Micro-fluidic chip

Abstract

The micro-fluidic chip comprises a base body, a first on-off switching piece, an electrochemical detection piece, a fluid driving piece, a first reagent storage piece and a second on-off switching piece, the base body is provided with an extraction chamber, a uniform mixing chamber, a reaction chamber, an electrochemical reaction tank and a waste liquid chamber, an outlet of the extraction chamber is communicated with an inlet of the uniform mixing chamber, and an outlet of the reaction chamber is communicated with an outlet of the uniform mixing chamber. An outlet of the uniform mixing chamber is communicated with an inlet of the reaction chamber through a first on-off switching piece, the outlet of the uniform mixing chamber is communicated with the waste liquid chamber through the first on-off switching piece, an outlet of the reaction chamber is communicated with an inlet of the electrochemical reaction tank through the first on-off switching piece, and an outlet of the electrochemical reaction tank is communicated with the waste liquid chamber; the electrochemical detection part is used for acquiring reaction information of the electrochemical reaction tank; the fluid driving part is connected with the base body and is communicated with the extraction chamber through the second on-off switching part; the first reagent storage part is arranged on the base body and is communicated with the extraction chamber through the second on-off switching part; and cross contamination is avoided.

Description

Technical Field

[0001] The utility model relates to the technical field of semiconductor manufacturing, in particular to a microfluidic chip. Background Art

[0002] With the development of molecular biology, the polymerase chain reaction (PCR) technology for rapid in vitro amplification of target DNA / RNA fragments has been widely used and has become the main method for rapid detection of pathogen genes. Real-time fluorescence PCR technology monitors the PCR process in real time by introducing fluorescent dyes or fluorescently labeled specific probes, achieving a leap from qualitative judgment to quantitative analysis. It can detect pathogen genes efficiently and specifically, greatly shortening the detection window period, which is conducive to early diagnosis and treatment and reducing mortality.

[0003] The existing rapid detection method for pathogen genes is: first, the sample is lysed, nucleic acid is released, nucleic acid is adsorbed, impurities are washed, and nucleic acid is eluted by an automatic nucleic acid extractor, and then the eluted nucleic acid is amplified by a PCR amplification instrument, and then the sample is tested by configuring a PCR reaction. However, in the above method, multiple devices need to be used in conjunction, and when the sample is transferred between the various devices, it is easy to cause sample contamination. Utility Model Content

[0004] In view of this, the utility model provides a microfluidic chip to solve the problem that the existing rapid pathogen gene detection method requires the use of multiple devices, and the sample is easily contaminated when transferred between the devices, and manual transfer is required.

[0005] The utility model provides a microfluidic chip, comprising:

[0006] A substrate, comprising an extraction chamber, a mixing chamber, a reaction chamber, an electrochemical reaction tank and a waste liquid chamber, wherein the outlet of the extraction chamber is connected to the inlet of the mixing chamber, the outlet of the mixing chamber is connected to the inlet of the reaction chamber via a first on-off switch, and the outlet of the mixing chamber is connected to the waste liquid chamber via the first on-off switch, the outlet of the reaction chamber is connected to the inlet of the electrochemical reaction tank via the first on-off switch, and the outlet of the electrochemical reaction tank is connected to the waste liquid chamber;

[0007] The first on-off switch is used to control the on-off between the mixing chamber and the reaction chamber, the on-off between the mixing chamber and the waste liquid chamber, and the on-off between the reaction chamber and the electrochemical reaction tank;

[0008] An electrochemical detection element connected to the substrate and having a detection end, the detection end extending into the electrochemical reaction tank, the electrochemical detection element being used to obtain reaction information of the electrochemical reaction tank;

[0009] A fluid driving member connected to the base body and communicated with the extraction chamber via a second on-off switching member;

[0010] a first reagent storage element, disposed on the substrate and connected to the extraction chamber via the second on-off switch;

[0011] The second on-off switch is used to control the on-off between the fluid driving member and the extraction chamber, and the on-off between the first reagent storage member and the extraction chamber;

[0012] The fluid driving member is capable of driving the flow of the fluid in the extraction chamber.

[0013] Beneficial effect: First, add the sample to the extraction chamber, and then use one or more of mechanical lysis, chemical lysis, and enzyme lysis to lyse the sample in the extraction chamber and release the nucleic acid, and then add magnetic bead liquid to the extraction chamber, the nucleic acid is adsorbed by the magnetic beads in the magnetic bead liquid, and the first reagent storage component is connected to the extraction chamber through the second on-off switch. The first reagent storage component can store cleaning liquid, and the cleaning liquid in the first reagent storage component is transported to the extraction chamber after passing through the first on-off switch, and then the first reagent storage component is disconnected from the extraction chamber through the second on-off switch, and then the fluid drive component is connected to the extraction chamber through the second on-off switch. The first on-off switch connects the mixing chamber to the waste liquid chamber, and the fluid driving component transports the cleaning fluid in the extraction chamber and the impurities therein to the mixing chamber. As the fluid driving component continues to work, the cleaning liquid in the extraction chamber is continuously transported to the mixing chamber until the cleaning fluid in the mixing chamber is completely transported to the waste liquid chamber, so as to clean the nucleic acid adsorbed by the magnetic beads. The second on-off switch disconnects the fluid driving component from the extraction chamber, and then adds elution liquid to the extraction chamber to elute the nucleic acid and the magnetic beads. Then, the dilution liquid is added to the extraction chamber, and then the fluid driving component is disconnected from the extraction chamber by the second on-off switch. The first on-off switch connects the mixing chamber to the reaction chamber, and the fluid driving member connects the dilution liquid in the extraction chamber and the nucleic acid therein to the mixing chamber for rapid mixing. The mixed liquid is driven by the fluid driving member and is transported to the reaction chamber. The fluid driving member is disconnected from the extraction chamber by the second on-off switch, and the mixing chamber is disconnected from the reaction chamber by the first on-off switch. At this time, the nucleic acid undergoes an amplification reaction (PCR) in the reaction chamber. After the nucleic acid is amplified, the fluid driving member is connected to the extraction chamber by the second on-off switch, and the mixing chamber is disconnected by the first on-off switch. The replacement component connects the mixing chamber with the reaction chamber, and connects the reaction chamber with the electrochemical reaction tank. The fluid driving component transports the liquid in the mixing chamber to the electrochemical reaction tank and performs an electrochemical reaction. The electrochemical detection component then detects and outputs the detection information, thereby realizing the detection of pathogen genes. By using the microfluidic chip, there is no need to use other equipment or transfer samples, thereby avoiding cross contamination caused by the transfer process. The detection of pathogen genes can be completed by the microfluidic chip alone, thereby reducing the use of other equipment, reducing the size of the detection equipment, and improving space utilization, thereby realizing the portability and compactness requirements of medical testing.

[0014] In an optional embodiment, the extraction chamber includes a lysis chamber, a washing chamber and an elution chamber which are connected in sequence, the inlet of the lysis chamber is connected to the second on-off switch, and the outlet of the elution chamber is connected to the mixing chamber.

[0015] Samples can complete different operations in different chambers, thereby providing a stable microenvironment for sample lysis, washing, and elution, ensuring efficient operation of nucleic acid extraction.

[0016] In an optional embodiment, the substrate further has:

[0017] An exhaust hole is provided on the base body, and the waste liquid chamber is connected with the outside through the exhaust hole.

[0018] The air pressure in the waste liquid chamber is balanced with that in the outside world to ensure that the liquid in the mixing chamber or the reaction tank can be smoothly transported to the waste liquid chamber.

[0019] In an optional embodiment, the substrate has a plurality of the reaction chambers;

[0020] The substrate has a plurality of electrochemical reaction tanks, and the outlet of any one of the electrochemical reaction tanks is connected to the waste liquid chamber;

[0021] The first on-off switching member includes a first valve seat and a first valve body, the first valve body is rotatably connected to the first valve seat; the first valve seat is arranged in the base, the first valve seat has a first channel, a second channel, a third channel, a plurality of fourth channels and a plurality of fifth channels, the outlet of the extraction chamber is connected to the first channel; the inlet of any one of the reaction chambers is connected to the second channel, the third channel is connected to the waste liquid chamber, the outlet of any one of the reaction chambers is connected to the corresponding fourth channel, and the inlet of any one of the reaction tanks is connected to the corresponding fifth channel; the valve body has a first communicating port, a second communicating port, a third communicating port, a plurality of fourth communicating ports and a plurality of fifth communicating ports, the first communicating port is connected to the second communicating port, and the first communicating port is connected to the first channel; any one of the fourth communicating ports is connected to the corresponding fifth communicating port;

[0022] The first valve body has a first switching state in which the second communication port is rotated under the action of an external force to communicate with the second channel, the third communication port is communicated with the third channel, any fourth communication port is communicated with the corresponding fourth channel, and any fifth communication port is communicated with the corresponding fifth channel;

[0023] The first valve body has a second switching state in which the first valve body rotates under the action of an external force to connect the second communication port with the third channel;

[0024] The first valve body has a third switching state in which the first valve body rotates under an external force to block the second communication port, the third communication port, any one of the fourth communication ports, and any one of the fifth communication ports.

[0025] In an optional embodiment, the first reagent storage unit includes a plurality of liquid capsule reagent packages;

[0026] The second on-off switch member includes a second valve seat and a second valve body, the second valve body is rotatably connected to the second valve seat; the second valve seat is arranged in the base, the second valve seat has a sixth channel, a seventh channel, and a plurality of eighth channels, the inlet of the extraction chamber is connected to the sixth channel, the fluid driving member is connected to the seventh channel, and any one of the liquid capsule reagent packages is connected to the corresponding eighth channel; the valve body has a sixth communication port and a seventh communication port, the sixth communication port is connected to the seventh communication port, and the sixth communication port is connected to the sixth channel;

[0027] The second valve body has a fourth switching state in which it rotates under the action of an external force to connect the seventh communication port with the seventh channel;

[0028] The second valve body has a fifth switching state in which it rotates under the action of an external force to connect the seventh communication port with one of the eighth channels and block the seventh channel.

[0029] The microfluidic chip can control the liquid flow route through the fluid driving element, the first on-off switching element, and the second on-off switching element, thereby realizing automated reagent loading, mixing, reaction, and waste liquid discharge, improving detection efficiency, reducing manual operation steps, reducing human errors, and improving detection consistency and reliability.

[0030] In an optional embodiment, an air inlet hole is formed on the base, the second valve seat has a ninth channel, the ninth channel is communicated with the air inlet hole, an eighth communication port and a ninth communication port are formed on the second valve body, the eighth communication port is communicated with the ninth communication port;

[0031] The second valve body has a sixth switching state in which the second valve body rotates under an external force to connect the eighth communication port with the seventh channel, the ninth communication port with the ninth channel, and blocks the seventh communication port.

[0032] The fluid driving component can be communicated with the outside world through the air inlet hole, so that the outside gas can enter the fluid driving component, thereby enabling the fluid driving component to work normally.

[0033] In an optional embodiment, the substrate further comprises:

[0034] a sample chamber, wherein the sample chamber is connected to the fluid driving member and the second on-off switching member respectively;

[0035] A sample hole is provided on the substrate, and the sample chamber is connected with the outside through the sample hole.

[0036] The sample is added into the sample chamber through the sample, and the fluid driving component drives the sample in the sample chamber to pass through the second on-off switch and the lysis chamber in sequence and then enter the washing chamber. The sample chamber and the extraction chamber are set separately to avoid the influence of the external environment on the extraction chamber.

[0037] In an optional embodiment, the mixing chamber is a spirally arranged flow channel and / or a flow channel with a varying inner diameter.

[0038] In an optional implementation, it also includes:

[0039] A second reagent storage element, disposed on the substrate;

[0040] The base body further includes a buffer chamber which is communicated with the second reagent storage part and the lifting chamber, respectively.

[0041] In an optional embodiment, the electrochemical detection element has multiple detection ends, any one of the detection ends has multiple electrodes, and any one of the electrodes can collect detection information. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] In order to more clearly illustrate the specific implementation methods of the utility model or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0043] Figure 1 It is a structural schematic diagram of the microfluidic chip provided by the utility model;

[0044] Figure 2 It is an exploded schematic diagram of the microfluidic chip provided by the utility model;

[0045] Figure 3 It is a front view of the second substrate in the microfluidic chip provided by the utility model;

[0046] Figure 4 It is a perspective view of the second substrate in the microfluidic chip provided by the utility model;

[0047] Figure 5 yes Figure 4 A partial enlarged schematic diagram of the middle circle A;

[0048] Figure 6 yes Figure 4 A partial enlarged schematic diagram of the middle circle B;

[0049] Figure 7It is a structural schematic diagram of the second valve body in the microfluidic chip provided by the utility model;

[0050] Figure 8 It is a structural schematic diagram of the first valve body in the microfluidic chip provided by the utility model;

[0051] Fig. 9 It is a structural schematic diagram of the electrochemical detection element in the microfluidic chip provided by the utility model;

[0052] Fig.10 It is a bottom view of the second substrate in the microfluidic chip provided by the utility model;

[0053] Figures 11 to 18 This is a work flow chart of the microfluidic chip provided by the utility model.

[0054] Description of reference numerals:

[0055] 1. substrate; 111. cracking chamber; 112. washing chamber; 113. elution chamber; 12. mixing chamber; 13. reaction chamber; 14. electrochemical reaction tank; 15. waste liquid chamber; 161. exhaust hole; 162. air inlet hole; 163. sample hole; 17. sample chamber; 18. buffer chamber; 191. first substrate; 192. second substrate; 193. buckle; 194. heat insulation tank;

[0056] 2. First on-off switch; 21. First valve seat; 211. First channel; 212. Second channel; 213. Third channel; 214. Fourth channel; 215. Fifth channel; 22. First valve body; 221. First communication port; 222. Second communication port; 223. Third communication port; 224. Fourth communication port; 225. Fifth communication port; 23. First valve cover;

[0057] 3. Electrochemical detection component; 31. Double-sided tape; 32. Gold finger; 33. Working electrode; 34. Reference electrode; 35. Counter electrode;

[0058] 4. Fluid driving part; 41. Sleeve; 42. Piston; 43. Sealing ring;

[0059] 5. Second on-off switch; 51. Second valve seat; 511. Sixth channel; 512. Seventh channel; 513. Eighth channel; 514. Ninth channel; 52. Second valve body; 521. Sixth communication port; 522. Seventh communication port; 523. Eighth communication port; 524. Ninth communication port; 53. Second valve cover

[0060] 61. First liquid capsule reagent pack; 62. Second liquid capsule reagent pack; 63. Third liquid capsule reagent pack; 64. Fourth liquid capsule reagent pack; 65. Fifth liquid capsule reagent pack; 66. Sixth liquid capsule reagent pack; 67. Liquid capsule reagent pack puncture needle;

[0061] 71. The seventh liquid capsule reagent pack; 72. The eighth liquid capsule reagent pack; 73. The ninth liquid capsule reagent pack. DETAILED DESCRIPTION

[0062] The technical solution of the utility model will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0063] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0064] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installation", "connection" and "connection" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0065] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0066] Example 1

[0067] This embodiment provides a microfluidic chip, such as Figures 1 to 10As shown, it includes a substrate 1, a first on-off switch 2, an electrochemical detection component 3, a fluid driving component 4, a first reagent storage component and a second on-off switch 5. The substrate 1 has an extraction chamber, a mixing chamber 12, a reaction chamber 13, an electrochemical reaction tank 14 and a waste liquid chamber 15. The outlet of the extraction chamber is connected to the inlet of the mixing chamber 12, the outlet of the mixing chamber 12 is connected to the inlet of the reaction chamber 13 through the first on-off switch 2, and the outlet of the mixing chamber 12 is connected to the waste liquid chamber 15 through the first on-off switch 2, the outlet of the reaction chamber 13 is connected to the inlet of the electrochemical reaction tank 14 through the first on-off switch 2, and the outlet of the electrochemical reaction tank 14 is connected to the waste liquid chamber 15; the first on-off switch 2 is used to control the mixing chamber 12 and the reaction chamber 13. The on-off switching element 5 is used to control the on-off switching between the reaction chamber 13, the on-off switching between the mixing chamber 12 and the waste liquid chamber 15, and the on-off switching between the reaction chamber 13 and the electrochemical reaction tank 14; the electrochemical detection element 3 is connected to the substrate 1 and has a detection end, the detection end extends into the electrochemical reaction tank 14, and the electrochemical detection element 3 is used to obtain the reaction information of the electrochemical reaction tank 14; the fluid driving element 4 is connected to the substrate 1 and is connected to the extraction chamber through the second on-off switching element 5; the first reagent storage element is arranged on the substrate 1 and is connected to the extraction chamber through the second on-off switching element 5; the second on-off switching element 5 is used to control the on-off switching between the fluid driving element 4 and the extraction chamber, and the on-off switching between the first reagent storage element and the extraction chamber; the fluid driving element 4 can drive the flow of the fluid in the extraction chamber.

[0068] Specifically, the base 1 includes a first substrate 191 and a second substrate 192. An extraction tank, a mixing tank, a reaction tank and a waste liquid tank are provided on one side surface of the first substrate 191. An electrochemical reaction tank 14 is provided on the side surface of the first substrate 191 opposite to the extraction tank. The first substrate 191 and the second substrate 192 can be made of any one of materials such as polymethyl methacrylate (PMMA), polycarbonate (PC), cycloolefin copolymer (COC) and polypropylene (PP). The extraction tank, mixing tank, reaction tank and waste liquid tank, and the electrochemical reaction tank 14 on the first substrate 191 can be processed by one or more combinations of hot pressing, injection molding, laser engraving and mechanical processing. The first substrate 191 and the second substrate 192 have a sealing connection on one side surface of the extraction tank. The plate 191 and the second substrate 192 are sealed by heat pressing, bonding or laser bonding. The extraction tank and the second substrate 192 are combined to form an extraction chamber, the mixing tank and the second substrate 192 are combined to form a mixing chamber 12, the reaction tank and the second substrate 192 are combined to form a reaction chamber 13, and the waste liquid tank and the second substrate 192 are combined to form a waste liquid chamber 15. As an alternative embodiment, corresponding grooves can be opened on the second substrate 192, and the first substrate 191 and the second substrate 192 are sealed and connected to form corresponding chambers. Of course, corresponding grooves can also be opened on the first substrate 191 and the second substrate 192, and the first substrate 191 and the second substrate 192 are sealed and connected, and the grooves on the first substrate 191 and the corresponding grooves on the second substrate 192 are combined to form corresponding chambers. The first on-off switch 2 and the second on-off switch 5 can be programmable or respond to external signals to control the flow direction of the fluid. The micro-scale rotary valve or the combination of multiple micro-valves, the first reagent storage part is a deformable or fixed volume container, the first reagent storage part may include one or more of the containers, the number of the containers can be adjusted as needed in this field, the first reagent storage part is used to store preset reagents or buffers, can be released at an appropriate time point, and quantitative step-by-step operation is realized. The mixing chamber 12 includes one or more first sub-chambers, and the mixing chamber 12 is used to achieve rapid and uniform mixing of samples or reagents. The fluid drive 4 can be a mechanical or air pressure driven micro pump, and the fluid drive 4 is used to realize the transmission and precise control of the volume of a tiny liquid on a micro scale. A flow channel is provided between the two connected chambers, and the chamber is also connected to the on-off switch connected to it through the flow channel, and of course the reaction tank is also connected to the chamber or the on-off switch connected to it through the flow channel. The reaction chamber 13 includes one or more second sub-reaction chambers 13 for different steps of biochemical reaction or nucleic acid expansion process, the extraction chamber liquid includes multiple third sub-reaction chambers 13 for different sample extraction steps, and the waste liquid chamber 15 is a separate waste liquid box or waste liquid collection container. The electrochemical detection element 3 is an electrochemical sensor, which is used to detect and record the changes of electrical signals generated during the reaction process in real time, so as to perform quantitative or qualitative analysis.This microfluidic chip can not only be used for the detection of nucleic acids, but also for the analysis of other drugs or samples.

[0069] The present embodiment provides a microfluidic chip. When using the microfluidic chip, first, a sample is added to an extraction chamber, and then the sample in the extraction chamber is lysed and nucleic acid is released by one or more of mechanical lysis, chemical lysis, and enzyme lysis methods. Then, a magnetic bead liquid is added to the extraction chamber, and the nucleic acid is adsorbed by the magnetic beads in the magnetic bead liquid. The first reagent storage component is connected to the extraction chamber through the second on-off switching component 5. The first reagent storage component can store a cleaning liquid. The cleaning liquid in the first reagent storage component is transported to the extraction chamber after passing through the first on-off switching component 2. Then, the first reagent storage component is disconnected from the extraction chamber through the second on-off switching component 5, and then the fluid driving component 4 is connected to the extraction chamber through the second on-off switching component 5. The first on-off switch 2 connects the mixing chamber 12 to the waste liquid chamber 15, and the fluid driving component 4 delivers the cleaning fluid in the extraction chamber and the impurities therein to the mixing chamber 12. As the fluid driving component 4 continues to work, the cleaning liquid in the extraction chamber is continuously delivered to the mixing chamber 12 until the cleaning fluid in the mixing chamber 12 is completely delivered to the waste liquid chamber 15, so as to clean the nucleic acid adsorbed by the magnetic beads. The fluid driving component 4 is disconnected from the extraction chamber by the second on-off switch 5, and then an elution liquid is added to the extraction chamber to elute the nucleic acid and the magnetic beads. Then, a dilution liquid is added to the extraction chamber, and then the fluid is disconnected by the second on-off switch 5. The driving member 4 is connected to the extraction chamber, and the mixing chamber 12 is connected to the reaction chamber 13 through the first on-off switching member 2. The diluted liquid in the extraction chamber and the nucleic acid therein are transported to the mixing chamber 12 for rapid mixing through the fluid driving member 4. The mixed liquid is transported to the reaction chamber 13 under the drive of the fluid driving member 4. The fluid driving member 4 is disconnected from the extraction chamber through the second on-off switching member 5, and the mixing chamber 12 is disconnected from the reaction chamber 13 through the first on-off switching member 2. At this time, the nucleic acid undergoes an amplification reaction (PCR) in the reaction chamber 13. After the nucleic acid is amplified, the fluid driving member 4 is connected to the extraction chamber through the second on-off switching member 5, and the nucleic acid is removed through the first on-off switching member 2. The first on-off switch 2 connects the mixing chamber 12 with the reaction chamber 13, and connects the reaction chamber 13 with the electrochemical reaction tank 14. The fluid driving component 4 transports the liquid in the mixing chamber 12 to the electrochemical reaction tank 14, and performs an electrochemical reaction. Then the electrochemical detection component 3 detects and outputs the detection information, thereby realizing the detection of pathogen genes. By using the microfluidic chip, there is no need to use other equipment or transfer the sample, thereby avoiding cross contamination caused by the transfer process. The detection of pathogen genes can be completed by the microfluidic chip alone, thereby reducing the use of other equipment, reducing the volume of the detection equipment, and improving space utilization, so as to meet the requirements of medical testing for portability and compactness.

[0070] The microfluidic chip provided in this embodiment is as follows: Figures 4 to 6 As shown, the extraction chamber includes a cracking chamber 111, a washing chamber 112 and an elution chamber 113 connected in sequence, and a flow channel is provided between the cracking chamber 111, the washing chamber 112 and the elution chamber 113 to achieve sequential connection, and the cracking chamber 111 is pre-filled with corresponding reagents. When the microfluidic chip is used for nucleic acid (pathogen) detection, the pre-filled proteinase K reagent in the cracking chamber 111 is a proteinase K reagent, which can be spherical freeze-dried, and the washing chamber 112 is used to wash the nucleic acid after combining with the magnetic beads to separate and purify the nucleic acid. The elution chamber 113 is used to make the nucleic acid fall off the magnetic beads, and the sample can complete different operations in different chambers, so as to provide a stable microenvironment for the cracking, washing and elution of the sample, and ensure the high-efficiency operation of nucleic acid extraction. When the microfluidic chip is used for the analysis of other drugs or samples, the extraction chamber can also be arranged into a plurality of third sub-chambers according to the needs of the drug, and the corresponding reagents are filled in the third sub-chambers as needed.

[0071] The microfluidic chip provided in this embodiment is as follows: Figure 4 As shown, the substrate 1 is also provided with an exhaust hole 161, which is provided on the first substrate 191. The cross-sectional shape of the exhaust hole 161 can be circular, square or other common shapes. The exhaust hole 161 is provided directly above the waste liquid chamber 15. Of course, the exhaust hole 161 can also be provided at other positions of the substrate 1. It is only necessary to connect the exhaust hole 161 with the waste liquid chamber 15, and the waste liquid chamber 15 is connected with the outside world through the exhaust hole 161, so that the air pressure of the waste liquid chamber 15 is balanced with that of the outside world, so as to ensure that the liquid in the mixing chamber 12 or the reaction tank can be smoothly transported to the waste liquid chamber 15.

[0072] The microfluidic chip provided in this embodiment is as follows: Figure 4 , Figure 5 , Figure 8As shown, the substrate 1 has a plurality of reaction chambers 13, the number of which can be adjusted as needed. In this embodiment, the number of reaction chambers 13 is four. The substrate 1 also has a plurality of electrochemical reaction tanks 14, the number of which can be the same as or different from the number of reaction chambers 13. Personnel skilled in the art can adjust the number of electrochemical reaction tanks 14 as needed. In this embodiment, the number of electrochemical reaction tanks 14 is four, and the outlet of any electrochemical reaction tank 14 is connected to the waste liquid pool through a flow channel. The four electrochemical reaction tanks 14 can also be connected to the waste liquid pool through a branch flow channel in the substrate 1. The first on-off switching valve includes a first valve seat 21 and a first valve body 22. The first on-off switching valve includes a first valve seat 21 and a first valve body 22. A valve seat 21 is provided in the base body 1, that is, the first valve seat 21 is a part of the second base plate 192. Of course, the first valve seat 21 can also be made separately. The first valve seat 21 is detachably connected to the base body 1 by means of threads, snaps or rivets. The first valve seat 21 has a groove. Part of the first valve body 22 extends into the groove and is rotatably connected to the first valve seat 21. Another part of the first valve body 22 extends out of the groove. The first on-off switching member 2 also includes a first valve cover 23. The first valve cover 23 is fixedly connected to the part of the first valve body 22 extending out of the groove. The first valve body 22 is rotatably connected to the first valve seat 21 by threads. The first valve body 22 is driven to rotate by rotating the first valve cover 23 to adjust the first on-off switching member 2. The first valve seat 21 has a first channel 211, a second channel 212, a third channel 213, four fourth channels 214, and four fifth channels 215. The number of channels is greater than or equal to the sum of the number of reaction chambers 13, the number of reaction tanks, the number of mixing chambers 12, and the number of waste liquid pools. The first channel 211 is located at the center of the first valve seat 21. The second channel 212, the four fourth channels 214, and the four fifth channels 215 are spaced around the first circumference. The center of the first circumference is the center point of the first channel 211, that is, the second channel 212, the four fourth channels 214, and the four fifth channels 215 surround the outer periphery of the first channel 211. One end of the first channel 211 is connected to the mixing chamber 12. The outlet of the chamber 12 is connected, and the substrate 1 has a branch flow channel, the branch flow channel has an inlet and four outlets, the inlet of the branch flow channel is connected to the second channel 212, and the four outlets of the branch flow channel are respectively connected to the inlets of the four reaction chambers 13. Of course, any reaction chamber 13 can also be directly connected to the second channel 212 through the corresponding flow channel, the waste liquid chamber 15 is connected to the third channel 213, and the outlet of any reaction chamber 13 is connected to one of the corresponding fourth channels 214, and the inlet of any reaction tank is connected to one of the corresponding fifth channels 215, and the reaction chamber 13 is paired with the fourth channel 214 one by one, and the reaction tank is paired with the sixth channel 511 one by one;The first valve body 22 is provided with a first communication port 221, a second communication port 222, a third communication port 223, four fourth communication ports 224 and four fifth communication ports 225 at the bottom thereof. The first communication port 221 is located at the center of the bottom surface of the first valve body 22, and the second communication port 222, the third communication port 223, the four fourth communication ports 224 and the four fifth communication ports 225 are spaced apart around a second circumference, the center of the second circumference being the center point of the first communication port 221, that is, the second communication port 222, the third communication port 223, the four fourth communication ports 224, the four fifth communication ports 225 are spaced apart around a second circumference. The through port 225 surrounds the outer circumference of the first communication port 221, the radius of the first circumference and the second circumference are the same, and the first valve body 22 is located directly above the first valve seat 21, the first communication port 221 is connected to the second communication port 222 through the flow channel, and the first communication port 221 is connected to the end of the first channel 211 away from the mixing chamber 12, any fourth communication port 224 is connected to the corresponding fifth communication port 225 through the flow channel, and is used to transport the liquid in the reaction chamber 13 to the corresponding reaction tank, and the first valve body 22 has the function of rotating under the action of an external force to transfer the second communication port 221 to the first channel 211. The first switching state is that the port 222 is connected to the second channel 212, the third connecting port 223 is connected to the third channel 213, any fourth connecting port 224 is connected to the corresponding fourth channel 214, and any fifth connecting port 225 is connected to the corresponding fifth channel 215. In this first switching state, the liquid in the mixing chamber 12 passes through the first channel 211, the first connecting port 221, the second connecting port 222, the second channel 212, and the branch flow channel in sequence and is respectively transported to the four reaction chambers 13, and the fluid in any reaction chamber 13 is sequentially After passing through the corresponding fourth channel 214, the fourth connecting port 224, and the fifth connecting port, the waste liquid is transported to the corresponding reaction tank, that is, the four reaction chambers 13 are connected to the four reaction tanks one by one; the first valve body 22 has a second switching state that rotates under the action of an external force to connect the second connecting port 222 with the third channel 213. In this state, the waste liquid in the mixing chamber 12 is transported to the waste liquid chamber 15 after passing through the first channel 211, the first connecting port 221, the second connecting port 222, and the third channel 213 in sequence, thereby avoiding the waste liquid from passing through the reaction chamber 13 and causing contamination;The first valve body 22 has a third switching state in which it rotates under the action of external force to block the second connecting port 222, the third connecting port 223, any one of the fourth connecting ports 224 and any one of the fifth connecting ports 225. In this state, the second connecting port 222 is located between two adjacent channels on the periphery of the first channel 211. At this time, the first connecting port 221, the third connecting port 223, the four fourth connecting ports 224 and the four fifth connecting ports 225 are all in a blocked state. The first on-off switching member 2 is a ten-way rotary valve. The first valve seat 21 and the first valve cover 23 can be machined or injection molded. On the one hand, the first valve body 22 needs to ensure good sealing with the first valve seat 21. On the other hand, the first valve body 22 needs to have a set rigidity so that it can be driven by external force without deformation. The first valve body 22 can be integrally processed and molded with soft glue and hard materials by using a rubber coating process or a two-color mold process, that is, the side of the first valve body 22 in contact with the first valve seat 21 is made of soft glue (such as polyurethane, thermoplastic elastomer material), and the other end is made of hard material (polycarbonate, polymethyl methacrylate material). In this way, the first on-off switching member 2 can be rotated under a relatively small external force. Of course, the first valve seat 21 and the first valve body 22 can also be integrally processed and molded with soft glue and hard materials by using a rubber coating process or a two-color mold process. ;

[0073] The microfluidic chip provided in this embodiment is as follows: Figure 1 , Figure 4 , Figure 6 , Figure 7As shown, the first reagent storage unit includes multiple liquid capsule reagent packs. Those skilled in the art can adjust the number of liquid capsule reagent packs as needed. In this embodiment, the first reagent storage unit includes a first liquid capsule reagent pack 61, a second liquid capsule reagent pack 62, a third liquid capsule reagent pack 63, a fourth liquid capsule reagent pack 64, a fifth liquid capsule reagent pack 65, and a sixth liquid capsule reagent pack 66. The first liquid capsule reagent pack 61, the second liquid capsule reagent pack 62, the third liquid capsule reagent pack 63, the fourth liquid capsule reagent pack 64, and the fifth liquid capsule reagent pack 65 can all store cleaning solutions, and of course other reagents can also be stored. The sixth liquid capsule reagent pack 66 can store detection solution, and of course other reagents can also be stored. The material of any sac reagent package includes one or more of aluminum foil and plastic. A corresponding groove is opened in the base 1. Any sac reagent package is sealed and connected to the corresponding groove by heat pressing or laser welding. A sac reagent package puncture needle 67 is provided at the bottom of any groove. When one of the sac reagent packages needs to be used, the sac reagent package is squeezed so that the sac reagent package is punctured by the sac reagent package puncture needle 67, and then the sac reagent package is squeezed to squeeze out the liquid in the sac reagent package. The second on-off switching valve includes a second valve seat 51 and a second valve body 52. ​​The second valve seat 51 is opened in the base body 1, that is, the second valve seat 51 is a part of the second substrate 192. Of course, the second valve seat 51 can also be made separately. The second valve seat 51 is detachably connected to the base body 1 by means of threads, snaps or rivets. The second valve seat 51 has a groove. Part of the second valve body 52 extends into the groove and is rotatably connected to the second valve seat 51, and another part of the second valve body 52 extends out of the groove. The second on-off switching member 5 also includes a second valve cover 53. The second valve cover 53 is fixedly connected to the part of the second valve body 52 extending out of the groove. The second valve body 52 is rotatably connected to the second valve seat 51 by threads. The second valve cover 53 is rotated to drive the second valve body 52 to rotate so as to adjust the state of the second on-off switching member 5. The second valve seat 51 has a sixth channel 511, a seventh channel 512, and six eighth channels 513. The number of channels is greater than or equal to the number of liquid capsule reagent packages. , the sum of the number of fluid driving members 4 and the number of extraction chambers, the sixth channel 511 is located at the center of the second valve seat 51, the seventh channel 512 and the six eighth channels 513 are spaced around the third circumference, the center of the third circumference is the center point of the sixth channel 511, that is, the seventh channel 512 and the six eighth channels 513 surround the outer circumference of the second channel 212, one end of the sixth channel 511 is connected to the inlet of the extraction chamber, one end of the seventh channel 512 is connected to the fluid driving member 4, the first liquid capsule reagent package 61, the second liquid capsule reagent package 62, the third liquid capsule reagent package 63, the fourth liquid capsule reagent package 64, the fifth liquid capsule reagent package 65, and the sixth liquid capsule reagent package 66 are respectively connected to the six eighth channels 513, and the first liquid capsule reagent package 61, the second liquid capsule reagent package 62, the third liquid capsule reagent package 63, the fourth liquid capsule reagent package 64, the fifth liquid capsule reagent package 65, and the sixth liquid capsule reagent package 66 are paired one by one with the six eighth channels 513;A sixth communication port 521 and a seventh communication port 522 are formed at the bottom of the second valve body 52. ​​The sixth communication port 521 is located at the center of the bottom surface of the second valve body 52. ​​The seventh communication port 522 is spaced apart from the sixth communication port 521. The radius of the third circle is equal to the distance between the sixth communication port 521 and the seventh communication port 522. The second valve body 52 is located directly above the second valve seat 51. The sixth communication port 521 is communicated with the seventh communication port 522 through a flow channel. The sixth communication port 521 is communicated with an end of the sixth channel 511 away from the extraction chamber. The second valve body 52 is capable of rotating under the action of an external force to connect the seventh communication port 522 with the seventh channel 511. 12 connected; in this fourth switching state, the fluid driving member 4 drives the gas therein to pass through the seventh channel 512, the seventh connecting port 522, the sixth connecting port 521, and the sixth channel 511 in sequence and then be delivered to the extraction chamber; the second valve body 52 has a fifth switching state that rotates under the action of external force to connect the seventh connecting port 522 with one of the eighth channels 513 and block the seventh channel 512. In this fifth switching state, the liquid in one of the liquid capsules passes through the eighth channel 513, the seventh connecting port 522, the sixth connecting port 521, and the sixth channel 511 in sequence and then is delivered to the extraction chamber. The second on-off switching member 5 is an eight-way rotary valve, and the second valve seat 51 and the second valve cover 53 can be formed by machining or injection molding. On the one hand, the second valve body 52 needs to ensure good sealing with the second valve seat 51, and on the other hand, the second valve body 52 needs to have a set rigidity so that it can be driven by an external force without deformation. The second valve body 52 can be formed by a two-body process of soft glue and hard material by a rubber coating process or a two-color molding process, that is, the two surfaces of the second valve body 52 in contact with the second valve seat 51 are made of soft glue (such as polyurethane, thermoplastic elastomer material), and the other two ends are made of hard material (polycarbonate, polymethyl methacrylate material). In this way, the second on-off switching member 5 can be rotated under a relatively small external force. Of course, the second valve seat 51 and the second valve body 52 can also be formed by a rubber coating process or a two-color molding process. The microfluidic chip can control the liquid flow route through the fluid driving element 4, the first on-off switching element 2, and the second on-off switching element 5, thereby realizing automated reagent loading, mixing, reaction, and waste liquid discharge, improving detection efficiency, reducing manual operation steps, reducing human errors, and improving detection consistency and reliability. ;

[0074] The microfluidic chip provided in this embodiment is as follows: Figure 4 , Figure 6 , Figure 7As shown, the base 1 is also provided with an air inlet 162, which is provided on the first substrate 191. A filter pad is also provided at the entrance of the air inlet 162 to prevent aerosols or particles in the air from entering each chamber. The cross-sectional shape of the air inlet 162 can be circular, square or other common shapes. The second valve seat 51 has a ninth channel 514. The seventh channel 512, six eighth channels 513, and the ninth channel 514 are distributed at intervals on the periphery of the sixth channel 511. The ninth channel 514 and the air inlet 162 are connected to each other. The second valve body 52 is connected to the air hole 162, and an eighth communication port 523 and a ninth communication port 524 are opened on the second valve body 52. ​​The eighth communication port 523 is connected to the ninth communication port 524, and the seventh communication port 522, the eighth communication port 523, and the ninth communication port 524 surround the outer periphery of the sixth communication port 521; the second valve body 52 has the function of rotating under the action of external force to connect the eighth communication port 523 with the seventh channel 512, and the ninth communication port 524 with the ninth channel 514, and block the sixth communication port 522. Switching state. In this sixth switching state, the external gas can be transported to the fluid driving member 4 through the air inlet 162, the ninth channel 514, the ninth connecting port 524, the eighth connecting port 523, and the seventh channel 512 in sequence. The fluid driving member 4 can be connected with the outside through the air inlet 162, so that the external gas can enter the fluid driving member 4, so that the fluid driving member 4 can work normally and avoid the gas storage capacity in the fluid driving member 4 being limited. The fluid driving member 4 can be a plunger pump. The fluid driving member 4 includes a sealing ring 43, a sleeve 41 and a piston 42. The sleeve 41 is sealed and connected to the base 1. The piston 42 is slidably connected to the sleeve 41 through the sealing ring 43. The sleeve 41 is connected to the extraction chamber and the ninth channel 514 through the flow channel respectively. The sealing ring 43 and the piston 42 can be made of one or more of fluororubber and silicone rubber. The sealing ring 43 and the piston 42 can be made by compression molding or injection molding. The material selection and processing technology of the sleeve 41 are the same as those of the first substrate 191 and the second substrate 192.

[0075] The microfluidic chip provided in this embodiment is as follows: Figure 6As shown, the matrix 1 also has a sample chamber 17 and a sample hole 163. The sample chamber 17 is arranged between the circulation driving member and the extraction chamber. The sample chamber 17 is connected with the fluid driving member 4 and the seventh channel 512 respectively. The gas in the fluid driving member 4 passes through the sample chamber 17, the seventh channel 512, the seventh connecting port 522, the sixth connecting port 521, and the sixth channel 511 in sequence and is transported to the extraction chamber. The sample hole 163 is opened on the first substrate 191. The cross-sectional shape of the sample hole 163 can be circular, square or other common shapes. The sample hole 163 is opened directly above the sample chamber 17. Of course, the sample hole 163 can also be set at other positions of the matrix 1. It is only necessary to connect the sample hole 163 with the sample chamber 17. The sample hole 163 is used to drip the sample into the sample chamber 17. After the sample is dripped into the sample chamber 17, the sample hole 163 needs to be blocked to avoid the influence of the outside world on the sample, which causes deviations in the detection results. The sample is added into the sample chamber 17, and the fluid driving component 4 drives the sample in the sample chamber 17 to pass through the second on-off switch 5 and the lysis chamber 111 in sequence and then enter the washing chamber 112. The sample chamber 17 is separated from the extraction chamber to avoid the influence of the external environment on the extraction chamber.

[0076] The microfluidic chip provided in this embodiment is as follows: Figure 5 As shown, the mixing chamber 12 is a spiral flow channel and / or a flow channel with a variable inner diameter. The mixing chamber 12 can be a combination of a serpentine flow channel, a raindrop-shaped flow channel, and a flow channel with a reduced inner diameter. The mixing chamber 12 can mix the liquid flowing through it, and the liquid in the mixing chamber 12 can also be driven by external magnetic force and physical vibration to achieve rapid and uniform mixing of the liquid therein.

[0077] The microfluidic chip provided in this embodiment is as follows: Figure 1 and Figure 4As shown, the second reagent storage unit and the first reagent storage unit include multiple liquid capsule reagent packs. Those skilled in the art can adjust the number of liquid capsule reagent packs as needed. In this embodiment, the first reagent storage unit includes a seventh liquid capsule reagent pack 71, an eighth liquid capsule reagent pack 72, and a ninth liquid capsule reagent pack 73. The material of any liquid capsule reagent pack includes one or more of aluminum foil and plastic. A corresponding groove is provided in the base 1. Any liquid capsule reagent pack is sealed and connected to the corresponding groove by hot pressing or laser welding. A liquid capsule reagent pack puncture needle 67 is provided at the bottom of any groove. When one of the liquid capsule reagent packs needs to be used, the liquid capsule reagent pack is squeezed so that the liquid capsule reagent pack is punctured by the liquid capsule reagent pack puncture needle 67, and then the liquid capsule reagent pack is squeezed to squeeze out the liquid in the liquid capsule reagent pack. The seventh liquid capsule reagent package 71 stores the magnetic bead solution, the eighth liquid capsule reagent package 72 stores the elution solution, and the ninth liquid capsule reagent package 73 stores the dilution solution. Of course, the seventh liquid capsule reagent package 71, the eighth liquid capsule reagent package 72, and the ninth liquid capsule reagent package 73 can store other reagent solutions. The seventh liquid capsule reagent package 71 and the eighth liquid capsule reagent package 72 are connected to the extraction chamber, and the ninth liquid capsule is connected to the mixing chamber 12 through the flow channel. The reagent can be transported to the corresponding chamber at the required time through the second reagent storage unit to achieve quantitative step-by-step operation.

[0078] The microfluidic chip provided in this embodiment is as follows: Fig. 9As shown, the electrochemical detection element 3 is provided with multiple detection ends. The number of detection ends can be adjusted as needed by those skilled in the art. In this embodiment, the electrochemical detection element 3 has four detection ends, and any detection end includes a three-electrode system consisting of a working electrode 33, a reference electrode 34 and a counter electrode 35. The working electrode 33 can be circular, rectangular or other irregular shapes. The number of working electrodes 33 corresponding to each detection end can be 24, 20, or 40. The electrochemical detection element 3 is an electrochemical reaction PCB. The electrochemical detection element 3 also has a gold finger 32, which is used to connect to the terminal, and the terminal is a computer. The corresponding capture probe is pre-modified on the electrode of the electrochemical reaction PCB. The electrochemical reaction PCB is attached to the substrate by double-sided tape 31, and is surrounded by any reaction tank to form a corresponding detection chamber; the reference electrode 34 and the counter electrode 35 are arranged on a single PCB board according to appropriate spacing and geometric shapes. Through the array electrode design method, each working electrode 33 can detect the target molecule, thereby realizing multi-index synchronous detection. A corrosion-resistant and conductive metal (such as gold) is used and covered on the electrode surface by a gold plating process. In the three-electrode system, each electrode can use gold as a material, which can greatly simplify the manufacturing process of the three-electrode system. In actual testing, the exposed gold electrode is combined with a specific capture probe by physical adsorption to enhance the detection ability of the target analyte. The electrode design in this embodiment can realize that multiple independent reaction units perform different electrochemical reactions on the same microfluidic chip at the same time, so that the nucleic acids of multiple viruses or pathogens can be detected at one time, greatly improving the detection efficiency and flux; each working electrode 33 can detect a specific target nucleic acid sequence, and through the optimized probe design and signal reading system, it can accurately distinguish different types of pathogen nucleic acids, which is conducive to the rapid clinical diagnosis of multiple infectious diseases; compared with the traditional single reaction system, the multi-electrode design separates each reaction zone in physical space, which helps to reduce the risk of cross-contamination between samples and improve the accuracy and reliability of the test results. Combined with microfluidic technology and automated control systems, the multi-electrode platform can achieve a high degree of automation from sample addition, pretreatment to nucleic acid amplification and detection, which is particularly suitable for large-scale screening and rapid response scenarios. Multiple tests can be completed in one experiment, saving reagents, consumables and time costs, which has significant advantages for situations where medical resources are tight or for large-scale epidemic prevention and control.

[0079] The microfluidic chip provided in this embodiment is as follows: Figure 4 As shown, the substrate 1 is also provided with a handle 193 or a grip for conveniently holding the microfluidic chip. The handle 193 is a through groove opened on the substrate 1. The cross-section of the through groove can be a rounded rectangle, a rectangle or other shapes. The handle can be a bump or a groove on the substrate 1 for convenient holding or pinching by the staff.

[0080] The microfluidic chip provided in this embodiment is as follows: Figure 4 As shown, the substrate 1 is further provided with an insulation groove 194, which is provided between the reaction chamber 13 and the reaction tank. Personnel skilled in the art can adjust the number of the insulation grooves 194 as needed. The insulation grooves 194 are used to reduce the heat transfer between the reaction chamber 13 and the reaction tank, and avoid the influence of the heat transfer between the reaction tank and the reaction chamber 13 on their respective reactions.

[0081] The size of the microfluidic chip provided in this embodiment is 170mm×80mm×36.5mm, the material of the first substrate 191 is polymethyl methacrylate, and the thickness is 12.5mm; the material of the second substrate 192 is polypropylene, and the thickness is 0.18mm; the mixing chamber 12 is a serpentine flow channel, and its minimum structural dimension is 200μm; the shape of the working electrode 33 is circular, and the number of working electrodes 33 corresponding to each electrochemical reaction cell is 24, and a total of 96 working electrodes 33. The diameter of the gold circle is 0.6mm.

[0082] In an alternative embodiment, the size of the microfluidic chip provided in this embodiment is 170mm×80mm×36.5mm, the material of the first substrate 191 is polycarbonate, and the thickness is 12.5mm; the material of the second substrate 192 is polycarbonate, and the thickness is 0.2mm. The first to ninth liquid capsule reagent packages 73 are attached to the corresponding positions on the first substrate 191 by photosensitive adhesive, and the mixing channel structure is raindrop-shaped, and its minimum structural size is 150μm. The shape of the working 33 33 electrode is circular, the number of working electrodes 33 is 20, and the total number of working electrodes is 80. The diameter of the gold circle is 0.8mm.

[0083] The working process of the microfluidic chip provided in this embodiment is as follows:

[0084] 1) If Fig.10 As shown, the sample is added into the sample chamber 17 from the sample hole 163, and then the sample hole 163 is sealed. The sealing method may be elastic mold sealing, or sealing by adhesive, or other conventional micropore sealing methods;

[0085] 2) If Fig.11As shown, the integrated microfluidic chip is placed in the detection instrument, which is a fully automatic nucleic acid analyzer, and then the detection instrument automatically completes all subsequent operation steps. The internal timing actions of the detection instrument are specifically described as follows: rotate the second on-off switch 5 to connect the sample chamber 17 with the lysis chamber 111, and then rotate the first on-off switch 2 to connect the elution chamber 113 with the waste liquid chamber 15, and then the fluid drive 4 works and fills the sample chamber 17 with gas, and the gas pushes the sample into the lysis chamber 111 111, and the proteinase K in the lysis chamber is mixed with the sample solution. Proteinase K is used to lyse the cells in the sample and release nucleic acids. At the same time, the reagent in the seventh liquid capsule reagent 71 package is squeezed, and the magnetic bead solution is stored in the reagent package, and the sample and the magnetic bead solution are pushed into the washing chamber 112, and the mixing device in the detection instrument is started to realize nucleic acid adsorption, and the mixing device is a magnetic device;

[0086] 3) If Fig.12 As shown, the second on-off switch 5 is rotated to make the first liquid capsule reagent package 61 communicate with the lysis chamber 111, and the first liquid capsule reagent package 61 stores a cleaning solution. The reagent in the first liquid capsule reagent package 61 is squeezed so that the cleaning solution in the first liquid capsule reagent package 61 passes through the second on-off switch 5 and the lysis chamber 111 in sequence and then enters the washing chamber 112, and then the second channel 212 switch is rotated to make the sample chamber 17 communicate with the lysis chamber 111, and the fluid driving member 4 works to continuously input gas into the sample chamber 17 to push the cleaning solution to flow through the washing chamber 112, the elution chamber 113, the mixing chamber 12, and the first on-off switch 2 in sequence, and then discharged into the sample waste liquid chamber 15, and then the mixing device is started to realize the first nucleic acid cleaning;

[0087] 4) Repeat step 3, respectively squeezing the reagents in the second liquid capsule reagent pack 62 and the third liquid capsule reagent pack 63, both of which contain cleaning solutions, to achieve the second and third nucleic acid cleaning;

[0088] 5) If Fig.13 and Fig.14 As shown, the magnetic beads are transferred to the elution chamber 113 by the mixing device, and the fluid drive works to discharge the waste liquid in the extraction chamber into the waste liquid chamber 15;

[0089] 6) If Fig.15 As shown, the eighth liquid capsule reagent package 72 stores an elution solution, and the reagent in the eighth liquid capsule reagent package 72 is squeezed so that the elution solution flows into the elution chamber 113 after passing through the buffer chamber 18, and the excess elution solution can be stored in the buffer chamber 18;

[0090] 7) Start the mixing device and turn on the heating module of the nucleic acid elution area on the detection instrument to elute the nucleic acid, wherein the elution temperature is 56° C. and the incubation time is 5 min;

[0091] 8) If Fig.16 As shown, the second on-off switch 5 is rotated to make the sample chamber 17 communicate with the lysis chamber 111, and the first on-off switch 2 is rotated to make the mixing chamber 12 communicate with the reaction chamber 13, and then the fluid driving member 4 works to squeeze the reagent in the ninth liquid capsule reagent package 73 at the same time, and the reagent package stores the diluent, so that the eluted sample and the diluent pass through the mixing chamber 12 at the same time and enter the reaction chamber 13;

[0092] 9) closing the second on-off switch 5 and the first on-off switch 2, starting the PCR thermal cycle, and realizing the PCR amplification process;

[0093] 10) As Fig.17 As shown, the second on-off switch 5 is rotated to make the sample chamber 17 communicate with the lysis chamber 111, and the first on-off switch 2 is rotated to make the mixing chamber 12 communicate with the reaction chamber 13. At this time, the outlet of the reaction chamber 13 is connected to the electrochemical reaction tank 14, and then the fluid driving member 4 works to push the amplified sample into the electrochemical reaction tank 14, and then nucleic acid hybridization is performed after standing.

[0094] 11) If Fig.18 As shown, the second on-off switch 5 is rotated to connect the fourth liquid capsule reagent pack 64 with the lysis chamber 111, and then the fourth liquid capsule reagent pack 64 is squeezed, and the fourth liquid capsule reagent pack 64 stores a cleaning solution; repeat the above steps, squeeze the fifth liquid capsule reagent pack 65, and the fifth liquid capsule reagent pack 65 stores a cleaning solution to complete the second cleaning, and then repeat step 10) to push the cleaning solution into the electrochemical reaction tank 14 to wash away the unhybridized products;

[0095] 12) Rotate the second on-off switch 5 to connect the sixth liquid capsule reagent pack 66 with the lysis chamber 111, and then squeeze the sixth liquid capsule reagent pack 66, which stores the detection solution; then repeat step 10) to push the detection solution into the electrochemical reaction cell, start the electrochemical detection and output the result.

[0096] Although the embodiments of the present invention are described in conjunction with the accompanying drawings, those skilled in the art may make various modifications and variations without departing from the spirit and scope of the present invention, and such modifications and variations are all within the scope defined by the appended claims.

Claims

1. A microfluidic chip, characterized in that: include: A substrate (1) comprises an extraction chamber, a mixing chamber (12), a reaction chamber (13), an electrochemical reaction tank (14) and a waste liquid chamber (15); the outlet of the extraction chamber is connected to the inlet of the mixing chamber (12); the outlet of the mixing chamber (12) is connected to the inlet of the reaction chamber (13) via a first on-off switch (2); the outlet of the mixing chamber (12) is connected to the waste liquid chamber (15) via the first on-off switch (2); the outlet of the reaction chamber (13) is connected to the inlet of the electrochemical reaction tank (14) via the first on-off switch (2); and the outlet of the electrochemical reaction tank (14) is connected to the waste liquid chamber (15); The first on-off switch (2) is used to control the on-off between the mixing chamber (12) and the reaction chamber (13), the on-off between the mixing chamber (12) and the waste liquid chamber (15), and the on-off between the reaction chamber (13) and the electrochemical reaction tank (14); An electrochemical detection element (3) connected to the substrate (1) and having a detection end, the detection end extending into the electrochemical reaction tank (14), the electrochemical detection element (3) being used to obtain reaction information of the electrochemical reaction tank (14); A fluid driving member (4) connected to the base (1) and communicated with the extraction chamber via a second on-off switching member (5); A first reagent storage element, arranged on the base (1) and connected to the extraction chamber via the second on-off switch (5); The second on-off switch (5) is used to control the on-off between the fluid driving member (4) and the extraction chamber, and the on-off between the first reagent storage member and the extraction chamber; The fluid driving member (4) is capable of driving the flow of the fluid in the extraction chamber.

2. The microfluidic chip according to claim 1, characterized in that: The extraction chamber comprises a lysis chamber (111), a washing chamber (112) and an elution chamber (113) which are connected in sequence, the inlet of the lysis chamber (111) is connected to the second on-off switch (5), and the outlet of the elution chamber (113) is connected to the mixing chamber (12).

3. The microfluidic chip according to claim 2, characterized in that: The substrate (1) also has: An exhaust hole (161) is provided on the base body (1), and the waste liquid chamber (15) is connected to the outside through the exhaust hole (161).

4. The microfluidic chip according to claim 3, characterized in that: The substrate (1) has a plurality of reaction chambers (13); The substrate (1) has a plurality of electrochemical reaction tanks (14), and the outlet of any one of the electrochemical reaction tanks (14) is connected to the waste liquid chamber (15); The first on-off switching member (2) comprises a first valve seat (21) and a first valve body (22), wherein the first valve body (22) is rotatably connected to the first valve seat (21); the first valve seat (21) is arranged in the base (1), the first valve seat (21) has a first channel (211), a second channel (212), a third channel (213), a plurality of fourth channels (214) and a plurality of fifth channels (215), the outlet of the mixing chamber (12) is connected to the first channel (211); the inlet of any one of the reaction chambers (13) is connected to the second channel (212), the waste liquid chamber (15) is connected to the third channel (213), and the waste liquid chamber (15) is connected to the third channel (213). The outlet of any one of the reaction chambers (13) is communicated with the corresponding fourth channel (214), and the inlet of any one of the reaction tanks is communicated with the corresponding fifth channel (215); the first valve body (22) has a first communication port (221), a second communication port (222), a third communication port (223), a plurality of fourth communication ports (224) and a plurality of fifth communication ports (225), the first communication port (221) is communicated with the second communication port (222), and the first communication port (221) is communicated with the first channel (211); any one of the fourth communication ports (224) is communicated with the corresponding fifth communication port (225); The first valve body (22) has a first switching state in which the second communication port (222) is rotated under the action of an external force to connect the second communication port (222) with the second channel (212), the third communication port (223) with the third channel (213), any fourth communication port (224) with the corresponding fourth channel (214), and any fifth communication port (225) with the corresponding fifth channel (215); The first valve body (22) has a second switching state in which it rotates under the action of an external force to connect the second communication port (222) with the third channel (213); The first valve body (22) has a third switching state in which it rotates under the action of an external force to block the second communication port (222), the third communication port (223), any one of the fourth communication ports (224) and any one of the fifth communication ports (225).

5. The microfluidic chip according to any one of claims 1 to 4, characterized in that: The first reagent storage unit includes a plurality of liquid capsule reagent packages; The second on-off switching member (5) comprises a second valve seat (51) and a second valve body (52), wherein the second valve body (52) is rotatably connected to the second valve seat (51); the second valve seat (51) is arranged in the base body (1), the second valve seat (51) has a sixth channel (511), a seventh channel (512), and a plurality of eighth channels (513), the inlet of the extraction chamber is connected to the sixth channel (511), the fluid driving member (4) is connected to the seventh channel (512), and any one of the liquid capsule reagent packages is connected to the corresponding eighth channel (513); the second valve body (52) has a sixth communication port (521) and a seventh communication port (522), the sixth communication port (521) is connected to the seventh communication port (522), and the sixth communication port (521) is connected to the sixth channel (511); The second valve body (52) has a fourth switching state in which it rotates under the action of an external force to connect the seventh communication port (522) with the seventh channel (512); The second valve body (52) has a fifth switching state in which it rotates under the action of an external force to connect the seventh communication port (522) with one of the eighth channels (513) and block the seventh channel (512).

6. The microfluidic chip according to claim 5, characterized in that: The base body (1) is provided with an air inlet hole (162); the second valve seat (51) has a ninth channel (514), the ninth channel (514) is communicated with the air inlet hole (162); the second valve body (52) is provided with an eighth communication port (523) and a ninth communication port (524), the eighth communication port (523) is communicated with the ninth communication port (524); The second valve body (52) has a sixth switching state in which it rotates under the action of an external force to connect the eighth communication port (523) with the seventh channel (512), the ninth communication port (524) with the ninth channel (514), and the seventh communication port (522) is blocked.

7. The microfluidic chip according to claim 1, characterized in that: The substrate (1) further comprises: a sample chamber (17), wherein the sample chamber (17) is respectively connected to the fluid driving member (4) and the second on-off switching member (5); A sample hole (163) is provided on the substrate (1), and the sample chamber (17) is connected to the outside through the sample hole (163).

8. The microfluidic chip according to claim 1, characterized in that: The mixing chamber (12) is a spirally arranged flow channel and / or a flow channel with a variable inner diameter.

9. The microfluidic chip according to claim 1, characterized in that: Also includes: A second reagent storage element, arranged on the substrate (1); The base body (1) further comprises a buffer chamber (18), wherein the buffer chamber (18) is communicated with the second reagent storage element and the extraction chamber respectively.

10. The microfluidic chip according to claim 1, characterized in that: The electrochemical detection element (3) has a plurality of detection ends, and any one of the detection ends has a plurality of electrodes.

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