A microfluidic chip, a microfluidic detection device and a detection method

Abstract

The application discloses a micro-fluidic chip, a micro-fluidic detection device and a detection method, relates to the technical field of biochemical reagent detection and analysis, and comprises a releasing agent sample tube and a micro-fluidic chip, and the outer wall of the bottom end of the releasing agent sample tube is movably connected with the inner wall of the top end of the micro-fluidic chip; a gas return filter element is arranged at the middle portion of the upper surface of the releasing agent sample tube; a sealing film is movably connected to the middle portion of the side surface of the micro-fluidic chip; four freeze-dried balls are arranged in the inside surrounded by the micro-fluidic chip and the sealing film; and air permeation devices and ventilation devices are arranged on the two sides of the top end of the micro-fluidic chip. The micro-fluidic detection device is provided with a circuit sensing control chip structure used in combination with the micro-fluidic chip, adopts a constant temperature amplification reaction technology, and adopts a Lamp colorimetric method to realize real-time detection of color changes of a reaction mixture, accurate analysis and reading of target data, and improvement of the accuracy and sensitivity of detection results, so that the missing detection and detection error phenomena are avoided.

Description

Technical Field

[0001] This invention relates to the field of biochemical reagent detection and analysis technology, specifically to a microfluidic chip, a microfluidic detection device, and a detection method. Background Technology

[0002] Microfluidic chips, also known as "lab-on-a-chip," are widely used in biotechnology, drug development, clinical diagnostics, environmental monitoring, and other fields. The technology integrates sample preparation, reaction, separation, and detection units of biological, chemical, and medical analysis processes onto a micrometer-scale chip to automatically complete the entire analysis process. Microfluidic chips can handle extremely small volumes of liquid reagents and perform biochemical analysis quickly, accurately, and efficiently.

[0003] For example, application number CN202110129528.9 discloses a pipetting drive device and method for microfluidic chips, and a detection device, which includes a base, a receiving tank, an inlet, an opening, a pressing protrusion, a pushing part, a guide groove, a slot, a piercing mechanism, a first support bracket, a piercing assembly, a squeezing mechanism, a second support bracket, a squeezing assembly, a stirring mechanism, a mounting bracket, a stirring assembly, a stirring drive mechanism, a squeezing drive mechanism, an imaging mechanism, a reflector assembly, a lens assembly, a camera, a positioning mechanism, a positioning component, a receiving groove, an elastic abutment component, an abutment top, a sealing mechanism, a valve pin, a heating mechanism, and a flexible heat-conducting component structure. This device automatically drives the stirring of the solution in the reaction chamber and squeezes the stirred solution out of the reaction chamber for processing, replacing manual operation, improving work efficiency and reducing costs. However, it suffers from the drawback of the drive mechanism consuming kinetic energy, making it environmentally unfriendly and energy-inefficient, and its applicability is limited to the reaction chamber. The present invention is inconvenient for portable home use, and the vent lacks the function of filtering and protecting against backflow, resulting in inaccurate test results due to contamination of the test reagent by gaseous or liquid impurities during the detection process. Application number CN201810761056.7 discloses a microfluidic detection chip and its preparation method, fixing device and centrifugal detection device, which includes a microfluidic detection chip, fixing device, centrifugal shaft, a fluorescent microsphere spotting area coated with antibody, a quality control antibody spotting area, a first spotting area of ​​detection antibody, a second spotting area of ​​detection antibody and a slanted groove structure. The chip is guaranteed not to deviate during high-speed centrifugation by the limiting of the central fixing plate. The centrifugal detection device is designed for easy assembly and simple operation. However, the pressure change in the closed space after centrifugation acceleration of the reagent reaction in the closed device can affect the acidity or alkalinity of the reaction solution, resulting in inaccurate test results.

[0004] Existing technologies have the following shortcomings: Due to the diversity, variability, and high infectivity of current pathogens, it is common for one family member to contract the disease and infect the whole family. Going to the hospital for testing is time-consuming, laborious, and costly. Pathogen nucleic acid testing is slow, and when there are many patients, there is also the risk of other viruses invading. Traditional home testing reagents and devices suffer from reagent oxidation and failure or low sensitivity of pathogen nucleic acid, resulting in missed detections, incorrect detections, and inaccurate and low-sensitivity test results. Therefore, a highly accurate, portable, and easy-to-use pathogen nucleic acid self-testing solution has become an urgent need for users.

[0005] The information disclosed in the background section is only intended to enhance the understanding of the background of this disclosure, and therefore may include information that does not constitute prior art known to those skilled in the art. Summary of the Invention

[0006] The purpose of this invention is to provide a microfluidic chip, a microfluidic detection device, and a detection method. This invention achieves convenient detection of nucleic acid primers for several pathogens and self-driving liquid transfer without the need for external force by incorporating a sealing film, a lyophilized sphere, and an optical path guiding structure on the microfluidic chip. The microfluidic detection device incorporates a circuit sensing chip, which enhances the optical path guiding and detection functions, thereby solving the problems mentioned in the background art.

[0007] To achieve the above objectives, the present invention provides the following technical solution: a microfluidic chip, a microfluidic detection device, and a detection method, comprising a release agent sample tube and a microfluidic chip, wherein the outer wall of the bottom end of the release agent sample tube is movably sleeved with the inner wall of the top end of the microfluidic chip, a return gas filter is provided in the middle of the upper surface of the release agent sample tube, a sealing membrane is movably connected to the middle of the side of the microfluidic chip, four freeze-dried spheres are provided inside the microfluidic chip and the sealing membrane, and a breathable device and a ventilation device are respectively provided on both sides of the top end of the microfluidic chip, wherein the breathable device and the ventilation device are identical in shape, size, and material;

[0008] The microfluidic chip includes a core board and a connector. The top of the core board is fixedly connected to the lower surface of the connector. A reaction chamber flow tube is located at the center of the top of the core board and inside the connector. A light guide structure is provided in the middle of the other side of the core board. One end of the light guide structure is provided with four spherical chambers. Upper locking blocks are fixedly connected to both ends of the bottom of the other side of the core board, and lower locking blocks are fixedly connected to both ends of the bottom of the side of the core board. The upper and lower locking blocks are symmetrical.

[0009] Preferably, the reaction chamber flow tube includes a puncture conduit, a liquid storage tank, a liquid flow channel, and a gas flow channel, wherein the liquid storage tank is fixedly connected to the puncture conduit and the four liquid flow channels, and the liquid flow channels and the gas flow channels are indirectly connected through a spherical chamber.

[0010] The light-guiding structure includes an incident convex lens, four light reflectors, four light guide pillars, and a focusing outlet. The four light reflectors are arranged sequentially in the inner cavity of the light-guiding structure, each corresponding to one of the four light guide pillars. The four light guide pillars are fixed to the upper surface of the core plate. They are used to refract light rays that are vertically incident from the lower surface of the incident convex lens through four refractions by the four light reflectors and exit from the lower part of the center of the focusing outlet. The tilt angles of the four light reflectors are horizontal, and from the outside to the inside, they are 45°, -45°, -45°, and -60° respectively. Specifically, the light rays that are vertically incident from the lower surface of the incident convex lens, together with the light guide pillars 3043, form a light path. After being reflected by the first light reflector, they become horizontal light rays. After being reflected by the second light reflector, they become vertically downward light rays. After being reflected again by the third light reflector, they become horizontal light rays and exit from the focusing outlet at a -30° angle.

[0011] Preferably, the release agent sample tube includes a tube body with a cavity inside. A tube cap is connected to the upper surface of the tube body via a connecting strip and a reservoir tube is fixedly connected to the lower surface of the tube body. An interface membrane tube is movably connected to the lower surface of the reservoir tube.

[0012] Preferably, the venting device includes a venting port and a first T-shaped filter element, and the venting port is movably sleeved with the first T-shaped filter element; the ventilation device includes a venting port and a second T-shaped filter element, and the venting port is movably sleeved with the second T-shaped filter element.

[0013] Preferably, the smoothness of the inner wall of the cavity of the puncture conduit, liquid flow channel, and gas flow channel is 0.001-0.005 mm, and all corners of the cavity are rounded. The radius of the rounded corner is equal to the diameter of the inner cavity, which is used to reduce the resistance of the inner wall of the cavity to gas and liquid and to avoid the loss of effective components of the lyophilized bulbs after reconstitution. The first T-type filter element and the second T-type filter element are made of the same material, ultra-high molecular weight polyethylene, which has the characteristics of double-layer sealing, high air permeability and hydrophobicity. It can block aerosols and automatically seals when it comes into contact with water to prevent water from flowing into the interior of the gas flow channel, thus playing a role in waterproof isolation protection, easy cleaning and anti-fouling. The sealing film is a transparent pressure-sensitive film, which is composed of a transparent pressure-sensitive adhesive layer and a transparent polypropylene film. A white PET film with a fluorosilicone coating on one side is used as a release film to seal and adhere to the surface of the core plate and cover the flow tube of the reaction chamber.

[0014] Preferably, the reaction chamber flow tube is connected to a venting device and a ventilation device for venting and depressurization. After the reaction chamber flow tube punctures the interface membrane tube, the release agent flows into the storage chamber through the puncture catheter. The gas in the gas channel and the liquid channel are discharged through the venting and ventilation ports, allowing the release agent in the storage chamber to automatically flow into the liquid channel and undergo a heating amplification reaction with the lyophilized bulb in the bulb chamber. The diameter of the bulb chamber is larger than the diameter of the lyophilized bulb, and it is used to place the lyophilized bulb inside the bulb chamber. The lyophilized bulb is an amplification reagent used to combine with the sampled mixed pathogen nucleic acid in the release agent for isothermal amplification reaction, resulting in a color change in the mixed solution.

[0015] A microfluidic detection device is used with a microfluidic chip, including a detection box that is movably sleeved with the microfluidic chip, and a circuit sensing chip is provided inside the detection box and on the lower surface of the microfluidic chip.

[0016] The detection box includes a box body and a shell, which are movably connected. A through slot is provided in the middle of the front of the box body. Inside the box body, from top to bottom, there are an upper limit plate, a screw, and a lower limit plate. The two ends of the screw are movably connected to the upper limit plate and the lower limit plate, respectively. Inner buckles are fixedly connected to all four sides of the inside of the box body, and outer buckles are fixedly connected to all four sides of the front of the shell, and the outer buckles and inner buckles are movably connected.

[0017] Preferably, the circuit sensing chip includes a circuit board with through holes on all four sides. On the upper surface of the circuit board, a light receiver, a thermal pad, and four LEDs are arranged in sequence, with the four LEDs evenly arranged in a fan shape surrounding the thermal pad. The thermal pad is semi-circular and surrounds the light receiver. Chip slots are provided on both sides of the center of the upper surface of the circuit board. The LEDs emit white light that directly illuminates the convex lens incident light reflector in the light guide structure. After four refractions in the light path, the light is focused onto the light receiver through the focusing port. The light is then detected by observing the color change of the mixed solution in the reaction chamber flow tube, further enabling the light receiver to identify the color of the reaction in the reaction chamber flow tube and accurately interpret the color change results.

[0018] Preferably, the surface of the light reflector is uniformly coated with a nano-reflective coating, which has excellent reflective performance, thermal conductivity, and light loss reduction performance. The material of the nano-reflective coating can be selected from any one of nano-sized titanium dioxide, silicon nitride, bismuth titanate, zinc oxide, and silver to achieve efficient light reflection, making the light path within the light guide structure clear and bright. The width and length of the chip slot are matched with the width and length of the upper card block, respectively, to insert and align the upper card block on the core board inside the chip slot, thereby fixing the position of the core board and allowing the thermal pad to heat the reaction chamber flow tube and the light receiver to align the emitted light with the light guide structure. The thermal pad is made of thermally conductive silicone, which has excellent thermal conductivity and corrosion resistance. The circuit board includes a power interface, a light color detection sensor, a light control module, a heating module, and a communication protocol. The working principle of the circuit board is as follows:

[0019] The power interface connects to an external power source to provide power to the light receiver, thermal pad, LED light, light color detection sensor, light control module, heating module, and communication protocol.

[0020] The light color detection sensor receives and identifies the light wavelength signal collected by the light receiver from the light path, and converts the identified photoelectric signal into a digital signal and transmits it to an external terminal device for interpretation through a communication protocol.

[0021] The light control module controls the switching on and off of the LED lights;

[0022] The heating module controls the temperature and switches on / off of the thermal pad.

[0023] The communication protocol connects to external terminals and controls the information input and output of the light and color detection sensor, light control module, and heating module.

[0024] A detection method, implemented using a microfluidic chip and a microfluidic detection device, includes the following steps:

[0025] Open the cap on the tube body, insert the cotton swab containing pathogen nucleic acid into the tube lumen, stir and mix it into the release agent, and then close the cap tightly. Insert the interface membrane tube at the bottom of the reservoir tube into the inside of the tube opening, and let the puncture catheter puncture the bottom of the interface membrane tube. Let the release agent containing pathogen nucleic acid in the tube lumen flow from the puncture catheter into the storage tank. The lumen is ventilated through the return air filter.

[0026] Lay the core plate flat, place the lyophilized amplification reagent bulbs into the bulb compartment, and tightly attach the sealing film to the surface of the core plate. The gas flow channel allows air to pass through the first T-type filter on the vent and the second T-type filter on the vent. The liquid in the liquid storage compartment flows into the liquid flow channel, and the gas flows out from the gas flow channel and is discharged through the vent and vent device, so that the release agent containing pathogen nucleic acid in the reaction chamber flow tube mixes with the lyophilized bulbs in the bulb compartment.

[0027] Install the upper limit plate, screw and lower limit plate, and install the circuit sensing chip inside the upper limit plate and lower limit plate. Place the installed part inside the box and snap the external buckle on the shell and the internal buckle inside the box together.

[0028] Insert the core board vertically into the slot, place it inside the built-in box on the upper surface of the circuit board, so that the upper card block is inserted into the chip card slot and limited, and connect the mobile terminal to the communication protocol inside the circuit board to control the operation of the light color detection sensor, light control module and heating module on the light receiver, LED light and heat conduction pad;

[0029] The Lamp colorimetric method is used. The reaction liquid path is heated and amplified by the microfluidic detection device, and the pH of the amplification reagent and the release agent mixed with pathogen nucleic acid changes, which causes the color of the reaction mixture to change. The color change data is analyzed in real time based on the light and color signal changes obtained by the external terminal to determine the presence of the pathogen nucleic acid of the detection target.

[0030] The technical effects and advantages provided by the present invention in the above technical solution are as follows:

[0031] This invention, through the design of a reaction chamber flow tube and optical path light guide structure for a microfluidic chip, combined with the use of release agent sample tubes, sealing films, and lyophilized bulbs, enables convenient detection of nucleic acid primers for several pathogens. It has a wide range of applications in home use, achieving functions such as reusability, easy cleaning, accurate detection, and prevention of water leakage and contamination. Furthermore, through the designed return gas filter, air permeation device, and ventilation device structure, it achieves a self-driven function of reaction pipetting without the need for external force, avoiding the consumption of mechanical kinetic energy and improving the convenience and ease of operation of the reaction.

[0032] This invention incorporates a circuit sensing chip and a microfluidic chip in a microfluidic detection device. It employs isothermal amplification reaction technology and Lamp colorimetry to detect color changes in the reaction mixture in real time, accurately analyze and read the target data, thereby improving the accuracy and sensitivity of the detection results and avoiding missed detections and errors. Attached Figure Description

[0033] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this invention. For those skilled in the art, other drawings can be obtained based on these drawings.

[0034] Figure 1 This is a schematic diagram of the assembly structure of the microfluidic chip and microfluidic detection device of the present invention.

[0035] Figure 2 This is a side view of the exploded structure of the microfluidic chip of the present invention.

[0036] Figure 3 This is a bottom view of the exploded structure of the microfluidic chip of the present invention.

[0037] Figure 4 This is a front view of the microfluidic chip structure of the present invention.

[0038] Figure 5 This is a longitudinal cross-sectional view of the optical path of the microfluidic chip structure of the present invention.

[0039] Figure 6 This is a schematic diagram of the release agent sample tube and return gas filter element of the present invention.

[0040] Figure 7 This is a schematic diagram of the sealing film structure of the present invention.

[0041] Figure 8 This is a schematic diagram of the circuit sensing and control chip structure of the present invention.

[0042] Figure 9 This is a schematic diagram of the overall exploded structure of the microfluidic detection device of the present invention.

[0043] Figure 10 This is a schematic diagram of a partial explosion of the microfluidic detection device of the present invention.

[0044] Figure 11 This is a rear view of the exploded structure of the microfluidic chip of the present invention.

[0045] Figure 12 This is a flowchart of the detection method of the present invention.

[0046] The attached figures are labeled as follows: 1. Release agent sample tube; 101. Tube body; 102. Tube lumen; 103. Connecting strip; 104. Tube cap; 105. Reservoir tube; 106. Interface membrane tube;

[0047] 2. Return air filter element;

[0048] 3. Microfluidic chip; 301. Core board; 302. Connecting port; 303. Reaction chamber flow tube; 3031. Puncture catheter; 3032. Liquid storage tank; 3033. Liquid flow channel; 3034. Gas flow channel; 304. Optical path light guide structure; 3041. Light entrance convex lens; 3042. Light reflector; 3043. Light guide column; 3044. Focusing outlet; 305. Sphere chamber; 306. Upper retaining block; 307. Lower retaining block;

[0049] 4. Sealing film; 5. Freeze-dried bulbs;

[0050] 6. Ventilation device; 601. Ventilation port; 602. First T-type filter element;

[0051] 7. Ventilation device; 701. Ventilation port; 702. Second T-type filter element;

[0052] 8. Detection box; 801. Box body; 802. Housing; 803. Upper limit plate; 804. Screw; 805. Lower limit plate; 806. Through slot; 807. External buckle; 808. Internal buckle;

[0053] 9. Circuit sensing and control chip; 901. Circuit board; 902. Through hole; 903. Photodetector; 904. Thermal pad; 905. LED light; 906. Chip slot. Detailed Implementation

[0054] Exemplary embodiments will now be described more fully with reference to the accompanying drawings. However, these exemplary embodiments can be implemented in many forms and should not be construed as limited to the examples set forth herein; rather, they are provided so that the description of this disclosure will be more complete and fully convey the concept of the exemplary embodiments to those skilled in the art.

[0055] Example 1

[0056] like Figure 1-7 As shown, the present invention provides a microfluidic chip, a microfluidic detection device and a detection method, including a release agent sample tube 1 and a microfluidic chip 3, wherein the outer wall of the bottom end of the release agent sample tube 1 is movably sleeved with the inner wall of the top end of the microfluidic chip 3, a return gas filter 2 is provided in the middle of the upper surface of the release agent sample tube 1, a sealing membrane 4 is movably connected to the middle of the side of the microfluidic chip 3, four freeze-dried balls 5 are provided inside the microfluidic chip 3 and the sealing membrane 4, and a breathable device 6 and a ventilation device 7 are respectively provided on both sides of the top end of the microfluidic chip 3, and the breathable device 6 and the ventilation device 7 are the same in shape, size and material;

[0057] The microfluidic chip 3 includes a core board 301 and a connector 302. The top of the core board 301 is fixedly connected to the lower surface of the connector 302. A reaction chamber flow tube 303 is provided at the center of the top of the core board 301 and inside the connector 302. A light guide structure 304 is provided in the middle of the other side of the core board 301. One end of the light guide structure 304 is provided with four spherical chambers 305. Upper locking blocks 306 are fixedly connected to both ends of the bottom of the other side of the core board 301, and lower locking blocks 307 are fixedly connected to both ends of the bottom of the side of the core board 301. The upper locking blocks 306 and lower locking blocks 307 are symmetrical in structure.

[0058] It should be noted that the reaction chamber flow tube 303 includes a puncture conduit 3031, a liquid storage chamber 3032, a liquid flow channel 3033, and a gas flow channel 3034. The liquid storage chamber 3032 is fixedly connected to the puncture conduit 3031 and the four liquid flow channels 3033. The liquid flow channels 3033 and the gas flow channels 3034 are indirectly connected through the ball chamber 305.

[0059] The light guide structure 304 includes an incident convex lens 3041, four light reflectors 3042, four light guide pillars 3043, and a focusing outlet 3044. The four light reflectors 3042 are arranged sequentially within the cavity of the light guide structure 304, each corresponding to one of the four light guide pillars 3043. The four light guide pillars 3043 are fixed to the upper surface of the core plate 301, and are used to direct light rays perpendicularly incident from the lower surface of the incident convex lens 3041 through four refractions by the four light reflectors 3042 before exiting from below the center of the focusing outlet 3044. The tilt angles of the four light reflectors 3042 are horizontal, and from the outside in, they are 45°, -45°, -45° and -60° respectively. Specifically, the light rays that enter vertically below the incident light convex lens 3041, together with the light guide column 3043, form an optical path. After being reflected by the first light reflector 3042, they become horizontal light rays. After being reflected by the second light reflector 3042, they become vertically downward light rays. After being reflected again by the third light reflector 3042, they become horizontal light rays. After being reflected by the third light reflector 3042, they form a -30° beam and exit from the focusing outlet 3043.

[0060] It should be noted that the release agent sample tube 1 includes a tube body 101, the tube body 101 has a cavity 102 inside, the upper surface of the tube body 101 is connected to a tube cap 104 via a connecting strip 103, the lower surface of the tube body 101 is fixedly connected to a reservoir tube 105, and the lower surface of the reservoir tube 105 is movably connected to an interface membrane tube 106.

[0061] It should be noted that the venting device 6 includes a venting port 601 and a first T-shaped filter element 602, and the venting port 601 is movably connected to the first T-shaped filter element 602; the ventilation device 7 includes a ventilation port 701 and a second T-shaped filter element 702, and the ventilation port 701 is movably connected to the second T-shaped filter element 702.

[0062] It should be noted that the smoothness of the inner wall of the cavity of the puncture catheter 3031, liquid flow channel 3033 and gas flow channel 3034 is 0.001-0.005 mm, and the corners of the cavity are all rounded. The radius of the rounded corner is equal to the diameter of the inner cavity. This is to reduce the resistance of the inner wall of the cavity to gas and liquid and to avoid the loss of effective ingredients of the lyophilized bulb 5 after reconstitution. The first T-type filter element 602 and the second T-type filter element 702 are made of the same material, ultra-high molecular weight polyethylene, which has the characteristics of double-layer sealing, high air permeability and hydrophobicity. It can block aerosols and automatically seals when it comes into contact with water to prevent water from flowing into the interior of the gas flow channel 3034. This provides waterproof isolation protection and facilitates cleaning and anti-fouling. The sealing film 4 is a transparent pressure-sensitive film, which is composed of a transparent pressure-sensitive adhesive layer and a transparent polypropylene film. A white PET film with a fluorosilicone coating on one side is used as a release film to seal and adhere to the surface of the core plate 301 and cover the reaction chamber flow tube 303.

[0063] It should be noted that the reaction chamber flow tube 303 is connected to the venting device 6 and the ventilation device 7 respectively for venting and depressurization. After the reaction chamber flow tube 303 punctures the interface membrane tube 106, the release agent flows into the storage chamber 3032 through the puncture catheter 3031. The gas in the gas channel 3034 and the liquid channel 3033 is discharged through the venting port 601 and the ventilation port 701, so that the release agent in the storage chamber 3032 automatically flows into the liquid channel 3033 and undergoes a heating amplification reaction with the lyophilized bulb 5 through the bulb chamber 305. The diameter of the bulb chamber 305 is larger than the diameter of the lyophilized bulb 5, which is used to place the lyophilized bulb 5 inside the bulb chamber 305. The lyophilized bulb 5 is an amplification reagent used to combine with the sampled mixed pathogen nucleic acid in the release agent for isothermal amplification reaction, and the mixed solution will produce a color change.

[0064] Example 2

[0065] like Figure 8-11 As shown, the present invention provides a microfluidic detection device, which further discloses the detection device for the microfluidic chip in Embodiment 1; it includes a detection box 8, which is movably sleeved with the microfluidic chip 3, and a circuit sensing chip 9 is provided inside the detection box 8 and located on the lower surface of the microfluidic chip 3;

[0066] The detection box 8 includes a box body 801 and a housing 802, which are movably connected. A through slot 806 is provided in the middle of the front of the box body 801. Inside the box body 801, from top to bottom, there are an upper limit plate 803, a screw 804 and a lower limit plate 805. The two ends of the screw 804 are movably connected to the upper limit plate 803 and the lower limit plate 805 respectively. Inner buckles 808 are fixedly connected to all four sides of the inside of the box body 801. Outer buckles 807 are fixedly connected to all four sides of the front of the housing 802, and the outer buckles 807 and the inner buckles 808 are movably sleeved together.

[0067] It should be noted that the circuit sensing and control chip 9 includes a circuit board 901. Through holes 902 are provided around the circuit board 901. On the upper surface of the circuit board 901, a light receiver 903, a heat-conducting pad 904, and four LEDs 905 are arranged in sequence. The four LEDs 905 are evenly arranged in a fan shape surrounding the heat-conducting pad 904. The heat-conducting pad 904 is semi-circular in shape and surrounds the light receiver 903. Chip slots 906 are provided on both sides of the middle of the upper surface of the circuit board 901. The LEDs 905 emit white light sources that directly shine on the convex lens incident light reflector 3041 in the light guide structure 304. After four refractions of the light path, the light is emitted from the focusing port 307 and focused on the light receiver 903. The light is then detected by the color reaction change of the mixed solution in the reaction chamber flow tube 303. This further enables the light receiver 903 to identify the color of the reaction in the reaction chamber flow tube 303 and to accurately interpret and indicate the color change result.

[0068] It should be noted that the surface of the light reflector 3042 is uniformly coated with a nano-reflective coating, which has excellent reflective performance, thermal conductivity, and light loss reduction performance. The material of the nano-reflective coating can be any one of nano-sized titanium dioxide, silicon nitride, bismuth titanate, zinc oxide, and silver to achieve a highly efficient light reflection effect, making the light path within the light guide structure 304 clear and bright. The width and length of the chip slot 906 are matched with the width and length of the upper card block 306, respectively, for inserting and aligning the upper card block 306 on the core board 301 into the chip slot. Inside the slot 906, the position of the core board 301 is fixed, allowing the thermal pad 904 to heat the reaction chamber flow tube 303 and align the light emitted by the photoreceiver 903 with the light guide structure 304. The thermal pad 904 is made of thermally conductive silicone, which has excellent thermal conductivity, flexibility, and uniform heat distribution. It is used to lay heating wires on the circuit board 901, and the thermal pad 904 is attached to the heating wires for uniform heat conduction. The circuit board 901 also includes a power interface, a light color detection sensor, a light control module, a heating module, and a communication protocol. The working principle of the circuit board 901 is as follows:

[0069] The power interface connects to an external power source to provide power to the light receiver 903, thermal pad 904, LED light 905, light color detection sensor, light control module, heating module, and communication protocol.

[0070] The light color detection sensor receives and identifies the light wavelength signal collected by the light receiver 903 from the light path, and converts the identified photoelectric signal into a digital signal and transmits it to an external terminal device for interpretation through a communication protocol.

[0071] The light control module controls the switching on and off of LED 905;

[0072] The heating module controls the temperature of the thermal pad 904 and switches it on and off.

[0073] The communication protocol connects to external terminals and controls the information input and output of the light and color detection sensor, light control module, and heating module.

[0074] The microfluidic detection device provided in this embodiment of the invention is implemented by the aforementioned microfluidic chip. For details on the specific use and process of the microfluidic detection device, please refer to the embodiments of the aforementioned microfluidic chip, which will not be repeated here.

[0075] Example 3

[0076] like Figure 12 As shown, the present invention provides a detection method, which further discloses the process of the microfluidic chip and microfluidic detection device in Embodiment 1; the method includes the following steps:

[0077] Open the tube cap 104 on the tube body 101, insert the cotton swab containing pathogen nucleic acid into the tube lumen 102 and stir it to mix it into the release agent. After tightly closing the tube cap 104, insert the interface membrane tube 106 at the bottom of the reservoir tube 105 into the inside of the connecting port 302, and make the puncture catheter 3031 puncture the bottom of the interface membrane tube 106. The release agent containing pathogen nucleic acid in the tube lumen 102 flows from the puncture catheter 3031 into the storage tank 3032, while the inside of the tube lumen 102 is ventilated by the return air filter 2.

[0078] The core plate 301 is laid flat, and the lyophilized amplification reagent bulbs 5 are placed into the bulb chamber 305. The sealing film 4 is tightly attached to the surface of the core plate 301. The gas flow channel 3034 is ventilated through the first T-type filter 602 on the vent 601 and the second T-type filter 702 on the vent 701. The liquid in the liquid storage chamber 3032 flows into the liquid flow channel 3033. The gas flows out from the gas flow channel 3034 and is discharged through the venting device 6 and the ventilation device 7, so that the release agent containing pathogen nucleic acid in the reaction chamber flow tube 303 mixes with the lyophilized bulbs 5 in the bulb chamber 305.

[0079] Install the upper limit plate 803, screw 804 and lower limit plate 805, and install the circuit sensing chip 9 inside the upper limit plate 803 and lower limit plate 805. Place the installed chip inside the box 801 and snap the outer buckle 807 on the housing 802 and the inner buckle 808 inside the box 801 together.

[0080] The core board 301 is vertically inserted into the through slot 806, and the inside of the built-in box 801 is placed on the upper surface of the circuit board 901, so that the upper card block 306 is inserted into the chip card slot 906 and limited. The mobile terminal is connected to the communication protocol in the circuit board 901 to control the operation of the light color detection sensor, light control module and heating module on the light receiver 903, LED light 905 and heat conduction pad 904.

[0081] The Lamp colorimetric method is used. The reaction liquid path is heated and amplified by the microfluidic detection device, and the pH of the amplification reagent and the release agent mixed with pathogen nucleic acid changes, which causes the color of the reaction mixture to change. The color change data is analyzed in real time based on the light and color signal changes obtained by the external terminal to determine the presence of the pathogen nucleic acid of the detection target.

[0082] The present invention provides a method for detecting a microfluidic chip, which is implemented by the microfluidic chip and the microfluidic detection device described above. The specific method and process of the detection method are detailed in the embodiments of the microfluidic chip and the microfluidic detection device described above, and will not be repeated here.

[0083] The above formulas are all dimensionless calculations. The formulas are derived from software simulations based on a large amount of collected data to obtain the most recent real-world results. The preset parameters in the formulas are set by those skilled in the art according to the actual situation.

[0084] The above embodiments can be implemented, in whole or in part, by software, hardware, firmware, or any other combination thereof. When implemented using software, the above embodiments can be implemented, in whole or in part, as a computer program product. The computer program product includes one or more computer instructions or computer programs. When the computer instructions or computer programs are loaded or executed on a computer, all or part of the processes or functions described in the embodiments of this application are generated. The computer can be a general-purpose computer, a special-purpose computer, a computer network, or other programmable device. The computer instructions can be stored in a computer-readable storage medium or transmitted from one computer-readable storage medium to another. For example, the computer instructions can be transmitted from one website, computer, server, or data center to another website, computer, server, or data center via wired or wireless (e.g., infrared, wireless, microwave, etc.) means. The computer-readable storage medium can be any available medium that a computer can access or a data storage device such as a server or data center that includes one or more sets of available media. The available medium can be a magnetic medium (e.g., floppy disk, hard disk, magnetic tape), an optical medium (e.g., DVD), or a semiconductor medium. A semiconductor medium can be a solid-state drive.

[0085] It should be understood that in the various embodiments of this application, the order of the above-mentioned processes does not imply the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of this application.

[0086] Those skilled in the art will recognize that the units and algorithm steps of the various examples described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, or a combination of computer software and electronic hardware. Whether these functions are implemented in hardware or software depends on the specific application and design constraints of the technical solution. Those skilled in the art can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.

[0087] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A microfluidic chip, characterized in that, The device includes a release agent sample tube (1) and a microfluidic chip (3). The outer wall of the bottom end of the release agent sample tube (1) is movably connected to the inner wall of the top end of the microfluidic chip (3). A return gas filter (2) is provided in the middle of the upper surface of the release agent sample tube (1). A sealing membrane (4) is movably connected to the middle of the side of the microfluidic chip (3). Four freeze-dried balls (5) are provided inside the microfluidic chip (3) and the sealing membrane (4). A ventilating device (6) and a ventilation device (7) are respectively provided on both sides of the top end of the microfluidic chip (3). The microfluidic chip (3) includes a core plate (301) and a connector (302). The top of the core plate (301) is fixedly connected to the lower surface of the connector (302). A reaction chamber flow tube (303) is provided at the center of the top of the core plate (301) and inside the connector (302). A light guide structure (304) is provided in the middle of the other side of the core plate (301). One end of the light guide structure (304) is provided with four spherical chambers (305). Upper locking blocks (306) are fixedly connected to both ends of the bottom of the other side of the core plate (301). Lower locking blocks (307) are fixedly connected to both ends of the bottom of the side of the core plate (301). The reaction chamber flow pipe (303) is connected to the ventilator (6) and the ventilation device (7) respectively; The reaction chamber flow tube (303) includes a puncture conduit (3031), a liquid storage chamber (3032), a liquid flow channel (3033), and a gas flow channel (3034). The liquid storage chamber (3032) is fixedly connected to the puncture conduit (3031) and the four liquid flow channels (3033). The liquid flow channels (3033) and the gas flow channels (3034) are indirectly connected through a spherical chamber (305). The light guide structure (304) includes an incident convex lens (3041), four light reflectors (3042), four light guide pillars (3043), and a focusing outlet (3044). The four light reflectors (3042) are arranged sequentially in the inner cavity of the light guide structure (304), each corresponding to one of the four light guide pillars (3043), and the four light guide pillars (3043) are fixedly mounted on the upper surface of the core plate (301).

2. A microfluidic chip according to claim 1, characterized in that, The release agent sample tube (1) includes a tube body (101), the inside of which is provided with a cavity (102). The upper surface of the tube body (101) is connected to a tube cap (104) via a connecting strip (103). The lower surface of the tube body (101) is fixedly connected to a reservoir tube (105), and the lower surface of the reservoir tube (105) is movably connected to an interface membrane tube (106).

3. A microfluidic chip according to claim 2, characterized in that, The ventilation device (6) includes a ventilation port (601) and a first T-shaped filter element (602), and the ventilation port (601) is movably connected to the first T-shaped filter element (602); the ventilation device (7) includes a ventilation port (701) and a second T-shaped filter element (702), and the ventilation port (701) is movably connected to the second T-shaped filter element (702).

4. A microfluidic chip according to claim 3, characterized in that, The smoothness of the inner wall of the cavity of the puncture catheter (3031), liquid flow channel (3033) and gas flow channel (3034) is 0.001-0.005 mm, and the corners of the cavity are all rounded, and the radius of the rounded corner is equal to the diameter of the inner cavity; the first T-type filter element (602) and the second T-type filter element (702) are made of the same material, which is ultra-high molecular weight polyethylene.

5. A microfluidic chip according to claim 1, characterized in that, The diameter of the sphere (305) is larger than the diameter of the lyophilized sphere (5), which is an amplification reagent.

6. A microfluidic detection device, used with a microfluidic chip according to any one of claims 1-5, characterized in that, It includes a detection box (8) and is movably connected to the microfluidic chip (3). The detection box (8) is provided with a circuit sensing chip (9) inside and on the lower surface of the microfluidic chip (3). The detection box (8) includes a box body (801) and a shell (802), and the box body (801) and the shell (802) are movably connected. A through slot (806) is provided in the middle of the front of the box body (801). Inside the box body (801), from top to bottom, there are an upper limit plate (803), a screw (804) and a lower limit plate (805). The two ends of the screw (804) are movably connected to the upper limit plate (803) and the lower limit plate (805) respectively. The inner buckle (808) is fixedly connected to the four sides of the inside of the box body (801). The outer buckle (807) is fixedly connected to the four sides of the front of the shell (802), and the outer buckle (807) and the inner buckle (808) are movably connected.

7. A microfluidic detection device according to claim 6, characterized in that, The circuit sensing chip (9) includes a circuit board (901), and through holes (902) are provided around the circuit board (901). On the upper surface of the circuit board (901), a light receiver (903), a heat-conducting pad (904) and four LEDs (905) are arranged in sequence. The four LEDs (905) are evenly arranged in a fan shape to surround the heat-conducting pad (904). The heat-conducting pad (904) is semi-circular in shape and surrounds the light receiver (903). Chip slots (906) are provided on both sides of the middle part of the upper surface of the circuit board (901).

8. A microfluidic detection device according to claim 7, characterized in that, The surface of the light reflector (3042) is uniformly coated with a nano-reflective coating. The material of the nano-reflective coating can be any one of nano-sized titanium dioxide, silicon nitride, bismuth titanate, zinc oxide, and silver. The width and length of the chip slot (906) are matched with the width and length of the upper card block (306), respectively. The thermal pad (904) is made of thermally conductive silicone. The circuit board (901) includes a power interface, a light color detection sensor, a light control module, a heating module, and a communication protocol.

9. A detection method, implemented using a microfluidic chip according to any one of claims 1-5 and a microfluidic detection device according to any one of claims 6-8, characterized in that, The steps include the following: Open the tube cap (104) on the tube body (101), insert the cotton swab containing pathogen nucleic acid into the tube lumen (102) and stir it to mix it into the release agent. After tightly closing the tube cap (104), insert the interface membrane tube (106) at the bottom of the reservoir tube (105) into the inside of the connecting port (302) and make the puncture catheter (3031) puncture the bottom of the interface membrane tube (106) so that the release agent containing pathogen nucleic acid in the tube lumen (102) flows from the puncture catheter (3031) into the storage tank (3032). The tube lumen (102) is ventilated through the return air filter (2). The core plate (301) is laid flat, and the lyophilized amplification reagent bulbs (5) are placed into the bulb chamber (305). The sealing film (4) is tightly attached to the surface of the core plate (301). The gas flow channel (3034) is ventilated through the first T-type filter (602) on the venting port (601) and the second T-type filter (702) on the venting port (701). The liquid in the liquid storage chamber (3032) flows into the liquid flow channel (3033). The gas flows out from the gas flow channel (3034) and is discharged through the venting device (6) and the ventilation device (7), so that the release agent containing pathogen nucleic acid in the reaction chamber flow tube (303) mixes with the lyophilized bulbs (5) in the bulb chamber (305). Install the upper limit plate (803), screw (804) and lower limit plate (805), and install the circuit sensing chip (9) inside the upper limit plate (803) and lower limit plate (805). Place the installed chip inside the box (801), and snap the external buckle (807) on the housing (802) and the internal buckle (808) inside the box (801) together. The core board (301) is vertically inserted into the through slot (806), and placed inside the inner box (801) on the upper surface of the circuit board (901), so that the upper card block (306) is inserted into the chip card slot (906) and limited. The mobile terminal is connected to the communication protocol in the circuit board (901) to control the operation of the light color detection sensor, light control module and heating module on the light receiver (903), LED light (905) and heat conduction pad (904). The Lamp colorimetric method is used. The reaction liquid path is heated and amplified by the microfluidic detection device, and the pH of the amplification reagent and the release agent mixed with pathogen nucleic acid changes, which causes the color of the reaction mixture to change. The color change data is analyzed in real time based on the light and color signal changes obtained by the external terminal to determine the presence of the pathogen nucleic acid of the detection target.

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