Process-editable nucleic acid extraction and transformation all-in-one machine and method using same

Abstract

Provided is a process-editable nucleic acid extraction and transformation all-in-one machine. A housing is provided on a frame and encloses an accommodating space; a sample module is configured for the changing and placement of sample test tubes; a tip rack is arranged on the frame and configured for the placement of tips; a centrifuge tube rack is configured for the placement of centrifuge test tubes; a dual-centrifugation module is configured to subject the test tubes to centrifugation; a multi-sample uncovering module is configured to open and close tube caps of the test tubes; a dual-robotic-arm module is configured to transfer the test tubes and transfer solutions by means of the tips; a step-by-step nucleic acid extraction module is configured to extract nucleic acid products from samples; and a dual-uniform-mixing module is configured to perform uniform mixing on the test tubes. By means of changing the sample module and editing a test process, the present invention can adapt to the testing of most of samples, such as stool samples, body fluid samples and swab samples. Compared with existing single-process products, the whole device integrates multiple test methods into one machine, thereby reducing instrument costs and saving on space.

Description

A nucleic acid extraction and conversion all-in-one machine with editable process and method thereof 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 with editable processes and a method thereof. Background Art

[0002] Nucleic acid research is an important topic in modern biological and medical research. As the carrier of genetic information, nucleic acid is the material basis of gene expression. In addition to playing a very important role in the normal growth, development and reproduction of organisms, it is also closely related to abnormal life conditions such as tumor occurrence, radiation damage, and genetic diseases.

[0003] CN202110643507.9 discloses an all-in-one nucleic acid extraction and PCR detection machine, which prevents the nucleic acid extraction module and the PCR detection module from being contaminated by external gases by setting a first filter at the air inlet; by setting a wind shield, a first air duct, a second air duct, and a changing air duct, the gas first passes through the nucleic acid extraction module and then passes through the PCR detection module, thereby preventing the aerosol of the PCR product from contaminating the nucleic acid extraction module and avoiding cross contamination. However, it can only detect a single nucleic acid sample, and the product function is single, which cannot meet the detection needs of multiple disease detection projects and different samples.

[0004] Therefore, we designed a nucleic acid extraction and conversion machine that integrates multiple detection modules and can detect multiple disease samples with editable processes. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A nucleic acid extraction and conversion integrated machine with editable processes, comprising: a frame, a casing disposed on the frame, and a receiving space formed by the casing;

[0007] A sample module, the sample module is arranged on the rack and is used to place sample tubes;

[0008] A TIP rack, the TIP rack is mounted on the rack and is used to place the TIP gun head;

[0009] A centrifuge tube rack, wherein the centrifuge tube rack is mounted on the rack and is used to place centrifuge test tubes;

[0010] a dual centrifuge module, the dual centrifuge module being disposed on the frame and being used to centrifuge the test tube;

[0011] a multiple cover opening module, the multiple cover opening module being arranged on the frame and being used for opening and closing the tube covers of the test tubes;

[0012] a dual robotic arm module, the dual robotic arm module being disposed on the rack and being used to transfer test tubes and solutions via the TIP gun head;

[0013] a step-by-step nucleic acid extraction module, the step-by-step nucleic acid extraction module being disposed on the rack and being used to extract nucleic acid products from the sample;

[0014] A dual mixing module, which is provided on the rack and is used to mix the test tubes;

[0015] Display control module; the display control module is used to control the operation of the dual centrifugation module, the dual mixing module, the multiple opening module, the dual robotic arm module and the step-by-step nucleic acid extraction module to perform nucleic acid sample processing.

[0016] Preferably, the multiple cover opening module includes a first cover opening component and a second cover opening component;

[0017] The first cover opening assembly includes: a first slide rail, on which a movable sample clamping portion is provided, and the sample clamping portion is suitable for clamping a sample test tube;

[0018] The first lifting platform is arranged above the first slide rail, and is provided with a first liquid-adding needle and a liftable first rotating clamping part, and the first rotating clamping part is suitable for opening or closing the tube cover of the sample tube.

[0019] The second cover opening assembly includes:

[0020] a second slide rail, wherein a movable centrifuge tube clamping portion is provided on the second slide rail, and the centrifuge tube clamping portion is suitable for clamping a tube body of a centrifuge tube;

[0021] The second lifting platform is arranged above the second slide rail, and a liquid adding needle and a liftable second rotating clamping part are provided on the second lifting platform, and the second rotating clamping part is suitable for opening or closing the tube cover of the centrifuge test tube.

[0022] Preferably, the dual mixing module includes:

[0023] An oscillating device, which is provided on the frame and is used for oscillating and mixing the sample tube;

[0024] Inversion mixing device: The inversion mixing device is arranged on the frame and is used for inverting and mixing the centrifugal test tube.

[0025] Preferably, the inversion mixing device comprises: a fixed bracket;

[0026] A rotating bracket is provided above the fixed bracket, the rotating bracket is provided with a plurality of centrifuge test tube holes, the edges of the centrifuge test tube holes are provided with a pair of elastic plates for fixing the centrifuge test tubes, and both ends of the rotating bracket pass through the fixed bracket and are rotatably connected to the fixed bracket;

[0027] A mixing motor is provided below the fixed support and is connected to the end of the rotating support via a transmission belt.

[0028] Preferably, the dual robotic arm module includes: a first robotic arm and a second robotic arm, the first robotic arm is provided with a first clamping part, the second robotic arm is provided with a second clamping part and a pipetting part, the second robotic arm has a second transfer activity area, the second clamping part transfers the test tube within the second transfer activity area, and the pipetting part transfers the solution through the TIP gun tip on the TIP rack within the second transfer activity area.

[0029] Preferably, the dual centrifugal module comprises a low-speed centrifugal device and a high-speed centrifugal device.

[0030] Preferably, the nucleic acid extraction and conversion integrated machine further comprises an anti-fouling and disinfection device, which comprises a UV lamp, an activated carbon filter and a directional ventilation device arranged on the top of the shell.

[0031] Preferably, the step-by-step nucleic acid extraction module includes a first nucleic acid extraction device and a second nucleic acid extraction device; the first nucleic acid extraction device includes a first detection kit and a first magnetic rod assembly, and the second nucleic acid extraction device includes a second detection kit and a second magnetic rod assembly.

[0032] Preferably, the nucleic acid extraction and conversion integrated machine further comprises:

[0033] a liquid adding module, the liquid adding module comprising a first liquid adding device and a second liquid adding device, the first liquid adding device being disposed on the frame and connected to the multiple cover opening module via a pipeline, and the second liquid adding device being disposed on the frame and connected to the step-by-step nucleic acid extraction module via a pipeline;

[0034] A temperature incubation module, which is provided on the rack and is used to store the test tube at a constant temperature;

[0035] The display control module is also used to control the operation of the liquid adding module and the temperature incubation module to process the nucleic acid sample.

[0036] A method for an all-in-one nucleic acid extraction and conversion machine with editable processes, the method comprising the following steps:

[0037] S1 Sample loading: A sample rack carrying multiple sample tubes is mounted on the sample frame. The first clamping portion of the first robotic arm grips and moves the sample tubes to the sample clamping portion. The sample clamping portion clamps and secures the sample tubes and moves along the first slide rail to the second transfer area.

[0038] S2 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;

[0039] S3 Transferring the Centrifuge Liquid: After the centrifugation of the sample tube is completed, the second robotic arm transfers the sample tube to the sample clamping part, which moves to just below the first rotating clamping part. The first rotating clamping part opens the tube cover of the sample tube. The first robotic arm clamps the first centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping part. The second rotating clamping part opens the tube cover of the first centrifuge tube. The pipetting part on the second robotic arm transfers the supernatant of the sample tube to the first centrifuge tube through the tip of the gun;

[0040] S4: Adding reaction reagents: The dosing needle adds reagents to the first centrifuge tube, the second rotating clamping part closes the centrifuge tube cover, and the first rotating clamping part closes the sample tube cover;

[0041] S5: Inversion and mixing, high-speed centrifugation: The first robotic arm moves the first centrifuge tube to the inversion and mixing device. After sufficient mixing, the first robotic arm moves the first centrifuge tube to the centrifuge tube clamping portion. The centrifuge tube clamping portion moves along the second slide rail to the second transfer activity area, and the second robotic arm moves the centrifuge tube to the high-speed centrifugation device for high-speed centrifugation.

[0042] S6: Transferring the centrifuge liquid: The first robotic arm clamps the second centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotating clamping part opens the cap of the second centrifuge tube, and the upper pipetting part of the second robotic arm transfers the supernatant of the first centrifuge tube to the second centrifuge tube through the tip of the gun;

[0043] S7: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0044] S8: 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 first nucleic acid extraction device or the second nucleic acid extraction device for nucleic acid extraction.

[0045] 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. Beneficial effects

[0046] 1. It is equipped with a sample module, dual centrifugation module, multiple opening modules, dual robotic arm module, step-by-step nucleic acid extraction module, and dual mixing module. Simply by replacing the sample module and editing the test process, it can be applied to most sample tests, such as stool samples, body fluid samples, and swab samples. Compared with existing single-process products, the entire device integrates multiple testing methods in one machine, reducing equipment costs and saving space.

[0047] 2. Without affecting the current nucleic acid extraction process, the next sample tube batch is replaced and the sample tube pretreatment operation is started at the same time, maximizing the utilization of the resources of each functional module of the system, reducing the waiting time in the original automatic processing process, and realizing parallel processing of multiple batches of samples, which greatly improves the detection capacity of the system, effectively reduces the detection labor cost, and shortens the sample detection time.

[0048] 3. 16 samples can be placed into the device of the present invention at one time. After a period of time after the samples enter, the nucleic acid template can be obtained, realizing full process automation, which is conducive to reducing the labor intensity of the experimenters and improving the stability and consistency of the experimental results. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] 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.

[0050] FIG1 is a schematic diagram of the overall structure of an embodiment of the present invention.

[0051] FIG2 is a schematic diagram of the internal structure of an embodiment of the present invention;

[0052] FIG3 is a second schematic diagram of the internal structure of an embodiment of the present invention;

[0053] FIG4 is a schematic diagram of the storage space partition according to an embodiment of the present invention;

[0054] FIG5 is a schematic structural diagram of a dual-manipulator module according to an embodiment of the present invention;

[0055] FIG6 is a schematic diagram of the overall structure of a multi-opening module according to an embodiment of the present invention;

[0056] FIG7 is a front structural diagram of a multiple cover opening module according to an embodiment of the present invention;

[0057] FIG8 is a schematic diagram of the overall structure of an inversion mixing device according to an embodiment of the present invention;

[0058] FIG9 is a schematic diagram of the overall structure of an inversion mixing device according to an embodiment of the present invention;

[0059] FIG10 is a schematic diagram of the overall structure of a sample module according to an embodiment of the present invention;

[0060] FIG11 is a schematic diagram of a comprehensive process of the method of the present invention;

[0061] In the figure: rack 1, lifting door 101, connecting arm 102, first transfer activity area 100, second activity area 200, overlapping area 300, sample module 2, sample card slot 20, sample frame 21, sample holder 22, TIP rack 3, centrifuge tube rack 4, dual centrifuge module 5, low-speed centrifuge device 51, high-speed centrifuge device 52, multi-capping module 6, first capping assembly 61, first slide rail 611, sample clamping part 612, first lifting platform 613, first rotary clamping part 615, second capping assembly 62, second slide rail 621, centrifuge tube clamping part 622, second lifting platform 623, liquid addition needle 624, second rotary clamping part 625, dual robotic arm module 7, first robotic arm 71, first clamping part 710, first X-axis arm 711, first Y-axis arm 712, first Z-axis arm 713, second robotic arm 72, second clamping part 720, pipetting part 721, second X-axis arm 722, second Y-axis arm 723, second Z-axis arm 724, step-by-step nucleic acid extraction module 8, first nucleic acid extraction device 81, first detection reagent kit 811, first magnetic rod assembly 812, second nucleic acid extraction device 82, second detection reagent kit 821, second magnetic rod assembly 822, double mixing module 9, oscillation device 91, inversion mixing device 92, centrifugal test tube hole 920, fixed bracket 921, rotating bracket 922, mixing motor 923, conveyor belt 924, elastic plate 925, display control module 10, liquid adding module 11, first liquid adding device 111, second liquid adding device 112, temperature incubation module 12, UV lamp 13, activated carbon filter 14, directional ventilation device 15. DETAILED DESCRIPTION

[0062] 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.

[0063] Embodiment: As shown in Figures 1 to 3: rack 1, sample module 2, TIP rack 3, centrifuge tube rack 4, dual centrifuge module 5, multiple opening module 6, dual robotic arm module 7, step-by-step nucleic acid extraction module 8, dual mixing module 9 and display control module 10,

[0064] A nucleic acid extraction and conversion integrated machine with editable processes, comprising: a frame 1, a casing provided on the frame, and a receiving space formed by the casing;

[0065] Sample module 2, the sample module is arranged on the rack and is used to place sample tubes;

[0066] TIP rack 3, the TIP is mounted on the rack and is used to place the TIP gun head;

[0067] Centrifuge tube rack 4, the centrifuge tube rack is mounted on the rack and is used to place centrifuge test tubes;

[0068] A dual centrifuge module 5, which is provided on the frame and is used for centrifuging test tubes;

[0069] A multiple cover opening module 6, which is provided on the frame and is used to open and close the tube covers of the test tubes;

[0070] A dual robotic arm module 7, which is provided on the frame and is used to transfer test tubes and transfer solutions through the TIP gun head;

[0071] A step-by-step nucleic acid extraction module 8, which is disposed on the frame and is used to extract nucleic acid products from the sample;

[0072] A dual mixing module 9, which is provided on the rack and is used to mix the test tubes;

[0073] The display control module 10 is used to control the operation of the dual centrifugation module 5, the dual mixing module 7, the multiple opening module 6, the dual robotic arm module 7 and the step-by-step nucleic acid extraction module 8 to process nucleic acid samples. The overall size of the nucleic acid extraction and conversion all-in-one machine is 1800mm long * 800mm wide * 1100mm high, and weighs 340KG. The casing is provided with a lifting door 101 and a multi-directional adjustable connecting arm 102. One end of the multi-directional adjustable connecting arm 102 is connected to the display control module 10. The display control module 10 is a 15.6-inch touch all-in-one machine with a processor inside to control the operation process of all modules and record intermediate test data.

[0074] The nucleic acid extraction and conversion machine also includes an anti-fouling and disinfection device, which includes a UV lamp 13, an activated carbon filter 14 and a directional ventilation device 15 arranged on the top of the shell.

[0075] As shown in Figures 1 to 4: the accommodating space is divided into a first transfer activity area 100, a second activity area 200 and an overlapping area 300; wherein, the sample module 2 and the centrifuge tube rack 4 are located in the first transfer activity area 100, the TIP rack 3, the step-by-step nucleic acid extraction module 8 and the dual centrifuge module 5 are located in the second transfer activity area, and the multiple lid opening module 6 is located in the overlapping area.

[0076] There are 16 sample tube slots on the sample tube. During the test, 16 samples can be placed in the device of the present invention at one time. After the sample enters, a nucleic acid template can be obtained after a period of time, realizing full process automation, which is beneficial to reduce the labor intensity of the experimenters and improve the stability and consistency of the experimental results.

[0077] As shown in Figures 2 and 3: In one embodiment of the present invention, 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 first nucleic acid extraction device 81 includes a first detection kit 811 and a first magnetic rod assembly 812; the second nucleic acid extraction device 82 includes a second detection kit 821 and a second magnetic rod assembly 822. The detection kits and magnetic rod assemblies in the first nucleic acid extraction device 81 and the second nucleic acid extraction device 82 can process samples or processed samples to extract nucleic acid products from the samples. The nucleic acid products extracted from different sampling samples can be used as a basis for diagnosing corresponding diseases. For example, if the sampling sample is a nucleic acid sample obtained by a throat swab specimen collection method or a nasal swab specimen collection method, the nucleic acid extraction module can perform cell lysis, inhibitor cleaning, nucleic acid elution and other steps on the nucleic acid sample to obtain a nucleic acid product, which can be used to detect the new coronavirus COVID-19. For another example, if the sample is a stool sample, the nucleic acid extraction module 8 can perform specific binding nucleic acid, inhibitor cleaning, nucleic acid elution and other steps on the processed stool sample to obtain a nucleic acid product, and the nucleic acid product of the stool sample can be used for the detection of colorectal cancer.

[0078] As shown in Figures 5 and 6, in one embodiment of the present invention, the multi-opening module 6 includes a first opening assembly 61 and a second opening assembly 62; the first opening assembly 61 includes: a first slide rail 611, and a movable sample clamping portion 612 is provided on the first slide rail 611, and the sample clamping portion is suitable for clamping a sample tube;

[0079] The first lifting platform 613 is disposed above the first slide rail and is provided with a liftable first rotating clamping portion 615 . The first rotating clamping portion 615 is adapted to open or close the tube cover of the sample tube.

[0080] The second opening cover 62 assembly includes:

[0081] A second slide rail 621 is provided with a movable centrifuge tube clamping portion 622, and the centrifuge tube clamping portion 622 is suitable for clamping the tube body of the centrifuge tube;

[0082] A second lifting platform 623 is positioned above the second slide rail 621 and is equipped with a dosing needle 624 and a second, retractable, rotating clamp 625. The second rotating clamp is adapted to open and close the cap of a centrifuge tube. The centrifuge tube can be a 2ml tube, and the sample tube can be a 50ml stool sample tube. The sample clamp 612 is larger than the second sample clamp 622, and the first rotating clamp 615 is larger than the second rotating clamp 625.

[0083] As shown in Figures 3 and 7: In one embodiment of the present invention, the dual-manipulator module 7 includes: a first manipulator 71 and a second manipulator 72. The first manipulator 71 is provided with a first clamping portion 710. The first manipulator 71 is located above the first transfer activity area 100. The first clamping portion 710 transfers the sample tube or the centrifuge tube within the first transfer activity area. The first manipulator is a manipulator that can move freely in three dimensions along the X, Y, and Z axes. The first manipulator 71 also includes a first X-axis arm 711, a first Y-axis arm 712, and a first Z-axis arm 713. The first clamping portion 710 moves in the left-right direction on the first X-axis arm 711, the first clamping portion 710 moves in the front-back direction on the first Y-axis arm 712, and the first clamping portion 710 moves in the up-down direction on the first Y-axis arm 713.

[0084] The first X-axis arm 711 is installed on the first Y-axis arm 713, and the first Y-axis arm 713 can drive the first X-axis arm 711 to move forward and backward. The first Z-axis arm 713 is installed on the first X-axis arm 711, and the first X-axis arm 711 can drive the first Z-axis arm 713 to move left and right. The first Z-axis arm 713 is connected to the first clamping part 710, and the first Z-axis arm 713 can drive the first clamping part 710 to move up and down, thereby enabling the first clamping part 710 to transfer sample tubes and centrifuge tubes within the first transfer activity area.

[0085] The second robotic arm 72 is provided with a second clamping part 720 and a pipetting part 721. The second robotic arm 72 is located in the second transfer activity area 200. The second clamping part 72 transfers the test tube in the second transfer activity area 200, and the pipetting part 721 transfers the solution through the TIP gun tip on the TIP rack 3 in the second transfer activity area; the second robotic arm 72 can be a three-axis robotic arm, and the second robotic arm 72 includes a second X-axis arm 722, a second Y-axis arm 723 and a second Z-axis arm 724. The second X-axis arm 722 extends in the left-right direction, the second Y-axis arm 723 moves in the front-back direction, and the second Y-axis arm 723 moves in the up-down direction. Among them, the second X-axis arm 722 can be connected to the frame 1, the second Y-axis arm 723 is installed on the second X-axis arm 722, the second X-axis arm 722 can drive the second Y-axis arm 723 to move left and right, the second Z-axis arm 724 is installed on the second Y-axis arm 723, the second Y-axis arm 723 can drive the second Z-axis arm 724 to move forward and backward, the second Z-axis arm 724 includes two Z-axis lifting arms, one of which is provided with a second clamping part 720, and the other Z-axis lifting arm is provided with a pipetting part 721, the second Z-axis arm 724 can drive the second clamping part 720 and the pipetting part 721 to move up and down independently, so that the second clamping part 721 transfers the test tube in the second transfer activity area, and the pipetting part 722 transfers the solution through the TIP gun head in the second transfer activity area.

[0086] The display control module 10 is used to control the operation of the dual centrifugation module 5, the dual mixing module 9, the multiple lid opening module 6, the dual robotic arm module 7 and the step-by-step nucleic acid extraction module 8 respectively to process nucleic acid samples.

[0087] During nucleic acid sample pre-treatment, the robotic arm module 7 transfers the nucleic acid sample tube from the sample module 2 to the multi-opening module 6, opens the nucleic acid sample tube through the multi-opening module 6, and the robotic arm module 7 transfers the solution in the nucleic acid sample tube to the nucleic acid extraction module 8 through the TIP gun head, thereby extracting the nucleic acid product through the nucleic acid extraction module. In this way, the full process automation of sample pre-treatment is realized, which is conducive to reducing the labor intensity of experimenters and improving the stability and consistency of experimental results.

[0088] As shown in FIG2 : In one embodiment of the present invention, the nucleic acid extraction and conversion integrated machine further includes:

[0089] A liquid adding module 11 is provided on the frame and is used to store reaction additive reagents. In one embodiment of the present invention, the liquid adding module 11 includes a first liquid adding device 111 and a second liquid adding device 112. The first liquid adding device 111 is provided in the first transfer activity area 100 and is connected to the various cover opening module 6 via a pipeline. The first liquid adding device 111 is located near the various cover opening module 6. Specifically, the first liquid adding device 111 provides liquid adding power for the liquid adding needle 624.

[0090] The second liquid adding device 112 is located in the second transfer area 200 and is connected to the step-by-step nucleic acid extraction module 8 via a pipeline.

[0091] The temperature incubation module 12 is located in the second transfer activity area 200 and is used to provide a constant temperature incubation environment for the test tube;

[0092] The display control module 10 is further used to control the operation of the first liquid adding device 111 , the second liquid adding device 112 and the temperature incubation module 12 to process the nucleic acid sample.

[0093] The nucleic acid extraction and conversion integrated machine also includes an anti-fouling and disinfection device, which includes a UV lamp 13, an activated carbon filter 14 and a directional ventilation device 15 arranged on the top of the shell.

[0094] In one embodiment of the present invention, the dual centrifuge module 5 is located in the second transfer activity area 200, and includes a low-speed centrifuge 51 and a high-speed centrifuge 52. The low-speed centrifuge 51 can have a centrifugal speed of 8000 rpm or less, and is used for centrifugal separation of sample tubes. The high-speed centrifuge 52 can have a centrifugal speed of 15000 rpm-30000 rpm, and is used for centrifugal stratification of centrifugal tubes.

[0095] As shown in FIG2 : In one embodiment of the present invention, the dual mixing module 9 includes

[0096] An oscillating device 91 is provided in the second transfer activity area and is used for oscillating and mixing the sample tube;

[0097] Inversion mixing device 92: The inversion mixing device is arranged in the first transfer activity area and is used to invert and mix the centrifuge tube.

[0098] As shown in FIG8-FIG9: In one embodiment of the present invention, the inversion mixing device 92 includes: a fixed bracket 921;

[0099] A rotating bracket 922 is provided above the fixed bracket 921. The rotating bracket 922 is provided with a plurality of centrifuge test tube holes 920, with a total of 16 centrifuge test tube holes arranged in a 4*4 grid. A pair of elastic plates 925 are provided at the edges of the centrifuge test tube holes for securing the centrifuge test tubes. Both ends of the rotating bracket 922 pass through the fixed bracket 921 and are rotatably connected to the fixed bracket 921.

[0100] The mixing motor 923 is provided below the fixed support 921 and is connected to the end of the rotating support 922 via a transmission belt 924. The mixing motor 923 drives the rotating support 922 to rotate via the transmission belt 924 to invert and mix the centrifuge tube.

[0101] As shown in Figure 10: The present invention relates to a sample module 2, which includes a sample frame 21 and a sample holder 22. The sample holder 22 is snapped into a slot 20 on the sample frame 22. In this embodiment, the sample holder 22 is a holder for a stool sample test tube. Of course, it can also be replaced with sample holders for other items, such as nucleic acid samples and body fluid samples. Different processing processes can be designed according to different samples, and different modules can be called.

[0102] FIG11 is a comprehensive flow chart of the method of the present invention, which is an operation method of an integrated nucleic acid extraction and conversion machine with editable flow. Example 1: When the sample to be detected is a stool sample, the method includes the following steps:

[0103] S1 Sample loading: A sample holder carrying multiple stool sample tubes is mounted on the sample frame. The stool sample tubes are 50ml tubes. The first clamping portion of the first robotic arm grips and moves the sample tubes to the sample clamping portion. The sample clamping portion clamps and secures the sample tubes and moves them along the first slide rail to the second transfer area.

[0104] S2 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;

[0105] S3 Transferring the Centrifuge Liquid: After the centrifugation of the sample tube is completed, the second robotic arm transfers the sample tube to the sample clamping part, which moves to just below the first rotating clamping part. The first rotating clamping part opens the tube cover of the sample tube. The first robotic arm clamps the first centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping part. The second rotating clamping part opens the tube cover of the first centrifuge tube. The pipetting part on the second robotic arm transfers the supernatant of the sample tube to the first centrifuge tube through the tip of the gun;

[0106] S4: Adding reaction reagents: The dosing needle adds reagents to the first centrifuge tube, the second rotating clamping part closes the centrifuge tube cover, and the first rotating clamping part closes the sample tube cover;

[0107] S5: Inversion and mixing, high-speed centrifugation: The first robotic arm moves the first centrifuge tube to the inversion and mixing device. After sufficient mixing, the first robotic arm moves the first centrifuge tube to the centrifuge tube clamping portion. The centrifuge tube clamping portion moves along the second slide rail to the second transfer activity area, and the second robotic arm moves the centrifuge tube to the high-speed centrifugation device for high-speed centrifugation.

[0108] S6: Transferring the centrifuge liquid: The first robotic arm clamps the second centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotating clamping part opens the cap of the second centrifuge tube, and the upper pipetting part of the second robotic arm transfers the supernatant of the first centrifuge tube to the second centrifuge tube through the tip of the gun;

[0109] S7: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0110] S8: 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 first nucleic acid extraction device or the second nucleic acid extraction device for nucleic acid extraction.

[0111] 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.

[0112] Example 1: When the test sample is a bladder cancer urine sample, steps S2 and S3 are omitted, and steps S4, S5, and S6 are repeated twice.

[0113] Sample loading: A sample holder carrying multiple 2ml bladder cancer urine sample tubes is mounted on the sample frame. The first gripping portion of the first robotic arm grips and moves the bladder cancer urine sample tubes to the sample gripping portion. The sample gripping portion secures the sample tubes and moves them along the first slide rail to the second transfer area.

[0114] Adding reaction reagent: adding reagent to the bladder cancer urine sample test tube with a liquid adding needle, and closing the cover of the bladder cancer urine sample test tube with a second rotating clamping part;

[0115] Inversion mixing and high-speed centrifugation: The first robotic arm 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 centrifuge tube clamping part. The centrifuge tube clamping part moves along the second slide rail to the second transfer activity area, and the second robotic arm moves the bladder cancer urine sample tube to the high-speed centrifuge device for high-speed centrifugation;

[0116] Transferring the centrifuge liquid: The first robotic arm clamps the first centrifuge tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotating clamping part opens the tube cover of the first centrifuge tube, and the upper pipetting part 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;

[0117] Adding the reaction reagent again: the adding needle adds the reagent to the first centrifuge tube, the second rotating clamping part closes the cap of the first centrifuge tube, and the first rotating clamping part closes the cap of the bladder cancer urine sample tube;

[0118] Inversion and mixing high-speed centrifugation: The first robotic arm moves the first centrifuge tube to the inversion and mixing device. After sufficient mixing, the first robotic arm moves the first centrifuge tube to the centrifuge tube clamping part. The centrifuge tube clamping part moves along the second slide rail to the second transfer activity area, and the second robotic arm moves the centrifuge tube to the high-speed centrifugation device for high-speed centrifugation.

[0119] 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;

[0120] The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0121] 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.

[0122] 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.

[0123] 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 test tube, the stool sample test tube is shaken and centrifuged to obtain a first supernatant. The first reagent may be a suspension that lyses cells and releases nucleic acids, and the second reagent may be a suspension that lyses cells and releases nucleic acids. A second centrifugation step: adding a third reagent to the first supernatant, shaking and centrifuging the first supernatant to obtain a second supernatant. The second reagent may be an inhibitor removal solution for removing PCR inhibitors such as polysaccharides and phenols. A third centrifugation step: adding a fourth reagent to the second supernatant, shaking, incubating the second supernatant at a controlled temperature, and centrifuging. The fourth reagent may be isopropanol to form a flocculent precipitate of nucleic acids. The supernatant from the third centrifugation step is discarded, and a fifth reagent is added to the remaining solution. The remaining solution after the addition of the fifth reagent is shaken, and the remaining solution after shaking is used to extract nucleic acid products. The fifth reagent may be a solubilizing and binding solution that provides conditions for magnetic beads to adsorb nucleic acids.

[0124] 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.

[0125] 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.

[0126] In addition, according to the stool sample pretreatment method of the embodiment of the present invention, the nucleic acid extraction and conversion device of the present invention can realize one-button intelligent operation through the display screen. The display screen can have an intelligent operation interface and can realize digital storage and recording of full-process experimental information, thereby improving the automation, informatization, and intelligent construction of the laboratory.

[0127] 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.

[0128] 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.

[0129] 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.

[0130] 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 with editable process, characterized in that: The nucleic acid extraction and conversion integrated machine comprises: a frame, a casing is arranged on the frame, and the casing surrounds and forms a containing space; A sample module, the sample module is arranged on the rack and is used to place sample test tubes; A TIP rack, the TIP rack is mounted on the rack and is used to place the TIP gun head; A centrifuge tube rack, the centrifuge tube rack is mounted on the rack and is used to place centrifuge test tubes; A dual centrifuge module, which is disposed on the frame and is used to centrifuge the test tube; A multiple cover opening module, which is arranged on the frame and used for opening and closing the tube covers of the test tubes; A dual robotic arm module, the dual robotic arm module is disposed on the rack and is used to transfer test tubes and transfer solutions through the TIP gun head; A step-by-step nucleic acid extraction module, which is disposed on the rack and is used to extract nucleic acid products from the sample; A dual mixing module, which is disposed on the rack and is used to mix the test tubes; Display control module; the display control module is used to control the operation of the dual centrifugation module, the dual mixing module, the multiple opening module, the dual robotic arm module and the step-by-step nucleic acid extraction module respectively to perform nucleic acid sample processing.

2. The nucleic acid extraction and conversion integrated machine with editable process as claimed in claim 1, characterized in that: The multiple cover opening module includes a first cover opening component and a second cover opening component; The first cover opening assembly comprises: a first slide rail, on which a movable sample clamping portion is disposed, and the sample clamping portion is suitable for clamping a sample test tube; A first lifting platform, the first lifting platform is arranged above the first slide rail, and a first rotating clamping portion that can be raised and lowered is arranged on the first lifting platform, and the first rotating clamping portion is suitable for opening or closing a tube cover of a sample test tube; The second cover opening component comprises: A second slide rail, wherein a movable centrifugal test tube clamping portion is provided on the second slide rail, and the centrifugal test tube clamping portion is suitable for clamping a tube body of a centrifugal test tube; The second lifting platform is arranged above the second slide rail, and a liquid adding needle and a second rotatable clamping part which can be lifted and lowered are arranged on the second lifting platform, and the second rotatable clamping part is suitable for opening or closing the tube cover of the centrifugal test tube.

3. The nucleic acid extraction and conversion integrated machine with editable process as claimed in claim 1, characterized in that: The dual mixing module comprises An oscillating device, which is disposed on the rack and is used for oscillating and mixing the sample test tube; Inversion mixing device: The inversion mixing device is arranged on the frame and is used for inverting and mixing the centrifugal test tube.

4. The nucleic acid extraction and conversion integrated machine with editable process as claimed in claim 3, characterized in that: The inversion mixing device comprises: a fixed support; A rotating bracket, the rotating bracket is arranged above the fixed bracket, the rotating bracket is provided with a plurality of centrifugal test tube holes, a pair of elastic plates for fixing centrifugal test tubes are arranged at the edges of the centrifugal test tube holes, and both ends of the rotating bracket pass through the fixed bracket and are rotatably connected to the fixed bracket; A mixing motor is arranged below the fixed support and is connected to the end of the rotating support through a transmission belt.

5. The nucleic acid extraction and conversion integrated machine with editable process according to claim 1, characterized in that: The dual robotic arm module includes: a first robotic arm and a second robotic arm, the first robotic arm is provided with a first clamping part, the second robotic arm is provided with a second clamping part and a pipetting part, the second robotic arm has a second transfer activity area, the second clamping part transfers the test tube in the second transfer activity area, and the pipetting part transfers the solution through the TIP gun tip on the TIP rack in the second transfer activity area.

6. The nucleic acid extraction and conversion machine with editable process as claimed in claim 1, characterized in that: The nucleic acid extraction and conversion integrated machine also includes an anti-fouling and disinfection device, which includes a UV lamp, an activated carbon filter and a directional ventilation device arranged on the top of the shell.

7. The nucleic acid extraction and conversion integrated machine with editable process as claimed in claim 1, characterized in that: The step-by-step nucleic acid extraction module comprises a first nucleic acid extraction device and a second nucleic acid extraction device; the first nucleic acid extraction device comprises a first detection kit and a first magnetic rod assembly, and the second nucleic acid extraction device comprises a second detection kit and a second magnetic rod assembly.

8. The nucleic acid extraction and conversion integrated machine with editable process as claimed in claim 7, characterized in that: The nucleic acid extraction and conversion integrated machine also includes: A liquid adding module, the liquid adding module comprising a first liquid adding device and a second liquid adding device, the first liquid adding device is arranged on the frame and connected to the multiple cover opening module through a pipeline, and the second liquid adding device is arranged on the frame and connected to the step-by-step nucleic acid extraction module through a pipeline; A temperature incubation module, which is disposed on the rack and is used to store the test tube at a constant temperature; The display control module is also used to control the operation of the liquid adding module and the temperature incubation module to process the nucleic acid sample.

9. A method for an all-in-one nucleic acid extraction and conversion machine with editable flow as described in any one of claims 1 to 8, characterized in that: The method comprises the following steps: S1 Sample loading: a sample rack carrying multiple sample test tubes is installed on the sample frame, the first clamping part on the first robot arm clamps and moves the sample test tube to the sample clamping part, the sample clamping part clamps and fixes the sample test tube, and moves along the first slide rail to the second transfer activity area; S2 shaking low-speed centrifugation: the second robot moves the sample tube to the shaking device for shaking, and the shaking of the sample tube is mixed, and then the second robot moves the sample tube to the low-speed centrifugation device for centrifugation; S3 transfer of centrifugal liquid: after the centrifugation of the sample test tube is completed, the second robot arm transfers the sample test tube to the sample clamping part, the sample clamping part moves to just below the first rotating clamping part, the first rotating clamping part opens the tube cover of the sample test tube, the first robot arm clamps the first 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 first centrifugal test tube, and the liquid transfer part on the second robot arm transfers the supernatant of the sample test tube to the first centrifuge test tube through the tip of the gun; S4: adding reaction reagents: the adding needle adds reagents to the first centrifugal test tube, the second rotating clamping part closes the cover of the centrifugal test tube, and the first rotating clamping part closes the cover of the sample test tube; S5 Inversion and mixing high-speed centrifugation: The first robot arm moves the first centrifugal test tube to the inversion and mixing device. After sufficient mixing, the first robot arm moves the first centrifugal test tube to the centrifugal test tube clamping part. The centrifugal test tube clamping part moves along the second slide rail to the second transfer activity area, and the second robot arm moves the centrifugal test tube to the high-speed centrifugal device for high-speed centrifugation; S6: Transferring the centrifugal liquid: the first mechanical 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 mechanical arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through the tip gun head; S7: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module; S8: When the incubation time 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 first nucleic acid extraction device or the second nucleic acid extraction device for nucleic acid extraction.

10. The method of the nucleic acid extraction and conversion integrated machine with editable process as claimed in 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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