Multi-task parallel nucleic acid extraction and conversion method and apparatus

Abstract

The present invention relates to a multi-task parallel nucleic acid extraction and conversion method and apparatus. The apparatus comprises a frame, a sample module, a TIP rack, a centrifuge tube rack, a dual-centrifugation module, a multi-test-tube cap opening module, a dual-mechanical-arm module, a stepwise nucleic acid extraction module, a dual-mixing module, and a display control module. A housing is arranged on the frame; the housing defines an accommodating space; the accommodating space is divided into a first transfer movement area, a second movement area and an overlapping area. According to the resource requirement of the current task process, replacement to the next batch of sample test tubes is implemented without affecting the process of the current nucleic acid extraction step, and a sample test tube preprocessing operation is started, thereby maximizing the utilization of resources of functional modules of a system, reducing the waiting time during previous automatic processing, implementing parallel processing of multiple batches of samples, greatly improving the detection capacity of the system, effectively reducing the labor costs of detection, shortening the sample detection time, and achieving positive significance for the popularization of early tumor detection.

Description

A multi-task parallel nucleic acid extraction and conversion method and device Technical Field

[0001] The present invention relates to the technical field of nucleic acid extraction, and in particular to a multi-task parallel nucleic acid extraction and conversion method and device 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] 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 sample pre-processing process is cumbersome and requires a large amount of manual operation, which puts a high labor intensity on experimenters. In addition, the experimental results may fluctuate due to differences in the experimental personnel's operations, affecting the stability of the test results.

[0004] The document with patent number CN202110643507.9 discloses an all-in-one nucleic acid extraction and PCR detection machine, including a casing, a nucleic acid extraction module, and a PCR detection module. The casing is a hollow shell, and the nucleic acid extraction module and the PCR detection module are arranged inside the casing. The casing is provided with an air inlet and an air outlet. The air inlet is provided with a first filter, and the air outlet is provided with a second filter. A wind shield is provided on the inner side of the casing near the air inlet, and the wind shield is provided between the air inlet and the PCR detection module. The wind shield extends from the air inlet to the nucleic acid extraction module; a first air duct is provided at the bottom of the nucleic acid extraction module, and a second air duct is provided at the bottom of the PCR detection module, and the first air duct is connected to the second air duct; a changing air duct and a fan are sequentially provided on the side of the second air duct away from the first air duct, and the side of the fan away from the changing air duct is connected to the second filter.

[0005] However, the above technical solution is still a single-threaded operation. After the sample is placed in the all-in-one machine, it takes a long time to get the results, and the overall efficiency is low. Summary of the Invention

[0006] 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:

[0007] A multi-task parallel nucleic acid extraction and conversion method, the method comprising:

[0008] S0: Installing a sample holder carrying multiple sample tubes onto the sample frame;

[0009] S1: The first clamping portion of the first robotic arm clamps and moves the sample tube to the sample clamping portion, which clamps and secures the sample tube and moves along the first slide rail to the second transfer area.

[0010] S2: 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;

[0011] S3: After the centrifugation of the sample tube is completed, the second robotic arm transfers the sample tube to the sample clamping portion, and the sample clamping portion moves to directly below the first rotating clamping portion, and the first rotating clamping portion opens the tube cover of the sample tube;

[0012] S4: 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 cap 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, the adding needle adds 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 sample tube;

[0013] S5: The first robotic arm moves the first centrifugal test tube to the inversion mixing device. After sufficient mixing, the first robotic arm moves the first centrifugal test tube to the centrifugal test tube clamping portion. The centrifugal test tube clamping portion moves along the second slide rail to the second transfer activity area, and the second robotic arm moves the first centrifugal test tube to the high-speed centrifugation device for high-speed centrifugation.

[0014] S6: 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 centrifugal test tube through the tip of the gun;

[0015] S7: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module;

[0016] S8: Repeat the above steps until all the sample tubes on the sample rack of this batch are transferred to the temperature incubation module, that is, the current number of sample tubes to be tested is 0;

[0017] S9: 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.

[0018] Preferably, according to the resource requirements of the current task process, without affecting the current nucleic acid extraction step process, when the display control module reads that the current number of sample tubes is 0, it prompts that the sample holder can be replaced;

[0019] When the next batch of sample holders are installed on the sample frame, the above steps S1-S8 are repeated to maximize the utilization of the resources of each functional module of the system.

[0020] A multi-tasking parallel nucleic acid extraction and conversion machine comprises: a frame, a sample module, a TIP rack, a centrifuge tube rack, a dual centrifuge module, a multi-capping module, a dual robotic arm module, a step-by-step nucleic acid extraction module, a dual mixing module, and a display control module. The frame is provided with a housing, which surrounds a storage space, and the storage space is divided into a first transfer activity area, a second activity area, and an overlapping area.

[0021] The dual-manipulator module includes: a first manipulator and a second manipulator, wherein the first manipulator is provided with a first clamping portion, the first manipulator is located above a first transfer activity area, and the first clamping portion transfers test tubes within the first transfer activity area; the second manipulator is provided with a second clamping portion and a pipetting portion, the second manipulator is located in a second transfer activity area, the second clamping portion transfers test tubes within the second transfer activity area, and the pipetting portion transfers solutions within the second transfer activity area through the TIP gun head;

[0022] The sample module and the centrifuge tube rack are located in the first transfer activity area, the TIP rack, the step-by-step nucleic acid extraction module, the dual centrifuge module, and the temperature incubation module are located in the second transfer activity area, and the multiple lid opening module is located in the overlapping area.

[0023] The step-by-step nucleic acid extraction module is disposed on the rack and is used to extract nucleic acid products from the sample;

[0024] The dual mixing module is provided on the rack and is used to mix the test tubes;

[0025] The display control module is used to respectively control the operation of the dual centrifugation module, the dual mixing module, the multiple lid opening module, the dual robotic arm module and the step-by-step nucleic acid extraction module to process nucleic acid samples.

[0026] Preferably, the nucleic acid extraction and conversion integrated machine further comprises:

[0027] A liquid adding module, the liquid adding module is arranged on the rack and is used to store reaction additive reagents;

[0028] A temperature incubation module, which is provided on the rack and is used to store the test tube at a constant temperature;

[0029] 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.

[0030] Preferably, the liquid adding module includes a first liquid adding device and a second liquid adding device, the first liquid adding device is arranged in the first transfer activity area and is connected to the multi-opening module through a pipeline, and the second liquid adding device is arranged in the second transfer activity area and is connected to the step-by-step nucleic acid extraction module through a pipeline.

[0031] Preferably, the multiple cover opening module includes a first cover opening component and a second cover opening component;

[0032] 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;

[0033] a first lifting platform, the first lifting platform being disposed above the first slide rail and having a first rotatable clamping portion that can be raised and lowered, the first rotatable clamping portion being adapted to open or close a tube cover of a sample tube;

[0034] The second cover opening assembly includes:

[0035] 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;

[0036] 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.

[0037] Preferably, the dual centrifugal module includes a low-speed centrifugal device and a high-speed centrifugal device, and both the low-speed centrifugal device and the high-speed centrifugal device are located in the second transfer activity area.

[0038] Preferably, the dual mixing module includes

[0039] an oscillating device, the oscillating device being provided in the second transfer activity area and being used for oscillating and mixing the sample tube;

[0040] Inversion mixing device: The inversion mixing device is arranged in the first transfer activity area and is used to invert and mix the centrifuge tube.

[0041] Preferably, the inversion mixing device comprises: a fixed bracket;

[0042] 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;

[0043] A mixing motor is provided below the fixed support and is connected to the end of the rotating support via a transmission belt.

[0044] 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; the second nucleic acid extraction device includes a second detection kit and a second magnetic rod assembly.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] 1. According to the resource requirements of the current task process, the next sample tube batch can be replaced without affecting the current nucleic acid extraction step process, and the sample tube pretreatment operation can be 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, shortens the sample detection time, and has positive significance for promoting the popularization of early tumor detection.

[0047] 2. 16 samples can be placed in the device of the present invention at one time. After the sample enters, the nucleic acid template can be obtained after a period of time, 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.

[0048] 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 a comprehensive process of the method of the present invention

[0051] FIG2 is a schematic diagram of the overall structure of an embodiment of the present invention.

[0052] FIG3 is a schematic diagram of the storage space partition according to an embodiment of the present invention;

[0053] FIG4 is a schematic diagram of the internal structure of 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 multiple cover opening module according to an embodiment of the present invention.

[0056] FIG. 7 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 a sample module according to an embodiment of the present invention.

[0059] 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, liquid addition needle 624, 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 71 1. 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. DETAILED DESCRIPTION

[0060] 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.

[0061] [Corrected 06.03.2024 in accordance with Rule 26] Example: As shown in Figures 1-9: A multi-tasking, parallel nucleic acid extraction and conversion machine comprises: a frame 1, a sample module 2, a tip rack 3, a centrifuge tube rack 4, a dual centrifuge module 5, a multi-capping module 6, a dual robotic arm module 7, a step-by-step nucleic acid extraction module 8, a dual mixing module 9, and a display control module 10. A housing is disposed on the frame, forming a surrounding storage space. The overall dimensions of the nucleic acid extraction and conversion machine are 1800 mm long, 800 mm wide, and 1100 mm high, and it weighs 340 kg. The housing is equipped with a lifting door 101 and a multi-directionally adjustable connecting arm 102. One end of the multi-directionally adjustable connecting arm 102 is connected to the display control module 10. The display control module 10 is a 15.6-inch touchscreen all-in-one machine with an internal processor that controls the operational processes of all modules and records intermediate experimental data.

[0062] The fecal sample pretreatment method includes the following steps: a first centrifugation step: shaking the fecal sample test tube and centrifuging at low speed to obtain a first supernatant after centrifugation. A second centrifugation step: adding a first reagent to the first supernatant, shaking the first supernatant with the first reagent added, and centrifuging at high speed to obtain a second supernatant after centrifugation. A third centrifugation step: adding a second reagent to the second supernatant, shaking, temperature-controlled incubating, and centrifuging the second supernatant with the second reagent added. The second reagent can be isopropanol that causes the nucleic acid to form a flocculent precipitate; discarding the supernatant after centrifugation in the third centrifugation step, adding a third reagent to the remaining liquid, shaking the remaining liquid with the third reagent added, and extracting the nucleic acid product from the remaining liquid after shaking. The third reagent can be a dissolving and binding solution that provides conditions for magnetic beads to adsorb nucleic acids.

[0063] 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.

[0064] [Corrected 06.03.2024 according to Rule 26] Example 1: FIG1 is a comprehensive flow chart of the method of the present invention, a method of a multi-tasking parallel nucleic acid extraction and conversion machine, when the sample to be tested is a fecal sample, the method comprises:

[0065] S0: Install the sample holder 22 carrying multiple sample tubes onto the sample frame 21;

[0066] S1: The first clamping portion 710 on the first robotic arm 71 clamps and moves the sample tube to the sample clamping portion 612. The sample clamping portion 612 clamps and secures the sample tube and moves along the first slide rail to the second transfer area 200.

[0067] S2: The second robotic arm 72 moves the sample tube to the shaking device 91 for shaking to mix the sample tube, and then the second robotic arm 72 moves the sample tube to the low-speed centrifugation device 51 for centrifugation;

[0068] S3: After the centrifugation of the sample tube is completed, the second robotic arm 72 transfers the sample tube to the sample clamping portion 612. The sample clamping portion 612 moves to directly below the first rotating clamping portion 615. The first rotating clamping portion 615 opens the tube cover of the sample tube.

[0069] S4: The first robotic arm 71 clamps the first centrifuge tube on the centrifuge tube rack 4 to the centrifuge tube clamping portion 622. The second rotating clamping portion 625 opens the cap of the first centrifuge tube. The pipetting portion 721 on the second robotic arm 72 transfers the supernatant of the sample tube to the centrifuge tube through the tip. The adding needle 624 adds reagent to the first centrifuge tube. The second rotating clamping portion 625 closes the cap of the first centrifuge tube. The first rotating clamping portion 615 closes the cap of the sample tube.

[0070] S5: The first robotic arm 71 moves the first centrifugal test tube to the inversion mixing device 92. After sufficient mixing, the first robotic arm 71 moves the first centrifugal test tube to the centrifugal test tube clamping portion 622. The centrifugal test tube clamping portion 622 moves along the second slide rail 621 to the second transfer activity area 200. The second robotic arm 72 then moves the centrifugal test tube to the high-speed centrifuge device 52 for high-speed centrifugation.

[0071] S6: The first robotic arm 71 clamps the second centrifugal test tube on the centrifuge tube rack to the centrifugal test tube clamping portion 622, the second rotating clamping portion 625 opens the cover of the second centrifugal test tube, and the pipetting portion 721 on the second robotic arm 72 transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through the tip of the pipette;

[0072] S7: The second robotic arm 72 transfers the second centrifugal test tube to the temperature incubation module;

[0073] S8: Repeat the above steps until all the sample tubes on the sample rack of this batch are transferred to the temperature incubation module. After all the sample tubes on the sample rack of this batch are transferred to the temperature incubation module, based on the resource requirements of the current task process and without affecting the current nucleic acid extraction step process, when the display control module reads that the current number of sample tubes is 0, the display control module issues a prompt, prompting the operator to replace the sample rack;

[0074] S9: Replace the next batch of sample holders onto the sample frame and repeat the above steps S1-S8;

[0075] Under normal circumstances, in a batch of 16 sample tubes, steps S1-S8 are completed in sequence, which takes 15-30 minutes. However, in the temperature incubation module, the incubation and cultivation of the centrifugal tubes require more than 60 minutes. Therefore, there will be a time difference in the middle. Calculated at full load processing speed, the original processing time is the number of batches n*(60+30)min. In order to save time and speed up the detection speed, this device separates the pretreatment step and the nucleic acid extraction step. The improved processing time is the number of batches n*30+60min. When n>2, the average waiting time for each batch can be reduced by 60min, which greatly reduces the waiting time in the original multi-batch sample processing process.

[0076] S10: 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.

[0077] According to the resource requirements of the current task process, the next sample tube batch can be replaced without affecting the current nucleic acid extraction step process, and the sample tube pretreatment operation can be 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 labor cost of detection, shortens the sample detection time, and has positive significance for promoting the popularization of early tumor detection.

[0078] 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.

[0079] 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.

[0080] 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.

[0081] 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.

[0082] As shown in Figures 3 to 5: 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 moves freely in three dimensions along the XYZ axis. 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 left and right on the first X-axis arm 711. The first clamping portion 710 moves left and right on the first X-axis arm 711. The holding portion 710 moves in the front-to-back direction on the first Y-axis arm 712, and the first clamping portion 710 moves in the up-and-down direction on the first Y-axis arm 713; 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 back and forth, and 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 clamping portion 710 is connected to the first Z-axis arm 713, and the first Z-axis arm 713 can drive the first clamping portion 710 to move up and down, thereby enabling the first clamping portion 710 to transfer the sample tube and the centrifuge tube within the first transfer activity area.

[0083] 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.

[0084] The display control module 10 is used to control the operation of the dual centrifugation module 5, the dual mixing module, the multiple lid opening module, the dual robotic arm module and the step-by-step nucleic acid extraction module to process nucleic acid samples.

[0085] As shown in FIG6-FIG7: In one embodiment of the present invention, the multi-opening module 6 includes a first opening component 61 and a second opening component 62;

[0086] The first cover opening assembly 61 includes: a first slide rail 611 , on which a movable sample clamping portion 612 is provided, the sample clamping portion being suitable for clamping a sample test tube;

[0087] 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.

[0088] The second opening cover 62 assembly includes:

[0089] 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;

[0090] 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.

[0091] 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.

[0092] As shown in FIG2 : In one embodiment of the present invention, the nucleic acid extraction and conversion integrated machine further includes:

[0093] 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.

[0094] 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.

[0095] 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;

[0096] 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.

[0097] 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.

[0098] 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 4000 rpm-5000 rpm or less, and is used for centrifugal separation of sample test tubes. The high-speed centrifuge 52 can have a centrifugal speed of 8000 rpm-10000 rpm, and the high-speed centrifuge is used for centrifugal stratification of centrifugal test tubes.

[0099] As shown in FIG2 : In one embodiment of the present invention, the dual mixing module 9 includes

[0100] An oscillating device 91 is provided in the second transfer activity area and is used for oscillating and mixing the sample tube;

[0101] 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.

[0102] As shown in FIG8-FIG9: In one embodiment of the present invention, the inversion mixing device 92 includes: a fixed bracket 921;

[0103] 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.

[0104] 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.

[0105] [Corrected 06.03.2024 according to Rule 26] As shown in Figure 9: 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 onto 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.

[0106] 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.

[0107] 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.

[0108] 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.

[0109] 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.

[0110] 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 multi-task parallel nucleic acid extraction and conversion method, characterized in that: The method comprises: S0: installing a sample holder carrying a plurality of sample tubes onto a sample frame; S1: 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: The second robot arm moves the sample test tube to the shaking device for shaking, and the sample test tube is shaken and mixed, and then the second robot arm moves the sample test tube to the low-speed centrifugation device for centrifugation; S3: After the centrifugation of the sample tube is completed, the second robot arm transfers the sample tube to the sample clamping part, and the sample clamping part moves to just below the first rotating clamping part, and the first rotating clamping part opens the tube cover of the sample tube; S4: The first robot arm clamps the first 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 first centrifugal test tube, the liquid transfer part on the second robot arm transfers the supernatant of the sample test tube to the first centrifugal test tube through the tip gun head, the liquid adding needle adds reagent to the first centrifugal test tube, the second rotating clamping part closes the tube cover of the first centrifugal test tube, and the first rotating clamping part closes the tube cover of the sample test tube; S5: The first robot arm moves the first centrifugal test tube to the inverted 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 first centrifugal test tube is moved to the high-speed centrifugal device for high-speed centrifugation by the second robot arm. S6: 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 pipetting 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; S7: The second robotic arm transfers the second centrifugal test tube to the temperature incubation module; S8: Repeat the above steps until all the sample tubes on the sample rack of this batch are transferred to the temperature incubation module, that is, the current number of sample tubes to be tested is 0; S9: 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.

2. A multi-task parallel nucleic acid extraction and conversion method as claimed in claim 1, characterized in that: According to the resource demand of the current task process, without affecting the current nucleic acid extraction step process, when the display control module reads that the number of sample tubes to be tested is 0, it prompts that the sample holder can be replaced; When the next batch of sample holders are installed on the sample frame, the above steps S1-S8 are repeated to maximize the utilization of the resources of each functional module of the system.

3. A multi-task parallel nucleic acid extraction and conversion machine, characterized in that: The nucleic acid extraction and conversion integrated machine is used to perform the method according to any one of claims 1-2, and the nucleic acid extraction and conversion integrated machine comprises: a rack, a sample module, a TIP rack, a centrifuge tube rack, a double centrifuge module, a multiple cover opening module, a double robotic arm module, a step-by-step nucleic acid extraction module, a double mixing module and a display control module, a housing is arranged on the rack, and the housing is surrounded to form a containing space, and the containing space is divided into a first transfer activity area, a second activity area and an overlapping area; The dual robot module comprises: a first robot and a second robot, wherein the first robot is provided with a first clamping part, the first robot is located above a first transfer activity area, the first clamping part transfers a sample test tube or a centrifuge test tube in the first transfer activity area, the second robot is provided with a second clamping part and a liquid transfer part, the second robot is located in a second transfer activity area, the second clamping part transfers a test tube in the second transfer activity area, and the liquid transfer part transfers a solution in the second transfer activity area through a TIP gun tip on the TIP rack; The sample module and the centrifuge tube rack are located in the first transfer activity area, the TIP rack, the step-by-step nucleic acid extraction module, the dual centrifuge module, and the temperature incubation module are located in the second transfer activity area, and the multiple lid opening module is located in the overlapping area; The step-by-step nucleic acid extraction module is disposed on the rack and is used to extract nucleic acid products from the sample; The dual mixing module is arranged on the rack and is used to mix the test tubes; The display control module is used to control the operation of the dual centrifugation module, the dual mixing module, the multiple cover opening module, the dual robotic arm module and the step-by-step nucleic acid extraction module respectively to perform nucleic acid sample processing.

4. A multi-task parallel nucleic acid extraction and conversion machine as claimed in claim 3, characterized in that: The nucleic acid extraction and conversion integrated machine also includes: A liquid adding module, which is arranged on the rack and is used to store reaction additive reagents; 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.

5. The multi-task parallel nucleic acid extraction and conversion integrated machine as claimed in claim 3, characterized in that: The liquid adding module includes a first liquid adding device and a second liquid adding device. The first liquid adding device is arranged in the first transfer activity area and connected to the multiple cover opening module through a pipeline. The second liquid adding device is arranged in the second transfer activity area and connected to the step-by-step nucleic acid extraction module through a pipeline.

6. The multi-task parallel nucleic acid extraction and conversion integrated machine as claimed in claim 3, 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 assembly 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.

7. The multi-task parallel nucleic acid extraction and conversion integrated machine according to claim 1, characterized in that: The dual centrifugal module comprises a low-speed centrifugal device and a high-speed centrifugal device, and both the low-speed centrifugal device and the high-speed centrifugal device are located in the second transfer activity area.

8. The multi-task parallel nucleic acid extraction and conversion integrated machine as claimed in claim 3, characterized in that: The dual mixing module comprises An oscillating device, which is disposed in the second transfer activity area and is used for oscillating and mixing the sample test tube; Inversion mixing device: The inversion mixing device is arranged in the first transfer activity area and is used for inverting and mixing the centrifuge test tube.

9. The multi-task parallel nucleic acid extraction and conversion integrated machine according to claim 8, 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.

10. The multi-task parallel nucleic acid extraction and conversion integrated machine as claimed in claim 3, 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; the second nucleic acid extraction device comprises a second detection kit and a second magnetic rod assembly.

Images