Vertical microbiological detection device and method

Abstract

The invention belongs to the technical field of microbiological detection, and particularly relates to a vertical microbiological detection device and method. The device comprises a longitudinal movement module, a transverse movement module, a negative pressure pumping module and a shell, the longitudinal movement module comprises a first rigid hollow pipe column, a second rigid hollow pipe column, a solid column, a fixing plate, a first tension spring and a second lead screw motor. The transverse movement module comprises a first lead screw motor, a third lead screw motor, a chip jig, a heating detection module, a guide rail, a second sliding block, a fixing frame and a push plate. The negative pressure pumping module comprises a first electromagnetic valve, a second electromagnetic valve and a stepping peristaltic pump; the detection device disclosed by the invention is matched with a micro-fluidic chip structure, different liquids in the chip can be driven to flow to a target chamber by only needing one stepping peristaltic pump, and the peristaltic pump does not discharge liquid but only discharges gas, so that pollution is prevented as much as possible in an experiment process, and the accuracy of an experiment result is ensured.

Description

Technical Field

[0001] This invention relates to the field of microbial detection technology, and in particular to a vertical microbial detection device and method. Background Technology

[0002] When using microfluidic chips to detect microorganisms, sample lysis, washing, elution, and reaction steps can be integrated onto a single microfluidic chip. However, to prevent contamination, the existing microfluidic chip chamber and channel designs typically require a separate pumping device to deliver each solution to the target chamber. This results in multiple pumping devices being used throughout the detection process, leading to a large, costly, and inconvenient mechanical structure, making it extremely difficult to move the device outdoors for testing. Therefore, it is essential to develop a small, more compact microbial detection device with high accuracy (using a single pump and ensuring no contamination). Summary of the Invention

[0003] In view of the shortcomings of the prior art, the purpose of this invention is to provide a vertical microbial detection device and method that is compact in structure, small in size, and has high detection accuracy.

[0004] The technical solution of the present invention is described in detail below.

[0005] A vertical microbial detection device includes a longitudinal motion module, a transverse motion module, a negative pressure module, and a housing; the longitudinal motion module is located above the transverse motion module, and initially, both the transverse and longitudinal motion modules are located inside the housing; wherein: The lateral motion module includes a mounting frame, a chip fixture and a pusher plate mounted on the mounting frame; the microfluidic chip is placed in the chip slot formed by the chip fixture; the mounting frame moves laterally in a linear motion under the drive of a motor to move the chip fixture to the outside of the housing to insert the microfluidic chip and to move it into the housing to enter the experimental state; the pusher plate moves laterally in a linear motion relative to the mounting frame under the drive of a motor to open or close the different solution storage chambers in the microfluidic chip. The longitudinal motion module includes a first rigid hollow tube column, a second rigid hollow tube column, a solid column, a fixing plate, and a first tension spring. A first guide plate and a second guide plate are respectively provided above and below the first rigid hollow tube column, the second rigid hollow tube column, and the solid column to guide their vertical movement. A clamp is fixedly mounted on each of the first rigid hollow tube column, the second rigid hollow tube column, and the solid column. One end of the clamp is fixedly connected to the first tension spring, and the other end of the first tension spring is fixedly connected to the second guide plate. A fixing plate is positioned near the clamp. When the fixing plate moves downwards linearly driven by a motor, the first tension spring remains in a stretched state while the fixing plate abuts against the clamp, ensuring that the first rigid hollow tube column, the second rigid hollow tube column, and the solid column provide sufficient clamping force when contacting the top surface of the microfluidic chip. This achieves a seal between the end faces of the first rigid hollow tube column, the second rigid hollow tube column, and the solid column in the microfluidic chip and the corresponding first negative pressure channel, second negative pressure channel, and lysis fluid chamber of the microfluidic chip. The negative pressure module is used to drive the flow of liquid stored in the microfluidic chip in an orderly manner.

[0006] In this invention, the microfluidic chip includes, from top to bottom, a first solution storage unit, a first slide valve, a second solution storage unit, a second slide valve, and a reaction unit. The first solution storage unit has a lysis liquid chamber, a washing liquid chamber, an elution liquid chamber, a reaction liquid chamber, a first negative pressure channel serving as a general suction port, and a second negative pressure channel serving as a sample loading suction port. The second solution storage unit has a waste liquid chamber, a premixing chamber, a third channel corresponding to the second negative pressure channel, and a fourth channel. The reaction unit has a reaction chamber. The first slide valve has several flow channels and through holes to achieve selective communication with the solutions in the solution chambers of the first and second solution storage units, as well as selective communication with the gas path. The second slide valve has through holes for selectively connecting the reaction liquid inlet of the reaction unit and the vent of the reaction chamber. A push plate moves laterally linearly relative to the fixed frame under the drive of a motor to push the first and second slide valves to control the opening or closing of the first solution storage unit, the second solution storage unit, and the reaction unit.

[0007] In this invention, the longitudinal motion module further includes a dual-position self-holding electromagnet, a shift fork sheet metal, a piercing component, and a second tension spring; one end of the second tension spring is fixedly mounted on the mounting base, and the other end is fixedly connected to the piercing component, which passes through the second guide plate, with its tip aligned with the cleaning fluid chamber; the dual-position self-holding electromagnet is fixedly mounted on the fixed plate, and its output shaft is fixedly connected to the shift fork sheet metal, which is rotatably connected to the fixed plate; during operation, the swing direction of its output shaft is adjusted by controlling the on / off state of the dual-position self-holding electromagnet, causing the shift fork sheet metal and the piercing component to engage or disengage, thereby controlling the piercing component to move closer to or further away from the bottom of the cleaning fluid chamber.

[0008] In this invention, the lateral motion module is driven by a first lead screw motor and a third lead screw motor to drive the linear motion of the fixed frame and the push plate, respectively. The fixed end of the first lead screw motor is fixedly mounted on the outer shell, and the driving end is movably connected to the fixed frame. The fixed end of the third lead screw motor is mounted on the fixed frame, and the driving end of the third lead screw motor is movably connected to the push plate. A guide rail is provided on the fixed frame, and a second slider is provided on the outer shell. During operation, the first lead screw motor drives the fixed frame to move linearly along the guide rail direction with the cooperation of the guide rail and the slider, causing the chip fixture to move out and into the outer shell. The third lead screw motor drives the push plate to push the first slide valve and the second slide valve to move linearly.

[0009] In this invention, the negative pressure pumping module includes a first solenoid valve, a second solenoid valve, and a stepperistaltic pump; the stepperistaltic pump is connected in parallel with the first solenoid valve and the second solenoid valve, the first solenoid valve is connected to one end of a first rigid hollow tube column, and the second solenoid valve is connected to one end of a second rigid hollow tube column, so that the first solenoid valve or the second solenoid valve can be selectively opened during operation to drive the liquid flow in an orderly manner.

[0010] The present invention also provides a method for operating the above-mentioned vertical microbial detection device, comprising the following steps: In the lateral motion module, the fixing frame moves laterally in a straight line to move the chip fixture to the outside of the shell to place the microfluidic chip and move it into the shell to enter the experimental state. The push plate moves laterally in a straight line relative to the fixing frame to push the first slide valve and the second slide valve to control the opening or closing of the first solution storage device, the second solution storage device, and the reaction device. In the longitudinal motion module, the fixed plate moves downward in a straight line to achieve a pressing and sealing between the end faces of the first rigid hollow column, the second rigid hollow column, and the solid column in the microfluidic chip and the first negative pressure channel, the second negative pressure channel, and the lysis liquid chamber corresponding to the microfluidic chip. In the negative pressure extraction module, the first or second solenoid valve is selectively opened, and the stepperistaltic pump drives the liquid flow in an orderly manner.

[0011] Compared with the prior art, the beneficial effects of the present invention are as follows: The detection device of this invention, in conjunction with a microfluidic chip structure, requires only a single stepperistaltic pump to drive different liquids within the chip to the target chamber. The peristaltic pump carries only gas, minimizing contamination during experiments and ensuring the accuracy of results. (Using a slide valve as the on / off component, the detection device controls the linear movement of the slide valve, allowing selective connection of corresponding chambers and channels. This eliminates the need for multiple on / off components, simplifying the overall structure and control process of the detection device.) Attached Figure Description

[0012] Figure 1 This is a schematic diagram of the structure of a microbial detection device. Figure 1 .

[0013] Figure 2 This is a schematic diagram of the structure of a microbial detection device. Figure 2 .

[0014] Figure 3 This is a schematic diagram of the longitudinal motion module structure. Figure 1 .

[0015] Figure 4 This is a schematic diagram of the longitudinal motion module structure. Figure 2 .

[0016] Figure 5 This is a schematic diagram of the longitudinal motion module.

[0017] Figure 6 This is a schematic diagram of the lateral motion module structure.

[0018] Figure 7 This is a schematic diagram of the clamping heating module structure.

[0019] Figure 8 This is a schematic diagram of the structure of synthetic stone.

[0020] Figure 9 This is a schematic diagram of the longitudinal motion module outside the housing.

[0021] Figure 10 This is a structural block diagram showing the gas-liquid path connection relationship when the lysis solution and cleaning solution pass through the membrane.

[0022] Figure 11 This is a structural block diagram showing the gas-liquid path connection during the elution and dehydration process.

[0023] Figure 12 This is a structural diagram showing the gas-liquid path connection during sample loading (nucleic acid and reaction solution entering the reaction chamber).

[0024] Figure 13 This is a structural diagram of the assembled microfluidic chip.

[0025] Figure 14 This is an exploded view of a microfluidic chip.

[0026] Figure 15 This is a structural diagram of the first solution storage device.

[0027] Figure 16 This is a structural diagram of the bottom surface of the first solution storage device.

[0028] Figure 17 This is a structural diagram of the first sealing gasket.

[0029] Figure 18 This is a structural diagram of the first slide valve.

[0030] Figure 19 This is a structural diagram of the second solution storage device.

[0031] Figure 20 It is the structural diagram of the second slide valve.

[0032] Figure 21 It is the structural diagram of the reaction part.

[0033] Reference numerals in the figure: 1 - longitudinal motion module, 2 - transverse motion module, 3 - stepping peristaltic pump, 4 - housing, 5 - microfluidic chip, 6 - first rigid hollow pipe column, 7 - second rigid hollow pipe column, 8 - solid column, 9 - first guide plate, 10 - fixing plate, 11 - fixture, 12 - first tension spring, 13 - second guide plate, 14 - second lead screw motor, 15 - lead screw nut, 16 - first slider, 17 - mounting seat, 18 - fork sheet metal, 19 - second tension spring, 20 - output shaft, 21 - limiting part, 22 - first lead screw motor, 23 - third lead screw motor, 24 - chip jig, 25 - heating and detection module, 26 - push plate, 27 - slide plate, 28 - second slider, 29 - fixing frame, 30 - pipe column fixing part, 31 - guide rail, 32 - piercing part, 33 - pressing plate, 34 - bearing, 35 - bearing fixing part, 36 - first heating block fixing part, 37 - second heating block fixing part, 38 - convex platform; 51 - first solution storage part, 52 - first slide valve, 53 - second solution storage part, 54 - second slide valve, 55 - reaction part, 56 - first gasket, 57 - second gasket, 58 - lysate cavity, 59 - cleaning solution cavity, 510 - eluent cavity, 511 - reaction solution cavity, 512 - first channel, 513 - second channel, 514 - groove, 515 - third gasket, 516 - protruding part, 517 - lysate outlet, 518 - cleaning solution outlet, 519 - eluent outlet, 520 - reaction solution outlet, 521 - extension part, 526 - third channel, 527 - fourth channel, 528 - waste liquid inlet, 529 - waste liquid cavity vent, 530 - solution inlet, 531 - premixing cavity vent, 532 - first through hole, 533 - first flow channel, 534 - second flow channel, 535 - third flow channel, 536 - fourth flow channel, 537 - nucleic acid adsorption flow channel, 538 - second through hole,​​​​​​​​​As shown; the microfluidic chip assembly includes: a first solution storage unit 51, a first slide valve 52, a second solution storage unit 53, a second slide valve 54, and a reaction unit 55; the first slide valve 52 is movably disposed between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 53, and the second slide valve 54 is movably disposed between the bottom surface of the second solution storage unit 53 and the top surface of the reaction unit 55; the first solution storage unit 51, the second solution storage unit 53, and the reaction unit 55 are fixedly connected, for example: the first solution storage unit 51, the second solution storage unit 53, and the reaction unit 55 are fastened together by bolts, so that the first slide valve 52 is fastened between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 53, and the second slide valve 54 is fastened between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 53, and the second slide valve 5 ...4 is fastened between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 54, and the second slide valve 55 is fastened between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 55, and the second slide valve 55 is fastened between the bottom surface of the first solution storage unit 51 and the top surface of the second solution storage unit 53, and the second slide valve 55 is Two slide valves 54 are fastened between the bottom surface of the second solution storage container 53 and the top surface of the reaction container 55. Under the action of external force, the first slide valve 52 can move linearly relative to the bottom surface of the first solution storage container 51 and the top surface of the second solution storage container 53, and the second slide valve 54 can move linearly relative to the bottom surface of the second solution storage container 53 and the top surface of the reaction container 55. In order to ensure the sealing between the first slide valve 52 and the second solution storage container 53, and between the second slide valve 54 and the reaction container 55, and to avoid leakage of the solution during the transfer process, a first sealing gasket 56 is provided between the first slide valve 52 and the top surface of the second solution storage container 53, and a second sealing gasket 57 is provided between the second slide valve 54 and the top surface of the reaction container 55.

[0036] The first solution storage unit 51 is provided with: a lysis buffer chamber 58, a washing buffer chamber 59 (due to the strong penetrating power of the washing buffer, the bottom of the washing buffer chamber and the inner wall of the washing buffer chamber are integrally formed to form the washing buffer chamber), an elution buffer chamber 510, a reaction buffer chamber 511, a first channel 512, and a second channel 513, wherein the first channel 512 and the second channel 513 are negative pressure channels; wherein, the washing buffer chamber 59 is pre-stored with washing buffer, the elution buffer chamber 510 is pre-stored with elution buffer, the reaction buffer chamber 511 is pre-stored with reaction buffer, and the lysis buffer chamber 58 is used to add lysis buffer (containing nucleic acid of the sample); in order to ensure the first solution storage unit 58... The pre-stored solution is stably stored in each chamber. The bottom surface of the first solution storage unit 51 is provided with a groove 514, and a third sealing gasket 515 is provided in the groove 514. The eluent outlet and reaction liquid outlet 520 on the first solution storage unit 51 extend outward to form an extension 521. The end of the extension 521 is provided with a protrusion 549. The groove 514 is also provided with a number of protrusions 516. The third sealing gasket 515 is provided with flow channel holes corresponding to the outlets of each chamber, the first channel 512, and the second channel 513 on the first solution storage unit 51, and is provided with a number of recesses that match the number of protrusions 516.

[0037] The second solution storage unit 53 is provided with: a waste liquid chamber, a premixing chamber, and a third channel 526 and a fourth channel 527 corresponding to the second negative pressure channel 513; the top surface of the waste liquid chamber is provided with a waste liquid inlet 528 and a waste liquid chamber vent 529; the top surface of the premixing chamber is provided with a solution inlet 530 and a premixing chamber vent 531; the third channel 526 penetrates the second solution storage unit 53, and the fourth channel 527 communicates with the premixing chamber; the third channel 526 and the premixing chamber outlet on the bottom surface of the second solution storage unit 53 are provided with sealing rings for sealing; the first sealing gasket 56 is provided with a first through hole 532 corresponding to and communicating with the third channel 526, the waste liquid inlet 528, the waste liquid chamber vent 529, the solution inlet 530, the premixing chamber vent 531, and the fourth channel 527.

[0038] The first slide valve 52 is provided with: a first flow channel 533 selectively connecting to the pyrolysis liquid outlet 517, the cleaning liquid outlet 518, and the eluent outlet 519 (one end of the first flow channel 533 is the pyrolysis liquid inlet, and the other end is the cleaning liquid (ethanol) and eluent (water) inlet); a second flow channel 534 selectively connecting to the reaction liquid outlet 520 (one end of the second flow channel 534 is the reaction liquid inlet, and the other end of the second flow channel 534 is the second through hole 538); a third flow channel 535 selectively connecting to the first negative pressure channel 512; and a fourth flow channel 536 selectively connecting to the outside atmosphere (the fourth flow channel 536 is...). The system includes an air inlet channel and a nucleic acid adsorption channel 537 for adsorbing nucleic acids. The nucleic acid adsorption channel 537 contains nucleic acid adsorbents for adsorbing nucleic acids. A second through-hole 538 can selectively connect to a waste liquid inlet 528 or a solution inlet 530. A third through-hole 539 can selectively connect to a waste liquid chamber vent 529 or a premixing chamber vent 531. A fourth through-hole 540 can selectively connect to a third channel 526. A fifth through-hole 541 can selectively connect to a fourth channel 527. To ensure the flow channels on the first slide valve 52 are sealed and leak-proof, and to create a strong negative pressure, so as to smoothly drive the solution in the first solution storage container 51... The flow path through the first slide valve 52 is sealed with a sealing membrane on the side of the first slide valve 52 with the flow path. To facilitate solution transfer and create a negative pressure channel, the following locations are sealed: both ends of the first flow path 533, the end of the second flow path 534 away from the second through hole 538, the end of the third flow path 535 away from the third through hole 539, and the end of the fourth flow path 536 away from the fifth through hole 541. The third through hole 539, fourth through hole 540, and fifth through hole 541 are not sealed with sealing membranes. In the initial state (in this state: lysis buffer passes through the membrane, ethanol (cleaning solution) passes through the membrane), under the action of external force (bolt tightening force), the second flow path... The end of channel 534 away from the second through hole 538 and the end of the third channel 535 away from the third through hole 539 are sealed; when the first slide valve 52 is in the second state (in this state: water passes through the membrane, and the reaction solution is mixed with nucleic acid), one end of the first channel 533 (lysate inlet) and the third through hole 539 are sealed; in order to simplify the structure of the slide valve, the first channel 533 is connected to the nucleic acid adsorption channel 537, the second through hole 538 is connected to both the nucleic acid adsorption channel 537 and the second channel 534, the third through hole 539 is connected to the third channel 535, and the fifth through hole 541 is connected to the fourth channel 536.

[0039] The reaction component 55 is provided with: a reaction chamber, a reaction liquid inlet 544, and a reaction chamber vent 545; the second sealing gasket 57 is provided with a sixth through hole corresponding to and communicating with the reaction liquid inlet 544 and the reaction chamber vent 545.

[0040] The second slide valve 54 is provided with a seventh through hole 547 that can selectively connect to the reaction liquid inlet 544 and an eighth through hole 548 that can selectively connect to the reaction chamber vent 545. In the initial state, the seventh through hole 547 is not connected to the reaction liquid inlet 544 and the eighth through hole 548 is not connected to the reaction chamber vent 545. The reaction liquid inlet 544 and the reaction chamber vent 545 are both closed. When it is necessary to load the sample, the second slide valve 54 is pushed by external force, and the seventh through hole 547 is connected to the reaction liquid inlet 544 and the eighth through hole 548 is connected to the reaction chamber vent 545.

[0041] The pathways for different steps in microfluidic chip detection include: Article 1: In the initial state (nucleic acid adsorption), during nucleic acid adsorption, the lysis liquid chamber 58, the first flow channel 533, the nucleic acid adsorption flow channel 537, the second through hole 538, the waste liquid chamber (waste liquid inlet 528, waste liquid chamber vent 529) (in the initial state, the second through hole 538 is connected to the waste liquid chamber), the third through hole 539 (in the initial state, the third through hole 539 is connected to the waste liquid chamber and the first negative pressure channel 512), and the first negative pressure channel 512 form a ventilation circuit; Article 2: The chip remains in its initial state (nucleic acid cleaning). During cleaning, the cleaning liquid chamber 59, the first flow channel 533, the nucleic acid adsorption flow channel 537, the second through hole 538, the waste liquid chamber (waste liquid inlet 528, waste liquid chamber vent 529), the third through hole 539, and the first negative pressure channel 512 form a ventilation circuit. Article 3: Before elution, the first slide valve 52 needs to be pushed linearly to the second state (nucleic acid elution and reaction solution entry) for the first time. The elution chamber 510, the first flow channel 533, the nucleic acid adsorption flow channel 537, the second through hole 538, the premixing chamber (solution inlet 530, premixing chamber vent 531), the third through hole 539, the third flow channel 535, and the first negative pressure channel 512 form a ventilation circuit; water and reaction solution enter the premixing chamber together to play a mixing role; Article 4 (This step involves drawing negative pressure into the premixing chamber): Before loading the sample, the second slide valve 54 needs to be pushed linearly to the first state, and the first slide valve 52 needs to be pushed again to the third state (loading the sample). The external environment, the fourth flow channel 536, the fifth through hole 541, the premixing chamber, the seventh through hole 547, the reaction liquid inlet 544, the reaction chamber, the reaction chamber vent 545, the eighth through hole 548, the third channel 526, the fourth through hole 540, and the second negative pressure channel 513 form a ventilation circuit. Article 5: Before the heating reaction, the first and second slide valves 52 and 54 need to be reset to their initial state to prevent nucleic acid spillage and contamination.

[0042] Furthermore, the present invention provides a microbial detection device, such as... Figures 1-9As shown, it includes a longitudinal motion module 1, a transverse motion module 2, a negative pressure suction module, and a housing 4; the longitudinal motion module 1 is located above the transverse motion module 2, and in the initial state, the transverse motion module 2 and the longitudinal motion module 1 are located inside the housing 4; The lateral motion module 2 includes: a first lead screw motor 22, a third lead screw motor 23, a chip fixture 24, a heating detection module 25, a push plate 26, a slide plate 27, a guide rail, a second slider 28, and a fixing frame 29. The first lead screw motor 22 is used to drive the lateral linear motion of the lateral motion module 2. The fixed end of the first lead screw motor 22 is fixedly mounted on the outer casing 4, and the driving end of the first lead screw motor 22 is movably connected to the fixing frame 29. The heating detection module 25 is fixedly connected to the chip fixture 24, and the chip fixture 24 is fixedly mounted on the fixing frame 29. After the microfluidic chip 5 is placed in the chip slot formed by the chip fixture 24, The heating detection module 25 is aligned with and attached to the reaction chamber of the chip. The guide rail and the fixing frame 29 are fixedly connected as a whole. The guide rail and the second slider 28 are slidably connected. The second slider 28 is fixedly mounted on the outer shell 4. During operation, the first lead screw motor 22 drives the fixing frame 29 to move laterally in a linear motion, which in turn drives the chip fixture 24 to move laterally in a linear motion. The fixed end of the third lead screw motor 23 is mounted on the fixing frame 29. The driving end of the third lead screw motor 23 is movably connected to the push plate 26. During operation, the third lead screw motor 23 drives the push plate 26 to move laterally in a linear motion, which in turn pushes the first slide valve 52 and the second slide valve 54 on the chip to specific positions.

[0043] The lateral movement module also includes: a chip clamping heating detection module; the chip clamping heating module includes: a bearing 34, a bearing fixing part 35, a heating block, a first heating block fixing part 36, a second heating block fixing part 37 (the heating block fixing parts 36 and 37 are preferably made of synthetic stone, which can effectively insulate against heat and prevent heat loss from the heating block, so that heat is not lost during the process of transferring to the chip, and the synthetic stone does not deform during the heating process), and a pressure plate 33; the bearing 34 is fixedly mounted on the bearing fixing part 35, and the bearing fixing part 35 is fixedly connected to the outer shell 4; the first heating block fixing part 36 and the second heating block fixing part 37 are movably connected by a spring, and after the microfluidic chip 5 is placed into the chip fixture 24, the reaction chamber falls exactly into the groove formed between the first heating block fixing part 36 and the second heating block fixing part 37; the heating block is fixedly mounted on the heating block fixing parts 36 and 37, and the pressure plate 33 is relatively tilted left and right and hinged to the chip fixture 24 (the two ends of the pressure plate 33 are connected to the two ends of the chip fixture 24). The first heating block fixing member 36 (with a front and rear misaligned hole hinge) has a boss 38 on the side that contacts the pressure plate 33. There are two bosses 38, and the plane on which the two bosses 38 are located is an inclined plane that matches the plane on which the pressure plate 33 is located. With this design, when the lateral movement module 2 moves from the outside of the outer shell 4 to the inside of the outer shell 4, the lower edge of the pressure plate 33 flips inward around the front and rear misaligned hole under the pressure of the bearing 34 to press the bosses 38 on the synthetic stone. The bosses 38 further press the chip. In addition, the two bosses 38 are located at the axial center position of the first heating block fixing member 36 in the vertical direction. Therefore, the pressure applied by the pressure plate 33 to the first heating block fixing member 36 is located at the center position of the first heating block fixing member 36, which controls the first heating block fixing member 36 to press the chip smoothly, so that the heating detection block on the first heating block fixing member 36 is accurately aligned with the reaction chamber. This process has high heating conduction efficiency, and the reaction chamber can reach the temperature required for the experiment, ensuring the accuracy of the experiment.

[0044] The longitudinal motion module 1 includes: a first rigid hollow tube column 6, a second rigid hollow tube column 7, a solid column 8 for connecting the pyrolysis chamber to the outside atmosphere, a first guide plate 9, and a second lead screw motor 14. The second guide plate 13 is fixedly mounted on the tube column fixing component 30, which is fixedly mounted on the outer shell 4. During detection, the end face of the first rigid hollow tube column 6 abuts against the first negative pressure channel 512 (the first negative pressure channel 512 is the main air extraction port) on the top surface of the microfluidic chip 5, and the end face of the second rigid hollow tube column 7 abuts against the second negative pressure channel 513 (the second negative pressure channel 513 is the sample extraction port) on the top surface of the microfluidic chip 5. In order to ensure that the first and second rigid hollow tube columns 6 and 7 and the solid column 8 are accurately aligned with the corresponding negative pressure channel positions without deviation, and to form a strong negative pressure between the negative pressure channel on the chip and the corresponding hollow tube column, the second rigid hollow tube column 7 and the solid column 8 both pass through the second guide plate 13.

[0045] The longitudinal motion module 1 also includes: clamps 11, on which clamps 11 are fixedly mounted on the first and second rigid hollow tubes 6, 7, and solid column 8. Each clamp 11 is connected to a first tension spring 12, one end of which is fixedly mounted on the clamp 11, and the other end of which is fixedly mounted on the second guide plate 13. The first tension spring 12 is always in a stretched state so that the first and second rigid hollow tubes 6, 7, and solid column 8 provide sufficient clamping force to the first negative pressure channel 512, the second negative pressure channel 513, and the lysis liquid chamber 58 on the top surface of the microfluidic chip 5, ensuring that the first and second rigid hollow tubes 6, 7, and solid column 8 are in contact with the first negative pressure channel 512 and the second negative pressure channel 513 on the top surface of the microfluidic chip 5 during operation. 13. The lysis fluid chambers 58 are sealed to prevent leakage, so as to form a strong negative pressure to drive the directional flow of the solution; in order to further prevent leakage between the end faces of the solid column 8, the first and second rigid hollow tubes 6 and 7 and the corresponding lysis fluid chambers 58, the first negative pressure channel 512, and the second negative pressure channel 513 of the microfluidic chip 5, and to form a strong negative pressure, a flexible layer (which can be silicone or rubber) is bonded to the end faces of the solid column 8, the first and second rigid hollow tubes 6 and 7, ensuring the sealing between the contact surfaces, so that the end faces of the first and second rigid hollow tubes 6 and 7 and the corresponding lysis fluid chambers, the first negative pressure channel 512, and the second negative pressure channel 513 of the microfluidic chip 5, and to form a strong negative pressure, so as to smoothly drive the directional flow of the solution.

[0046] The longitudinal motion module 1 also includes: a fixed plate 10; the screw nut 15 of the second screw motor 14 is fixedly connected to the first slider 16, the first slider 16 is fixedly connected to the fixed plate 10, the second screw motor 14 rotates to drive the fixed plate 10 to reciprocate linearly along the guide rail 31, the guide rail 31 is fixedly set on the outer shell 4, so that the fixed plate 10 abuts or separates from the clamp 11; specifically, in the initial state, the first tension spring 12 is in the stretched state, and the clamp 11 abuts with the end face of the fixed plate 10; after the experiment starts, the second screw motor 14 rotates to drive the fixed plate 10 to move vertically downwards in a linear motion. During this process, the stretched tension spring resets and releases elastic potential energy to drive the clamp 11 and the first and second rigid hollow tube columns 6 and 7 and the solid column 8 to move vertically in a linear motion until the first and second rigid hollow tube columns 6 and 7 abut with the top surface of the chip, and the clamp 11 and the end face of the fixed plate 10 are always in contact.

[0047] The longitudinal motion module 1 also includes: a mounting base 17, a dual-position self-holding electromagnet, a shift fork sheet metal 18, a piercing element 32, and a second tension spring 19; the mounting base 17 is fixedly connected to the guide rail 31, the output shaft (iron core) 20 of the dual-position self-holding electromagnet is fixedly connected to the shift fork sheet metal 18, the second lead screw motor 14 is fixedly mounted on the mounting base 17, one end of the second tension spring 19 is fixedly mounted on the mounting base 17, and the other end is fixedly mounted on the piercing element 32, the piercing element 32 passing through the second lead screw motor 19. The guide plate 13 and the dual-position self-holding electromagnet are fixedly mounted on the fixed plate 10. The tip of the piercing part 32 is aligned with the cleaning fluid chamber 9, i.e., the ethanol chamber. The shift fork sheet metal 18 is rotatably connected to the fixed plate 10. The dual-position self-holding electromagnet and the shift fork sheet metal 18 are connected through the output shaft 20. By controlling the on and off of the dual-position self-holding electromagnet, the swing direction of the output shaft 20 is adjusted, so that the shift fork sheet metal 18 and the piercing part 32 are engaged or disengaged, thereby controlling the piercing part 32 to move closer to or further away from the bottom of the ethanol chamber.

[0048] The negative pressure module includes: a stepperistaltic pump 3, a first solenoid valve, and a second solenoid valve (both the first and second solenoid valves are used for gas flow; when sequentially extracting lysis buffer, washing buffer, and eluent, the first solenoid valve is open and the second solenoid valve is closed; during sample loading, the first solenoid valve is closed and the second solenoid valve is open); the stepperistaltic pump, the first solenoid valve, and the second solenoid valve are all fixedly mounted on the outer casing, and the stepperistaltic pump 3 is connected in parallel with the first and second solenoid valves; the first solenoid valve is connected to one end of the first rigid hollow tube column 6, and the second solenoid valve is connected to one end of the second rigid hollow tube column 7; during detection, the end face of the first rigid hollow tube column 6 abuts against the first negative pressure channel 512 on the top surface of the microfluidic chip 5, and the end face of the second rigid hollow tube column 7 abuts against the second negative pressure channel 513 on the top surface of the microfluidic chip 5; to ensure that the first and second rigid hollow tube columns 7 and the solid column 8 are accurately aligned with their corresponding positions without deviation, both the second rigid hollow tube column 7 and the solid column 8 pass through the second guide plate 13 ( Figures 10-12 ).

[0049] The testing method using the above-mentioned vertical microbial detection device is as follows: 1. Inserting the chip: The first lead screw motor 22 controls the horizontal movement module 2 to move to the left, moving the chip slot to the outside of the housing (please refer to...). Figure 9 The chip (containing the lysed sample solution) is placed into the chip slot. The first lead screw motor 22 controls the lateral movement module 2 to move to the right, moving the chip slot into the housing and back to its initial position (please refer to...). Figure 1 and Figure 2 At this time, the distance between the end face of the solid column 8 and the top surface of the chip is the first distance (e.g., 6mm), and the solid column 8 is aligned with the top surface of the pyrolysis chamber. The distance between the end face of the first rigid hollow tube column 6 and the end face of the second rigid hollow tube column 7 and the top surface of the chip is the second distance (e.g., 3mm). The first distance is greater than the second distance. II. Extraction of lysis buffer (please refer to...) Figure 10 The second lead screw motor 14 controls the longitudinal motion module 1 to move downward in a straight line, driving the first rigid hollow tube 6 and the second rigid hollow tube 7 to move downward a second distance (e.g., 3mm). At this time, the end faces of the first rigid hollow tube 6 and the second rigid hollow tube 7 are in contact with the top surface of the chip. The distance between the end face of the solid column 8 and the top surface of the chip is the second distance (e.g., 3mm, so that the pyrolysis chamber 58 is connected to the outside atmosphere, so that the negative pressure can drive the pyrolysis liquid smoothly). The first solenoid valve is controlled to open and the second solenoid valve is closed, and the stepper peristaltic pump 3 is turned on to draw the pyrolysis liquid through the membrane. After the pyrolysis liquid is drawn, the stepper peristaltic pump 3 is turned off, and the second lead screw motor 14 controls the solid column 8 to move downward a second distance (e.g., 3mm). At this time, the end face of the solid column 8 is in contact with the top surface of the chip (which is equivalent to closing the vent to the atmosphere on the top surface of the pyrolysis chamber, so that the pyrolysis chamber 58 is not connected to the outside atmosphere. Since the liquid flow resistance is greater than the air flow resistance, it is ensured that the ethanol cleaning liquid can be smoothly driven by the negative pressure in the next step, and will not drive the air). 3. Piercing the bottom of the cleaning fluid chamber 9: At this time, the left station of the dual-station self-holding electromagnet is energized, and the electromagnet mechanism is not associated with the piercing part 32 that pierces the bottom of the ethanol chamber (i.e., the fork sheet metal 18 and the piercing part 32 are separate). The piercing part 32 is in the initial state, and the top surface of the piercing part 32 abuts against the limiting part 21. At this time, the needle has not yet pierced the bottom of the cleaning fluid chamber 9, i.e., the bottom of the ethanol chamber. Control the right station of the dual-station self-holding electromagnet to be energized, so that the fork sheet metal 18 is close to the piercing part 32 (the fork sheet metal 18 is close to the piercing part 32 and associated with the piercing part 32). Start the second lead screw motor 14. The motor drives the fixed plate 10, the fork sheet metal 18, and the piercing part 32 to move downward in a straight line until the bottom of the ethanol chamber is pierced. At the same time, the solid column 8 further abuts against the top surface of the chip. IV. The puncture component 32 retracts, and the ethanol chamber is opened to the atmosphere: The second lead screw motor 14 is activated. The second lead screw motor 14 controls the longitudinal motion module 1 to move linearly upwards, driving the fixed plate 10, the shift fork sheet metal 18, and the puncture component 32 to a suitable position (approximately 2mm below the position in step three; this is to prevent air leakage caused by the separation of the hollow tube from the top surface of the chip; at this position, the solid column 8, the first and second rigid hollow tube columns 6 and 7 are still in contact with the top surface of the chip, and the tip of the puncture component 32 leaves the bottom surface of the ethanol chamber, opening the ethanol chamber to the atmosphere). The left position of the dual-station self-holding electromagnet is energized, and the shift fork sheet metal 18 separates from the needle. At this time, the needle returns to its initial position and contacts the upper limit under the action of the tension spring, initiating the ethanol extraction program. Please refer to [reference needed]. Figure 10 (After the ethanol is completely drawn out, the ethanol chamber outlet is closed by controlling the slide valve to move.) The stepperistaltic pump 3 is turned on to draw the cleaning solution through the membrane. After the cleaning solution is completely drawn out, the stepperistaltic pump 3 is turned off. 5. After the ethanol is completely drawn off, start the third lead screw motor 23. The lead screw nut drives the push plate 26 (the push plate 26 is fixedly connected to the drive end of the third lead screw motor 23) to move linearly close to the first slide valve 52, pushing the first slide valve 52 on the chip to slide linearly to the first position. At this time, the lysis liquid chamber 58 and the washing liquid chamber 59 are not connected to the premixing chamber, while the elution liquid chamber 510 and the reaction liquid chamber 511 are connected to the premixing chamber. Control the second lead screw motor 14 to control the longitudinal motion module 1 to move linearly downward, driving the fixed plate 10, the shift fork sheet metal 18, and the puncture part 32 to move linearly downward. The two extrusion rods respectively extrude the walls of the elution liquid chamber and the reaction liquid chamber (the elution liquid chamber 510 and the reaction liquid chamber 511 are equipped with rubber stoppers), causing the rubber stoppers to squeeze the solution on the walls into the premixing chamber (function: to prevent the solution from remaining on the walls and to ensure that it can completely enter the premixing chamber, thus ensuring the accuracy of the experimental results). At the same time, please refer to Figure 11 Turn on the stepperistaltic pump 3 to draw negative pressure into the premixing chamber. Under the combined drive of positive and negative pressure, the eluent and reaction liquid flow into the premixing chamber. After the eluent and reaction liquid are drawn out, turn off the stepperistaltic pump 3. The second lead screw motor 14 controls the longitudinal motion module 1 to move upward linearly, driving the fixed plate 10, the shift fork sheet metal 18, the puncture part 32, and the two extrusion rods to move upward linearly to the position of step four. VI. Sample Loading (Please refer to) Figure 12 The third lead screw motor 23 is started, and the push plate 26 is controlled to push the first slide valve 52 on the chip to slide linearly to the second position and the second slide valve 54 to the first position. At this time, the premixed chamber outlet is connected to the reaction liquid inlet 544, and the reaction chamber vent 545 is connected to the sample suction port. The stepperistaltic pump 3 is started, and the first solenoid valve is closed and the second solenoid valve is opened. The stepperistaltic pump 3 draws negative pressure on the reaction chamber. The reaction liquid and nucleic acid mixture flow into the reaction chamber under the drive of negative pressure. After the suction is completed, the stepperistaltic pump 3, the first solenoid valve, and the second solenoid valve are closed. 7. Start the third lead screw motor 23 to control the first slide valve 52 and the second slide valve 54 on the chip to return to the initial position. At this time, the reaction liquid inlet 544 and the reaction chamber vent are both in a sealed state (to prevent the amplification product from overflowing to the outside of the chip during heating and to avoid contamination). Under heating conditions, the nucleic acid and the reaction liquid undergo a color reaction. By observing the color change, the positive or negative of the sample can be determined. 8. Removing the chip: Control the first, second, and third lead screw motors 22, 14, and 23 to return to the zero position. The first lead screw motor 22 controls the horizontal movement module 2 to move to the left, moving the chip slot to the outside of the housing and removing the chip from the chip slot. The first lead screw motor 22 then controls the horizontal movement module 2 to move to the right, moving the chip slot to the inside of the housing and returning to the initial position.

[0050] Compared to traditional equipment, the detection device of this invention requires only one stepperistaltic pump 3. By controlling the position change of the slide valve, the connection between different solution storage chambers and corresponding flow channels is realized, simplifying the device structure and reducing the cost of the equipment. Moreover, the stepperistaltic pump 3 is only used for air extraction and not liquid extraction, avoiding liquid residue after passing through the peristaltic pump and minimizing the occurrence of contamination. Furthermore, the stepperistaltic pump 3 is connected in parallel with the first solenoid valve and the second solenoid valve. During operation, the first solenoid valve or the second solenoid valve is selectively opened, realizing the sequential and directional flow of different solutions to the target chamber, further simplifying the device structure. The first solenoid valve and the second solenoid valve are only used for air extraction and not liquid extraction, avoiding liquid residue after passing through the solenoid valve and minimizing the occurrence of contamination.

Claims

1. A vertical microbial detection device, characterized in that, It includes a longitudinal motion module, a transverse motion module, a negative pressure suction module, and a housing; the longitudinal motion module is located above the transverse motion module, and initially, both the transverse and longitudinal motion modules are located inside the housing; wherein: The lateral motion module includes a mounting frame, a chip fixture and a pusher plate mounted on the mounting frame; the microfluidic chip is placed in the chip slot formed by the chip fixture; the mounting frame moves laterally in a linear motion under the drive of a motor to move the chip fixture to the outside of the housing to insert the microfluidic chip and to move it into the housing to enter the experimental state; the pusher plate moves laterally in a linear motion relative to the mounting frame under the drive of a motor to open or close the different solution storage chambers in the microfluidic chip. The longitudinal motion module includes a first rigid hollow tube column, a second rigid hollow tube column, a solid column, a fixing plate, and a first tension spring. A first guide plate and a second guide plate are respectively provided above and below the first rigid hollow tube column, the second rigid hollow tube column, and the solid column to guide their vertical movement. A clamp is fixedly mounted on each of the first rigid hollow tube column, the second rigid hollow tube column, and the solid column. One end of the clamp is fixedly connected to the first tension spring, and the other end of the first tension spring is fixedly connected to the second guide plate. A fixing plate is positioned near the clamp. When the fixing plate moves downwards linearly driven by a motor, the first tension spring remains in a stretched state while the fixing plate abuts against the clamp, ensuring that the first rigid hollow tube column, the second rigid hollow tube column, and the solid column provide sufficient clamping force when contacting the top surface of the microfluidic chip. This achieves a seal between the end faces of the first rigid hollow tube column, the second rigid hollow tube column, and the solid column in the microfluidic chip and the corresponding first negative pressure channel, second negative pressure channel, and lysis fluid chamber of the microfluidic chip. The negative pressure module is used to drive the flow of liquid stored in the microfluidic chip in an orderly manner.

2. The vertical microbial detection device according to claim 1, characterized in that, The microfluidic chip includes, from top to bottom, a first solution storage device, a first slide valve, a second solution storage device, a second slide valve, and a reaction device; The first solution storage unit is provided with a pyrolysis liquid chamber, a washing liquid chamber, an elution liquid chamber, a reaction liquid chamber, a first negative pressure channel serving as a general suction port, and a second negative pressure channel serving as a sample loading suction port. The second solution storage unit is provided with a waste liquid chamber, a premixing chamber, a third channel corresponding to the second negative pressure channel, and a fourth channel. The reaction unit is provided with a reaction chamber. The first slide valve is provided with several flow channels and through holes to achieve selective communication with the solutions in the solution chambers of the first and second solution storage units, as well as selective communication with the gas path. The second slide valve is provided with through holes for selectively connecting the reaction liquid inlet of the reaction unit and the gas vent of the reaction chamber. Driven by a motor, the push plate moves laterally in a straight line relative to the fixed frame to push the first slide valve and the second slide valve to control the opening or closing of the first solution storage device, the second solution storage device, and the reaction device.

3. The vertical microbial detection device according to claim 2, characterized in that, The longitudinal motion module also includes a dual-position self-holding electromagnet, a shift fork sheet metal, a piercing component, and a second tension spring. One end of the second tension spring is fixedly mounted on the mounting base, and the other end is fixedly connected to the piercing component. The piercing component passes through the second guide plate, and the tip of the piercing component is aligned with the cleaning fluid chamber. The dual-position self-holding electromagnet is fixedly mounted on the fixed plate, and the output shaft of the dual-position self-holding electromagnet is fixedly connected to the shift fork sheet metal. The shift fork sheet metal is rotatably connected to the fixed plate. During operation, the swing direction of its output shaft is adjusted by controlling the on / off state of the dual-position self-holding electromagnet, causing the shift fork sheet metal and the piercing component to engage or disengage, thereby controlling the piercing component to move closer to or further away from the bottom of the cleaning fluid chamber.

4. The vertical microbial detection device according to claim 2, characterized in that, A sealing membrane is provided on the surface of the first slide valve where the flow channel is located; rubber plugs are provided on the elution chamber and the reaction chamber; two extrusion rods are provided on the fixed plate in the longitudinal motion module, which extrude the solution on the walls of the elution chamber and the reaction chamber by extruding the rubber plugs on the elution chamber and the reaction chamber respectively, so as to extrude the solution on the walls into the premixing chamber.

5. The vertical microbial detection device according to claim 2, characterized in that, In the longitudinal motion module, the motor is a second lead screw motor, which is connected to the first slider and the fixed plate. The second lead screw motor is mounted on the mounting base, which is fixedly connected to the guide rail set on the outer shell. During operation, the second lead screw motor rotates to drive the fixed plate to move linearly downward along the guide rail.

6. The vertical microbial detection device according to claim 2, characterized in that, In the lateral motion module, a chip clamping heating detection module is set on the fixed frame. The chip clamping heating detection module is fixedly connected to the chip fixture. After the microfluidic chip is placed in the chip slot formed by the chip fixture, the heating detection module is aligned with the reaction chamber of the chip and fits into it.

7. The vertical microbial detection device according to claim 6, characterized in that, The chip clamping heating detection module includes a first heating block fixing component, a second heating block fixing component, and a pressure plate. The first and second heating block fixing components are mounted on the outer shell via bearings and bearing fixing components. Heating blocks are fixedly installed on the first and second heating block fixing components, and the first and second heating block fixing components are movably connected by a spring. After the microfluidic chip is placed into the chip fixture, the reaction chamber falls exactly into the groove formed between the first and second heating block fixing components. The pressure plate is hinged to the chip fixture with relative left and right tilts. The side of the first heating block fixing component that contacts the pressure plate is provided with a boss that is parallel to and abuts against the pressure plate. There are two bosses, and the plane on which the two bosses are located is an inclined plane that matches the plane on which the pressure plate is located.

8. The vertical microbial detection device according to claim 2, characterized in that, In the lateral motion module, the linear motion of the fixed frame and the push plate is driven by the first lead screw motor and the third lead screw motor, respectively. The fixed end of the first lead screw motor is fixedly mounted on the outer shell, and the driving end is movably connected to the fixed frame. The fixed end of the third lead screw motor is mounted on the fixed frame, and the driving end of the third lead screw motor is movably connected to the push plate. A guide rail is provided on the fixed frame, and a second slider is provided on the outer shell. During operation, the first lead screw motor drives the fixed frame to move linearly along the guide rail direction with the cooperation of the guide rail and the slider, causing the chip fixture to move out and into the outer shell. The third lead screw motor drives the push plate to push the first slide valve and the second slide valve to move linearly.

9. The vertical microbial detection device according to claim 1, characterized in that, The negative pressure extraction module includes a first solenoid valve, a second solenoid valve, and a stepperistaltic pump. The stepperistaltic pump is connected in parallel with the first and second solenoid valves. The first solenoid valve is connected to one end of a first rigid hollow tube, and the second solenoid valve is connected to one end of a second rigid hollow tube, so that the first or second solenoid valve can be selectively opened during operation to drive the liquid flow in an orderly manner.

10. A method of operating the vertical microbial detection device according to any one of claims 1-9, characterized in that, Includes the following steps: In the lateral motion module, the fixing frame moves laterally in a straight line to move the chip fixture to the outside of the shell to place the microfluidic chip and move it into the shell to enter the experimental state. The push plate moves laterally in a straight line relative to the fixing frame to push the first slide valve and the second slide valve to control the opening or closing of the first solution storage device, the second solution storage device, and the reaction device. In the longitudinal motion module, the fixed plate moves downward in a straight line to achieve a pressing and sealing between the end faces of the first rigid hollow column, the second rigid hollow column, and the solid column in the microfluidic chip and the first negative pressure channel, the second negative pressure channel, and the lysis liquid chamber corresponding to the microfluidic chip. In the negative pressure extraction module, the first or second solenoid valve is selectively opened, and the stepperistaltic pump drives the liquid flow in an orderly manner.

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