Nucleic acid extraction and conversion integrated machine and use method therefor

Abstract

A nucleic acid extraction and conversion integrated machine and a method therefor. The machine comprises: a machine frame, a sample tube rack, a TIP rack, a centrifuge tube rack, a dual-centrifuge module, a multi-lid opening module, a dual-mechanical-arm module, a stepwise nucleic acid extraction module, an inverted mixing device, and a display and control module. Corresponding nucleic acid extraction processes can be edited on the basis of different samples, and the machine is applicable to the detection of most samples, thereby saving on an occupied space. Centrifuge tubes can be accurately picked up according to numbers, and a basket of a centrifuge stops at a specified position after a zero trigger triggers a photoelectric switch, so that mechanical arms can accurately grab or put the centrifuge tubes in the basket. During a centrifugal operation, the basket is in an inclined state similar to that of a fixed-angle rotor, so that the working area on a plane is reduced while a centrifugal effect is guaranteed. The basket in the design can have a vertical state under any working condition when the centrifuge stops, so that the mechanical arms can vertically and accurately grab the centrifuge tubes.

Description

A nucleic acid extraction and conversion integrated machine and its use method Technical Field

[0001] The present invention relates to the technical field of nucleic acid extraction, and in particular to an all-in-one nucleic acid extraction and conversion machine and a method for using the same. Background Art

[0002] At present, the operational procedures for sample nucleic acid extraction and library construction are complex and have a low degree of automation. Experimenters often need to perform repetitive operations for a long time, resulting in low work efficiency. At the same time, the experimental methods for each specification are different, the sample pre-processing process is cumbersome, and multiple independent experimental equipment takes up a large space, which may occupy a single laboratory. In addition, there are many nucleic acid testing projects, including respiratory specimen testing, blood testing, and fecal testing. The testing processes for different testing projects are different, and experimenters need to classify the tests according to different processes, which requires a high level of professionalism. Therefore, it is necessary to design an automatic nucleic acid detection all-in-one machine.

[0003] The document with patent number CN202010424015.6 discloses a dual-extraction mode nucleic acid extraction workstation and a nucleic acid extraction method. The dual-extraction mode nucleic acid extraction workstation includes a workbench provided with a liquid storage tank and a plate position. The liquid storage tank is used to place the reagents required for DNA extraction, and the plate position is used to place a lysis plate, a binding plate, a washing plate, and an elution plate; a centrifugal device for separating different products in the sample lysed in the lysis plate under centrifugal action; a magnetic rod device, provided on the workbench, for adsorbing magnetic beads used in DNA extraction; a robotic arm device, provided on the workbench, capable of adding the reagents in the liquid storage tank to the lysis plate, and capable of shifting the lysis plate, the binding plate, or the washing plate. The present invention can reduce the difficulty of nucleic acid extraction and ensure smooth nucleic acid extraction.

[0004] However, the above technical solution is still a single-threaded operation and cannot select the process according to the type of nucleic acid sample. Moreover, the overall equipment is relatively discrete and occupies a large space, which is not conducive to the miniaturization of the equipment. Summary of the Invention

[0005] The present invention aims to solve at least one of the above-mentioned technical problems in the prior art to a certain extent, and the present invention adopts the following technical solution:

[0006] A nucleic acid extraction and conversion integrated machine, comprising a whole machine frame and a lifting door,

[0007] A workbench and a filtration system are provided inside the entire machine frame. The filtration system is arranged above the workbench and is used to absorb and process the reaction gas during the nucleic acid extraction process.

[0008] The whole machine frame is further provided with a dual-manipulator module, the range of movement of which covers the workbench, and the dual-manipulator module includes: a first manipulator arm and a second manipulator arm;

[0009] A display control module is provided on the outside of the whole machine frame for editing the nucleic acid extraction process. The display control module is connected to control the dual-manipulator module to transfer the sample tubes so that the sample tubes complete the nucleic acid sample processing on the workbench according to the editing steps;

[0010] The working platform includes a sample placement area, a cover opening and liquid adding area, and a nucleic acid separation and extraction area;

[0011] The sample placement area is equipped with a sample tube rack, an inversion mixing device and a centrifuge tube rack, and the lid opening and liquid adding area is equipped with a variety of lid opening modules;

[0012] The nucleic acid separation and extraction area is equipped with a TIP gun head rack, a step-by-step nucleic acid extraction module, a dual centrifuge module, a shaking device and a temperature incubation module, wherein the dual centrifuge modules are both equipped with an automatic vertical positioning device.

[0013] Preferably, the dual centrifugal module includes a low-speed centrifugal device and a high-speed centrifugal device, the automatic vertical positioning device includes a centrifugal motor and a centrifugal platform, the centrifugal platform is provided with a high-speed centrifugal frame and a hanging basket, both ends of the rotating shaft of the centrifugal motor extend out of the centrifugal motor, the upper end of the rotating shaft is connected to the high-speed centrifugal frame, and the lower end of the rotating shaft is provided with a zero position control device;

[0014] The zero position control device is fixed on the centrifuge positioning plate. The zero position control device includes a clutch, a zero position monitoring device and a positioning motor. One end of the clutch is fixedly connected to the lower end of the rotating shaft, and the other end of the clutch is fixedly connected to the positioning motor. The zero position monitoring device includes a zero position trigger and a zero position detection component. The zero position trigger is fixed on the clutch. The zero position monitoring device and the positioning motor are synchronously controlled.

[0015] Preferably, a shock-absorbing support device is provided between the centrifugal platform and the centrifuge positioning plate, the shock-absorbing support device includes a shock-absorbing plate and a plurality of support frames, the centrifugal motor is fixed on the bottom surface of the shock-absorbing plate, and a plurality of shock absorbers are also provided on the shock-absorbing plate, the shock-absorbing plate is rectangular, and the shock absorber is a rubber shock absorber, the height of the rubber shock absorber is greater than the shock-absorbing plate, and the upper surface of the rubber shock absorber is against the bottom surface of the centrifugal platform.

[0016] Preferably, the clutch is an electromagnetic clutch, which includes an electromagnetic generating end and a clutch suction cup end. The positioning motor is connected to the electromagnetic generating end, and the lower end of the rotating shaft is connected to the clutch suction cup end.

[0017] Preferably, the zero-position trigger component includes a fixed ring and a trigger piece, the fixed ring is sleeved on the lower end of the rotating shaft, and the zero-position detection component is a photoelectric switch. When the trigger piece rotates past the photoelectric switch, the photoelectric switch controls the positioning motor to rotate at a fixed speed.

[0018] Preferably, the high-speed centrifuge frame is provided with a plurality of hanging baskets, and the high-speed centrifuge frame is provided with a plurality of receiving slots for storing the hanging baskets; in each receiving slot, a swing shaft is provided on the two side walls of the receiving slot, the hanging basket and the swing shaft rotate in cooperation, the hanging basket includes an angle adjustment part and a vertical positioning part, the end of the high-speed centrifuge frame close to the angle adjustment part is provided with a limit plate, and the weight of the vertical positioning part is greater than the weight of the angle adjustment part.

[0019] Preferably, the limiting plate is provided with a vertical surface and an inclined surface. In a stationary state, the bottom of the angle adjustment portion abuts against the vertical surface; during high-speed rotation separation, the upper portion of the angle adjustment portion abuts against the inclined surface.

[0020] Preferably, the inclined surface and the horizontal plane form an angle of 30-45 degrees, the limiting plate is further provided with a limiting block, the angle adjustment portion is provided with a corresponding limiting groove, and the vertical positioning portion is provided with a counterweight plate, and the weight of the counterweight plate is greater than the weight of two centrifuge test tubes.

[0021] The method of using the nucleic acid extraction and conversion all-in-one machine comprises the following steps:

[0022] S1 Sample loading: Open the lift door, install the sample rack carrying multiple sample tubes onto the sample frame, close the lift door, enter the sample type and sample quantity in the display control module, and start the nucleic acid extraction process;

[0023] S2 Sample Addition: The first robotic arm grabs and moves the sample tube to the multi-capping module. Depending on the sample type, it chooses whether to open the bottle cap and add the first reagent;

[0024] S3 shaking low-speed centrifugation: The second robotic arm moves the sample tube to the shaking device for shaking to mix the sample tube, and then the second robotic arm moves the sample tube to the low-speed centrifugation device for centrifugation;

[0025] S4 transfers the centrifuged liquid: After the centrifugation of the sample tube is completed, the second robotic arm transfers the sample tube to the multi-capping module and opens the cap of the sample tube. The second robotic arm transfers the supernatant of the sample tube to the first centrifuge tube through the TIP gun head;

[0026] S5: Adding reaction reagents: The multi-capping module adds the second reagent to the first centrifuge tube and closes the cap of the sample tube;

[0027] S6 inversion mixing and high-speed centrifugation: The first robotic arm moves the first centrifuge tube to the inversion mixing device. After sufficient mixing, the first robotic arm moves the first centrifuge tube to the multi-capping module, and the second robotic arm moves the first centrifuge tube to the high-speed centrifugation device for high-speed centrifugation;

[0028] S7: Transferring the centrifugal liquid: The second robotic arm moves the first centrifuge tube to the second lid opening module and opens the lid of the first centrifuge tube. The upper pipetting portion of the second robotic arm extracts the supernatant of the first centrifuge tube through the TIP gun head. The first robotic arm clamps the second centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping portion. The second rotating clamping portion opens the lid of the second centrifuge tube. The upper pipetting portion of the second robotic arm transfers the supernatant of the first centrifuge tube to the second centrifuge tube through the TIP gun head.

[0029] S8: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0030] S9: When the incubation timer in one of the centrifuge tubes is completed, the extraction step is performed: the second robotic arm transfers the second centrifuge tube to the step-by-step nucleic acid extraction module for nucleic acid extraction.

[0031] Preferably, according to the detection process of different types of cancer, the display control module deletes or repeats at least one of steps S2, S3, S4, S5 or S6 multiple times; or, according to the detection process of different types of cancer, the execution order of the above S2, S3, S4, S5, S6 is readjusted.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] 1. All instruments used for nucleic acid extraction are integrated into a single machine framework. This means that the corresponding nucleic acid extraction process can be edited according to different samples. This makes it suitable for most sample tests, including respiratory specimen testing, blood testing, and stool testing. Compared with existing single-process products, the entire device integrates multiple testing methods into one machine, reducing equipment costs and saving space. Within the same space, this nucleic acid extraction and conversion all-in-one machine can accommodate more devices and conduct more tests.

[0034] 2. It can accurately pick up centrifuge test tubes according to their numbers. The dual centrifuge modules are equipped with an electromagnetic clutch, a positioning motor, and a zero-position monitoring device. After the centrifuge motor stops, the electromagnetic clutch works to connect the positioning motor and the centrifuge motor. The positioning motor drives the rotating shaft of the centrifuge motor to rotate. After the zero-position trigger triggers the photoelectric switch, it serves as the centrifuge positioning zero point. Then, it rotates a certain angle to ensure that the centrifuge basket stops after reaching the specified position. At this time, the robotic arm can accurately grab or place the centrifuge tube in the basket.

[0035] 3. When the centrifugal motor is working, the hanging basket is in a tilted state of 30 degrees. The tilt angle is limited by the structure of the limit plate. The upper part of the angle adjustment part of the hanging basket is against the inclined surface to limit the hanging basket from swinging. During centrifugal operation, the hanging basket is in a tilted state similar to that of a fixed-angle rotor, which reduces the working area on the plane while ensuring the centrifugal effect.

[0036] 4. Since the vertical positioning part of the hanging basket is provided with a counterweight plate and a weighted design is carried out, the bottom of the hanging basket angle adjustment part is against the vertical surface of the limit plate, so that the hanging basket can reliably restore the vertical state when the centrifuge stops. Therefore, the hanging basket of this design can achieve a vertical state under any working conditions when the centrifuge stops, so that the manipulator can accurately grab the centrifuge test tube vertically. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0038] FIG1 is a schematic diagram of a comprehensive process of the method of the present invention

[0039] FIG2 is a schematic diagram of the overall structure of an embodiment of the present invention.

[0040] FIG3 is a schematic diagram of a fully open lift door according to an embodiment of the present invention;

[0041] FIG4 is a diagram of the internal structure layout of an embodiment of the present invention;

[0042] FIG5 is a perspective view of the internal structure of an embodiment of the present invention.

[0043] FIG6 is a schematic diagram of the overall structure of a multiple cover opening module according to an embodiment of the present invention.

[0044] FIG7 is a schematic diagram of the dual-manipulator module structure according to an embodiment of the present invention.

[0045] FIG8 is a schematic structural diagram of a low-speed centrifugal device according to an embodiment of the present invention.

[0046] FIG9 is a schematic diagram of the overall structure of an automatic vertical positioning device according to an embodiment of the present invention.

[0047] FIG10 is a schematic structural diagram of the centrifugal platform according to an embodiment of the present invention.

[0048] FIG. 11 is a schematic diagram of a partially disassembled structure of a high-speed centrifuge frame according to an embodiment of the present invention.

[0049] FIG12 is an enlarged view of the structure of section A in FIG11 of the present invention.

[0050] FIG13 is a schematic diagram of the static and moving states of a hanging basket according to an embodiment of the present invention.

[0051] FIG14 is a schematic structural diagram of a hanging basket according to an embodiment of the present invention.

[0052] FIG15 is a schematic bottom view of a hanging basket according to an embodiment of the present invention.

[0053] FIG16 is a schematic structural diagram of a shock-absorbing support device according to an embodiment of the present invention.

[0054] FIG17 is a schematic diagram showing the principle of a zero-position monitoring device according to an embodiment of the present invention.

[0055] FIG18 is a cross-sectional view of the internal power structure of the automatic vertical positioning device according to an embodiment of the present invention.

[0056] FIG19 is a schematic structural diagram of a zero-position triggering member according to an embodiment of the present invention.

[0057] In the figure: the whole machine frame 1, the lifting door 101, the sample placement area 100, the cover opening and liquid adding area 200, the nucleic acid separation and extraction area 300, the display control module 102, the filtration system 103, the sample tube rack 11, the temperature incubation module 12, the liquid adding device 13, the centrifuge tube rack 14, the dual centrifuge module 2, the low-speed centrifuge device 21, the high-speed centrifuge device 22, the automatic vertical positioning device 3, the high-speed centrifuge rack 31, the receiving tank 310, the swing shaft 3101, the limit plate 311, the vertical surface 3111, the inclined surface 3112, the limit block 312, the limit slot 3120, the connecting shaft 32, the hanging basket 33, the fixed slot 300, the angle adjustment part 331, the centrifuge tube slot 3310, the vertical positioning part 332, the counterweight plate 3321, centrifugal platform 34, centrifugal motor 35, electromagnetic clutch 6, electromagnetic generating end 361, clutch suction cup end 362, centrifuge positioning plate 37, positioning motor 371, zero position trigger 38, fixing ring 381, trigger plate 382, ​​photoelectric switch 383, shock-absorbing support device 39, shock-absorbing plate 391, support frame 392, rubber shock absorber 393, multiple cover opening module 4, first cover opening assembly 41, second cover opening assembly 42, TIP gun head rack 5, centrifuge tube rack 14, dual robotic arm module 7, first robotic arm 71, second robotic arm 72, step-by-step nucleic acid extraction module 8, first nucleic acid extraction device 81, second nucleic acid extraction device 82, inversion mixing device 91, and oscillation device 92. DETAILED DESCRIPTION

[0058] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0059] Example: As shown in Figures 1 to 19:

[0060] As shown in Figures 2 to 5: the nucleic acid extraction and conversion integrated machine includes: a whole machine frame 1 and a lifting door 101. A workbench 10 and a filtration system 103 are provided inside the whole machine frame 1. The filtration system 103 includes a UV lamp, an activated carbon filter and a directional ventilation device arranged on the top of the shell.

[0061] The filtration system 103 is disposed above the workbench 10 and is used to absorb and process the reaction gas during the nucleic acid extraction process;

[0062] The whole machine frame is further provided with a dual-manipulator module 7, the movable range of the dual-manipulator module 7 covers the workbench, and the dual-manipulator module 7 is located below the filtering system 103. The dual-manipulator module 7 includes: a first manipulator 71 and a second manipulator 71;

[0063] The working platform includes a sample placement area 100, a cover opening and liquid adding area 200, and a nucleic acid separation and extraction area 300;

[0064] As shown in FIG. 2 to FIG. 5 , the sample placement area is provided with a sample tube rack 11 , an inversion mixing device 91 and a centrifuge tube rack 14 .

[0065] The sample tube rack 11 is arranged on the workbench and is used to place various sample tubes. The sample tubes can be replaced in batches. There are 16 sample tube slots on the sample tube rack 11. During the experiment, 16 samples can be placed in the device of the present invention at one time. After the samples enter, a nucleic acid template can be obtained after a period of time, realizing full process automation, which is beneficial to reducing the labor intensity of the experimenters and improving the stability and consistency of the experimental results.

[0066] The inversion mixing device is used to fully mix the centrifuge tubes added with reagents, and the centrifuge tube rack 14 stores empty centrifuge tubes for the dual-manipulator module 7 to pick up.

[0067] The lid opening and liquid adding area 200 is provided with a multi-lid opening module 4, which is used to open and close the lids of various test tubes, including 2ml centrifuge test tubes and 50ml stool sample test tubes.

[0068] The nucleic acid separation and extraction area 300 is provided with a TIP gun head rack 5, a step-by-step nucleic acid extraction module 8, a dual centrifugation module 2, a shaking device 92 and a temperature incubation module 12. The temperature incubation module 12 is used to provide a suitable temperature for nucleic acid sequence amplification to increase the reaction rate.

[0069] The shaking device 92 is used to shake the centrifugal test tube to disperse the substances in the test tube.

[0070] The TIP gun tip rack 4 is used to place the TIP gun tip, which can temporarily store intermediate liquid for liquid transfer.

[0071] The dual centrifuge module 2 is used to centrifuge the test tubes. Depending on the size of the test tubes and the reaction requirements, a low-speed centrifuge device 21 or a high-speed centrifuge device 22 is used.

[0072] As shown in FIG7 , a dual-manipulator module 7 is provided on the workbench and is used to transfer test tubes and transfer solutions through the TIP gun head;

[0073] The step-by-step nucleic acid extraction module 8 includes a first nucleic acid extraction device 81 and a second nucleic acid extraction device 82 . The step-by-step nucleic acid extraction module is disposed on the workbench and is used to extract nucleic acid products from the sample.

[0074] The overall size of the nucleic acid extraction and conversion machine is 1800mm long * 800mm wide * 1100mm high, and the weight is 340KG. A lifting door 101 is provided on the whole machine frame 1.

[0075] A display control module 102 is provided on the outside of the entire machine frame. The display control module 102 is a 15.6-inch touch-sensitive integrated machine with a processor inside to control the operation process of all modules and record intermediate test data. The display control module 102 is connected to control the dual robotic arm module 7 to transfer sample tubes, so that the sample tubes complete nucleic acid sample processing on the workbench 10 according to the editing steps.

[0076] As shown in Figure 6, in one embodiment of the present invention, the multi-opening module 4 includes a first opening component 41 and a second opening component 42; the first opening component 41 and the second opening component 42 are used to open sample tubes and centrifuge tubes, respectively. The test tubes include 2ml centrifuge tubes and 50ml stool sample tubes.

[0077] As shown in Figures 3 and 7, in one embodiment of the present invention, the dual-arm module 7 includes a first robotic arm 71 and a second robotic arm 72. The two robotic arms can independently complete different steps, thereby improving experimental efficiency.

[0078] In one embodiment of the present invention, the nucleic acid extraction and conversion integrated machine further comprises:

[0079] A liquid adding device 13 is provided on the workbench and is used to store reaction additive reagents; in one embodiment of the present invention, the liquid adding device 13 provides liquid adding power for the multiple cover opening module 4;

[0080] In one embodiment of the present invention, the dual centrifugal module 2 is located in the nucleic acid separation and extraction area 300, and includes a low-speed centrifugal device 21 and a high-speed centrifugal device 22. The centrifugal speed of the low-speed centrifugal device 21 can be 8000 rpm or less, and is used for centrifugal separation of sample test tubes. The centrifugal speed of the high-speed centrifugal device 22 can be 15000 rpm-30000 rpm, and the high-speed centrifugal device is used for centrifugal stratification of centrifugal test tubes.

[0081] As shown in FIG9-19 : As a further improvement of the present invention, an automatic vertical positioning device 3 is provided in each of the dual centrifugal modules 2 .

[0082] As shown in Figure 9: the automatic vertical positioning device 3 includes a centrifugal motor 35 and a centrifugal platform 34, and a high-speed centrifugal frame 31 and a hanging basket 33 are provided on the centrifugal platform 34. The upper end of the connecting shaft 32 is connected to the high-speed centrifugal frame 31, and the high-speed centrifugal frame 31 is driven by the connecting shaft 32 of the centrifugal motor 35 to rotate, thereby driving the hanging basket 33 and the centrifugal test tubes in the hanging basket to rotate. This device can already meet the traditional manual operation process, but in a fully automated integrated nucleic acid extraction machine, it is necessary to meet the requirements of automatic positioning. In order to be able to accurately place the test tubes, and to make each centrifugal test tube numbered in a fixed order, complete centrifugal separation in sequence, and then grab them in sequence, it is necessary to add this control system.

[0083] The hanging baskets 33 can be set to three, four or five. The lower end of the connecting shaft 32 is driven to rotate by a motor. The upper end of the connecting shaft 32 is clamped to the high-speed centrifugal frame 31 and the position is limited by a top cover.

[0084] As shown in Figure 10, the high-speed centrifuge frame 31 is provided with four receiving slots 310 for storing the hanging baskets 33. Connecting devices are provided on both sides of the receiving slots 310. The hanging baskets 33 and the receiving slots 310 need to be snap-fitted and rotatably matched. Therefore, in each receiving slot 310, a swing shaft 3101 is provided on the two side walls of the receiving slot 310. Fixed slots 300 are provided on both sides of the hanging basket 33. The two swing shafts 3101 are respectively snap-fitted into the corresponding fixed slots 300. Centrifuge tube slots 3310 are provided on the four corners of each hanging basket 33.

[0085] As shown in Figure 11, the hanging basket 33 and the swing shaft 3101 rotate together. The hanging basket 33 includes an angle adjustment part 331 and a vertical positioning part 332. The traditional hanging basket is designed to be symmetrical on both sides. When there are only 1, 2, or 3 centrifuge test tubes in the hanging basket or the weights of the four test tubes are different, the hanging basket will tilt. The hanging basket 33 can only be kept horizontally when four centrifuge test tubes of equal weight are extracted or placed at one time. However, the robotic arm of the nucleic acid all-in-one machine is designed to handle a single test tube at a time, so the hanging basket is very easy to tilt, which is very unfavorable for the automatic operation of the nucleic acid all-in-one machine, and the gripper of the robotic arm cannot accurately grip the test tube.

[0086] In order to enable the hanging basket 33 to always maintain a vertical state, the weight of the vertical positioning portion 332 needs to be greater than the weight of the angle adjustment portion 331, so that the hanging basket 33 can be maintained in a vertical state at any time.

[0087] As a further improvement of this embodiment, a counterweight plate 3321 is provided in the vertical positioning portion 332, and the weight of the counterweight plate 3321 is greater than the weight of two centrifuge test tubes. Since the vertical positioning portion 332 of the hanging basket 33 is provided with the counterweight plate 3321, a weighted design is implemented, and the bottom of the angle adjustment portion 331 of the hanging basket 33 rests on the vertical surface of the limit plate 311.

[0088] The hanging basket can reliably restore to a vertical state when the centrifuge stops. Therefore, the hanging basket of this design can achieve a vertical state under any working conditions when the centrifuge stops.

[0089] At the same time, in order to ensure that the volume of the entire centrifuge is reduced when rotating at high speed, as an embodiment of the present invention, a limit plate 311 is provided at the end of the high-speed centrifuge frame 31 close to the angle adjustment portion 331, so that the rotation angle of the hanging basket can be limited.

[0090] As a further optimization of this embodiment, the counterweight plate 3321, the angle adjustment portion 331 and the vertical positioning portion 332 are an integrally formed structure.

[0091] During the startup process, first, the bottom of the angle adjustment part 331 of the hanging basket 33 rests on the vertical surface of the limit plate 311. Under the action of centrifugal force, the hanging basket 33 and the swing shaft 3101 rotate. Due to the heavy weight of the liquid inside the test tube, the bottom of the hanging basket will form centrifugal force and be thrown out. If there is no restriction, the hanging basket will reach a horizontal state, and the centrifuge must maintain a certain distance from other equipment. However, this will increase the safety distance of the entire centrifuge and occupy a larger space, which is not conducive to the miniaturization of the nucleic acid integrated machine.

[0092] The limiting plate 311 is provided with a vertical surface 3111 and an inclined surface 3112. In a stationary state, the bottom of the angle adjustment portion 331 abuts against the vertical surface 3111; during high-speed rotation separation, the upper portion of the angle adjustment portion 331 abuts against the inclined surface 3112.

[0093] The inclined surface 3112 and the horizontal surface may be designed to form an angle of 30-45 degrees, and in this embodiment, the angle is 30 degrees.

[0094] As shown in FIG. 12 , a limiting block 312 is further provided on the limiting plate 311 , and a corresponding limiting groove 3120 is provided on the angle adjustment portion 331 .

[0095] When the centrifuge is working, the basket is in a tilted state of 30 degrees. The tilt angle is limited by the structure of the limit plate. The upper part of the angle adjustment part of the basket rests on the inclined surface to limit the basket from swinging further. During centrifugal operation, the basket is in a tilted state similar to that of a fixed-angle rotor, which reduces the working area on the plane while ensuring the centrifugal effect.

[0096] Since the vertical positioning part of the hanging basket is provided with a counterweight plate and a weighted design is adopted, the hanging basket can reliably restore to a vertical state when the centrifuge stops. In addition, when the robot arm places or grabs the centrifuge tubes from the four holes of the hanging basket, if the two centrifuge tubes on the outside of the hanging basket are grabbed first, the two centrifuge tubes on the inside of the hanging basket will generate a counterclockwise deflection torque with the center of the swing axis, causing the hanging basket to deflect toward the outside of the centrifuge rack and not be vertical. Similarly, in the process of placing the centrifuge tubes, if the two centrifuge tubes on the inside of the hanging basket are placed first, the same effect will be produced. Therefore, the hanging basket of this design can achieve a vertical state under any working conditions when the centrifuge stops.

[0097] As shown in Figures 13-19: As a further improvement to this embodiment, the connecting shaft 32 of the centrifugal motor 35 is elongated and extends from the bottom end of the centrifugal motor 35. The upper end of the connecting shaft 32 is connected to the high-speed centrifuge frame 31, and the lower end of the connecting shaft 32 is provided with a zero-position control device. The zero-position control device is used to control the high-speed centrifuge frame 31 to ensure that the centrifugal test tube is always in the same position on the centrifuge platform 34 before and after the centrifugal operation.

[0098] The zero position control device is fixed on the centrifuge positioning plate 37. The zero position control device includes a clutch, a zero position monitoring device and a positioning motor 371. One end of the clutch is fixedly connected to the lower end of the connecting shaft 32, and the other end of the clutch is fixedly connected to the positioning motor 371.

[0099] The positioning motor 371 can be a stepper motor, a servo motor or other motor with a circular positioning function, which can accurately control the rotation angle according to the electronic control signal to improve the position accuracy of the high-speed centrifugal frame 31.

[0100] As shown in Figure 2-3: A shock-absorbing support device 39 is provided between the centrifugal platform 34 and the centrifuge positioning plate 37. The shock-absorbing support device 39 includes a shock-absorbing plate 391 and four support frames 392. The lower end of each support frame 392 is fixedly connected to the centrifuge positioning plate 37, and the upper end of each support frame 392 is fixedly connected to the shock-absorbing plate 391.

[0101] The centrifugal motor 35 is fixed to the bottom surface of the damping plate 391. The damping plate 391 may be provided with multiple dampers. These are arranged at equal intervals and angles on the upper surface of the damping plate 391 to reduce vibration of the centrifugal motor 35 and the impact of the high torque generated when the clutch is engaged, thereby reducing the impact of the high torque generated when the positioning motor 371 and the centrifugal motor 35 are engaged.

[0102] The shock absorbing plate 391 is rectangular, and shock absorbers are provided on the four corners of the shock absorbing plate 391 , and the shock absorbers are rubber shock absorbers 393 .

[0103] The height of the rubber shock absorber 393 is greater than that of the shock absorbing plate 391 , and the upper surface of the rubber shock absorber 393 abuts against the bottom surface of the centrifugal platform 34 .

[0104] The clutch is an electromagnetic clutch, and the electromagnetic clutch 6 includes an electromagnetic generating end 361 and a clutch suction cup end 362 . The motor main shaft of the positioning motor 371 is connected to the electromagnetic generating end 361 , and the lower end of the connecting shaft 32 is connected to the clutch suction cup end 362 .

[0105] The zero position monitoring device includes a zero position triggering member 38 and a zero position detecting member. The zero position triggering member 38 is fixed on the clutch. The zero position monitoring device and the positioning motor 371 are synchronously controlled.

[0106] As shown in FIG5 , the zero-position trigger member 38 includes a fixing ring 381 and a trigger plate 382 . The fixing ring 42 is sleeved on the clutch suction cup end 362 . The fixing ring 42 is provided with a through hole for placing screws for tightening and fixing.

[0107] The zero position detection component is a photoelectric switch 42, and the trigger plate 43 does not contact the photoelectric switch most of the time. When the centrifugal motor 35 stops being powered off, the clutch and the photoelectric switch 42 are started. When the trigger plate 43 rotates past the photoelectric switch 42, the photoelectric switch 42 controls the positioning motor 371 to rotate at a fixed speed.

[0108] The connecting shaft 32 of the centrifugal motor 35 is fixedly connected to the suction cup end 31 of the electromagnetic clutch, and the electromagnetic generating end 32 of the electromagnetic clutch is connected to the motor main shaft of the positioning motor 371. A zero-position trigger 38 is also installed on the connecting shaft 32 of the centrifugal motor 35, and a zero-position detection component is fixedly installed next to it; after the centrifugal motor 35 stops, the electromagnetic clutch works, and the positioning motor 371 and the centrifugal motor 35 are rotated synchronously and connected, and the positioning motor 371 drives the rotating shaft of the centrifuge motor 1 to rotate. After the zero-position trigger 38 triggers the photoelectric switch 42, it serves as the centrifuge positioning zero point, and then the positioning motor drives the high-speed centrifuge frame 31 to rotate a certain angle to ensure that the hanging basket 22 of the centrifuge stops after reaching the specified position. At this time, the robot arm can accurately grab or put the centrifuge tube in the hanging basket 22;

[0109] A shock-absorbing support device 39 is provided to reduce the vibration of the centrifugal motor 35 and the influence of the high torque generated at the moment of clutch operation, when the positioning motor 371 and the centrifugal motor 3 are combined.

[0110] Specific embodiment 1: When the sample to be detected is a stool sample, the method comprises the following steps:

[0111] S1 Sample loading: Open the lift door 101, install the sample tubes carrying multiple stool samples on the sample tube rack 11, close the lift door 101, input the stool sample type and sample quantity 16 in the display control module 10, and start the nucleic acid extraction process;

[0112] S2: Add liquid to sample: The first robotic arm 71 grabs and moves the stool sample tube to the first opening module 41, opens the bottle cap according to the sample type, and adds the first reagent;

[0113] S3: low-speed centrifugation with shaking: the second robotic arm 72 moves the sample tube to the shaking device 92 for shaking to mix the sample tube, and then the second robotic arm 72 moves the sample tube to the low-speed centrifugation device 21 for centrifugation;

[0114] S4: After the centrifugation of the sample tube is completed, the second robotic arm 72 transfers the sample tube to the second opening module 42 and opens the cap of the sample tube. The second robotic arm 72 transfers the supernatant of the sample tube to the first centrifuge tube through the TIP gun head 5.

[0115] S5: Adding reaction reagents: The second opening module 42 adds the second reagent to the first centrifuge tube and then closes the lid of the sample tube;

[0116] S6: Inversion mixing and high-speed centrifugation: The first robotic arm 71 moves the first centrifuge tube to the inversion mixing device 91. After sufficient mixing, the first robotic arm 71 moves the first centrifuge tube to the second cover opening module 42, and the second robotic arm 72 moves the first centrifuge tube to the high-speed centrifuge device 22 for high-speed centrifugation;

[0117] S7: Transferring the centrifugal liquid: The second robotic arm 72 moves the first centrifuge tube to the second cover-opening module 42 and opens the cover of the first centrifuge tube. The upper pipetting portion of the second robotic arm extracts the supernatant of the first centrifuge tube through the TIP gun tip. The first robotic arm 71 grabs the second centrifuge tube on the centrifuge tube rack and moves it to the second cover-opening module 42, opens the cover of the second centrifuge tube, and the upper pipetting portion of the second robotic arm transfers the supernatant of the first centrifuge tube to the second centrifuge tube through the TIP gun tip.

[0118] S8: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module 12;

[0119] S9: When the incubation timer in one of the centrifuge tubes is completed, the extraction step is performed: the second robotic arm transfers the second centrifuge tube to the step-by-step nucleic acid extraction module 8 for nucleic acid extraction.

[0120] According to the detection process of different cancer types, the display control module deletes or repeats at least one of steps S2, S3, S4, S5 or S6 multiple times; or, according to the detection process of different cancer types, readjusts the execution order of the above S2, S3, S4, S5, and S6.

[0121] Specific embodiment 2: When the test sample is a bladder cancer urine sample, steps S2 and S3 are canceled, and steps S4, S5 and S6 are repeated twice.

[0122] Sample loading: A sample holder carrying multiple bladder cancer urine sample tubes is mounted on the sample frame. The bladder cancer urine sample tubes are 2 ml tubes. The clamping portion on the first robotic arm clamps and moves the bladder cancer urine sample tubes to the second opening module 42.

[0123] Adding reaction reagents: The second opening module 42 opens the bladder cancer urine sample bottle cap, the adding needle adds reagents to the bladder cancer urine sample test tube, and the clamping part of the second opening module 42 closes the bladder cancer urine sample test tube cap;

[0124] Inversion mixing and high-speed centrifugation: The first robotic arm 71 moves the bladder cancer urine sample tube to the inversion mixing device. After sufficient mixing, the first robotic arm moves the bladder cancer urine sample tube to the second opening module 42, and the second robotic arm 72 moves the bladder cancer urine sample tube to the high-speed centrifugation device 22 for high-speed centrifugation;

[0125] Transferring the centrifuge liquid: The first robotic arm 71 grabs the first centrifuge tube on the centrifuge tube rack and brings it to the second cover-opening module 42, and opens the cover of the first centrifuge tube. The upper pipetting unit of the second robotic arm transfers the supernatant of the bladder cancer urine sample tube to the first centrifuge tube through the tip of the gun;

[0126] Adding reaction reagents again: the adding needle adds reagents to the first centrifugal test tube, and the second opening module 42 closes the caps of the first centrifugal test tube and the bladder cancer urine sample test tube;

[0127] Inversion mixing and high-speed centrifugation: The first robotic arm 71 moves the first centrifuge tube to the inversion mixing device. After sufficient mixing, the first robotic arm moves the first centrifuge tube to the nucleic acid separation and extraction area, and the second robotic arm moves the centrifuge tube to the high-speed centrifugation device for high-speed centrifugation;

[0128] Transferring the centrifugal liquid: The first robotic arm clamps the second centrifugal test tube on the centrifuge tube rack to the centrifuge test tube clamping part, the second rotating clamping part opens the tube cover of the second centrifugal test tube, and the upper pipetting part of the second robotic arm transfers the supernatant of the first centrifugal test tube to the second centrifuge test tube through the tip of the gun;

[0129] The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0130] When the incubation timer in one of the centrifugal test tubes is completed, the extraction step is performed: the second robotic arm transfers the second centrifugal test tube to the first nucleic acid extraction device or the second nucleic acid extraction device for nucleic acid extraction.

[0131] At the same time, according to the resource requirements of the current task process, the next new parallel step can be started at the same time without affecting the current task step process, maximizing the utilization of the resources of each functional module of the system, and realizing parallel processing of multiple batches of samples, which greatly improves the detection capacity of the system, effectively reduces the labor cost of detection, shortens the sample detection time, and has positive significance for promoting the popularization of early tumor detection.

[0132] The stool sample pretreatment method includes the following steps: a first centrifugation step: After adding a first reagent and a second reagent to a stool sample tube, the stool sample tube is shaken and centrifuged to obtain a first supernatant after centrifugation. The first reagent can be a suspension that lyses cells and releases nucleic acids, and the second reagent can be a suspension that lyses cells and releases nucleic acids.

[0133] In some embodiments of the present invention, in the first centrifugation step, the first supernatant after centrifugation is obtained by repeatedly centrifuging and removing the supernatant, and the number of repetitions may be two to ensure the centrifugal effect in the first centrifugation step. In some embodiments of the present invention, in the second centrifugation step, the second supernatant after centrifugation is obtained by repeatedly centrifuging and removing the supernatant, and the number of repetitions may be two to ensure the centrifugal effect in the second centrifugation step.

[0134] During the pretreatment of stool samples, the nucleic acid product of the stool sample is obtained through the first centrifugation step, the second centrifugation step, the third centrifugation step and the extraction step in sequence, thereby realizing the full process automation of the stool sample pretreatment, which is conducive to reducing the labor intensity of the experimenters and improving the stability and consistency of the experimental results.

[0135] The present invention features a modular design and flexible configuration, allowing for the addition or removal of modules as needed. This provides high compatibility and, by replacing different sample modules and fixtures, compatibility with a variety of sample tubes and centrifuge tubes. The present invention also offers excellent safety and stability, with integrated material placement detection at the workstation, ensuring accurate and precise placement of consumables and proper opening and closing of tube caps. Furthermore, the stool sample pretreatment method according to an embodiment of the present invention allows for accurate and efficient testing, enabling 24-hour uninterrupted operation and completing a batch every 1.5 hours.

[0136] Traditional manual labor takes 6 hours to test 16 samples, while the device of the present invention is easy to operate, and one person can operate and observe 5 devices of the present invention at the same time, which greatly improves the detection speed and is more suitable for large-scale and time-sensitive detection needs.

[0137] The various technical features of the above-described embodiments can be combined arbitrarily, and the execution order of the method steps is not restricted. In order to make the description concise, not all possible combinations of the various technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0138] The above-described embodiments merely represent several implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present application. It should be noted that a person of ordinary skill in the art may make various modifications and improvements without departing from the spirit of the present application, and these modifications and improvements fall within the scope of protection of the present application. Therefore, the scope of protection of the present application shall be determined by the appended claims.

Claims

1. A nucleic acid extraction and conversion machine, characterized in that: Including the whole machine frame and lifting door, A workbench and a filtration system are provided inside the whole machine frame. The filtration system is arranged above the workbench and is used to absorb and process the reaction gas in the nucleic acid extraction process; The whole machine frame is also provided with a dual-mechanical arm module, the activity range of the dual-mechanical arm module covers the workbench, and the dual-mechanical arm module includes: a first mechanical arm and a second mechanical arm; A display control module is provided outside the whole machine frame for editing the nucleic acid extraction process, and the display control module is connected to control the dual-manipulator module to transfer the sample tubes so that the sample tubes complete the nucleic acid sample processing on the workbench according to the editing steps; The working platform includes a sample placement area, a cover opening and liquid adding area, and a nucleic acid separation and extraction area; The sample placement area is provided with a sample tube rack, an inversion mixing device and a centrifuge tube rack, and the lid opening and liquid adding area is provided with a variety of lid opening modules; The nucleic acid separation and extraction area is provided with a TIP gun head rack, a step-by-step nucleic acid extraction module, a dual centrifuge module, a shaking device and a temperature incubation module, wherein the dual centrifuge modules are both provided with an automatic vertical positioning device.

2. A nucleic acid extraction and conversion integrated machine as claimed in claim 1, characterized in that: The dual centrifugal module includes a low-speed centrifugal device and a high-speed centrifugal device, the automatic vertical positioning device includes a centrifugal motor and a centrifugal platform, the centrifugal platform is provided with a high-speed centrifugal frame and a hanging basket, both ends of the rotating shaft of the centrifugal motor extend out of the centrifugal motor, the upper end of the rotating shaft is connected to the high-speed centrifugal frame, and the lower end of the rotating shaft is provided with a zero position control device; The zero position control device is fixed on the centrifuge positioning plate, the zero position control device includes a clutch, a zero position monitoring device and a positioning motor, one end of the clutch is fixedly connected to the lower end of the rotating shaft, the other end of the clutch is fixedly connected to the positioning motor, the zero position monitoring device includes a zero position trigger and a zero position detection member, the zero position trigger is fixed on the clutch, and the zero position monitoring device and the positioning motor are synchronously controlled.

3. A nucleic acid extraction and conversion integrated machine according to claim 2, characterized in that: A shock-absorbing support device is provided between the centrifugal platform and the centrifuge positioning plate, the shock-absorbing support device includes a shock-absorbing plate and a plurality of support frames, the centrifugal motor is fixed on the bottom surface of the shock-absorbing plate, a plurality of shock absorbers are also provided on the shock-absorbing plate, the shock-absorbing plate is rectangular, the shock absorbers are rubber shock absorbers, the height of the rubber shock absorbers is greater than the shock-absorbing plate, and the upper surface of the rubber shock absorbers is against the bottom surface of the centrifugal platform.

4. A nucleic acid extraction and conversion integrated machine according to claim 3, characterized in that: The clutch is an electromagnetic clutch, which includes an electromagnetic generating end and a clutch suction cup end. The positioning motor is connected to the electromagnetic generating end, and the lower end of the rotating shaft is connected to the clutch suction cup end.

5. The nucleic acid extraction and conversion integrated machine according to claim 2, characterized in that: The zero position triggering member includes a fixing ring and a triggering piece. The fixing ring is sleeved on the lower end of the rotating shaft. The zero position detecting member is a photoelectric switch. When the triggering piece rotates past the photoelectric switch, the photoelectric switch controls the positioning motor to rotate at a fixed speed.

6. The nucleic acid extraction and conversion integrated machine according to claim 5, characterized in that: The high-speed centrifuge frame is provided with a plurality of hanging baskets, and the high-speed centrifuge frame is provided with a plurality of receiving slots for storing the hanging baskets; in each receiving slot, a swing shaft is provided on the two side walls of the receiving slot, the hanging basket and the swing shaft are rotatably matched, the hanging basket includes an angle adjustment part and a vertical positioning part, the end of the high-speed centrifuge frame close to the angle adjustment part is provided with a limit plate, and the weight of the vertical positioning part is greater than the weight of the angle adjustment part.

7. The nucleic acid extraction and conversion integrated machine according to claim 6, characterized in that: The limiting plate is provided with a vertical surface and an inclined surface. In a static state, the bottom of the angle adjustment part abuts against the vertical surface; during high-speed rotation separation, the upper part of the angle adjustment part abuts against the inclined surface.

8. The nucleic acid extraction and conversion integrated machine according to claim 7, characterized in that: The inclined surface and the horizontal plane form an angle of 30-45 degrees, the limiting plate is further provided with a limiting block, the angle adjustment portion is provided with a corresponding limiting groove, and the vertical positioning portion is provided with a counterweight plate, the weight of which is greater than the weight of the two centrifuge test tubes.

9. A method for using a nucleic acid extraction and conversion machine, using the nucleic acid extraction and conversion machine according to any one of claims 1 to 8, characterized in that: The method comprises the following steps: S1 Sample loading: Open the lift door, install the sample rack carrying multiple sample tubes onto the sample frame, close the lift door, input the sample type and sample quantity in the display control module, and start the nucleic acid extraction process; S2 Sample addition: The first robotic arm grabs and moves the sample tube to the multi-cap opening module, and chooses whether to open the bottle cap and add the first reagent according to the sample type; S3 shaking low-speed centrifugation: the second robot moves the sample test tube to the shaking device for shaking, and the shaking of the sample test tube is mixed, and then the second robot moves the sample test tube to the low-speed centrifugation device for centrifugation; S4 transfers the centrifuge liquid: After the centrifugation of the sample tube is completed, the second robot transfers the sample tube to the multi-capping module and opens the cap of the sample tube. The second robot transfers the supernatant of the sample tube to the first centrifuge tube through the TIP gun head; S5: adding reaction reagents: the multiple cover opening module adds the second reagent to the first centrifugal test tube and then closes the cover of the sample test tube; S6 inversion mixing and high-speed centrifugation: The first robot arm moves the first centrifugal test tube to the inversion mixing device. After sufficient mixing, the first robot arm moves the first centrifugal test tube to the multiple opening module, and the second robot arm moves the first centrifugal test tube to the high-speed centrifugation device for high-speed centrifugation; S7: Transferring the centrifugal liquid: the first robot arm clamps the second centrifugal test tube on the centrifugal tube rack to the centrifugal test tube clamping part, the second rotating clamping part opens the tube cover of the second centrifugal test tube, and the upper liquid transfer part of the second robot arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through the TIP gun head; S8: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module; S9: When the incubation timer in one of the centrifugal test tubes is completed, an extraction step is performed: the second robotic arm transfers the second centrifugal test tube to the step-by-step nucleic acid extraction module for nucleic acid extraction.

10. The method for using the nucleic acid extraction and conversion integrated machine according to claim 9, characterized in that: According to the detection process of different cancer types, the display control module deletes or repeats at least one of steps S2, S3, S4, S5 or S6 multiple times; or, according to the detection process of different cancer types, readjusts the execution order of the above S2, S3, S4, S5, S6.

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