional view of a switching valve according to an embodiment of the present disclosure is shown, wherein, Figure 10 This is an exploded view of the switching valve. Figure 11 This is a cross-sectional view of the valve in its normally open position. Figure 12 This is a cross-sectional view of the valve in its fully open position. Figure 13 A cross-sectional view of the valve in the closed position;

[0073] Figure 14 A cross-sectional perspective view of the detection chip in the first freeze-drying chamber according to an embodiment of the present disclosure is shown;

[0074] Figure 15 A partial view of the first and second freeze-drying chambers is shown.

[0075] Figure 16 A schematic diagram of the liquid inlet and liquid outlet channels of the first freeze-drying chamber is shown.

[0076] Figure 17 An exploded perspective view of a push rod according to an embodiment of the present disclosure is shown;

[0077] Figure 18 A perspective view of the detection chip at the mixing structure according to an embodiment of the present disclosure is shown;

[0078] Figure 19 It shows Figure 18 Another perspective view of the detection chip at the mixing structure, in which the push rod and the first gas isolation membrane have been removed;

[0079] Figures 20A-20C A cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, illustrating the steps of mixing magnetic beads with reagents;

[0080] Figures 21A-21C Another cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, illustrating the steps of collecting magnetic beads and emptying waste liquid;

[0081] Figure 22 Another partial view of a detection chip according to an embodiment of the present disclosure is shown, illustrating the transfer steps to the amplification chamber;

[0082] Figure 23 A heating groove for a detection chip according to an embodiment of the present disclosure is shown;

[0083] Figure 24 An amplification cavity of a detection chip according to an embodiment of the present disclosure is shown;

[0084] Figure 25 A partial cross-sectional view of a detection chip according to an embodiment of the present disclosure is shown, which illustrates the amplification chamber and the venting chamber;

[0085] Figure 26 It shows Figure 24 The diagram shows the amplification chamber used for amplification and optical detection.

[0086] Figure 27 A perspective view of a detection chip according to at least one embodiment of the present disclosure is shown;

[0087] Figure 28 It shows Figure 27 Exploded perspective view of the detection chip;

[0088] Figure 29A It shows Figure 27 A top view of the substrate of the detection chip;

[0089] Figure 29B It shows Figure 27 A bottom view of the substrate of the detection chip;

[0090] Figure 29C It shows Figure 29B A magnified view of a portion of the flow channel converging structure;

[0091] Figure 30 It shows Figure 27 An exploded perspective view of the detection chip and the sealing cap.

[0092] Figure 31 It shows Figure 27 Another top view of the substrate of the detection chip, showing the sealing groove and the snap-fit;

[0093] Figure 32A and 32B The top view and bottom view of the sealing cap without the film are shown respectively;

[0094] Figure 32C and 32D Top view and bottom view of the sealing cap with the adhesive film are shown respectively;

[0095] Figures 33A-33C Schematic diagrams of the elastic membrane of the sealing cap according to at least one embodiment are shown respectively;

[0096] Figure 33D-33F A schematic diagram of the reagent release process of a storage container according to another embodiment is shown;

[0097] Figures 34A to 34F A schematic diagram of a driving component for a detection chip according to at least one embodiment is shown;

[0098] Figure 35A and Figure 35B A schematic diagram of the amplification cavity of a detection chip according to at least one embodiment is shown;

[0099] Figures 36 to 43 A schematic diagram of liquid flow is shown in the operation method of the detection chip according to at least one embodiment;

[0100] Figure 44 A detection apparatus according to an embodiment of the present disclosure is shown. Detailed Implementation

[0101] To make the objectives, technical solutions, and advantages of the embodiments of this disclosure clearer, the technical solutions of the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this disclosure. All other embodiments obtained by those skilled in the art based on the described embodiments of this disclosure without creative effort are within the scope of protection of this disclosure.

[0102] Unless otherwise defined, the technical or scientific terms used in this disclosure shall have the ordinary meaning understood by one of ordinary skill in the art to which this disclosure pertains. The terms "first," "second," and similar terms used in this disclosure do not indicate any order, quantity, or importance, but are merely used to distinguish different components. Terms such as "comprising" or "including" mean that the element or object preceding the word encompasses the elements or objects listed following the word and their equivalents, without excluding other elements or objects. Terms such as "upper," "lower," "left," and "right" are used only to indicate relative positional relationships, and these relative positional relationships may change accordingly when the absolute position of the described object changes.

[0103] In the design of microfluidic chips, the goal is typically to integrate as many analytical and detection functions as possible onto the chip to reduce reliance on external operations, thereby achieving automation and integration. Microfluidic chips are mostly single-use products, eliminating the need for complex cleaning and waste liquid treatment systems and avoiding contamination caused by such systems. To achieve integration, reagent storage components can be incorporated into the microfluidic chip to store various reagents required for analysis and detection. However, typical microfluidic chips with reagent storage capabilities have complex chip structures or fabrication processes, resulting in high costs as consumables. Furthermore, microfluidic chips capable of multiplex detection are even more complex to manufacture and therefore more expensive.

[0104] At least one embodiment of this disclosure provides a detection chip, including a substrate and at least one storage container. The substrate has a first surface and includes at least one cavity with a first opening in the first surface. The at least one storage container is movably accommodated in the at least one cavity through the first opening. The at least one storage container includes a second opening and a first sealing membrane sealing the second opening. The at least one cavity includes a puncture structure disposed in the cavity for puncturing the first sealing membrane when the storage container moves to contact the puncture structure, thereby allowing reagents in the storage container to enter the cavity. This embodiment, through the separate storage container, allows samples and reaction reagents to be pre-stored in a closed storage container, isolated from the outside environment for long-term preservation. It can be combined with the substrate of the detection chip as needed, thereby reducing the manufacturing cost of microfluidic chips and increasing their application range. During use, the closed space within the storage container is broken, thereby quantitatively releasing the reagents.

[0105] At least one embodiment of this disclosure provides a detection chip that may, as needed, include one or more of the following: an elution chamber, a first cleaning chamber, a second cleaning chamber, a third cleaning chamber, a sample chamber, a first mixing chamber, a second mixing chamber, a waste liquid chamber, a venting chamber, and an amplification chamber. One or more of these chambers may be configured to house the storage container described above. This detection chip can integrate all or part of the detection process, thereby facilitating the design and operation of the detection process, reducing detection time, and minimizing contamination during the detection process.

[0106] At least one embodiment of this disclosure also provides a detection apparatus including a chip positioning structure for mounting the detection chip as described above, for use with the detection chip as described above.

[0107] The detection chip of the present disclosure embodiment will now be described in general with reference to the accompanying drawings.

[0108] Figure 1 A perspective view of a detection chip according to at least one embodiment of the present disclosure is shown. Figure 2 It shows Figure 1 An exploded perspective view of the detection chip. Figure 3A It shows Figure 1 A top view of the substrate of the detection chip. Figure 3B It shows Figure 1 A bottom view of the substrate of the detection chip.

[0109] like Figure 1 and Figure 2 As shown, the detection chip according to at least one embodiment of the present disclosure includes a substrate 100, a first flexible layer 20, an adhesive layer 30, a sealing layer 31, at least one storage container 40, a mixing structure 50, a sample container 60, and a cap 601.

[0110] like Figure 3A and Figure 3B As shown, the substrate 100 has multiple cavities, multiple mixing cavities, a first freeze-drying cavity 102 for storing lysed freeze-dried material, a second freeze-drying cavity 103 for storing amplified freeze-dried material, a waste liquid cavity 190, a venting cavity 400, and multiple substrate grooves 100c. The substrate 100 can be injection molded from a polymer material such as PMMA (polymethyl methacrylate), PC (polycarbonate), PP (polypropylene), or PS (polystyrene).

[0111] At least one storage container 40 may store samples, eluents, washing solutions or magnetic beads.

[0112] Multiple cavities include a sample chamber 110 and multiple liquid storage chambers, which cooperate with corresponding storage containers 40. These liquid storage chambers include, for example, an eluent chamber 180, a first washing fluid chamber 150, a second washing fluid chamber 160, a third washing fluid chamber 170, and a magnetic bead chamber 120. One or more of the sample chamber 110, eluent chamber 180, first washing fluid chamber 150, second washing fluid chamber 160, third washing fluid chamber 170, and magnetic bead chamber 120 are configured to accommodate the corresponding storage container 40. In this embodiment, each of the sample chamber 110, eluent chamber 180, first washing fluid chamber 150, second washing fluid chamber 160, third washing fluid chamber 170, and magnetic bead chamber 120 contains a corresponding storage container 40. Each corresponding storage container 40 contains a corresponding reagent. For example, the sample chamber 110 can contain a storage container storing the sample and lysis buffer, the eluent chamber 180 can contain a storage container storing the eluent, the first washing fluid chamber 150 can contain a storage container storing the first washing fluid, the second washing fluid chamber 160 can contain a storage container storing the second washing fluid, the third washing fluid chamber 170 can contain a storage container storing the third washing fluid, and the magnetic bead chamber 120 can contain a storage container storing magnetic beads. The corresponding reagents can be released by operating the storage container 40, and the release process will be described in detail below.

[0113] For example, the surface of these magnetic beads has been modified so that when the detection chip is used for detection, such as for detecting specific nucleic acid fragments, molecular structures such as nucleic acid fragments can bind to the magnetic beads during the extraction process to achieve the extraction function. For example, the aforementioned nucleic acid fragments and other molecular structures are obtained after lysing the sample to be detected. For details regarding the surface modification of magnetic beads 001, please refer to conventional design guidelines; further details are omitted here.

[0114] The multiple mixing chambers include a first mixing chamber 130 and a second mixing chamber 140, which constitute part of the mixing structure 50 of the detection chip.

[0115] like Figure 3B As shown, a plurality of substrate grooves 100c are formed in the second surface 100b of the substrate 100 facing the sealing layer 31.

[0116] An adhesive layer 30 is disposed between the substrate 100 and the sealing layer 31, and configured to bond the substrate 100 to the sealing layer 31. The adhesive layer 30 exposes at least a plurality of substrate recesses 100c in the substrate 100 and separates a plurality of fluid channels from each other. For example, the adhesive layer 30 includes a plurality of intermediate through grooves, each positioned to correspond to one of the plurality of substrate recesses 100c, and bonded to the sealing layer 31 to form a plurality of fluid channels.

[0117] The adhesive layer 30 can be a double-sided adhesive layer, an adhesive layer, or a plastic film layer attached between the substrate 100 and the sealing layer 31 by means of hot pressing, ultrasonic welding, etc.

[0118] Those skilled in the art will recognize that, in other embodiments, the detection chip may not have the adhesive layer 30. For example, the sealing layer 31 may be attached to the substrate 100 by means of hot pressing, ultrasonic welding, or laser welding.

[0119] The sealing layer 31 is attached to the second surface 100b of the substrate 100. A description of exemplary structures of cavities according to embodiments of this disclosure can be found below in conjunction with examples... Figures 5A-5C The sealing layer 31 can be made of polymer materials such as PE (polyethylene), PP, PS, or PC. For example, in at least one example, the surface properties of the sealing layer 31 can be modified by surface modification, making it difficult for enzymes, nucleic acid molecules, etc. in the detection chip to adhere to the sealing layer 31, thereby giving the sealing layer 31 good biocompatibility.

[0120] The mixing structure 50 includes a pusher 502, a first gas isolation membrane 501, and a first mixing chamber 130 and a second mixing chamber 140 that are interconnected. The first gas isolation membrane 501, the pusher 502, and the cover 601 are attached to the first surface 100a of the substrate 100 at the first mixing chamber 130, the second mixing chamber 140, and the sample chamber 110, respectively, to at least fluid seal the corresponding chambers. Figure 18 In the illustrated embodiment, the first gas isolation membrane 501 and the push rod 502 can be provided independently of each other. The first mixing chamber 130 and the second mixing chamber 140 are interconnected. The first gas isolation membrane 501 seals the opening of the first mixing chamber 130, and the push rod 502 is slidably and sealingly installed in the second mixing chamber 140. The reciprocating motion of the push rod 502 in the second mixing chamber 140 causes the reagent to be transported between the first mixing chamber 130 and the second mixing chamber 140.

[0121] The detection chip according to embodiments of this disclosure allows for the integration of one or more, or even all, of steps such as sample loading, lysis, mixing, cleaning, amplification, and detection. Therefore, it simplifies the operation of the detection chip, improves its automation level, and reduces the time required for related detections.

[0122] As described above, the substrate 100 includes a sample chamber 110, an elution chamber 180, a first cleaning chamber 150, a second cleaning chamber 160, a third cleaning chamber 170, a magnetic bead chamber 120, a first mixing chamber 130, a second mixing chamber 140, a waste liquid chamber 190, an amplification chamber 200, and a gas permeation chamber 400.

[0123] The substrate 100, the adhesive layer 30, and the sealing layer 31 together form multiple fluid channels. The multiple flow channels include multiple flow paths and multiple switching valves respectively arranged on some or all of the flow paths. The multiple flow paths include the first flow path 1, the second flow path 2, the third flow path 3, the fourth flow path 4, the fifth flow path 5, the sixth flow path 6, the seventh flow path 7, the eighth flow path 8, the ninth flow path 9, and the tenth flow path 10, etc.

[0124] The first flow path 1 is configured to connect the sample chamber 110, the first lyophilization chamber 102, and the first mixing chamber 130. The second flow path 2 is configured to connect the first mixing chamber 130 and the magnetic bead chamber 120. The third flow path 3 is configured to connect the first mixing chamber 130 and the first washing solution chamber 150. The fourth flow path 4 is configured to connect the first mixing chamber 130 and the second washing solution chamber 160. The fifth flow path 5 is configured to connect the first mixing chamber 130 and the third washing solution chamber 170. The sixth flow path 6 is configured to connect the first mixing chamber 130 and the elution solution chamber 180. The seventh flow path 7 is configured to connect the first mixing chamber 130 and the second mixing chamber 140. The eighth flow path 8 is configured to connect the amplification chamber 200, the second lyophilization chamber 103, and the second mixing chamber 140. The ninth flow path 9 is configured to connect the amplification chamber 200 and the gas permeation chamber 400. The tenth flow path 10 is configured to connect the second mixing chamber 140 and the waste liquid chamber 190.

[0125] Furthermore, the first flow path 1, the second flow path 2, the third flow path 3, the fourth flow path 4, the fifth flow path 5, the sixth flow path 6, the seventh flow path 7, the eighth flow path 8, the ninth flow path 9, and the tenth flow path 10 are respectively equipped with a first switching valve V1, a second switching valve V2, a third switching valve V3, a fourth switching valve V4, a fifth switching valve V5, a sixth switching valve V6, a seventh switching valve V7, an eighth switching valve V8, a ninth switching valve V9, and a tenth switching valve V10. These switching valves are configured to control the connection and disconnection of at least a portion of the corresponding fluid channels, thereby correspondingly closing and opening the flow path in which they are located.

[0126] For example, in at least one use case, the detection chip can operate as follows.

[0127] S1, sample addition procedure.

[0128] Open cap 601, add the sample to be tested into sample chamber 110, and then close cap 601. For example, the volume of the sample to be tested can be 10 μL - 1000 μL. For example, sample lysis buffer can be pre-stored in sample chamber 110. The sample and lysis buffer are mixed in sample chamber 110, thereby lysing biological materials such as cells in the sample to release nucleic acid molecules.

[0129] S2, Sample and Lysis-Freeze Release Step.

[0130] The first switch valve V1 is opened, and the sample is squeezed from the sample chamber 110 to flow through the first flow path 1 to the first lyophilization chamber 102, where the lysed lyophilized sample stored in the first lyophilization chamber 102 dissolves. The sample then flows to the first mixing chamber 130, and the first switch valve V1 is then closed. The lysed lyophilized sample contains proteinase K, internal control plasmids, and other contents, which can further enhance the lysis effect.

[0131] S3, magnetic bead release step.

[0132] The second switching valve V2 is opened, and by squeezing the magnetic bead chamber 120, 50 μL of magnetic bead solution (i.e., liquid containing magnetic beads) flows through the second flow path 2 to the first mixing chamber 130. Then the second switching valve V2 is closed. Thus, the lysis buffer, sample, and magnetic bead solution are mixed in the first mixing chamber 130.

[0133] S4, pyrolysis and first mixing step.

[0134] Open the seventh switching valve V7, and use the external operating part to control the reciprocating motion of the push rod 502, so that the lysis buffer, sample, and magnetic bead solution are transported back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. By thoroughly mixing the sample, lysis buffer, and magnetic beads, the sample can be effectively lysed to release nucleic acid fragments, and the magnetic beads can efficiently adsorb nucleic acid fragments. The uniform mixture terminates in the first mixing chamber 130, and then the V7 flow path switching valve is closed.

[0135] The lysis and first mixing steps promote thorough mixing of the sample, lysis buffer, and magnetic beads, allowing nucleic acid molecules to be fully released and adsorbed by the magnetic beads. For example, the reciprocating motion of the push rod 502 controlled by the external operating part causes the mixture of sample, lysis buffer, and magnetic beads to move back and forth between the first mixing chamber 130 and the second mixing chamber 140 1-100 times to achieve the desired mixing effect.

[0136] S5, the first magnetic bead collection and the second mixing step.

[0137] When the magnet is brought close to the bottom of the first mixing chamber 130 (i.e., the side near the sealing layer 31) from the outside of the detection chip, the seventh switch valve V7 is opened. Through the reciprocating motion of the push rod 502, the mixture of sample, lysis buffer and magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. Finally, the mixture without magnetic beads stays in the second mixing chamber 140, while the magnetic beads with nucleic acid molecules are attracted by the magnet and stay in the first mixing chamber 130. Then the magnet is removed and the seventh switch valve V7 is closed.

[0138] After the first magnetic bead collection and the second mixing steps, the magnetic beads with adsorbed nucleic acid molecules are separated from other components in the mixture. For example, the number of mixing cycles (i.e., the number of round trips) can be set to 1-100.

[0139] S6, the first step of emptying the waste liquid.

[0140] Open the tenth switch valve V10, and the external operating part squeezes the push rod 502 downward, so that the mixture without magnetic beads flows as waste liquid from the second mixing chamber 140 through the tenth flow path 10 to the waste liquid chamber 190, and then close the tenth switch valve V10.

[0141] S7, First cleaning fluid release step.

[0142] Open the third switch valve V3, and by squeezing the first cleaning liquid chamber 150, the first cleaning liquid flows from the first cleaning liquid chamber 150 through the third flow path 3 to the first mixing chamber 130 and redissolves the magnetic beads remaining in the first mixing chamber 130. Then close the third switch valve V3.

[0143] S8, the first washing and third mixing steps.

[0144] Open the seventh switch valve V7. The external operating part controls the movement of the push rod 502, causing the first cleaning fluid in the first mixing chamber 130 to flow through the seventh flow path 7 to the second mixing chamber 140. Through the reciprocating motion of the push rod 502, the mixture of the first cleaning fluid and the magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7, ultimately causing the mixture to settle in the first mixing chamber 130. Then, close the seventh switch valve V7. For example, the number of mixing cycles (i.e., the number of reciprocating cycles) can be set to 1-100.

[0145] The surface of magnetic beads that have adsorbed nucleic acid molecules may retain impurities such as components from the sample and lysis buffer. These impurities will affect subsequent amplification reactions and detection. The first washing step, as well as the subsequent second and third washing steps, will remove these impurities. For example, the first washing solution in the first washing step can be used to clean protein molecules from the surface of the nucleic acid.

[0146] S9, the second magnetic bead collection and the fourth mixing step.

[0147] The magnet is brought close to the bottom of the first mixing chamber 130 from the outside of the detection chip. The seventh switch valve V7 is opened. Through the reciprocating motion of the push rod 502, the mixture of the first cleaning solution and the magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. Finally, the mixture without magnetic beads stays in the second mixing chamber 140, while the magnetic beads with nucleic acid molecules are attracted by the magnet and stay in the first mixing chamber 130. Then the magnet is removed and the seventh switch valve V7 is closed.

[0148] After the second magnetic bead collection and the fourth mixing step, the magnetic beads with adsorbed nucleic acid molecules are separated from other components in the mixture. For example, the number of mixing cycles (i.e., the number of round trips) can be set to 1-100.

[0149] S10, the second step of emptying the waste liquid.

[0150] Open the tenth switch valve V10, and the external operating part squeezes the push rod 502 downward, so that the mixture without magnetic beads flows as waste liquid from the second mixing chamber 140 through the tenth flow path 10 to the waste liquid chamber 190, and then close the tenth switch valve V10.

[0151] S11, Second cleaning fluid release step.

[0152] Open the fourth switch valve V4, and by squeezing the second cleaning liquid chamber 160, the second cleaning liquid flows from the second cleaning liquid chamber 160 through the fourth flow path 4 to the first mixing chamber 130 and redissolves the magnetic beads remaining in the first mixing chamber 130. Then close the fourth switch valve V4.

[0153] S12, the second washing and fifth mixing steps.

[0154] Open the seventh switch valve V7. Through the reciprocating motion of the push rod 502, the mixture of the second cleaning fluid and the magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7, eventually causing the mixture to settle in the first mixing chamber 130. Then, close the seventh switch valve V7. For example, the number of mixing cycles (i.e., the number of reciprocating cycles) can be set to 1-100.

[0155] For example, the second cleaning solution in the second cleaning step can be used to clean small molecule impurities and salt ions from the surface of the magnetic beads.

[0156] S13, the third magnetic bead collection and the sixth mixing step.

[0157] When a magnet is brought close to the bottom of the first mixing chamber 130 from the outside of the detection chip, the seventh switch valve V7 is opened. The reciprocating motion of the push rod 502 causes the mixture of the second cleaning solution and the magnetic beads to move back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. Ultimately, the mixture without magnetic beads remains in the second mixing chamber 140, while the magnetic beads with attached nucleic acid molecules remain in the first mixing chamber 130 due to the attraction of the magnet. Then, the magnet is removed and the seventh switch valve V7 is closed. For example, the number of mixing cycles (i.e., the number of reciprocating cycles) can be set to 1-100.

[0158] S14, the third step of emptying the waste liquid.

[0159] Open the tenth switch valve V10, and the external operating part squeezes the push rod 502 downward, so that the mixture without magnetic beads flows as waste liquid from the second mixing chamber 140 through the tenth flow path 10 to the waste liquid chamber 190, and then close the tenth switch valve V10.

[0160] S15, Third cleaning fluid release step.

[0161] Open the fifth switch valve V5, and by squeezing the third cleaning liquid chamber 170, the third cleaning liquid flows from the third cleaning liquid chamber 170 through the fifth flow path 5 to the first mixing chamber 130 and redissolves the magnetic beads remaining in the first mixing chamber 130. Then close the fifth switch valve V5.

[0162] S16, the third washing and the seventh mixing steps.

[0163] Open the seventh switch valve V7. Through the reciprocating motion of the push rod 502, the mixture of the first cleaning fluid and the magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7, eventually causing the mixture to settle in the first mixing chamber 130. Then, close the seventh switch valve V7. For example, the number of mixing cycles (i.e., the number of reciprocating cycles) can be set to 1-100.

[0164] For example, the third cleaning solution in this third cleaning step can be used to clean small molecules and salt ions remaining on the surface of the magnetic beads. For example, this third cleaning solution is the same as the second cleaning solution. Since the second and third cleaning solutions are the same, the second cleaning solution chamber 160 and the third cleaning solution chamber 170 are designed to share a part of the fourth flow path 4 to reduce the size of the detection chip and simplify the layout of the flow path of the detection chip.

[0165] S17, fourth magnetic bead collection and eighth mixing step.

[0166] When a magnet is brought close to the bottom of the first mixing chamber 130 from the outside of the detection chip, the seventh switch valve V7 is opened. The reciprocating motion of the push rod 502 causes the mixture of the second cleaning solution and the magnetic beads to move back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. Ultimately, the mixture without magnetic beads remains in the second mixing chamber 140, while the magnetic beads with attached nucleic acid molecules remain in the first mixing chamber 130 due to the attraction of the magnet. Then, the magnet is removed and the seventh switch valve V7 is closed. For example, the number of mixing cycles (i.e., the number of reciprocating cycles) can be set to 1-100.

[0167] S18, the fourth step of emptying the waste liquid.

[0168] Open the tenth switch valve V10, and the external operating part squeezes the push rod 502 downward, so that the mixture without magnetic beads flows as waste liquid from the second mixing chamber 140 through the tenth flow path 10 to the waste liquid chamber 190, and then close the tenth switch valve V10.

[0169] S19, Magnetic bead drying step.

[0170] The bottom of the first mixing chamber 130 is heated and the temperature is maintained at a constant temperature, for example, within the range of 30-70°C, for a certain period of time to dry the surface of the magnetic beads.

[0171] S20, eluent release step.

[0172] Open the sixth switch valve V6, and squeeze the eluent chamber 180 to make the eluent flow from the eluent chamber 180 through the sixth flow path 6 to the first mixing chamber 130 and redissolve the magnetic beads remaining in the first mixing chamber 130. Then close the sixth switch valve V6.

[0173] S21, elution and ninth mixing step.

[0174] Open the seventh switch valve V7. Through the reciprocating motion of the push rod 502, the mixture of eluent and magnetic beads moves back and forth between the first mixing chamber 130 and the second mixing chamber 140 through the seventh flow path 7. Finally, the mixture is stationed in the first mixing chamber 130. Then, close the seventh switch valve V7 and stop heating.

[0175] As the mixture is heated, nucleic acid molecules will detach from the surface of the magnetic beads. Furthermore, by thoroughly mixing the eluent with the magnetic beads, nucleic acid molecules can be effectively detached from the surface of the beads. Finally, the liquid remaining in the first mixing chamber 130 will be the eluent containing nucleic acid molecules, i.e., the sample to be amplified. For example, the number of mixing cycles (i.e., the number of round trips) can be set to 1-100.

[0176] S22, the fifth magnetic bead collection and the ninth mixing step.

[0177] When a magnet is brought close to the bottom of the first mixing chamber 130 from the outside of the detection chip, the seventh switching valve V7 is opened. The reciprocating motion of the push rod 502 causes the sample to be amplified to move back and forth between the first mixing chamber 130 and the second mixing chamber 140 via the seventh flow path 7. Ultimately, the sample without magnetic beads remains in the second mixing chamber 140, while the magnetic beads without nucleic acid molecules remain in the first mixing chamber 130 due to the magnet's attraction. Then, the magnet is removed and the seventh switching valve V7 is closed. For example, the number of mixing cycles (i.e., the number of round trips) can be set to 1-100.

[0178] S23, the transfer step to amplification chamber 200.

[0179] Open the eighth switch valve V8 and the ninth switch valve V9. Actuate the push rod 502 downwards to allow the sample to flow through the eighth flow path 8 to the second lyophilization chamber 103 to rehydrate the amplified lyophilized sample, and then flow to the amplification chamber 200. After filling the amplification chamber 200, it flows through the ninth flow path 9 to the gas permeation chamber 400. Then, close the eighth switch valve V8 and the ninth switch valve V9. This transfer step helps to fill the amplification chamber 200 with the sample, and excess eluent containing nucleic acid molecules will remain in the eighth flow path 8, the ninth flow path 9, or the gas permeation chamber 400.

[0180] S24, amplification step.

[0181] The amplification chamber is cyclically heated to amplify the sample inside, and then the optical properties of the amplified sample are detected to obtain the detection results.

[0182] The above operation can be performed by operating the detection chip through a detection device. For example, after step S1, the detection chip can be placed in the appropriate position of the detection device, and then the relevant components of the detection device can operate the detection chip.

[0183] The detection chip according to the embodiments of this disclosure can process the entire process of original sample from sample addition to detection, and has a high degree of integration.

[0184] Furthermore, as described above, in the above embodiments, the first mixing chamber is used as an intermediate chamber, which enables the effective integration and relative isolation of multiple processing steps.

[0185] Those skilled in the art will understand that the detection chip according to embodiments of this disclosure can be operated in a different order than described 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 already undergone lysis treatment, the steps related to lysis cannot be omitted.

[0186] Furthermore, those skilled in the art should understand that in some examples, the detection chip may omit one or more of the following: sample chamber 110, eluent chamber 180, first cleaning fluid chamber 150, second cleaning fluid chamber 160, third cleaning fluid chamber 170, magnetic bead chamber 120, first mixing chamber 130, second mixing chamber 140, waste liquid chamber 190, and venting chamber 400, to meet the needs of different applications.

[0187] The structure and working principle of the storage container 40 will be explained below with reference to the accompanying drawings.

[0188] Figure 4A An exploded perspective view of a storage container according to at least one embodiment of the present disclosure is shown. Figure 4B It shows Figure 4B A cross-sectional perspective view of the storage container. Figure 5A A perspective view of a cavity illustrating a puncture structure according to at least one embodiment of the present disclosure is shown. Figure 5B It shows Figure 5A A magnified view of a section of the punctured structure. Figures 6A-6C A schematic diagram of the reagent release process from the storage container is shown.

[0189] like Figures 4A-4B , Figures 5A-5C As shown, the cavity has a first opening on the first surface 100a of the substrate 100. It should be noted that "cavity" in the following text refers to one or more of the sample cavity 110, eluent cavity 180, first cleaning fluid cavity 150, second cleaning fluid cavity 160, third cleaning fluid cavity 170, and magnetic bead cavity 120. For example, in this embodiment, the sample cavity 110, eluent cavity 180, first cleaning fluid cavity 150, second cleaning fluid cavity 160, third cleaning fluid cavity 170, and magnetic bead cavity 120 each have a first opening on the first surface 100a of the substrate 100. A sealing layer 31 is attached to the second surface 100b of the substrate 100. A substrate groove 100c for forming a flow path is provided on the surface of the second surface 100b of the substrate 100. For example, this flow path can be a first flow path 1, a second flow path 2, a third flow path 3, etc. A storage container 40 is movably accommodated in the cavity through the first opening. Here, the term "movably" means that relative movement can occur between two elements, including but not limited to sliding, rotation, and helical motion. For example, storage container 40 can be slidably accommodated in a cavity through a first opening. Storage container 40 includes a second opening 4012 and a first sealing membrane 403 that seals the second opening 4012.

[0190] The cavity includes a puncture structure 70 disposed in the cavity, which is used to puncture the first sealing membrane 403 when the storage container 40 moves so that the first sealing membrane 403 comes into contact with the puncture structure 70, so that the reagent in the storage container 40 can enter the cavity.

[0191] The storage container 40 also includes a third opening 4011 and a second sealing membrane 402 sealing the third opening 4011. A liquid storage space for containing reagents is defined between the first sealing membrane 403 and the second sealing membrane 402 via the sidewall of the storage container 40. Therefore, a certain amount of biochemical reagent can be sealed and stored in the liquid storage space. When it is necessary to release the quantitative biochemical reagent, the storage container 40 can be actuated to cause the puncture structure 70 to puncture the first sealing membrane 403, thereby releasing the reagent from the liquid storage space into the detection chip. The first sealing membrane is a flexible film made of polymer materials such as PE, PP, or PS. The second sealing membrane is a flexible film made of polymer materials such as PE, PP, or PS, or a composite aluminum film.

[0192] In at least one example, the storage container 40 may also include a removable lid 601, thereby allowing the storage container 40 to be configured as a sample container 60. The structure of the sample container 60 will be discussed later. Figures 7A-7E As described in the text.

[0193] The detection chip may further include at least one first seal 404 disposed between the storage container 40 and the cavity of the substrate to seal the gap between the storage container 40 and the cavity. For example, the columnar outer periphery of the storage container 40 may have at least one groove 405, in which at least one first seal 404 is installed. Optionally, the columnar inner periphery of the cavity may have a groove (not shown), in which the first seal 404 is installed. Optionally, the at least one first seal 404 may be an elastic ring for sealing, such as a silicone ring.

[0194] The storage container 40 is coaxially arranged with the cavity. The puncture structure 70 is located at the end of the cavity opposite to the first opening, for example, in... Figure 5A The bottom of the middle.

[0195] Exemplarily, the storage container 40 may include a drainage channel 406 and an inner wall 407 having a first angle at its end facing the second opening 4012, the drainage channel 406 connecting the inner wall 407 and the second opening 4012. Optionally, the first angle ranges from 5 degrees to 60 degrees, for example, 30 degrees. The angled inner wall 407 helps guide reagents within the storage space through the drainage channel 406 into the detection chip, preventing reagent residue in the storage space.

[0196] The inner diameter of the drainage channel 406 ranges from 0.1 to 10 mm. The inner diameter of the drainage channel must be larger than the size of the puncture structure to ensure that the puncture structure can be smoothly inserted into the drainage channel to facilitate the release of reagents from the storage container.

[0197] The cavity also includes a fluid guiding channel 1001, which is located at the puncture structure 70 and connects the cavity to the fluid channel. The inner diameter of the fluid guiding channel 1001 ranges from 0.1 to 5 mm, for example, it can be equal to the inner diameter of the drainage channel 406 to ensure uniform hydraulic pressure within the flow channel.

[0198] A needle-shaped protrusion 701 is provided at one end of the puncture structure 70 facing the storage container 40, and the center point 702 of the needle-shaped protrusion coincides with the orthographic projection of the center of the bottom surface of the cavity. The size of the needle-shaped protrusion 701 is smaller than the size of the second opening 4012.

[0199] like Figure 5A and 5B As shown, in an exemplary embodiment, the needle-like protrusion 701 is approximately pyramidal in shape, that is, its outer contour resembles a pyramid and has three lateral edges. The opening of the liquid channel 1001 in the cavity is located between two lateral edges of the pyramid and is connected to the base and one side of the pyramid.

[0200] Figure 5C A partially enlarged top view of another embodiment of the puncture structure is shown. According to the concept of this disclosure, the needle-like protrusion 701' can also be other pyramidal shapes with multiple side edges, such as having four side edges (e.g., Figure 5C (As shown). The opening of the liquid guiding channel 1001 in the cavity is located between two of the four side edges and is connected to the base and one side of the pyramid. This disclosure is not limited thereto; those skilled in the art can also provide other numbers of side edges and position the opening of the liquid guiding channel between two of the side edges.

[0201] In another embodiment, not shown, the needle-like protrusion may also be formed as a cone, with the opening of the liquid channel in the cavity connected to the bottom surface of the cone.

[0202] Optionally, the area of ​​the opening of the liquid guiding channel in the cavity is smaller than the area of ​​the bottom surface of the needle-shaped protrusion. This arrangement helps to improve the efficiency of liquid entering the fluid channel and prevents liquid from stagnating in other parts of the cavity.

[0203] The bottom of the cavity is provided with a flange, configured to contact the storage container 40 and confine the storage container 40 within the cavity. Figure 6B The extreme position shown. At this extreme position, the puncture structure 70 punctures the first sealing membrane 403.

[0204] Combination Figures 6A-6C The procedure for releasing reagents from storage container 40 is described. The storage container is held in its installed position due to friction between the first seal 404 and the inner wall of the cavity. When reagent release is required, it can be done via a push rod (e.g., Figure 6AThe push rod 90 applies a force F1 to the second sealing membrane 402 of the storage container 40. This force F1 overcomes friction, causing the storage container 40 to move downwards. When it moves downwards to its limit position, i.e., when the bottom of the storage container 40 contacts the flange of the cavity, as... Figure 6B As shown, the puncture structure 70 punctures the first sealing membrane 403, allowing the reagent in the storage space to enter the fluid channel through the liquid guiding channel 1001. A force F2 is further applied to the second sealing membrane 402, which is greater than the elastic force of the second sealing membrane 402. This causes the second sealing membrane 402 to undergo elastic deformation and compress the air in the storage space, pressurizing the reagent and pushing it completely into the fluid channel, thus completing the reagent release process.

[0205] This separate storage container allows for the independent storage of the required biochemical reagents, enabling separate production and preparation without storing all the reagents in the detection chip at once. As a result, the storage container and the main body of the detection chip can be manufactured independently, simplifying the manufacturing process and facilitating quality control during production, thereby improving product yield.

[0206] For example, the storage container can pre-store reagents, which are at least fluidly sealed in the reservoir space and are permitted to be sold together with the detection chip. When using the detection chip, the user does not need to add reagents to the reservoir chamber to begin using the chip. That is, the storage container can be actuated by an external operating mechanism to release the reagents, thereby allowing the pre-stored reagents to be released. Therefore, the use of the detection chip is convenient. The storage container according to embodiments of this disclosure allows for long-term storage of reagents, for example, up to one year. Furthermore, the substrate 100, sealing layer 31, first flexible layer 20, and second sealing film 402 can be designed to isolate air and liquid, thus isolating the reagents stored in the storage container from air and external substances, and maintaining stability during the transportation of the detection chip.

[0207] The following will combine Figures 7A to 7E The sample container 60 is described.

[0208] As described above, the substrate 100 includes a sample container 60, which is housed within the sample cavity 110 of the substrate. The sample container 60 has all the structures of the aforementioned storage container 40, such as the first sealing membrane 603, the second sealing membrane 602, the first seal 604, and the groove 605. The difference is that the sample container 60 also includes a removable cap 601 and mounting features corresponding to the cap 601.

[0209] The sample container 60 can be sealed and installed inside the sample chamber 110.

[0210] Figure 7A An exploded perspective view of a sample container according to at least one embodiment of the present disclosure is shown. Figure 7BIt shows Figure 7A A perspective view of the lid of the sample container. Figure 7C It shows Figure 7A A perspective view of the sample container with the lid removed. Figure 7D A cross-sectional view of the assembled sample container is shown. Figure 7E A perspective view shows the sample container installed in the sample chamber of the detection chip.

[0211] like Figures 7A-7E As shown, the lid 601 can be opened and closed. The lid 601 can be opened to add a sample to the sample container 60 and then the lid 601 can be closed.

[0212] The cover 601 includes a through hole 6011, a snap-fit ​​protrusion 6013 surrounding the through hole 6011, and a sealing groove 6012 surrounding the snap-fit ​​protrusion 6013. The through hole 6011 penetrates the cover 601 to form an opening, and a second sealing film 602 is bonded to the upper end of the through hole 6011 to seal the through hole 6011. The sample container 60 includes a side wall 607 and a snap-fit ​​receiving portion 606 formed at the lower end of the side wall 607. Figure 7D As shown, the thickness of the sidewall 607 gradually decreases in the direction toward the cover 601. Furthermore, the snap-fit ​​protrusion 6013 engages with the snap-fit ​​receiving portion 606, and the sealing groove 6012 accommodates the sidewall 607 inserted therein. Thus, the cover 601 can seal an opening of the sample container 60.

[0213] In addition, the cover 601 may include a flip-up protrusion 6014 to facilitate the user in opening the cover 601.

[0214] The switching valve in the flow path of the embodiment will be described below with reference to the accompanying drawings.

[0215] As described above, the first flow path 1, the second flow path 2, the third flow path 3, the fourth flow path 4, the fifth flow path 5, the sixth flow path 6, the seventh flow path 7, the eighth flow path 8, the ninth flow path 9, and the tenth flow path 10 are respectively equipped with a first switching valve V1, a second switching valve V2, a third switching valve V3, a fourth switching valve V4, a fifth switching valve V5, a sixth switching valve V6, a seventh switching valve V7, an eighth switching valve V8, a ninth switching valve V9, and a tenth switching valve V10 to control the opening and closing of the corresponding flow paths.

[0216] The following describes a switching valve according to at least one embodiment of the present disclosure, wherein the first to tenth switching valves V10 can be designed as such switching valves.

[0217] Figure 8 A schematic perspective view of a portion of a switching valve according to an embodiment of the present disclosure is shown. Figure 9 It shows Figure 8 A perspective view of the substrate groove 100c of the substrate 100. Figures 10-13 A cross-sectional view of a switching valve according to an embodiment of the present disclosure is shown, the cross-sectional view being along... Figure 8 The midline LL was used as the cutoff point. Among them, Figure 10 This is an exploded view of the switching valve. Figure 11 This is a cross-sectional view of the valve in its normally open position. Figure 12 This is a cross-sectional view of the valve in its fully open position. Figure 13 This is a cross-sectional view of the valve in the closed position.

[0218] like Figures 8-13 As shown, in at least one example, the switching valve is formed by a substrate groove 100c of the substrate 100, an intermediate through-groove 510 in the adhesive layer 30, and a sealing layer 31. The substrate groove 100c has a break region that divides the substrate groove 100c into two sub-sub ...

[0219] When there is no fluid flow and the sealing layer is not subjected to external force, there is a certain gap between the sealing layer 31 and the substrate 100, and the switch valve is in the open state (see...). Figure 11 The grooves of the sub-base on both sides of the switching valve are connected to each other through this gap. When the fluid used for detection is pushed and flows to the switching valve, under the action of fluid pressure, the sealing layer 31 will deform, causing the gap between the sealing layer 31 and the base 100 at the valve action area 511 to widen, so that the fluid can flow fully through the switching valve in the fully open state (see...). Figure 12 When the switch valve is in the open position, this gap is, for example, in the range of 0.001-1 mm. When the valve is pressed in the actuating area 511 (e.g., by a push rod or air pump), the sealing layer 31 will deform to fit against the base 100, making the gap zero, thereby closing the switch valve (see...). Figure 13 ).

[0220] For example, the width of the base groove 100c can be in the range of 0.1-3 mm, and its depth can be in the range of 0.2-2 mm. For example, the valve actuating region 511 can be circular, with a diameter in the range of 0.2-6 mm. The projection of the valve actuating region 511 onto the surface of the base 100 overlaps with the projection of the disconnected area of ​​the base groove 100c and a portion of the projection of the two sub-base grooves. For example, the area of ​​the overlapping region can be, for example, 5%-50%, or, for example, 10%-30% of the area of ​​the valve actuating region 511. Such a range of overlapping area areas facilitates the complete closure and full opening of the valve.

[0221] The first freeze-drying chamber and the second freeze-drying chamber of the embodiment will be described below with reference to the accompanying drawings.

[0222] As described above, the substrate 100 is formed with a first freeze-drying chamber 102 for storing lysed freeze-dried material and a second freeze-drying chamber 103 for storing amplified freeze-dried material.

[0223] Figure 14 A cross-sectional perspective view of the detection chip in the first freeze-drying chamber according to an embodiment of the present disclosure is shown. Figure 15 A partial view of the first freeze-drying chamber 102 and the second freeze-drying chamber 103 is shown. Figure 16 A schematic diagram of the liquid inlet channel 1023 and the liquid outlet channel 1024 of the first freeze-drying chamber 102 is shown.

[0224] The structure of the freeze-drying chamber is described below using the first freeze-drying chamber 102 as an example. The structure of the second freeze-drying chamber 103 is similar to that of the first freeze-drying chamber 102, the only difference being the location of the flow channels and the type of freeze-drying bulbs, so it will not be described in detail here. The first freeze-drying chamber 102 is located in the first flow path 1, and the second freeze-drying chamber 103 is located in the eighth flow path 8. The first freeze-drying chamber 102 contains lysing freeze-dried material for sample lysis, and the second freeze-drying chamber 103 contains amplification freeze-dried material for amplification.

[0225] The first freeze-drying chamber 102 includes a liquid inlet chamber 1021, a freeze-drying chamber 1022, a liquid inlet channel 1023, a liquid outlet channel 1024, and a freeze-drying bulb 1025. The flow direction of the fluid in the first freeze-drying chamber 102 is as follows: Figure 14 As indicated by the arrow in the diagram. The first freeze-drying chamber 102 is integrally formed as a cavity protruding from the substrate 100. The liquid inlet chamber 1021 is located at the upper end of the first freeze-drying chamber 102 and is sealed by the first flexible layer 20. The lower end of the first freeze-drying chamber 102 is sealed by the sealing layer 31.

[0226] The inner diameter and height of the liquid inlet chamber 1021 and the freeze-drying chamber 1022 are different, and the inner diameter and height of the freeze-drying chamber 1022 are both larger than those of the liquid inlet chamber 1021 to accommodate the freeze-dried bulbs 1025. The inner diameter of the freeze-drying chamber 1022 ranges from 0.1 to 10 mm, which is larger than the diameter of the freeze-dried bulbs 1025. The height of the freeze-drying chamber 1022 can be adaptively set according to the number of freeze-dried bulbs 1025 it can accommodate, and can be configured to accommodate 1 to 10 freeze-dried bulbs 1025.

[0227] When the reagent flows through the first lyophilization chamber 102, it first enters the inlet chamber 1021 through the inlet channel 1023, then enters the lyophilization chamber 1022 and comes into contact with the lyophilized bulb 1025 to rehydrate the lyophilized bulb 1025, and finally enters the first flow path 1 through the outlet channel 1024.

[0228] The mixing structure of the embodiment will now be described with reference to the accompanying drawings.

[0229] Figure 17 An exploded perspective view of a push rod according to an embodiment of the present disclosure is shown. Figure 18 A perspective view of the detection chip at the mixing structure according to an embodiment of the present disclosure is shown. Figure 19 It shows Figure 18 Another perspective view of the detection chip at the mixing structure, where the push rod and the first gas isolation membrane have been removed. Figures 20A-20C A cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, illustrating the steps of mixing magnetic beads with reagents. Figures 21A-21C Another cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, illustrating the steps of collecting magnetic beads and emptying waste liquid. Figure 22 Another partial view of a detection chip according to an embodiment of the present disclosure is shown, illustrating the transfer steps to the amplification chamber.

[0230] like Figure 17-19 As shown, the mixing structure 50 includes a first mixing chamber 130 and a second mixing chamber 140, which are connected to each other via a seventh flow path 7. The first mixing chamber 130 has two openings, one on the second surface 100b of the substrate 100 and the other on the first surface 100a of the substrate 100. The second mixing chamber 140 has two openings, one on the second surface 100b of the substrate 100 and the other on the first surface 100a of the substrate 100. The first and second mixing chamber openings on the second surface 100b are covered by a sealing layer 31, the first mixing chamber opening on the first surface 100a is covered by a first gas isolation membrane 501, and the second mixing chamber opening on the first surface 100a is covered by a push rod 502.

[0231] The push rod 502 includes an operating part 5021, a groove 5022, and a sealing element 5023. The push rod 502 is cylindrical in shape, and its outer diameter matches the opening of the second mixing chamber 140. One end of the push rod 502 has the operating part 5021, which protrudes from one side surface of the push rod 502 and is configured to engage with an external operating part to control the movement of the push rod 502. At least one groove 5022 is provided on the cylindrical outer periphery of the push rod 502, and the sealing element 5023 is installed in this groove 5022 to seal the gap between the outer periphery of the push rod 502 and the opening of the second mixing chamber.

[0232] The first mixing chamber 130 is in gaseous communication with the detection chip and is liquid-isolated through the first gas isolation membrane 501 and the opening of the first mixing chamber. The first gas isolation membrane 501 can be a composite membrane, which includes a polyester substrate and a hydrophobic ePTFE membrane.

[0233] Initially, for example, push rod 502 is located at its lower limit position in the second mixing chamber 140. During the mixing step, for example, transferring fluid from the first mixing chamber 130 to the second mixing chamber 140, the seventh switch valve V7 can be opened, and the push rod 502 can be actuated upwards via an external operating part. When push rod 502 moves upwards, the first mixing chamber 130 is isolated from the external gas, and the reagent in the first mixing chamber 130 flows into the second mixing chamber 140 via the seventh flow path 7. At this time, air enters the interior of the first mixing chamber 130 through the first gas isolation membrane 501 to replenish the volume of the reagent and maintain the pressure inside the first mixing chamber 130 always equal to the external atmospheric pressure. Conversely, the push rod 502 moves downward so that the reagent in the second mixing chamber 140 enters the first mixing chamber 130 through the seventh flow path 7. The air in the first mixing chamber 130 is discharged through the first gas isolation membrane 501, and the pressure in the first mixing chamber 130 is always equal to the external atmospheric pressure.

[0234] Figures 20A-20C A cross-sectional view of a mixing system according to an embodiment of the present disclosure is shown, illustrating the steps of mixing magnetic beads 001 with reagents.

[0235] like Figure 20A As shown, magnet 900 is moved away from the first mixing chamber 130, and reagent is placed in the first mixing chamber 130. The seventh switch valve V7 is opened, causing the actuator push rod 502 to move upwards, allowing the reagent to flow from the first mixing chamber 130 through the seventh flow path 7 to the second mixing chamber 140, as shown. Figure 20B As shown. Figure 20C As shown, the reagent flows from the second mixing chamber 140 to the first mixing chamber 130 via the seventh flow path 7 by the downward movement of the actuating push rod 502.

[0236] Figures 21A-21C Another cross-sectional view of the mixing structure according to an embodiment of the present disclosure is shown, illustrating the steps of collecting and emptying the waste liquid from the magnetic beads 001.

[0237] like Figure 21A As shown, when magnet 900 approaches the first mixing chamber 130, the seventh switch valve V7 is opened, and push rod 502 is actuated upwards to transfer the reagent. Due to the attraction of magnet 900, during the transfer of reagent from the first mixing chamber 130 to the second mixing chamber 140, magnetic bead 001 remains in the first mixing chamber 130, while the reagent is transferred to the second mixing chamber 140. At this time, the seventh switch valve V7 is closed, and the tenth switch valve V10 is opened, actuating push rod 502 downwards to discharge the reagent into waste liquid chamber 190. The embodiments of this disclosure do not limit the type of magnet 900; for example, it can be a permanent magnet or an electromagnet.

[0238] The waste liquid chamber of this disclosure embodiment will now be described in conjunction with the accompanying drawings.

[0239] like Figure 3A As shown, the substrate 100 includes a waste liquid chamber 190, which has a waste liquid chamber opening on a second surface 100b and a waste liquid chamber opening on a first surface 100a of the substrate 100. The waste liquid chamber opening on the second surface 100b of the substrate 100 is covered by a sealing layer 31, and the waste liquid chamber opening on the first surface 100a of the substrate 100 is covered by a first flexible layer 20 for collecting and storing waste liquid. A waste liquid chamber through-hole 311 may be provided in a first portion 310 of the first flexible layer 20, which is aligned with the waste liquid chamber 190 in the substrate 100. In addition, the detection chip also includes a second gas isolation membrane 201, which is attached to the waste liquid chamber through-hole 311 of the first flexible layer 20 and may be a waterproof and breathable membrane that prevents liquid from passing through while allowing gas to pass through. Therefore, the waste liquid chamber 190 is in gaseous communication with the detection chip and liquidally isolated from it through the second gas isolation membrane 201 and the waste liquid chamber through-hole 311. Thus, waste liquid can flow into the waste liquid chamber 190 while gas flows out of the waste liquid chamber 190, while the waste liquid in the waste liquid chamber 190 will not flow out and contaminate the detection environment. The second gas isolation membrane 201 can be a composite membrane comprising a polyester substrate and a hydrophobic ePTFE membrane. Furthermore, in the illustrated embodiment, the second gas isolation membrane 201 is attached to the first flexible layer 20 from the side of the first flexible layer 20 away from the substrate 100. However, those skilled in the art will understand that the second gas isolation membrane 201 can also be attached to the first flexible layer 20 from the other side of the first flexible layer 20.

[0240] The amplification cavity of the present disclosure embodiment will now be described with reference to the accompanying drawings.

[0241] Figure 22 Another partial view of a detection chip according to an embodiment of the present disclosure is shown, illustrating the transfer steps to the amplification chamber. Figure 23 A heating groove for a detection chip according to an embodiment of the present disclosure is shown. Figure 24 An amplification cavity of a detection chip according to an embodiment of the present disclosure is shown. Figure 25 A partial cross-sectional view of a detection chip according to an embodiment of the present disclosure is shown, which illustrates the amplification chamber and the venting chamber. Figure 26 It shows Figure 24 The diagram shows amplification and optical detection performed in the amplification chamber.

[0242] An amplification chamber 200 is formed, configured to store amplification reagents. For example, the amplification reagents may be lyophilized powders pre-stored in the amplification chamber 200, including enzymes, probes, primers, and other reagents required for amplification.

[0243] like Figure 22-26 As shown, one end 200a of the amplification chamber 200 is closed, while the other end 200b has an opening. This opening can be elongated. One end of the opening of the amplification chamber 200 is connected to the eighth flow path 8, and the other end of the opening is connected to the ninth flow path 9. The eighth flow path 8 and the ninth flow path 9, respectively, connecting to the two ends of the elongated opening, help to fill the amplification chamber 200 with the reagent to be amplified (e.g., an eluent containing nucleic acid molecules) and prevent the generation of air bubbles, thereby enabling accurate quantification of the volume of the reagent to be detected entering the amplification chamber 200. Therefore, the reagent to be amplified can flow from the second mixing chamber 140 through the eighth flow path 8 and one end of the opening into the amplification chamber 200, and after filling the amplification chamber 200, flow through the other end of the opening and the eighth flow path 8 into the venting chamber 400, which can expel air to the outside of the detection chip.

[0244] In embodiments according to this disclosure, by rationally designing the shape and size of the amplification chamber 200, the reagent to be detected entering the amplification chamber 200 can actively wet the volume of the amplification chamber 200 under capillary action, reducing the generation of air bubbles within the amplification chamber 200. For example, the volume of the amplification chamber 200 can be in the range of 10-100 μL, such as 50 μL. By filling the rigid amplification chamber 200 with reagent of a fixed volume, the volume of the reagent to be detected entering the amplification chamber 200 can be quantified. For example, the width of the opening can be in the range of 0.1-3 mm, such as 1.5 mm. A smaller opening width facilitates the flow of the reagent to be detected into the amplification chamber 200 by means of capillary force.

[0245] For example, the sealing layer 31 can be made of a material transparent to the detection light. A heating module is positioned above the heating recess 300, and an optical module can be positioned below the sealing layer 31 corresponding to the amplification chamber 200. The optical module emits detection light, which includes light incident into the amplification chamber 200 to excite the reagents in the amplification chamber 200 to emit fluorescence signals, and also includes the fluorescence signals emitted by the reagents in the amplification chamber 200. A second substrate 400 made of a transparent material facilitates optical detection of the amplification reaction occurring in the amplification chamber 200. For example, the wall thickness of the amplification chamber 200 can be in the range of 0.1-3 mm, such as 0.5 mm. A thinner wall thickness can improve the heating efficiency of the heating module for the reagents in the amplification chamber 200.

[0246] The first flexible layer 20 may be provided with a venting cavity through-hole 203, which is aligned with the venting cavity 400 in the substrate 100. Furthermore, the detection chip also includes a third gas isolation membrane 101, which is attached to the venting hole 401 at the bottom of the venting cavity 400 and may be a waterproof and breathable membrane that prevents liquid permeation while allowing gas permeation. Therefore, the venting cavity 400 is in gaseous communication with and liquid-isolated from the external environment of the detection chip through the third gas isolation membrane 101, the venting hole 401, and the venting cavity through-hole 203. The waterproof performance of the venting cavity 400 prevents aerosols and volatile vapors from escaping from the venting cavity 400 during amplification, while the breathability of the venting cavity 400 allows air inside the detection chip to be expelled when the test reagent enters the amplification chamber 200. The third gas isolation membrane 101 may be a composite membrane comprising a polyester substrate and a hydrophobic ePTFE membrane. Furthermore, in the illustrated embodiment, the third gas barrier membrane 101 is attached to the first flexible layer 20 from the side of the first flexible layer 20 away from the substrate 100. However, those skilled in the art will understand that the third gas barrier membrane 101 can also be attached to the first flexible layer 20 from the other side of the first flexible layer 20.

[0247] The following is a perspective view of a detection chip according to another embodiment of the present disclosure, described with reference to the accompanying drawings. In the following description of the embodiments, the same or similar reference numerals denote the same or similar elements. The following description focuses primarily on the differences between this embodiment and the foregoing embodiments; the same or similar elements will be omitted from the description, and reference can be made to the structure and function of the foregoing embodiments.

[0248] Figure 27 A perspective view of a detection chip according to at least one embodiment of the present disclosure is shown; Figure 28 It shows Figure 27 An exploded perspective view of the detection chip. Figure 29A It shows Figure 27 A top view of the substrate of the detection chip. Figure 29B It shows Figure 27 A bottom view of the substrate of the detection chip. Figure 29C It shows Figure 29B A magnified view of a portion of the flow channel converging structure is shown.

[0249] like Figure 27 and Figure 28 As shown, the detection chip according to at least one embodiment of the present disclosure includes a substrate 100', a sealing cap 20', a connecting layer 30', a sealing layer 31', at least one storage container 40', a mixing structure 50', a driving component 60, and an amplification portion 70.

[0250] In this embodiment, the structure of the storage container 40' is similar to that of the previous embodiment, and will not be described again here.

[0251] The substrate 100' has multiple cavities, multiple mixing cavities, a sample cavity 1002', a waste liquid cavity 1010', a venting cavity 400', a magnetic bead trapping cavity 1012', and multiple substrate grooves. The substrate 100' can be injection molded from polymer materials such as PMMA (polymethyl methacrylate), PC (polycarbonate), PP (polypropylene), or PS (polystyrene), which will not be described in detail here.

[0252] The multiple cavities include multiple liquid storage cavities and a drive cavity 1011'. These liquid storage cavities cooperate with corresponding storage containers 40', and the drive cavity 1011' is used to quantitatively inject elution buffer into the amplification cavity 70. The amplification cavity 70 is a quantitative cavity; a fixed volume of elution buffer enters the quantitative cavity through its volume, and excess reagent overflows from the quantitative cavity as waste liquid. Furthermore, PCR amplification lyophilized powder can be pre-loaded inside the amplification cavity 70. The multiple liquid storage cavities include, for example, a lysis buffer cavity 1003' for storing lysis buffer, an elution buffer cavity 1007' for elution buffer, a first washing buffer cavity 1004' for a first washing buffer, a second washing buffer cavity 1005' for a second washing buffer, a third washing buffer cavity 1006' for a third washing buffer, and a magnetic bead cavity 1001' for magnetic beads, etc. One or more of the pyrolysis fluid chamber 1003', elution fluid chamber 1007', first cleaning fluid chamber 1004', second cleaning fluid chamber 1005', third cleaning fluid chamber 1006' and magnetic bead chamber 1001' are configured to accommodate the corresponding storage container 40. In this embodiment, each of the lysis buffer chamber 1003', eluent chamber 1007', first washing buffer chamber 1004', second washing buffer chamber 1005', third washing buffer chamber 1006', and magnetic bead chamber 1001' contains a corresponding storage container 40'. Each corresponding storage container 40' contains a corresponding reagent. For example, the lysis buffer chamber 1003' can contain a storage container storing lysis buffer, the eluent chamber 1007' can contain a storage container storing eluent, the first washing buffer chamber 1004' can contain a storage container storing the first washing solution, the second washing buffer chamber 1005' can contain a storage container storing the second washing solution, the third washing buffer chamber 1006' can contain a storage container storing the third washing solution, and the magnetic bead chamber 1001' can contain a storage container storing magnetic beads. The corresponding reagent can be released by operating the storage container 40'. The structure and release process of the storage container 40' are the same as and similar to those of the storage container 40 in the previous embodiment, and will not be described again here.

[0253] Multiple mixing chambers, including mixing chamber 1009' and power chamber 1008', constitute part of the mixing structure 50' of the detection chip.

[0254] A magnetic bead capturing chamber 1012' is disposed between the power chamber 1008' and the mixing chamber 1009'. Magnetic beads pass through the magnetic bead capturing chamber 1012' and are captured by an external magnet. For example... Figure 29B As shown, the magnetic bead capturing cavity 1012' is a concave spindle shape. The depth of the concavity is consistent with the depth of the flow channel.

[0255] The magnetic bead capturing chamber 1012' is designed in a spindle shape. Its middle area is large, which can accommodate more magnetic beads and avoid clogging the flow channel. Its two ends are small, which can prevent liquid flow interruption. At the same time, the liquid can converge and avoid forming magnetic bead clumps that cannot be mixed evenly.

[0256] Unlike the previous embodiments, the detection chip in this embodiment includes a sealing cap 20', which is connected to the substrate 100' and seals the first opening of the substrate 100'. As previously described, the cavity has a first opening on the first surface 100a of the substrate 100, and the storage container 40' is movably and sealingly accommodated within the first opening. The sealing cap 20' can be detachably connected to the substrate 100', for example, by snap-fit, or it can be connected to the substrate 100' by adhesive or welding.

[0257] In this embodiment, a flow channel converging structure 1013' is provided between the power cavity 1008' and the plurality of cavities in the substrate 100'. For example... Figure 29C As shown, the central part of the flow channel converging structure 1013' is circular, and multiple trapezoidal structures are evenly distributed around the central part.

[0258] The function of the flow channel converging structure 1013' is to prevent liquid flow interruption and cross-contamination. When the power chamber 1008' draws liquid from the storage chamber, the reagent enters the flow channel converging structure 1013' and wets the entire structure under the action of capillary force, thus avoiding flow interruption. On the other hand, due to the capillary force, the surrounding reagents will tend to converge towards the flow channel converging structure 1013'. Even if a small amount of reagent crosses into other channels, it can eventually converge and be drawn away by the power chamber 1008'.

[0259] The power chamber 1008' passes through the power port 2040' of the sealing cover 20. Figure 32A It is connected to an external air supply system to apply positive and negative pressure to the power chamber 1008' to drive the fluid movement in the fluid channel.

[0260] The substrate 100', adhesive layer 30', and sealing layer 31' together form multiple fluid channels. The multiple flow channels include multiple flow paths and multiple switching valves respectively arranged on some or all of the flow paths. The multiple flow paths include the first flow path 1', the second flow path 2', the third flow path 3', the fourth flow path 4', the fifth flow path 5', the sixth flow path 6', the seventh flow path 7', the eighth flow path 8', the ninth flow path 9', the tenth flow path 10', the eleventh flow path 11', the twelfth flow path 12', and the thirteenth flow path 13', etc.

[0261] The first flow path 1' connects the amplification chamber and the venting chamber 400', the second flow path 2' connects the mixing chamber 1009' and the amplification chamber, the third flow path 3' connects the mixing chamber 1009' and the power chamber 1008', the fourth flow path 4' connects the mixing chamber 1009' and the waste liquid chamber 1010', the fifth flow path 5' connects the elution liquid chamber 1007' and the flow channel converging structure 1013', the sixth flow path 6' connects the sample chamber 1002' and the flow channel converging structure 1013', and the seventh flow path 7' connects the magnetic bead chamber 1001' and the flow channel converging structure 1013'. The eighth flow path 8' connects the power chamber 1008' and the flow channel converging structure 1013'; the ninth flow path 9' connects the magnetic bead chamber 1001' and the flow channel converging structure 1013'; the tenth flow path 10' connects the third cleaning fluid chamber 1006' and the flow channel converging structure 1013'; the eleventh flow path 11' connects the second cleaning fluid chamber 1005' and the flow channel converging structure 1013'; the twelfth flow path 12' connects the first cleaning fluid chamber 1004' and the flow channel converging structure 1013'; and the thirteenth flow path 13' connects the drive chamber 1011' and the amplification chamber.

[0262] Furthermore, the first flow path 1', second flow path 2', third flow path 3', fourth flow path 4', fifth flow path 5', sixth flow path 6', seventh flow path 7', eighth flow path 8', ninth flow path 9', tenth flow path 10', eleventh flow path 11', twelfth flow path 12', and thirteenth flow path 13' are respectively equipped with a first switching valve V1', a second switching valve V2', a third switching valve V3', a fourth switching valve V4', a fifth switching valve V5', a sixth switching valve V6', a seventh switching valve V7', an eighth switching valve V8', a ninth switching valve V9', a tenth switching valve V10', an eleventh switching valve V11', a twelfth switching valve V12', and a thirteenth switching valve V13. These switching valves are configured to allow the sealing layer 31 to approach and separate from the portion of the switching valve, thereby correspondingly closing and opening the flow path in which they are located. The structure of the switching valves is the same as in the aforementioned embodiment and will not be described again here.

[0263] For example, unlike the previous embodiments, the sealing cap 20' in this embodiment is configured to close at least one cavity, such as each cavity, thereby providing a closed environment that isolates the cavity from the outside world, preventing contamination and protecting the first surface 100a of the substrate 100'.

[0264] Figure 30 It shows Figure 27 An exploded perspective view of the detection chip and the sealing cap. Figure 31 It shows Figure 27 Another top view of the substrate of the detection chip shows the sealing groove and the snap-fit.

[0265] For example, such as Figure 30 and 31 As shown, the sealing cap 20' can snap onto the base 100' (i.e., a snap-fit ​​connection). For example, the sealing cap 20' includes a cap body 201', a snap-fit ​​assembly, and a sealing ring.

[0266] For example, the snap-fit ​​assembly includes at least one first snap-fit ​​2010' disposed on the outer peripheral portion of the cover body 201' and a second snap-fit ​​2011' disposed on the central portion of the cover body 201'. A sealing ring is disposed on the inner side of the outer peripheral portion of the cover body 201'.

[0267] Correspondingly, a first sealing groove 1020' along the outer periphery of the power cavity 1008' and a second sealing groove 1021' along the outer periphery of the plurality of cavities can be provided on the first surface 100a of the base 100'. At least one third snap fastener 1010' is also provided on the outer periphery of the base 100' for engaging with at least one first snap fastener 2010' of the cover body 201', and a fourth snap fastener 1011' is also provided on the outer periphery of the power cavity 1008' of the base 100' for engaging with a second snap fastener 2011'. For example, the number of first and third snap fasteners is equal, for example, 1-100. Using snap fasteners can achieve a simple and reliable connection.

[0268] like Figure 30 As shown, the sealing ring may include a first sealing ring 2020' and a second sealing ring 2021'. The first sealing ring 2020' corresponds to the position of the first sealing groove 1020' and is used to seal the power chamber 1008'; the second sealing ring 2021' corresponds to the position of the second sealing groove 1021' and is used to seal the space of the remaining first surface of the substrate 100'.

[0269] Optionally, the cover body 201' and the snap-fit ​​assembly are formed of a rigid material, while the sealing ring is formed of a flexible material. For example, the cover body 201', the snap-fit ​​assembly, and the sealing ring are integrally formed by two-color injection molding or assembled using an overmolding process. The rigid material is an injection-moldable material such as ABS, PC, or acrylic, used to make the snaps in the snap-fit ​​assembly and to provide rigidity for the sealing cover. The flexible material is silicone or TPU, used as the sealing ring. The second snap 2011' corresponds to the fourth snap 1011', making the center of the sealing cover 20' rigid.

[0270] Figure 32A and 32B Top view and bottom view of the sealing cap without the film adhering to it are shown respectively. Figure 32C and 32D Top view and bottom view of the sealing cap with the adhesive film are shown respectively. Figures 33A-33C Schematic diagrams of the elastic membrane of the sealing cap according to at least one embodiment are shown.

[0271] The various membrane structures disposed in the sealing cap 20' are described in detail below. For example... Figures 32A to 32D As shown, the sealing cap 20' is provided with a sample port 2030', a power port 2040' and a vent 2050'.

[0272] The sample port 2030' is used to add samples to the detection chip, and its upper end is covered by a separating membrane 1030'. The separating membrane 1030' can adhere to the upper end of the sample port 2030', and the separating membrane 1030' is completely airtight, thus gas-sealing the sample port 2030'. During operation, the operator tears open the separating membrane 1030', adds the sample, and then replaces the separating membrane 1030'.

[0273] Preferably, the release liner is a single-sided adhesive with adhesive properties. Optionally, the diameter of the release liner 1030' is 1-5 mm, preferably 3 mm.

[0274] The power port 2040' is connected to an external equipment air supply system, allowing positive and negative pressure to be applied to the power chamber 1008'. A primary isolation membrane 1040' is bonded to the inner surface of the power port 2040'; this isolation membrane 1040' is a breathable membrane. The isolation membrane 1040' can be a waterproof and breathable membrane, gauze, cotton pads, or other sheet materials, preventing liquid leakage while allowing air to pass through. Optionally, the diameter of the isolation membrane 1040' is 1-5 mm, preferably 2 mm.

[0275] The vent 2050' is the outlet through which the overall gas inside the detection chip communicates with the atmosphere. The vent 2050' can be connected, for example, to a venting cavity. A separating membrane 1050' is bonded to the vent 2050'; the separating membrane 1050' is a breathable membrane. The separating membrane 1050' is preferably a glass fiber film, which helps to adsorb nucleic acid aerosols and prevent aerosol leakage and environmental pollution. Optionally, the diameter of the separating membrane 1050' is 1-5 mm, preferably 3 mm.

[0276] The storage container 40' is loaded into the cavity of the base 100' and sealed by the snap-fit ​​structure of the sealing cap 20'. During operation, an external device compresses the storage container 40' via a first operating part, puncturing the second sealing membrane 402 of the second opening of the storage container 40'. This allows the storage container 40' to initially connect with the outside atmosphere, while simultaneously propelling the storage container 40' forward in a piston-like motion, ultimately puncturing the first sealing membrane 403. Thus, unlike the previous embodiments, in this embodiment, the second sealing membrane 402 is punctured first. The first operating part used to puncture the second sealing membrane 402 will be described in detail later.

[0277] To completely seal the detection chip, the sealing cap 20' directly separates the first operating part from the storage container 40'. The sealing cap 20' has an elastic membrane (e.g., a silicone membrane) 2060' at the first opening, specifically at the upper end of the storage container 40'. In this embodiment, the elastic membrane 2060' is gas-conducting silicone. When the first operating part punctures the second sealing membrane 402, because the contact area of ​​the elastic membrane 2060' is silicone with a thickness of 0.5-2 mm, it will not be punctured, ensuring that the upper end of the storage container 40' is not directly exposed to air after being punctured.

[0278] Optionally, the surface of the elastic membrane 2060' facing the first opening is provided with a protruding structure. For example... Figures 33A to 33C As shown, the protruding structure can be a cross-shaped, straight, or circular protrusion with a width of 0.5mm-5mm in the middle. The purpose is to make the silicone in this area not contact two smooth surfaces with the second sealing membrane 402, but to have an uneven structure, thereby forming an air passage to facilitate gas to enter the punctured hole of the second sealing membrane 402 through the air passage.

[0279] The following is combined Figure 33D-33F The operation of releasing reagents from the storage container 40 according to another embodiment is described. The storage container is held in the installed position due to friction between the first seal 404 and the inner wall of the cavity. When reagent release is required, it can be done via a push rod (e.g., Figure 33D The push rod 90 is operated by the cap body 201. The end of the push rod 90 facing the cap body 201 may have a pointed tip for indirectly piercing the second sealing membrane 402. During operation, the push rod 90 moves towards the cap body 201, for example... Figure 33DThe push rod 90 moves downwards until it contacts the elastic membrane 2060. The push rod 90 continues to move downwards, and the elastic membrane 2060 deforms under the force of the push rod 90 until it contacts the second sealing membrane 402 of the storage container 40. Further, as the push rod 90 moves downwards, since the second sealing membrane 402 is a flexible film or composite aluminum film, it is easily ruptured under external forces. Therefore, at the contact point between the elastic membrane 2060 and the second sealing membrane 402, the second sealing membrane 402 is punctured, thereby connecting the inner cavity of the storage container 40 to the atmosphere. Figure 33E As shown.

[0280] The push rod 90 continues to move downwards, and the storage container 40 also moves downwards. When it moves downwards to its limit position, that is, when the bottom of the storage container 40 contacts the flange of the cavity, as... Figure 33F As shown, the puncture structure 70 punctures the first sealing membrane 403, and the reagent in the liquid storage space begins to enter the fluid channel through the liquid guiding channel 1001.

[0281] This separate storage container allows for the independent storage of the required biochemical reagents, enabling separate production and preparation without storing all the reagents in the detection chip at once. As a result, the storage container and the main body of the detection chip can be manufactured independently, simplifying the manufacturing process and facilitating quality control during production, thereby improving product yield.

[0282] Figures 34A to 34F A schematic diagram of a driving component for a detection chip according to at least one embodiment is shown.

[0283] The drive assembly 60' is movably housed in the drive cavity 1011'. The drive assembly 60' includes a drive cover 6020' and an elastic element 6030'. The elastic element 6030' is disposed between the drive cover 6020' and the drive cavity 1011', and biases the drive cover 6020' away from the drive cavity 1011'. It should be noted that, in this disclosure, "biasing" refers to applying pressure in one direction, which is typically used in assemblies with elastic elements. Under the action of the elastic force of the elastic element, pressure is applied in one direction to the component connected to the elastic element to hold the component in a predetermined position.

[0284] Exemplarily, the drive assembly 60' further includes at least one second seal 6011', disposed between the drive cover 6020' and the drive cavity 1011' to seal the gap between them. Accordingly, the columnar outer periphery of the drive cover 6020' is provided with at least one groove 6010'. Alternatively, the columnar inner periphery of the drive cavity 1011' is provided with at least one groove. The at least one second seal 6011' is correspondingly mounted in the at least one groove. The at least one second seal 6011' can be, for example, a sealing ring.

[0285] For example, a sliding structure may also be provided between the drive cavity 1011' and the drive cover 6020'. Figures 34A to 34F An alternative embodiment of the sliding structure is illustrated by way of example. For instance, the drive cover 6020' may include a snap fastener 6021' disposed on the outer periphery of the column, corresponding to the groove structure 1021' on the inner wall of the drive cavity 1011', for engagement, such that the groove structure 1021' limits the vertical movement of the two during piston movement. The groove structure 1021' is disposed on the upper side of the drive cavity 1011' and is rectangular in shape, with dimensions matching the snap fastener 6021'. Other movable structures may be used by those skilled in the art, and this disclosure is not limited thereto.

[0286] Elastic element 6030' Figures 34A to 34F The drive chamber 1011' is shown to be installed inside the drive chamber 1011'. When the drive chamber 1011', elastic element 6030', and drive cover 6020' are assembled together, the elastic element 6030' is compressed due to the positional constraints of the snap-fit ​​6021' and the groove structure 1021'. At this time, when an external device presses the drive cover 6020', gas is expelled from the drive chamber 1011'. When the external device's pressure is removed, the drive cover 6020' automatically rebounds under the action of the elastic element 6030', allowing external gas to enter the drive chamber 1011'.

[0287] Figure 35A and Figure 35B A schematic diagram of the amplification cavity of a detection chip according to at least one embodiment is shown. Figure 35A This is an exploded perspective view of the amplification chamber. Figure 35B This is a top view of the amplification matrix in the amplification chamber.

[0288] The amplification chamber 70' can be detachably mounted to the substrate 100', and includes an upper membrane 701', an amplification substrate 702', and a lower membrane 703'. The upper membrane 701' and the lower membrane 703' are respectively bonded to the upper and lower surfaces of the amplification substrate 702' to be assembled together, and the upper membrane 701' and the lower membrane 703' are composed of a transparent film material, preferably PC or PP material, with a thickness in the range of 0.01-0.2 mm.

[0289] The lower membrane 703' includes two connecting ports 7031'. After the amplification cavity 70' is assembled, the two connecting ports 7031' are connected to the first flow path and the thirteenth flow path of the substrate 100', respectively.

[0290] The amplification substrate 702' is a 0.01-2 mm thick sheet with a cut-out structure. The amplification substrate 702' has interconnected reaction regions 7022' and at least one defoaming region 7021'. The reaction region 7022' is the PCR amplification area and is shaped like a spindle. The defoaming region 7021' is located upstream and / or downstream of the reaction region 7022'. In this embodiment, the amplification substrate 702' includes two defoaming regions 7021', located upstream and downstream of the reaction region 7022', respectively. For example, the defoaming region 7021' is a circle with a diameter of 1-3 mm, which can intercept air bubbles in the reagent.

[0291] Figures 36 to 43 A schematic diagram of liquid flow is shown illustrating an operation method of a detection chip according to at least one embodiment. For example, in at least one application scenario, the detection chip may operate as follows.

[0292] S1, sample addition procedure.

[0293] Tear open the isolation membrane 1030', add the sample to be tested into the sample chamber 1002', and then replace the isolation membrane 1030'.

[0294] S2, magnetic bead release step.

[0295] Open the eighth switch valve V8 and the ninth switch valve V9, and use an external instrument to apply negative pressure to the power chamber 1008', so that 200 μl of magnetic beads flow from the magnetic bead chamber 1001' through the ninth flow path 9' and the eighth flow path 8' to the power chamber 1008'. Then close the eighth switch valve V8 and the ninth switch valve V9. S3, lysis buffer release step.

[0296] Open the eighth switch valve V8 and the seventh switch valve V7, apply negative pressure to the power chamber 1008' using external instruments, extract 500 μl of lysis liquid from the lysis liquid chamber 1003' and transport it to the power chamber 1008 through the seventh flow path 7' and the eighth flow path 8', and then close the eighth switch valve V8 and the seventh switch valve V7.

[0297] S4, Sample extraction step.

[0298] Open the eighth switch valve V8 and the sixth switch valve V6, apply negative pressure to the power chamber 1008' using an external instrument, extract 500 μl of sample from the sample chamber and transport it to the power chamber 1008' through the sixth flow path 6' and the eighth flow path 8', and then close the eighth switch valve V8 and the sixth switch valve V6.

[0299] S5, magnetic bead adsorption step.

[0300] Open the third switch valve V3. Using an external instrument, alternate between negative and positive pressure on the power chamber 1008', causing the lysis buffer, sample, and magnetic bead solution to travel back and forth between the power chamber 1008' and the mixing chamber 1009' via the third flow path 3'. Mix 1-100 times until a homogeneous mixture is achieved. Thoroughly mixing the sample, lysis buffer, and magnetic beads effectively lyses the sample and releases nucleic acid fragments, while the magnetic beads efficiently adsorb the nucleic acid fragments. The homogeneous mixture terminates at the power chamber 1008'; then close the third switch valve V3.

[0301] S6, First magnetic bead collection step.

[0302] Using an external instrument, bring the magnet close to the magnetic bead trapping chamber 1012' and open the third switch valve V3. Apply alternating negative and positive pressure to the power chamber 1008' using the external instrument. The sample, lysis buffer, and magnetic bead mixture are transported back and forth between the power chamber 1008' and the mixing chamber 1009' via the third flow path 3', mixing 1-100 times. The magnetic beads are attracted by the magnet and thus retained in the magnetic bead trapping chamber 1012', while the waste liquid ultimately remains in the power chamber 1008'. Then close the third switch valve V3.

[0303] S7, the first step of emptying the waste liquid.

[0304] Open the fourth switch valve V4, and use external instruments to apply positive pressure to the power chamber 1008'. The waste liquid in the power chamber 1008' is transported to the waste liquid chamber 1010' through the third flow path 3' and the fourth flow path 4'. Close the fourth switch valve V4. Use external instruments to move the magnet away from the bottom of the magnetic bead trapping chamber 1012'.

[0305] S7, First cleaning fluid release step.

[0306] Open the eighth switch valve V8 and the twelfth switch valve V12, apply negative pressure to the power chamber 1008' using external instruments, and extract 500 μl of the first cleaning fluid from the first cleaning fluid chamber 1004' and transport it to the power chamber 1008' through the twelfth flow path 12' and the eighth flow path 8'. Then close the eighth switch valve V8 and the twelfth switch valve V12.

[0307] S8, the first washing and mixing step.

[0308] Open the third switch valve V3. Using an external instrument, alternate negative and positive pressure are applied to the power chamber 1008'. The mixture formed by the first cleaning solution and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009', mixing 1-100 times until a homogenized mixture is achieved. By thoroughly mixing the first cleaning solution and the magnetic beads, residual sample, lysis buffer, and other impurities on the surface of the magnetic beads can be effectively washed away. The homogenized mixture terminates at the power chamber 1008', then the third switch valve V3 is closed.

[0309] The surface of magnetic beads that have adsorbed nucleic acid molecules may retain impurities such as components from the sample and lysis buffer. These impurities will affect subsequent amplification reactions and detection. The first washing step, as well as the subsequent second and third washing steps, will remove these impurities. For example, the first washing solution in the first washing step can be used to clean protein molecules from the surface of the nucleic acid.

[0310] S9, the second magnetic bead collection and mixing step.

[0311] An external instrument brings a magnet close to the magnetic bead capture chamber 1012' and opens the third switch valve V3. Using the external instrument, alternating negative and positive pressures are applied to the power chamber 1008'. The mixture formed by the first cleaning fluid and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009' through the third flow path 3', mixing 1-100 times. The magnetic beads are attracted by the magnet and thus retained in the magnetic bead capture chamber 1012', while the waste liquid remains in the power chamber 1008'. Then, the third switch valve V3 is closed.

[0312] After the second magnetic bead collection and mixing step, the magnetic beads with adsorbed nucleic acid molecules are separated from other components in the mixture.

[0313] S10, the second step of emptying the waste liquid.

[0314] Open the fourth switch valve V4 and use external instruments to apply positive pressure to the power chamber 1008'. The waste liquid in the power chamber 1008' is transported to the waste liquid chamber 1010' through the third flow path 3' and the fourth flow path 4'. Then close the fourth switch valve V4. The external instruments move the magnet away from the bottom of the magnetic bead trapping chamber 1012'.

[0315] S11, Second cleaning fluid release step.

[0316] Open the eighth switch valve V8 and the eleventh switch valve V11, apply negative pressure to the power chamber 1008' using external instruments, extract 500 μl of the second cleaning fluid from the second cleaning fluid chamber 1005' and transport it to the power chamber 1008' through the eleventh flow path 11' and the eighth flow path 8', and then close the eighth switch valve V8 and the eleventh switch valve V11.

[0317] S12, the second washing and mixing step.

[0318] Open the third switch valve V3. Using an external instrument, alternate negative and positive pressure are applied to the power chamber 1008'. The mixture formed by the second cleaning solution and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009', mixing 1-100 times until a homogenized state is achieved. By thoroughly mixing the second cleaning solution and the magnetic beads, residual sample, lysis buffer, and other impurities on the surface of the magnetic beads can be effectively washed away. The homogenized mixture terminates at the power chamber 1008', then the third switch valve V3 is closed.

[0319] For example, the second cleaning solution in the second cleaning step can be used to clean small molecule impurities and salt ions from the surface of the magnetic beads.

[0320] S13, the third magnetic bead collection and mixing step.

[0321] An external instrument brings a magnet close to the magnetic bead trapping chamber 1012' and opens the third switch valve V3. The external instrument applies alternating negative and positive pressure to the power chamber 1008'. The mixture formed by the second cleaning fluid and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009' through the third flow path 3', mixing 1-100 times. The magnetic beads are attracted by the magnet and remain in the magnetic bead trapping chamber 1012', while the waste liquid remains in the power chamber 1008'. Then, the third switch valve V3 is closed.

[0322] S14, the third step of emptying the waste liquid.

[0323] Open the fourth switch valve V4 and use external instruments to apply positive pressure to the power chamber 1008'. The waste liquid in the power chamber 1008' is transported to the waste liquid chamber 1010' through the third flow path 3' and the fourth flow path 4'. Then close the fourth switch valve V4. The external instruments move the magnet away from the bottom of the magnetic bead trapping chamber 1012'.

[0324] S15, Third cleaning fluid release step.

[0325] Open the eighth switch valve V8 and the tenth switch valve V10, apply negative pressure to the power chamber 1008' using external instruments, extract 500 μl of the third cleaning fluid from the third cleaning fluid chamber 1006' and transport it to the power chamber 1008' through the tenth flow path 10' and the eighth flow path 8', and then close the eighth switch valve V8 and the tenth switch valve V10.

[0326] S16, the third cleaning and mixing step.

[0327] Open the third switch valve V3. Using an external instrument, alternate negative and positive pressure are applied to the power chamber 1008'. The mixture formed by the third cleaning solution and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009', mixing 1-100 times until a homogenized mixture is achieved. By thoroughly mixing the third cleaning solution and the magnetic beads, residual sample, lysis buffer, and other impurities on the surface of the magnetic beads can be effectively washed away. The homogenized mixture is terminated at the power chamber 1008', then the third switch valve V3 is closed.

[0328] For example, the third cleaning solution in this third cleaning step can be used to clean small molecules and salt ions remaining on the surface of the magnetic beads. For example, this third cleaning solution is the same as the second cleaning solution.

[0329] S17, the fourth step of collecting and mixing magnetic beads.

[0330] An external instrument brings a magnet close to the magnetic bead capture chamber 1012' and opens the third switch valve V3. The external instrument applies alternating negative and positive pressure to the power chamber 1008'. The mixture formed by the third cleaning fluid and the magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009' through the third flow path 3', mixing 1-100 times. The magnetic beads are attracted by the magnet and remain in the magnetic bead capture chamber 1012', while the waste liquid remains in the power chamber 1008'. Then, the third switch valve V3 is closed.

[0331] S18, the fourth step of emptying the waste liquid.

[0332] Open the fourth switch valve V4 and use external instruments to apply positive pressure to the power chamber 1008'. The waste liquid in the power chamber 1008' is transported to the waste liquid chamber 1010' through the third flow path 3' and the fourth flow path 4'. Then close the fourth switch valve V4. The external instruments move the magnet away from the bottom of the magnetic bead trapping chamber 1012'.

[0333] S19, Eluent Release Step.

[0334] Open the eighth switch valve V8 and the fifth switch valve V5, apply negative pressure to the power chamber 1008' using external instruments, extract 100 μl of eluent from the eluent chamber 1007' and transport it to the power chamber 1008' through the fifth flow path 5' and the eighth flow path 8', and then close the eighth switch valve V8 and the fifth switch valve V5.

[0335] S20, elution and mixing steps.

[0336] Open the third switch valve V3. Using an external instrument, alternate negative and positive pressure are applied to the power chamber 1008'. The eluent and magnetic beads form a mixture that flows back and forth between the power chamber 1008' and the mixing chamber 1009' via the third flow path 3', mixing 1-100 times until a homogenized mixture is achieved. By thoroughly mixing the eluent and magnetic beads, nucleic acid fragments can be effectively eluted from the surface of the magnetic beads. The homogenized mixture terminates at the power chamber 1008'; then close the third switch valve V3.

[0337] S21, the fifth step of collecting and mixing magnetic beads.

[0338] The external instrument brings the magnet close to the magnetic bead trapping chamber 1012' and opens the third switch valve V3. The external instrument applies alternating negative and positive pressure to the power chamber 1008'. The mixture of eluent and magnetic beads is transported back and forth between the power chamber 1008' and the mixing chamber 1009' through the third flow path 3', mixing 1-100 times. The magnetic beads are attracted by the magnet and remain in the magnetic bead trapping chamber 1012', while the waste liquid remains in the power chamber 1008'. Then, the third switch valve V3 is closed. The external instrument moves the magnet away from the bottom of the magnetic bead trapping chamber 1012'.

[0339] S22, the transfer step of amplification cavity 70'.

[0340] Open the first switch valve V1 and the thirteenth switch valve V13, and use an external instrument to squeeze the drive chamber 1011' to expel the gas. Then close the first switch valve V1 and the thirteenth switch valve V13.

[0341] Open the second switch valve V2, the external instrument removes the pressure on the drive chamber 1011', the drive chamber 1011' rebounds, and the eluent in the mixing chamber 1009' is drawn into the drive chamber 1011'. Then close the second switch valve V2.

[0342] Reopen the first switch valve V1 and the thirteenth switch valve V13, and use an external instrument to squeeze the drive chamber 1011' to push the eluent into the amplification chamber 70', where it reconstitutes with the lyophilized powder to form the amplification reagent. Then close the first switch valve V1 and the thirteenth switch valve V13.

[0343] S23, Amplification step.

[0344] The amplification chamber is cyclically heated to amplify the sample inside, and then the properties of the amplified sample, such as optical properties, are detected to obtain the detection results.

[0345] The above operation process can be implemented by operating the detection chip through a detection device. For example, after step S1, the detection chip can be placed in the appropriate position of the detection device, and then the relevant components of the detection device can operate the detection chip.

[0346] The detection chip according to the embodiments of this disclosure can process the entire process of original sample from sample addition to detection, and has a high degree of integration.

[0347] Those skilled in the art will understand that the detection chip according to embodiments of this disclosure can be operated in a different order than described 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 already undergone lysis treatment, the steps related to lysis cannot be omitted.

[0348] The detection chip of this embodiment can be installed at an appropriate position in the detection device of this embodiment so as to cooperate with the detection device to complete the detection process.

[0349] Figure 44 A detection apparatus according to an embodiment of the present disclosure is shown.

[0350] The detection chip 1000 includes a first chip positioning structure 1060 for positioning the detection chip 1000 at a suitable position in the detection device 2000, such as fixing the detection chip 1000, so that it can be used for transmission and detection. For example, the first chip positioning structure 1060 may include a chip positioning hole provided in the substrate 100, which may be a circular through hole with a diameter in the range of 0.1-10 mm, for cooperating with a positioning pin on the detection device 2000, which serves as a second chip positioning structure 2001, to position the detection chip 1000 onto the detection device 2000. Alternatively, the first chip positioning structure may include elongated chip positioning grooves located on both sides of the detection chip 1000 for matching with a fixing device on the detection device 2000 to fix the chip onto the detection device 2000.

[0351] The detection device 2000 may include the push rod 90 as described above as a first operating part 2010 for changing the volume of the cavity. For example, if necessary, there may be multiple push rods 90, which are respectively disposed above different cavities to change the volume of the cavity. Alternatively, the detection device 2000 may include push rods 90 that are movable above different cavities.

[0352] The detection device 2000 may include a second operating portion 2020 that engages with the push rod 502 to operate the push rod 502.

[0353] The detection device 2000 may include a third operating portion 2030, such as a push rod, for pressing the first flexible layer 20 to operate the switching valve. The push rod may include multiple push rods or be configured to be movable.

[0354] The specific implementation of the second operating part 2020 is not limited. For example, it can be a combination of a hydraulic device, a propulsion control mechanism (e.g., a control circuit or control chip), a cylinder with a slot, and a limiting mechanism, with the limiting mechanism engaging with the push rod 502. Alternatively, it can be a combination of a motor, a propulsion control mechanism, a cylinder with a slot, and a limiting mechanism, with the limiting mechanism engaging with the push rod 502, or any other arbitrary implementation, depending on actual needs. Similarly, the third operating part 2030 can also adopt a similar structure, simply replacing the cylinder with a slotless cylinder to serve as a protrusion. The third operating part 2030 can be, for example, a combination of a pneumatic control device, an air compressor, and a gas delivery pipe (or gas circuit board), or any other arbitrary implementation, depending on actual needs. The embodiments of this disclosure do not limit this.

[0355] In addition, the detection device 2000 may also include a heating device for heating the amplification chamber 200, a detection device for detecting the amplification reaction in the amplification chamber 200, a heating module for heating the first mixing chamber 130 and the second mixing chamber 140, a magnet 900 for fixing the magnetic beads, etc.

[0356] It should be noted that, in the embodiments of this disclosure, the detection device 2000 may also include more components and units, not limited to the chip positioning structure, operating part, and heating device described above. For example, the detection device 2000 may also include a power supply, a central processing unit (CPU), an optical detection unit, a temperature control unit, etc., thereby giving the detection device 2000 more complete and richer functions. For example, the optical detection unit may include a light source, a photoelectric detection device, etc., coupled to the CPU, and the light source may be infrared light, visible light, etc., depending on the detection principle. The embodiments of this disclosure do not limit these additional components and units.

[0357] The scope of this disclosure is not limited by the embodiments described above, but by the appended claims and their equivalents.

Claims

1. A detection chip, comprising: A matrix having a first surface and including at least one cavity and at least one fluid channel leading to said at least one cavity, the cavity having a first opening in the first surface; At least one storage container, the at least one storage container including a second opening and a first sealing film sealing the second opening; The at least one cavity includes a puncture structure disposed in the cavity for puncturing the first sealing membrane, so that the reagent in the storage container can enter the fluid channel. The storage container is separately disposed from the substrate, and the at least one storage container is slidably accommodated in the at least one cavity through a first opening.

2. The detection chip according to claim 1, wherein, The at least one storage container further includes a third opening and a second sealing film that seals the third opening. The first sealing membrane and the second sealing membrane define a liquid storage space for containing reagents.

3. The detection chip according to claim 2, wherein, The storage container also includes a removable lid, which is installed at the third opening and covers the second sealing film.

4. The detection chip according to claim 1 further comprises: A sealing cap is connected to the substrate and seals the first opening of the substrate.

5. The detection chip according to claim 4, wherein, The sealing cap engages with the substrate.

6. The detection chip according to claim 5, wherein, The sealing cap includes a cap body and a snap-fit ​​assembly. The latching assembly includes at least one first latch disposed on the outer peripheral portion of the cover body and a second latch disposed on the central portion of the cover body.

7. The detection chip according to claim 6, wherein, The sealing cap also includes a sealing ring, which is disposed on the inner side of the outer peripheral portion of the cap body. The cover body and the snap-fit ​​assembly are made of rigid material, while the sealing ring is made of flexible material.

8. The detection chip according to claim 7, wherein, The cover body, the buckle assembly, and the sealing ring are integrally formed by two-color injection molding or assembled by a rubber coating process.

9. The detection chip according to claim 6, wherein, The sealing cap has a covering film at the first opening, and the covering film is an elastic film.

10. The detection chip according to claim 9, wherein, The surface of the covering film facing the first opening has a protruding structure.

11. The detection chip of claim 10, wherein, The protruding structure includes a central protrusion and a plurality of peripheral protrusions distributed circumferentially around the central protrusion. The central protrusion is in the shape of a line, a cross, or a circle, and the plurality of peripheral protrusions are in the shape of a line or a circle.

12. The detection chip according to claim 1, further comprising: At least one first seal is disposed between the storage container and the cavity to seal the gap between the storage container and the cavity.

13. The detection chip according to claim 12, wherein, The storage container has at least one groove on its outer surface, or the cavity has at least one groove on its inner surface. The at least one first seal is installed in the at least one groove. The at least one first seal is an elastic ring.

14. The detection chip according to claim 1 or 2, wherein, The storage container includes a drainage channel and an inner wall having an inclined first angle at the end facing the second opening, the drainage channel connecting the inner wall and the second opening.

15. The detection chip according to claim 14, wherein, The first angle ranges from 5 degrees to 60 degrees.

16. The detection chip according to claim 14, wherein, The cavity further includes a liquid guiding channel, and the matrix further includes a fluid channel. The fluid channel is located at the puncture structure and connects the cavity and the fluid channel.

17. The detection chip according to claim 16, wherein, The puncture structure has a needle-shaped protrusion at one end facing the storage container. The center point of the needle-shaped protrusion coincides with the orthographic projection of the center of the bottom surface of the cavity. The size of the needle-shaped protrusion is smaller than the size of the second opening.

18. The detection chip according to claim 17, wherein, The needle-shaped protrusion is pyramidal in shape and has multiple side edges. The opening of the liquid guiding channel in the cavity is located between two of the multiple side edges and is connected to the bottom surface and one side surface of the pyramid.

19. The detection chip according to claim 17, wherein, The needle-shaped protrusion is a cone, and the opening of the liquid guiding channel in the cavity is connected to the bottom surface of the cone.

20. The detection chip according to claim 18 or 19, wherein, The area of ​​the opening of the fluid channel in the cavity is smaller than the area of ​​the bottom surface of the needle-shaped protrusion.

21. The detection chip according to claim 4, wherein, The bottom of the cavity is provided with a flange, which is configured to contact the storage container and confine the storage container to an extreme position, whereby the puncture structure punctures the first sealing film.

22. The detection chip according to claim 21, further comprising: Multiple fluid channels; as well as A mixing structure wherein at least one of the plurality of cavities is in communication with the mixing structure via at least one of the plurality of fluid channels. The mixing structure includes a pusher, a first gas isolation membrane, and a first mixing chamber and a second mixing chamber that are interconnected. The first gas isolation membrane seals the opening of the first mixing chamber. The push rod can be slidably and sealingly installed inside the second mixing chamber or disposed separately on the opening side of the second mixing chamber relative to the detection chip. The reciprocating motion of the push rod in the second mixing chamber enables the reagent to be transported between the first mixing chamber and the second mixing chamber.

23. The detection chip of claim 22, wherein, The push rod includes an operating part disposed at one end of the push rod and protruding from the surface of the push rod.

24. The detection chip of claim 23, wherein, The push rod also includes at least one groove and a sealing ring, the at least one groove being disposed on the outer surface of the push rod, and the sealing ring being installed in the at least one groove.

25. The detection chip according to claim 22, wherein, The mixing structure includes an air vent, a first air-permeable membrane, a second air-permeable membrane, a power chamber, and a mixing chamber communicating with the power chamber. The power chamber and the plurality of cavities are connected by multiple fluid channels.

26. The detection chip of claim 25, wherein, The vent is located on the sealing cover.

27. The detection chip of claim 25, wherein, The first breathable membrane covers the vent, and the substrate communicates with the outside atmosphere through the vent and the first breathable membrane.

28. The detection chip of claim 25, wherein, The second breathable membrane covers the power chamber, which is connected to an external air source system through the second breathable membrane to receive positive or negative pressure applied by the external air source system.

29. The detection chip according to claim 25, further comprising: A magnetic bead capturing chamber is disposed between the power chamber and the mixing chamber and is connected to both the power chamber and the mixing chamber respectively. The area or size of the middle region of the magnetic bead capturing cavity is larger than the area or size of the two end regions.

30. The detection chip of claim 25, wherein, A flow channel converging structure is provided between multiple cavities.

31. The detection chip according to claim 30, wherein, A flow channel converging structure is provided between the power cavity and the plurality of cavities.

32. The detection chip according to claim 31, wherein, The central part of the flow channel converging structure is circular, and multiple trapezoidal structures are evenly distributed around the central part in a circumferential direction.

33. The detection chip according to claim 22, wherein, The substrate has a second surface opposite to the first surface, and the plurality of fluid channels are disposed in the second surface.

34. The detection chip according to claim 33, wherein, The plurality of fluid channels include a plurality of substrate grooves formed in the second surface. The detection chip also includes a sealing layer on the second surface to at least cover the plurality of substrate grooves to form a fluid channel.

35. The detection chip according to claim 34, further comprising: The adhesive layer on the second surface, The adhesive layer is disposed between the substrate and the sealing layer and configured to bond the substrate and the sealing layer to each other. The adhesive layer exposes at least the plurality of substrate grooves in the substrate and separates the plurality of fluid channels from each other.

36. The detection chip according to claim 33, wherein, The fluid channel also includes multiple flow paths and multiple switching valves, with the multiple switching valves located in the multiple flow paths respectively. Each of the switching valves is configured to control the connection and disconnection of at least a portion of the corresponding fluid passage. The substrate has a substrate groove on its second surface for forming at least one of the plurality of flow paths.

37. The detection chip of claim 36, wherein, The second surface has a partition portion in the membrane valve section, the partition portion having a platform portion that separates the substrate grooves belonging to the same flow path.

38. The detection chip according to claim 37, wherein, The platform portion is flush with the rest of the second surface or has a smaller recess depth compared to the substrate groove.

39. The detection chip according to claim 22, further comprising: The sample chamber is sealed with an isolation membrane, which is an adhesive single-sided adhesive.

40. The detection chip according to claim 22, further comprising: The amplification chamber is a quantitative chamber.

41. The detection chip according to claim 40, wherein, The amplification chamber has an interconnected reaction zone and a debubbling zone.

42. The detection chip of claim 41, wherein, The defoaming zone is located upstream and / or downstream of the reaction zone.

43. The detection chip of claim 41 or 42, wherein, The reaction zone is shaped like a spindle, and the defoaming zone is shaped like a circle.

44. The detection chip according to claim 40, further comprising: The driving cavity is in fluid communication with the amplification cavity and the mixing structure.

45. The detection chip according to claim 44, further comprising: A drive assembly movably housed in the drive cavity, the drive assembly including a drive cover and an elastic element disposed between the drive cover and the drive cavity, and biasing the drive cover.

46. ​​The detection chip according to claim 45, wherein, The drive assembly further includes at least one second seal disposed between the drive cover and the drive cavity to seal the gap between the drive cover and the drive cavity. The outer surface of the drive cover is provided with at least one groove, or the inner surface of the drive cavity is provided with at least one groove. The at least one second seal is correspondingly installed in the at least one groove.

47. The detection chip according to claim 45 or 46, wherein, A sliding structure is provided between the drive cavity and the drive cover.

48. The detection chip according to any one of claims 1-13, 15-19, 21-42, 44-46, further comprising: The first chip positioning structure is configured to position and install the detection chip into the detection device.

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