A fully automatic nucleic acid detection analysis system and method

Abstract

The application discloses a kind of full-automatic nucleic acid detection analysis system and method, it belongs to medical examination testing instrument, molecular diagnostic detection instrument field, its technical key points are in at, including: consumable operation executor, it is connected with the rotating liquid transfer device of consumable and can drive rotating liquid transfer device rotation and lift and provide the power of liquid suction and liquid discharge for rotating liquid transfer device;Carriage assembly, for carrying and positioning consumable;Function warehouse processing component, for heating, magnetic separation, mixing operation of the magnetic bead lysis liquid warehouse of consumable;Reaction tube temperature control component, for the temperature adjustment of amplification reaction for consumable reaction tube;Optical assembly, for the fluorescence optical detection of solution in the reaction tube of consumable.The application aims to provide a kind of full-automatic nucleic acid detection analysis system and method, can realize sample processing, amplification, detection integrated operation.

Description

Technical Field

[0001] This invention relates to the technical field of medical examination and testing instruments and molecular diagnostic testing instruments, specifically to a fully automated nucleic acid detection and analysis system and method. Background Technology

[0002] The research team proposed an integrated sample processing and detection device in CN223134456U. Its design features include: after sample addition, sample processing is achieved through aspiration, vertical movement, and 360-degree rotation of a rotary pipette; temperature control of the PCR tube and nucleic acid amplification reaction are performed; and fluorescence detection is used to detect the amplification results, completing a fully enclosed nucleic acid extraction and detection process and reducing the probability of sample contamination.

[0003] For consumables, automated equipment is generally required to operate them in order to achieve a fully automated nucleic acid testing process.

[0004] Therefore, it is necessary to develop a fully automated device for the aforementioned consumables. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of the prior art by providing a fully automated nucleic acid detection and analysis system.

[0006] Another objective of this invention is to provide a fully automated nucleic acid detection and analysis method.

[0007] The technical solution of the present invention is as follows: A fully automated nucleic acid detection and analysis system, comprising: Consumable operation actuator, which is connected to the rotary pipette of consumables and can drive the rotary pipette to rotate and lift, as well as provide power for the rotary pipette to aspirate and dispense liquid; Carrier assembly, used to support and position consumables; Functional compartment processing component, which is used to perform heating, magnetic separation and mixing operations on the magnetic bead pyrolysis liquid compartment of consumables; The reaction tube temperature control assembly is used to adjust the temperature of the reaction tubes used for amplification reactions; Optical components used for fluorescence optical detection of solutions in reaction tubes of consumables.

[0008] Furthermore, the consumable operation actuator includes: The consumable connection unit includes: a gripper, a fixing block, a connecting block, and a bottom pneumatic connector; both the gripper and the bottom pneumatic connector are located at the lower end of the fixing block. The gripper is used to hold and position the consumable, and the bottom pneumatic connector is used to connect to the fluid interface of the consumable; the top of the fixing block is fixed to the bottom of the connecting block. A rotating unit is used to drive the consumable connection unit to rotate; the connecting block is connected to the rotating unit. The lifting unit is used to drive the consumable connection unit, the rotating unit, and the gas suction unit to perform lifting movements. The gas suction unit is used to provide negative / positive pressure to suction and dispense liquids from consumables via a bottom pneumatic connector.

[0009] Furthermore, the lifting unit includes: a lifting drive motor and a lifting platform; the lifting drive motor is capable of driving the lifting platform to move up and down; The rotating unit includes: a rotary drive motor, a drive wheel, a belt, a driven wheel, a consumable connection unit rotating shaft, and a bearing housing; the housing of the rotary drive motor is fixed to the lifting platform; the output shaft of the rotary drive motor is connected to the drive wheel; the drive wheel drives the driven wheel through the belt; the bearing housing is fixedly disposed on the lower surface of the lifting platform; the consumable connection unit rotating shaft is rotatably disposed in the bearing housing, with its upper end connected to the driven wheel and its lower end fixedly connected to the consumable connection unit. The gas suction unit includes a plunger pump, a front pneumatic pipe, a quick-connect air pipe, a pneumatic connector clamp, a rotary connector, a pneumatic connecting pipe, and a lower pneumatic connecting pipe connected in sequence; the plunger pump is fixed on the upper surface of the lifting platform; the center of the rotating shaft of the consumable connection unit is provided with a hollow part, which serves as a pneumatic connecting pipe. The pneumatic coupling clamp is fixed to the lifting platform, and the lower pneumatic connection joint is connected to the bottom pneumatic joint through a pipeline.

[0010] Furthermore, it also includes: a clamping assembly for clamping the reaction tube of the consumable onto the reaction tube temperature control assembly, comprising: Lifting and pressing motor; The lifting plate moves up and down driven by the lifting and pressing motor. A clamping plate is located below the lifting plate, and its lower surface can contact the outside of the consumable cover. Multiple springs are arranged between the lifting plate and the pressing plate.

[0011] Furthermore, the consumables include: a rotary pipette and a sample tray; the sample tray is equipped with multiple functional compartments: a waste liquid compartment, a magnetic bead lysis buffer compartment, a reagent tube, a reaction tube, and a washing liquid compartment; when the rotary pipette is rotated, the biased pipette at the end of the rotary pipette can sequentially reach and access each functional compartment on the sample tray and can transfer the solution in any functional compartment to other functional compartments.

[0012] Furthermore, the carrier assembly includes: Carrier platform; Consumables placement rack, which is used to hold consumables; The horizontal drive mechanism has its fixed end fixed to the carrier platform and its moving end connected to the consumables placement carrier. The lifting drive mechanism can drive the carrier platform to move up and down so that the consumables placed on the carrier can come into contact with or separate from the functional compartment processing components.

[0013] Furthermore, the functional warehouse processing components include: The heat-insulating support is provided with a support position corresponding to the magnetic bead pyrolysis liquid chamber and the reaction tube of the consumables, and an optical fiber hole assembly is provided below the support position corresponding to the reaction tube. An ultrasonic unit is installed at a position corresponding to the bearing position of the magnetic bead pyrolysis liquid chamber of the consumable, and is used to mix the solution in the magnetic bead pyrolysis liquid chamber. A heating unit is located at a position corresponding to the bearing position of the magnetic bead pyrolysis liquid chamber of the consumable, and is used to heat the solution in the magnetic bead pyrolysis liquid chamber; A magnet unit, located on the side of the heat-insulating support, has a movable magnet for adsorbing or releasing magnetic beads in the magnetic bead pyrolysis liquid chamber.

[0014] Furthermore, the heating unit includes: a heating block sheet metal bracket, a heating block, a heating film, and a retaining spring; the heating block sheet metal bracket is fixedly connected to the heat insulation support, and the heating block is connected to the heating block sheet metal bracket via the retaining spring; the heating film is provided on the back of the heating block; The ultrasonic unit includes: an ultrasonic mounting base, an ultrasonic device, and a connecting spring; the ultrasonic device is connected to the ultrasonic mounting base via the connecting spring. The magnet unit includes: a magnet motor mounting base, a magnet rotation drive motor, a magnet fixing base, and a magnet; the housing of the magnet rotation drive motor is fixed on the magnet motor mounting base; the magnet rotation drive motor is used to drive the magnet fixing base to rotate; a magnet is installed and fixed at the end of the magnet fixing base.

[0015] Furthermore, the functional compartment processing components also include: a base plate, an ultrasonic fixing seat, and a magnet motor mounting seat, all of which are fixed on the base plate.

[0016] Furthermore, the reaction tube temperature control assembly includes: a heating / cooling element, a heat sink, and a fan; a heating / cooling element is installed below each fiber optic port assembly; A radiator is installed below the heat-insulating support, and a fan is installed on the side of the radiator; The heating and cooling elements are connected to the fiber optic hole assembly and the heat sink, respectively.

[0017] It also includes: rack assembly; the rack assembly includes: a lower horizontal plate, an upper horizontal plate, and vertical support columns connecting the upper horizontal plate and the lower horizontal plate; The fixed ends of the lifting drive motors of the optical components and lifting units, the lifting and pressing motors of the pressing components, and the lifting drive mechanisms of the carrier components are all fixedly mounted on the upper horizontal plate. The base plate of the functional compartment processing component is fixedly mounted on the lower horizontal plate.

[0018] A fully automated nucleic acid detection and analysis method, which is a non-disease diagnostic method, includes the following steps: S1: Consumable loading and initial positioning: Place the consumables onto the carrier assembly, and then the load assembly carries the consumables to the top of the functional compartment processing assembly; S2: Consumables are placed in the functional compartment processing component, and the reaction tube of the consumables is in close contact with the reaction tube temperature control component; S3: The consumables operation actuator is connected to the rotary pipette of the consumables; S4, Nucleic acid extraction and purification steps; S5: Preparation of the amplification reaction system; S6: Nucleic acid amplification and real-time fluorescence detection; S7: Unload consumables.

[0019] The beneficial effects of this application are as follows: First, this application is the first to develop a fully automated nucleic acid detection and analysis system for rotary pipetting consumables such as CN223134456U.

[0020] Secondly, rotary pipettes need to have the following functions: aspiration, vertical movement, and 360° rotation. These three functions facilitate the transfer of substances between different compartments of the consumable. How to simultaneously satisfy these functions was one of the key issues in the research and development process.

[0021] The consumable connection unit consists of two components: a gripper and a bottom pneumatic connector. The gripper is used to securely connect to the rotary pipette, and the bottom pneumatic connector is connected to the pneumatic connector of the rotary pipette.

[0022] The consumable connection unit has a hollow section at the center of its rotating shaft, which serves as a pneumatic connection pipe. A rotary connection joint is installed in the pneumatic connection pipe, which solves the motion conflict problem between the stationary gas suction unit and the rotating consumable connection unit.

[0023] The lifting unit drives the lifting movement of the consumable connection unit, the rotating unit, and the gas suction unit, thus solving the problem of vertical movement.

[0024] The above design simultaneously satisfies the functions of "suction, up-and-down movement, and 360° rotation", and these three functions do not conflict with each other.

[0025] Third, the magnetic bead pyrolysis liquid chamber requires heating, mixing, and magnetic separation. Existing technologies perform these three processes at different workstations, but this application only has one workstation for these three processes. Therefore, how to integrate these functions together is a problem.

[0026] The functional compartment processing component integrates three functions into one location by "setting an ultrasonic unit and a heating unit at the position corresponding to the bearing position of the magnetic bead pyrolysis liquid compartment of the consumable, and setting a rotatable magnet unit that can be close to the side of the magnetic bead pyrolysis liquid compartment".

[0027] Fourth, the clamping assembly is also a core design element. Consumables are placed in the carrier assembly and then into the functional compartment processing assembly. At this point, the clamping assembly ensures that the reaction tube of the consumable is in close contact with the reaction tube temperature control assembly, thereby improving heating / cooling efficiency. Attached Figure Description

[0028] The present invention will be further described in detail below with reference to the embodiments shown in the accompanying drawings, but this does not constitute any limitation on the present invention.

[0029] Figure 1 This is a three-dimensional structural diagram of the fully automated nucleic acid detection and analysis system of this application.

[0030] Figure 2 This is a three-dimensional structural diagram of the consumable operation actuator of this application.

[0031] Figure 3 This is a three-dimensional structural diagram of the consumable operation actuator of this application from another perspective.

[0032] Figure 4 This is a three-dimensional structural schematic diagram of the consumable connection unit of this application.

[0033] Figure 5 This is a three-dimensional structural schematic diagram of the lifting unit of this application.

[0034] Figure 6 This is a three-dimensional structural schematic diagram of the rotating unit of this application.

[0035] Figure 7 This is a side view of the rotating unit of this application.

[0036] Figure 8 This is a three-dimensional structural diagram of the gas suction unit of this application.

[0037] Figure 9 This is a three-dimensional structural schematic diagram of the gas extraction unit of this application from another perspective.

[0038] Figure 10 This is a three-dimensional structural schematic diagram of the clamping component of this application.

[0039] Figure 11 This is a three-dimensional structural schematic diagram of the clamping component of this application from another perspective.

[0040] Figure 12 This is a schematic diagram showing the relationship between the clamping components and consumables in this application.

[0041] Figure 13 This is a three-dimensional structural schematic diagram of the carrier assembly of this application.

[0042] Figure 14 This is a three-dimensional structural schematic diagram of the carrier assembly of this application from another perspective.

[0043] Figure 15 This is a three-dimensional structural diagram of the functional module processing component of this application.

[0044] Figure 16 This is a three-dimensional structural diagram of the thermal insulation support of this application.

[0045] Figure 17 This is a schematic diagram showing the location of the fiber optic port assembly in this application.

[0046] Figure 18 This is a three-dimensional structural diagram of the heat-insulating support and heating unit of this application.

[0047] Figure 19 This is a three-dimensional structural schematic diagram of the heating unit of this application.

[0048] Figure 20 This is a three-dimensional structural schematic diagram of the ultrasonic unit of this application.

[0049] Figure 21 This is a three-dimensional structural schematic diagram of the magnet unit of this application.

[0050] Figure 22 This is a schematic diagram showing the positional relationship between the functional compartment processing components and consumables in this application.

[0051] Figure 23 This is a three-dimensional structural schematic diagram of the reaction tube temperature control component of this application.

[0052] Figure 24 This is a three-dimensional structural schematic diagram of the reaction tube temperature control component of this application from another perspective.

[0053] Figure 1-24 The annotations in the accompanying drawings are explained as follows: 1000 fully automated nucleic acid detection and analysis system; The frame assembly 100, the lower horizontal plate 101, the upper horizontal plate 102, and the vertical support column 103; Consumables operation actuator 200; Consumable connection unit 210, gripper 211, fixing block 212, connecting block 213, bottom pneumatic connector 214; Lifting unit 220, lifting drive motor 221, lifting platform 222, vertical guide rail 223, lifting drive motor mounting base 224; Rotating unit 230, rotating drive motor 231, drive wheel 232, belt 233, driven wheel 234, consumable connection unit rotating shaft 235, bearing seat 236; Gas suction unit 240, plunger pump 241, front pneumatic pipe, quick-connect air pipe 242, pneumatic connection pipe, lower pneumatic connection joint 243, pneumatic joint clamp 244, rotary connection joint 245, connector 246; The clamping assembly 300, the lifting clamping motor 301, the lifting plate 302, the spring 303, the clamping plate 304, and the vertical guide rod 305 are included. Carrier assembly 400, carrier platform 401, carrier horizontal electric telescopic mechanism 402, consumable placement carrier 403, carrier platform lifting drive mechanism 404, load platform vertical guide rail 405; Functional compartment processing component 500, heat preservation support 510, support 511, fiber optic hole assembly 512, base plate 520, ultrasonic unit 530, ultrasonic fixing seat 531, ultrasonic device 532, connecting spring 533, heating unit 540, heating block sheet metal bracket 541, heating block 542, heating film 543, retaining ring 544, magnet unit 550, magnet motor mounting seat 551, magnet rotation drive motor 552, magnet fixing seat 553, magnet 554; The reaction tube temperature control assembly 600, the heating and cooling element 601, the radiator 602, and the fan 603; Optical components 700. Detailed Implementation

[0054] <Research and Development Needs and Challenges> The consumables described in CN223134456U include: a rotary pipette and a sample disk; the sample disk is provided with multiple functional compartments: a waste liquid compartment, a magnetic bead lysis buffer compartment, a reagent tube, a reaction tube, and a washing liquid compartment; by rotating the rotary pipette, the biased pipette at its end can sequentially reach and access each functional compartment on the sample disk and can transfer the solution in any functional compartment to other functional compartments.

[0055] Consumable requirements: First, the rotary pipette needs to have the following functions: aspiration, up-and-down movement, and 360° rotation. These three functions can facilitate the transfer of substances in the various compartments of the consumable: (1) the solution in the magnetic bead pyrolysis solution compartment is transferred to the waste liquid compartment; (2) the solution in the washing compartment is transferred to the magnetic bead pyrolysis solution compartment.

[0056] Second, the consumables are based on the magnetic bead method for extracting and purifying nucleic acids. The magnetic bead method for nucleic acid extraction and purification requires the following steps: lysis, mixing, magnetic separation, liquid transfer, washing, and elution. For these steps, lysis, washing, and elution are accomplished using the solution (i.e., via the rotary pipette of the consumables). However, mixing and magnetic separation require the use of other mechanisms.

[0057] The main challenges in developing operating equipment to complement rotary pipetting consumables lie in: First, how to simultaneously satisfy the functions of "suction, up and down movement, and 360° rotation".

[0058] Second, the magnetic bead pyrolysis liquid chamber requires heating, mixing, and magnetic separation. Existing technologies perform these three processes at different workstations, but this application only has one workstation for these three processes. Therefore, how to integrate these functions together is a problem.

[0059] 2.1 How to achieve mixing. The existing technique of shaking is unsuitable because shaking the magnetic bead pyrolysis liquid chamber will also affect other functional chambers.

[0060] 2.2 How is the magnetic separation function implemented?

[0061] 2.3 Pyrolysis and elution require specific temperatures (55-65°C).

[0062] <Example 1> Figure 1 The diagram illustrates the three-dimensional structure of a fully automated nucleic acid detection and analysis system 1000. The fully automated nucleic acid detection and analysis system 1000 (which uses the consumables described in CN223134456U) includes: (1) Rack assembly 100, which includes: a lower horizontal plate 101, an upper horizontal plate 102 and a vertical support column 103 connecting the upper horizontal plate and the lower horizontal plate.

[0063] (2) Consumable operation actuator 200, which is connected to the rotary pipette of the consumable and can drive the rotary pipette to rotate and lift, as well as provide power for the rotary pipette to aspirate and dissipate liquid.

[0064] (3) The clamping component 300 is used to clamp the consumables to ensure that the reaction tube of the consumables is tightly attached to the reaction tube temperature control component 600.

[0065] (4) Carrier assembly 400, which is used to carry consumables and transport them over the functional compartment processing assembly 500.

[0066] (5) Functional chamber processing component 500, which is used to heat, magnetically separate and mix the magnetic bead pyrolysis liquid chamber of consumables.

[0067] (6) Reaction tube temperature control assembly 600, which is used to heat and cool the reaction tube (i.e., the temperature cycle required for the amplification program).

[0068] (7) Optical component 700. The light source of optical component 700 is the light emitted by the consumable solution, and then the excitation light of the solution in the consumable is directed onto the photoelectric sensor of optical component 700. It adopts existing technology, such as CN118392842B, which will not be described in detail here.

[0069] <Construction of Consumable Operation Actuator 200> Figure 2 and Figure 3 The three-dimensional structure of the consumable operation actuator is illustrated in different diagrams. The consumable operation actuator 200 includes: a consumable connection unit 210, a lifting unit 220, a rotating unit 230, and a gas suction unit 240.

[0070] Figure 4 The three-dimensional structure of the consumable connection unit 210 is illustrated. The consumable connection unit 210 includes: a gripper 211, a fixing block 212, a connecting block 213, and a bottom pneumatic connector 214; the gripper 211 and the bottom pneumatic connector 214 are both disposed at the lower end of the fixing block 212, and the top of the fixing block 212 is fixed to the bottom of the connecting block 213.

[0071] Figure 5 The three-dimensional structure of the lifting unit 220 is illustrated. The lifting unit 220 includes: a lifting drive motor 221, a lifting platform 222, and a vertical guide rail 223; the lifting platform 222 is provided with a threaded hole, and the output shaft of the lifting drive motor 221 is provided with a threaded section, the threaded section of the output shaft of the lifting drive motor 221 passing through the threaded hole provided in the lifting platform 222; the vertical guide rail 223 passes through the guide hole provided in the lifting platform 222; that is, when the lifting drive motor 221 rotates, the lifting platform 222 can move up and down along the vertical guide rail 223.

[0072] The lifting unit 220 further includes a lifting drive motor mounting base 224, on which the housing of the lifting drive motor 221 is fixedly mounted; the lifting drive motor mounting base 224 is fixed on the upper horizontal plate 102.

[0073] The bottom of the vertical guide rail 223 is fixedly installed on the upper horizontal plate 102.

[0074] Figure 6The three-dimensional structure of the rotation unit 230 is shown. The rotation unit 230 includes: a rotation drive motor 231, a drive wheel 232, a belt 233, and a driven wheel 234; the housing of the rotation drive motor 231 is fixed to the lifting platform 222; the output shaft of the rotation drive motor 231 is connected to the drive wheel 232; the drive wheel 232 drives the driven wheel 234 through the belt 233, that is, when the rotation drive motor 231 rotates, the driven wheel 234 is driven to rotate through the drive wheel 232 and the belt 233.

[0075] As Figure 7 shown, the rotation unit 230 further includes: a consumable connection unit rotating shaft 235 and a bearing seat 236; the bearing seat 236 is fixedly arranged on the lower surface of the lifting platform 222; the consumable connection unit rotating shaft 235 is rotatably arranged in the bearing seat 236 (that is, the bearing seat 236 limits the vertical displacement of the consumable connection unit rotating shaft 235); the driven wheel 234 is fixedly connected to the upper part of the consumable connection unit rotating shaft 235; the lower part of the consumable connection unit rotating shaft 235 is fixedly connected to the connection block 213 of the consumable connection unit 210. That is, when the driven wheel 234 rotates, the jaws 211 of the consumable connection unit 210 are driven to rotate through the consumable connection unit rotating shaft 235.

[0076] Figure 8 and Figure 9 The structure of the gas suction unit 240 is shown from different angles. The gas suction unit 240 includes: a plunger pump 241, a front pneumatic pipeline, a quick connector for the air pipe 242, a pneumatic connection pipeline, a lower pneumatic connection joint 243, a pneumatic joint clamp 244, a rotary connection joint 245, and a connecting member 246; The plunger pump 241 is fixed on the upper surface of the lifting platform 222; A hollow part is provided at the center of the consumable connection unit rotating shaft 235, and the hollow part of the consumable connection unit rotating shaft 235 serves as the pneumatic connection pipeline; The lower pneumatic connection joint 243 is fixedly installed at the bottom of the pneumatic connection pipeline, and the lower pneumatic connection joint 243 is connected to the bottom pneumatic joint 214 through a pipeline; the bottom pneumatic joint 214 is connected to the pump connection interface of the consumable; The connecting member 246 is fixedly connected to the lifting platform 222, and it is in a "U" shape, and it includes a horizontally upper plate 2461 and a vertically supporting plate 2462; The top of the pneumatic connection pipe is connected to a rotary connector 245 (the pneumatic rotary connector is used to keep the upper quick-connect air pipe connector 242 stationary, while the lower bottom pneumatic connector 214 can rotate freely). The top of the rotary connector 245 is connected to the lower side of the pneumatic connector clamp 244, and the quick-connect air pipe connector 242 is connected to the upper side of the pneumatic connector clamp 244. The outer shell of the pneumatic connector clamp 244 is fixedly connected to the horizontal upper plate 2461; The plunger pump 241 and the quick-connect air pipe 242 are connected through a front pneumatic pipe.

[0077] The consumable operation actuator 200 uses a rotary drive motor 231 and a lifting drive motor 221 to lift and rotate the consumable connection unit 210, thereby driving the gripper 211, the bottom pneumatic connector 214 and the rotary pipette of the consumable to connect (the gripper 211 clamps the rotary pipette, and the bottom pneumatic connector 214 is connected to the air pump adapter of the rotary pipette). Then, it drives the rotary pipette to lift and rotate, and the plunger pump provides the power for liquid suction and discharge. Through the above, the liquid in the consumable is transferred between the various compartments.

[0078] The gripper 211 is combined with the rotary pipette of the consumables, and the gripper and the rotary pipette are engaged to make the two work together; at the same time, the bottom pneumatic connector 214 is fully engaged with the consumable connector to ensure a seal.

[0079] <Construction of clamping assembly 300> Figure 10 and Figure 11 The structure of the clamping assembly 300 is illustrated from different angles. The clamping assembly 300 includes: a lifting clamping motor 301, a lifting plate 302, a spring 303, a clamping plate 304, and a vertical guide rod 305; The housing of the lifting and pressing motor 301 is fixed to the upper horizontal plate 102; The lifting and pressing motor 301 drives the lifting plate 302 to move up and down (the position of the lifting plate 302 is adjusted by a screw and nut between the lifting and pressing motor 301 and the lifting plate). The pressing plate 304 is provided below the lifting plate 302, and a plurality of springs 303 are provided between the lifting plate 302 and the pressing plate 304. The bottom of the vertical guide rod 305 is fixed to the upper surface of the upper horizontal plate 102, and its upper part passes through the guide hole provided in the lifting plate 302.

[0080] like Figure 12As shown, the clamping assembly 300 is used to clamp the consumables. That is, the consumables are placed in the consumables placement carrier 403. The lifting plate 302, the spring 303, and the clamping plate 304 descend together. When the clamping plate 304 contacts the consumables, the lifting plate 302 continues to descend, the spring 303 is compressed, and the clamping plate 304 clamps the consumables.

[0081] <Construction of the frame assembly 400> Figure 13 and Figure 14 The structure of the carrier assembly 400 is illustrated from different angles. The carrier assembly 400 includes: a carrier platform 401, a carrier horizontal electric telescopic mechanism 402, a consumable placement carrier 403, a carrier platform lifting drive mechanism 404, and a load platform vertical guide rail 405.

[0082] The fixed end of the lifting drive mechanism 404 of the carrier platform is fixed to the upper horizontal plate 102, and its moving end is fixed to the carrier platform 401; the vertical guide rail 405 of the load platform is fixed to the lower surface of the upper horizontal plate 102, and the carrier platform 401 is provided with a guide hole for the vertical guide rail 405 of the load platform to pass through. The carrier platform 401 is horizontally arranged between the lower horizontal plate 101 and the upper horizontal plate 102; the fixed end of the carrier horizontal electric telescopic mechanism 402 is fixed to the upper surface of the carrier platform 401, and its moving end is fixedly connected to the consumables placement carrier 403, so that the consumables placement carrier 403 can move horizontally. The consumables placement rack 403 is used to place consumables.

[0083] When using the clamping assembly 300 and the carrier assembly 400, the working process is as follows: In the first step, the horizontal electric telescopic mechanism 402 of the carrier rack drives the consumable placement carrier rack 403 to extend outward, and the consumables are manually placed on the consumable placement carrier rack 403; In the second step, the consumable placement carrier 403 retracts back to directly above the reaction tube temperature control component 600, then lowers down to fit against the reaction tube temperature control component 600. Then, the consumable pressing component, driven by a motor, presses down on the consumable, making the consumable and the reaction tube temperature control component 600 completely fit together. The consumable pressing component forms an elastic pressing structure through 4 springs, pressing the consumable tightly onto the reaction tube temperature control component 600.

[0084] <Construction of Functional Warehouse Processing Component 500> Figure 15 The structure of the functional compartment processing component 500 is illustrated. The functional compartment processing component 500 includes: an insulated support 510, a base plate 520, an ultrasonic unit 530, a heating unit 540, and a magnet unit 550; Figure 16 The three-dimensional structure of the heat-insulating support 510 is illustrated. The heat-insulating support 510 includes: 5 supports 511 corresponding to the reaction tubes of the consumables, and each support 511 has a fiber optic hole assembly 512 below it. The fiber optic hole assembly 512 is used to install the emitting optical fiber and the excitation optical fiber of the optical component 700.

[0085] Figure 18 The three-dimensional structure of the heat preservation support 510 and the heating unit 540 is shown. Figure 19 The three-dimensional structure of the heating unit 540 is illustrated. The heating unit 540 includes: a sheet metal support for the heating block 541, a heating block 542, a heating film 543, and a retaining spring 544; The sheet metal bracket 541 of the heating block is fixedly connected to the heat insulation support 510; The heating block 542 is connected to the sheet metal bracket 541 of the heating block via a snap ring 544; The heating block 542 has a heating film 543 on its back side, and its front side is in contact with the magnetic bead pyrolysis liquid chamber of the consumable.

[0086] The heating unit 540 forms an elastic structure through the retaining spring 544 to ensure the elastic fit between the heating block and the magnetic bead pyrolysis liquid chamber. After the heating film 543 is energized, it heats the heating block 542, and the heating block 542 then heats the magnetic bead pyrolysis liquid chamber.

[0087] Figure 20 The three-dimensional structure of the ultrasonic unit 530 is illustrated. The ultrasonic unit 530 includes: an ultrasonic fixing base 531, an ultrasonic device 532, and a connecting spring 533; the ultrasonic fixing base 531 is fixedly connected to the base plate 520, and the ultrasonic device 532 forms an elastic structure through the connecting spring 533 to ensure the elastic fit between the ultrasonic device 532 and the consumable magnetic bead compartment.

[0088] Figure 21 The three-dimensional structure of the magnet unit 550 is illustrated. The magnet unit 550 includes: a magnet motor mounting base 551, a magnet rotation drive motor 552, a magnet fixing base 553, and a magnet 554. The magnet motor mounting base 551 is fixed to the base plate 520. The housing of the magnet rotation drive motor 552 is fixed to the magnet motor mounting base 551. The drive shaft of the magnet rotation drive motor 552 drives the magnet fixing base 553 to rotate. The magnet 554 is fixedly mounted at the end of the magnet fixing base 553. The magnet rotation drive motor 552 controls the magnet to rotate and approach the magnetic bead pyrolysis liquid chamber to attract the magnetic beads.

[0089] <Construction of the reaction tube temperature control assembly 600> Figure 23 and Figure 24The three-dimensional structure of the reaction tube temperature control assembly 600 is illustrated from different angles. The reaction tube temperature control assembly 600 includes: a heating / cooling element 601 (TEC), a heat sink 602, and a fan 603; a heating / cooling element 601 (TEC) is disposed below each of the fiber optic port assemblies 512. A heat sink 602 is disposed below the insulation support 510, and a fan 603 is disposed to the side of the heat sink 602. The heating / cooling element 601 (TEC) is connected to both the fiber optic port assembly 512 and the heat sink 602.

[0090] When the reaction tube needs to be heated, the side of the heating element 601 facing the fiber optic hole assembly 512 is heated, and the side facing the heat sink 602 is cooled; when the reaction tube needs to be cooled, the side of the heating element 601 facing the fiber optic hole assembly 512 is cooled, and the side facing the heat sink 602 is heated.

[0091] It should be noted that it also includes: an ambient temperature sensor placed on the heat sink 602. The ambient temperature sensor is used to detect the ambient temperature of the heat sink. When the ambient temperature of the heat sink is detected to be higher than the set value, the fan 603 works to reduce the temperature of the heat sink in order to ensure the working efficiency of the heating and cooling chip 601 (TEC).

[0092] The fully automated nucleic acid detection and analysis method of this application includes the following steps: S1: Consumable loading and initial positioning; The consumable placement carrier is moved to the outside by the horizontal drive mechanism of the carrier assembly (i.e., the horizontal electric telescopic mechanism of the carrier assembly), and the operator places the consumables on the consumable placement carrier; then, the horizontal electric telescopic mechanism 402 of the carrier assembly drives the consumable placement carrier 403 to carry the consumables and move it above the reaction tube temperature control assembly and the functional compartment processing assembly.

[0093] S2: Consumables compression; The lifting drive mechanism of the carrier assembly 400 drives the consumable placement carrier 403 to descend, so that the bottom of the consumable enters the functional compartment processing assembly; Subsequently, the lifting and pressing motor 301 of the pressing assembly 300 is started, and the pressing plate 304 descends and elastically presses the consumable, ensuring that the reaction tube of the consumable is in close contact with the reaction tube temperature control assembly.

[0094] S3: The consumable operation actuator is connected to the rotary pipette of the consumable; the lifting unit descends, the gripper is fixedly connected to the rotary pipette with a snap-fit ​​connection, and the bottom pneumatic connector is connected to the pneumatic connector of the rotary pipette; S4, Nucleic acid extraction and purification steps; S41: Binding and lysis: The sample is transferred from the sample compartment of the consumable to the magnetic bead lysis buffer compartment and mixed with the magnetic beads and lysis buffer pre-loaded in the compartment; the ultrasonic unit 530 of the functional compartment processing component 500 is activated to mix the mixture and allow the nucleic acid to be adsorbed onto the magnetic beads.

[0095] S42: Magnetic separation and waste liquid removal: The magnet unit 550 of the functional chamber processing component 500 is activated, so that the magnet is brought close to the magnetic bead pyrolysis liquid chamber to adsorb the magnetic beads; the consumable operation actuator 200 transfers the waste liquid to the waste liquid chamber of the consumable.

[0096] S43: Washing: The magnet of the magnet unit 550 moves away from the magnetic bead pyrolysis liquid tank, the consumable operation actuator 200 transfers the washing liquid from the washing liquid tank of the consumable to the magnetic bead pyrolysis liquid tank, and starts the ultrasonic unit 530 to clean; repeat the magnetic separation and waste liquid removal operation (i.e. step S42) at least once.

[0097] S44: Elution: After washing, the consumable operation actuator 200 transfers the eluent from the eluent chamber of the consumable to the magnetic bead lysis buffer chamber; the heating unit 540 of the functional chamber processing component 500 is activated to heat and promote nucleic acid elution; finally, magnetic separation is performed, and the consumable operation actuator 200 aspirates the eluent containing purified nucleic acid to complete the extraction.

[0098] S5: Preparation of the amplification reaction system; The consumable actuator 200 drives the rotary pipette to transfer the elution buffer containing purified nucleic acid and the amplification reaction premix solution drawn from the reagent compartment of the consumable into the reaction tube of the consumable to form the reaction system to be amplified.

[0099] S6: Nucleic Acid Amplification and Real-Time Fluorescence Detection S61: Temperature control cycle: The reaction tube temperature control component 600 performs cyclic temperature control on the reaction tube according to a preset nucleic acid amplification program, including denaturation, annealing and extension temperatures.

[0100] S62: Synchronous optical detection: In the predetermined stage of each temperature-controlled cycle, the optical component 700 is activated, and its light source emits excitation light into the solution inside the tube through the optical path below the reaction tube, and receives the fluorescence signal generated by the solution; the system processes the fluorescence signal in real time and generates an amplification curve.

[0101] S7: Results Analysis and Consumable Unloading Procedures.

[0102] The system software automatically analyzes the amplification curve, determines the test results, and generates a report. Subsequently, the consumable operation actuator and the rotary pipette of the consumable are released, and the consumable connection unit 210 of the consumable operation actuator rises. The clamping assembly 300 is released from clamping and rises. The lifting drive mechanism of the carrier assembly 400 drives the consumable placement carrier 403 to rise. The horizontal electric telescopic mechanism 402 of the carrier drives it to extend to the outside of the equipment for the operator to remove the used consumables.

[0103] The above-described embodiments are preferred embodiments of the present invention and are only used to facilitate the illustration of the present invention. They are not intended to limit the present invention in any way. Any person skilled in the art who makes local modifications or alterations to the technical content disclosed in the present invention without departing from the scope of the technical features of the present invention shall still fall within the scope of the technical features of the present invention.

Claims

1. A fully automated nucleic acid detection and analysis system, characterized in that, include: Consumable operation actuator, which is connected to the rotary pipette of consumables and can drive the rotary pipette to rotate and lift, as well as provide power for the rotary pipette to aspirate and dispense liquid; Carrier assembly, used to support and position consumables; Functional compartment processing component, which is used to perform heating, magnetic separation and mixing operations on the magnetic bead pyrolysis liquid compartment of consumables; The reaction tube temperature control assembly is used to adjust the temperature of the reaction tubes used for amplification reactions; Optical components used for fluorescence optical detection of solutions in reaction tubes of consumables.

2. The fully automated nucleic acid detection and analysis system according to claim 1, characterized in that, The consumables operation actuator includes: The consumable connection unit includes: a gripper, a fixing block, a connecting block, and a bottom pneumatic connector; both the gripper and the bottom pneumatic connector are located at the lower end of the fixing block. The gripper is used to hold and position the consumable, and the bottom pneumatic connector is used to connect to the fluid interface of the consumable; the top of the fixing block is fixed to the bottom of the connecting block. A rotating unit is used to drive the consumable connection unit to rotate; the connecting block is connected to the rotating unit. The lifting unit is used to drive the consumable connection unit, the rotating unit, and the gas suction unit to perform lifting movements. The gas suction unit is used to provide negative / positive pressure to suction and dispense liquids from consumables via a bottom pneumatic connector.

3. The fully automated nucleic acid detection and analysis system according to claim 2, characterized in that, The lifting unit includes: a lifting drive motor and a lifting platform; the lifting drive motor is capable of driving the lifting platform to move up and down. The rotating unit includes: a rotary drive motor, a drive wheel, a belt, a driven wheel, a consumable connection unit rotating shaft, and a bearing housing; the housing of the rotary drive motor is fixed to the lifting platform; the output shaft of the rotary drive motor is connected to the drive wheel; the drive wheel drives the driven wheel through the belt; the bearing housing is fixedly disposed on the lower surface of the lifting platform; the consumable connection unit rotating shaft is rotatably disposed in the bearing housing, with its upper end connected to the driven wheel and its lower end fixedly connected to the consumable connection unit. The gas suction unit includes a plunger pump, a front pneumatic pipe, a quick-connect air pipe, a pneumatic connector clamp, a rotary connector, a pneumatic connecting pipe, and a lower pneumatic connecting pipe connected in sequence; the plunger pump is fixed on the upper surface of the lifting platform; the center of the rotating shaft of the consumable connection unit is provided with a hollow part, which serves as a pneumatic connecting pipe. The pneumatic coupling clamp is fixed to the lifting platform, and the lower pneumatic connection joint is connected to the bottom pneumatic joint through a pipeline.

4. The fully automated nucleic acid detection and analysis system according to claim 1, characterized in that, Also includes: A clamping assembly for clamping the consumable reaction tube onto the reaction tube temperature control assembly, comprising: Lifting and pressing motor; The lifting plate moves up and down driven by the lifting and pressing motor. A clamping plate is located below the lifting plate, and its lower surface can contact the outside of the consumable cover. Multiple springs are arranged between the lifting plate and the pressing plate.

5. The fully automated nucleic acid detection and analysis system according to claim 1, characterized in that, The carrier assembly includes: Carrier platform; Consumables placement rack, which is used to hold consumables; The horizontal drive mechanism has its fixed end fixed to the carrier platform and its moving end connected to the consumables placement carrier. The lifting drive mechanism can drive the carrier platform to move up and down so that the consumables placed on the carrier can come into contact with or separate from the functional compartment processing components.

6. The fully automated nucleic acid detection and analysis system according to claim 1, characterized in that, The functional warehouse processing components include: The heat-insulating support is provided with a bearing position corresponding to the magnetic bead pyrolysis liquid chamber and the reaction tube of the consumables, and an optical fiber hole assembly is provided below the bearing position corresponding to the reaction tube. An ultrasonic unit is installed at a position corresponding to the bearing position of the magnetic bead pyrolysis liquid chamber of the consumable, and is used to mix the solution in the magnetic bead pyrolysis liquid chamber. A heating unit is located at a position corresponding to the bearing position of the magnetic bead pyrolysis liquid chamber of the consumable, and is used to heat the solution in the magnetic bead pyrolysis liquid chamber; A magnet unit, located on the side of the heat-insulating support, has a movable magnet for adsorbing or releasing magnetic beads in the magnetic bead pyrolysis liquid chamber.

7. The fully automated nucleic acid detection and analysis system according to claim 6, characterized in that, The heating unit includes: a sheet metal support for the heating block, a heating block, a heating film, and a retaining spring; the sheet metal support for the heating block is fixedly connected to the heat insulation support, and the heating block is connected to the sheet metal support for the heating block via the retaining spring; the heating film is provided on the back of the heating block; The ultrasonic unit includes: an ultrasonic mounting base, an ultrasonic device, and a connecting spring; the ultrasonic device is connected to the ultrasonic mounting base via the connecting spring. The magnet unit includes: a magnet motor mounting base, a magnet rotation drive motor, a magnet fixing base, and a magnet; the housing of the magnet rotation drive motor is fixed on the magnet motor mounting base; the magnet rotation drive motor is used to drive the magnet fixing base to rotate; a magnet is installed and fixed at the end of the magnet fixing base.

8. The fully automated nucleic acid detection and analysis system according to claim 7, characterized in that, The reaction tube temperature control assembly includes: a heating / cooling element, a heat sink, and a fan; a heating / cooling element is installed below each fiber optic port assembly. A radiator is installed below the heat-insulating support, and a fan is installed on the side of the radiator; The heating and cooling elements are connected to the fiber optic hole assembly and the heat sink, respectively.

9. The fully automated nucleic acid detection and analysis system according to claim 1, characterized in that, Also includes: The rack assembly includes consumables such as a rotary pipette and a loading disk. The loading disk has multiple functional compartments: a waste liquid compartment, a magnetic bead lysis buffer compartment, a reagent tube, a reaction tube, and a washing liquid compartment. When the rotary pipette is rotated, the biased pipette at the end of the rotary pipette can sequentially reach and access each functional compartment on the loading disk and can transfer the solution in any functional compartment to other functional compartments.

10. A fully automated nucleic acid detection and analysis method, wherein the method is a non-disease diagnostic method, characterized in that, The detection and analysis are performed using the fully automated nucleic acid detection and analysis system as described in claim 1. Includes the following steps: S1: Consumable loading and initial positioning: Place the consumables onto the carrier assembly, and then the load assembly carries the consumables to the top of the functional compartment processing assembly; S2: Consumables are placed in the functional compartment processing component, and the reaction tube of the consumables is in close contact with the reaction tube temperature control component; S3: The consumables operation actuator is connected to the rotary pipette of the consumables; S4, Nucleic acid extraction and purification steps; S5: Preparation of the amplification reaction system; S6: Nucleic acid amplification and real-time fluorescence detection; S7: Unload consumables.

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