CN117106570A - Nucleic acid extraction and transformation integrated machine capable of editing flow and method thereof - Google Patents
Nucleic acid extraction and transformation integrated machine capable of editing flow and method thereof Download PDFInfo
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- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 123
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 123
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 109
- 238000000605 extraction Methods 0.000 title claims abstract description 89
- 238000000034 method Methods 0.000 title claims abstract description 34
- 230000009466 transformation Effects 0.000 title claims abstract description 12
- 238000012360 testing method Methods 0.000 claims abstract description 161
- 238000002156 mixing Methods 0.000 claims abstract description 59
- 238000001514 detection method Methods 0.000 claims abstract description 34
- 239000007788 liquid Substances 0.000 claims description 44
- 238000012546 transfer Methods 0.000 claims description 36
- 238000005119 centrifugation Methods 0.000 claims description 32
- 239000003153 chemical reaction reagent Substances 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 239000006228 supernatant Substances 0.000 claims description 19
- 238000011534 incubation Methods 0.000 claims description 16
- 239000000047 product Substances 0.000 claims description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 10
- 238000000703 high-speed centrifugation Methods 0.000 claims description 8
- 206010028980 Neoplasm Diseases 0.000 claims description 7
- 201000011510 cancer Diseases 0.000 claims description 6
- 238000000464 low-speed centrifugation Methods 0.000 claims description 5
- 238000009423 ventilation Methods 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 2
- 230000003373 anti-fouling effect Effects 0.000 claims 1
- 230000000249 desinfective effect Effects 0.000 claims 1
- 230000002550 fecal effect Effects 0.000 abstract description 14
- 210000001124 body fluid Anatomy 0.000 abstract description 3
- 239000010839 body fluid Substances 0.000 abstract description 3
- 206010005003 Bladder cancer Diseases 0.000 description 11
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 11
- 201000005112 urinary bladder cancer Diseases 0.000 description 11
- 210000002700 urine Anatomy 0.000 description 11
- 230000000694 effects Effects 0.000 description 7
- 230000009977 dual effect Effects 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
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- 208000019155 Radiation injury Diseases 0.000 description 1
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- 230000006037 cell lysis Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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Abstract
The application discloses a nucleic acid extraction and transformation all-in-one machine capable of editing a process, which is characterized in that a machine frame is provided with a machine shell, an accommodating space is formed by surrounding the machine shell, a sample module is used for replacing and placing a sample test tube, a TIP is mounted on the machine frame and used for placing a TIP gun head, a centrifuge tube rack is used for placing a centrifuge test tube, a double-centrifuge module is used for centrifuging the test tube, a multiple cover opening module is used for opening and closing a tube cover of the test tube, a double-mechanical arm module is used for transferring the test tube and transferring a solution through the TIP gun head, and a step-by-step nucleic acid extraction module is used for extracting nucleic acid products of a sample; the double mixing module is used for mixing test tubes uniformly, and can be suitable for most sample detection such as fecal samples, body fluid samples, swab samples and the like by replacing the sample module and editing the test flow.
Description
Technical Field
The application relates to the technical field of nucleic acid extraction, in particular to a nucleic acid extraction and transformation integrated machine capable of editing a flow and a method thereof.
Background
Nucleic acid research is an important subject in modern biology and medicine research, nucleic acid is taken as a carrier of genetic information, is a material basis of gene expression, has very important roles in life activities such as normal growth, development and reproduction of organisms, and has close relation with abnormal conditions of life, such as tumorigenesis, radiation injury, genetic diseases and the like.
CN202110643507.9 discloses an integrated machine for nucleic acid extraction and PCR detection, which prevents the nucleic acid extraction module and PCR detection module from being polluted by external air by arranging a first filter at the air inlet; through setting up deep bead, first wind channel, second wind channel, diversion wind channel, make gaseous at first through nucleic acid extraction module, later through PCR detection module to prevent that the aerosol of PCR product from polluting nucleic acid extraction module, avoid cross contamination, but he can only detect single nucleic acid sample, and the product function is single, can't satisfy multiple disease detection item, the detection demand of different samples.
Therefore, a nucleic acid extraction and conversion integrated machine integrating multiple detection modules and capable of detecting editable flows of multiple disease samples is designed
Disclosure of Invention
In order to solve the technical problems, the application adopts the following technical scheme:
a nucleic acid extraction conversion all-in-one machine of editable flow, the nucleic acid extraction conversion all-in-one machine comprising: the machine comprises a rack, wherein a shell is arranged on the rack, and the shell surrounds an accommodating space;
the sample module is arranged on the rack and used for placing a sample test tube;
the TIP frame is arranged on the rack and used for placing the TIP gun head;
the centrifuge tube rack is arranged on the rack and used for placing the centrifuge test tubes;
the double-centrifugal module is arranged on the rack and is used for centrifuging the test tube;
the multiple cover opening modules are arranged on the rack and used for opening and closing tube covers of the test tubes;
the double mechanical arm module is arranged on the rack and used for transferring test tubes and transferring solutions through the TIP gun head;
the step-by-step nucleic acid extraction module is arranged on the rack and is used for extracting nucleic acid products of a sample;
the double mixing module is arranged on the rack and used for mixing the test tubes uniformly;
a display control module; the display control module is used for respectively controlling the double centrifugal modules, the double mixing module, the multiple uncovering module, the double mechanical arm module and the step-by-step nucleic acid extraction module to operate so as to process nucleic acid samples.
Preferably, the multiple door module includes a first door assembly and a second door assembly;
the first door assembly includes: the first slide rail is provided with a movable sample clamp
A holding portion adapted to hold a sample tube;
the first lifting platform is arranged above the first sliding rail, and is provided with a first liquid adding needle head and a first rotary clamping part capable of lifting, and the first rotary clamping part is suitable for opening or closing a tube cover of a sample test tube.
The second door assembly includes:
the second slide rail is provided with a movable centrifugal test tube clamping part, and the centrifugal test tube clamping part is suitable for clamping a tube body of a centrifugal test tube;
the second lifting platform is arranged above the second sliding rail, and is provided with a liquid adding needle head and a liftable second rotary clamping part, and the second rotary clamping part is suitable for opening or closing a tube cover of the centrifugal test tube.
Preferably, the double mixing module includes:
the vibration device is arranged on the rack and is used for uniformly vibrating and mixing the sample test tubes;
reversing and evenly mixing device: the reversing and evenly mixing device is arranged on the frame and is used for reversing and evenly mixing the centrifugal test tube.
Preferably, the reverse mixing device comprises: a fixed bracket;
the rotating bracket is arranged above the fixed bracket, a plurality of centrifugal test tube holes are formed in the rotating bracket, a pair of elastic plates used for fixing the centrifugal test tubes are arranged at the edges of the centrifugal test tube holes, and two ends of the rotating bracket penetrate through the fixed bracket and are in rotary connection with the fixed bracket;
the mixing motor is arranged below the fixed support and is connected with the end part of the rotating support through a conveying belt.
Preferably, the dual-mechanical arm module comprises: the device comprises a first mechanical arm and a second mechanical arm, wherein a first clamping part is arranged on the first mechanical arm, a second clamping part and a liquid transferring part are arranged on the second mechanical arm, the second mechanical arm is provided with a second transferring active area, the second clamping part transfers test tubes in the second transferring active area, and the liquid transferring part transfers solution in the second transferring active area through a TIP gun head on a TIP frame.
Preferably, the dual centrifugal module includes a low-speed centrifugal device and a high-speed centrifugal device.
Preferably, the nucleic acid extraction and conversion integrated machine further comprises an anti-pollution sterilizing device, wherein the anti-pollution sterilizing device comprises a UV lamp, an activated carbon filter and a directional ventilation device which are arranged at the top of the shell.
Preferably, the step-wise 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 bar assembly, and the second nucleic acid extraction device comprises a second detection kit and a second magnetic bar assembly.
Preferably, the nucleic acid extraction and transformation integrated machine further comprises:
the liquid adding module comprises a first liquid adding device and a second liquid adding device, the first liquid adding device is arranged on the rack and is connected with the multiple cover opening modules through pipelines, and the second liquid adding device is arranged on the rack and is connected with the step-by-step nucleic acid extraction modules through pipelines;
the temperature incubation module is arranged on the rack and used for controlling the temperature of the test tube for preservation; the display control module is also used for controlling the operation of the liquid adding module and the temperature incubation module so as to process the nucleic acid sample.
A method of an integrated nucleic acid extraction and transformation machine capable of editing a process, the method comprising the steps of:
s1 sample feeding: mounting a sample support carrying a plurality of sample tubes on a sample frame, clamping and moving the sample tubes to the sample clamping parts by a first clamping part on a first mechanical arm, clamping and fixing the sample tubes by the sample clamping parts, and moving the sample tubes to a second transferring active area along a first sliding rail;
s2, oscillating low-speed centrifugation: the second mechanical arm moves the sample test tube to the vibration device for vibration, uniformly mixes the vibration of the sample test tube, and then moves the sample test tube to the low-speed centrifugal device for centrifugation;
s3, transferring centrifugate: after the centrifugation of the sample tube is completed, the second mechanical arm transfers the sample tube to the sample clamping part, the sample clamping part moves to the position right below the first rotary clamping part, the first rotary clamping part opens the tube cover of the sample tube, the first mechanical arm clamps the first centrifugal tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotary clamping part opens the tube cover of the first centrifugal tube, and the pipetting part on the second mechanical arm transfers the supernatant of the sample tube to the first centrifugal tube through the tip gun head;
s4, adding a reaction reagent: the liquid adding needle adds reagent into the first centrifugal test tube, the second rotary clamping part closes the centrifugal test tube cover, and the first rotary clamping part closes the sample test tube cover;
s5, reversely mixing and high-speed centrifuging: the first mechanical arm moves the first centrifugal test tube to the reversing and mixing device, after full mixing, the first mechanical arm moves the first centrifugal test tube to the centrifugal test tube clamping part, the centrifugal test tube clamping part moves to the second transferring movable area along the second sliding rail, and the centrifugal test tube is moved to the high-speed centrifugal device through the second mechanical arm for high-speed centrifugation;
s6, transferring centrifugate: the first mechanical arm clamps a second centrifugal test tube on the centrifugal tube rack to a centrifugal test tube clamping part, the second rotary clamping part opens a second centrifugal test tube cover, and the upper pipetting part of the second mechanical arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through a tip gun head;
s7, transferring a second centrifugal test tube to a temperature incubation module by a second mechanical arm;
s8, after incubation timing in one of the centrifugal test tubes is completed, carrying out an extraction step: the second mechanical 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.
Preferably, the display control module deletes or repeats at least one of the steps S2, S3, S4, S5 or S6 multiple times according to the detection flow of different cancer species; or, the execution sequence of the above S2, S3, S4, S5, S6 is readjusted according to the detection flow of different cancer species.
The beneficial effects are that:
1. the sample module, the double-centrifugation module, the multiple uncovering module, the double-mechanical arm module, the step-by-step nucleic acid extraction module and the double-mixing module are arranged, so that the device can be suitable for detecting most samples, such as fecal samples, body fluid samples, swab samples and the like, only by replacing the sample module and editing the test flow, and compared with the existing single-flow products, the whole device integrates multiple detection modes in one machine, thereby reducing the cost of instruments and saving the space;
2. under the condition that the current nucleic acid extraction step flow is not affected, the next sample test tube batch is replaced, meanwhile, the sample test tube pretreatment operation is started, the resources of each functional module of the system are utilized to the greatest extent, the waiting time in the original automatic processing process is shortened, the parallel processing of multiple batches of samples can be realized, the detection productivity of the system is greatly improved, the detection labor cost is effectively reduced, and the sample detection time is shortened.
And thirdly, 16 samples can be placed into the device at one time, a nucleic acid template can be obtained after the samples enter for a period of time, full-flow automation is realized, further the labor intensity of experimental staff is reduced, and the stability and consistency of experimental results are improved.
Drawings
In order to more clearly illustrate the embodiments of the application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic overall structure of an embodiment of the present application.
FIG. 2 is a schematic diagram of an internal structure of an embodiment of the present application;
FIG. 3 is a schematic diagram showing an internal structure of a second embodiment of the present application;
FIG. 4 is a schematic view illustrating the partitioning of an accommodating space according to an embodiment of the present application;
FIG. 5 is a schematic diagram of a dual-robot module according to an embodiment of the application;
fig. 6 is a schematic view of the overall structure of a multiple-lid module according to an embodiment of the present application;
fig. 7 is a schematic front view of a multiple lid module according to an embodiment of the present application;
fig. 8 is a schematic view of the overall structure of an inverted mixing device according to an embodiment of the present application;
FIG. 9 is a schematic view showing the overall structure of an inverted mixing device according to an embodiment of the present application;
FIG. 10 is a schematic diagram showing the overall structure of a sample module according to an embodiment of the present application;
FIG. 11 is a schematic diagram of the overall flow of the method of the present application;
in the figure: a frame 1, a lifting door 101, a connecting arm 102, a first transferring active area 100, a second active area 200, overlap zone 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, multiple lid opening module 6, first lid opening assembly 61, first slide rail 611, sample clamping portion 612, first lifting table 613, first rotational clamping portion 615, second lid opening assembly 62, second slide rail 621, centrifuge tube clamping portion 622, second lifting table 623, filling needle 624, second rotational clamping portion 625, dual robotic arm module 7, first robotic arm 71, first clamping portion 710, first X-axis arm 711, first Y-axis arm 712, first Z-axis arm 713, second robotic arm 72, second clamping portion 720 pipetting unit 721, second X-axis arm 722, second Y-axis arm 723, second Z-axis arm 724, step-wise nucleic acid extraction module 8, first nucleic acid extraction device 81, first detection kit 811, first magnetic bar assembly 812, second nucleic acid extraction device 82, second detection kit 821, second magnetic bar assembly 822, double mixing module 9, oscillation device 91, inverted mixing device 92, centrifugal test tube hole 920, fixed support 921, rotating support 922, mixing motor 923, conveyor belt 924, elastic plate 925, display control module 10, liquid feeding module 11, first liquid feeding device 111, second liquid feeding device 112, temperature incubation module 12, UV lamp 13, activated carbon filter 14, directional ventilation device 15.
Detailed Description
The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
Examples: as shown in fig. 1-3: a frame 1, a sample module 2, a TIP frame 3, a centrifuge tube rack 4, a double-centrifuge module 5, a multi-cover opening module 6, a double-mechanical arm module 7, a step-by-step nucleic acid extraction module 8, a double-mixing module 9 and a display control module 10,
a nucleic acid extraction conversion all-in-one machine of editable flow, the nucleic acid extraction conversion all-in-one machine comprising: the device comprises a rack 1, wherein a shell is arranged on the rack, and the shell surrounds an accommodating space;
the sample module 2 is arranged on the rack and is used for placing a sample test tube;
the TIP frame 3 is arranged on the rack and used for placing a TIP gun head;
the centrifuge tube rack 4 is arranged on the rack and used for placing the centrifuge test tubes;
the double-centrifugal module 5 is arranged on the rack and is used for centrifuging the test tube;
the multiple cover opening modules 6 are arranged on the rack and used for opening and closing tube covers of the test tubes;
the double mechanical arm module 7 is arranged on the rack and is used for transferring test tubes and transferring solutions through the TIP gun head;
a step-wise nucleic acid extraction module 8 provided to the rack and configured to extract a nucleic acid product of a sample;
the double mixing module 9 is arranged on the rack and is used for mixing test tubes uniformly;
the display control module 10 is used for respectively controlling the double centrifugal modules 5, the double mixing module 7, the multiple cover opening module 6, the double mechanical arm module 7 and the step-by-step nucleic acid extraction module 8 to operate so as to process nucleic acid samples. The whole size of the nucleic acid extraction and transformation integrated machine is 1800mm long by 800mm wide by 1100mm high and 340KG in weight. The shell is provided with a lifting door 101 and a multidirectional adjusting connecting arm 102, one end of the multidirectional adjusting connecting arm 102 is connected with a display control module 10, the display control module 10 is a 15.6-inch touch integrated machine, a processor is arranged in the shell, the operation flow of all modules is controlled, and experimental intermediate data are recorded.
The nucleic acid extraction and conversion integrated machine further comprises an anti-pollution sterilizing device, wherein the anti-pollution sterilizing device comprises a UV lamp 13, an activated carbon filter 14 and a directional ventilation device 15 which are arranged at the top of the shell
As shown in fig. 1-4: the accommodation space is divided into a first transfer active region 100, a second active region 200, and an overlap region 300; wherein the sample module 2 and the centrifuge tube rack 4 are located in a first transfer active area 100, the TIP rack 3, the step-by-step nucleic acid extraction module 8 and the dual centrifugation module 5 are located in a second transfer active area, and the multiple lid opening module 6 is located in the overlap area.
The sample tube is provided with 16 sample tube grooves, 16 samples can be placed into the device of the application once in test, and a nucleic acid template can be obtained after the samples enter for a period of time, so that the full-flow automation is realized, the labor intensity of experimental staff is reduced, and the stability and consistency of experimental results are improved.
As shown in fig. 2-3: in one embodiment of the present application, the step-wise 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 bar assembly 812; the second nucleic acid extraction device 82 includes a second detection kit 821 and a second magnetic rod assembly 822. The detection kit and the magnetic bar assembly in the first nucleic acid extraction device 81 and the second nucleic acid extraction device 82 can process a sample or a processed sample to extract a nucleic acid product of the sample, the nucleic acid product extracted based on different sampling samples can be used for diagnosing a corresponding disease, for example, the sampling samples are nucleic acid samples obtained by a throat swab sample collection method or a nose swab sample collection method, and the nucleic acid extraction module can process steps of cell lysis, inhibitor cleaning, nucleic acid elution and the like on the nucleic acid samples to obtain the nucleic acid product, wherein the nucleic acid product of the nucleic acid sample can be used for detecting novel coronavirus COVID-19. For another example, the sample is a stool sample, and the nucleic acid extraction module 8 may perform steps such as specific binding of nucleic acid, inhibitor washing, and nucleic acid elution on the stool sample after the treatment to obtain a nucleic acid product, and the nucleic acid product of the stool sample may be used for detecting intestinal cancer.
As shown in fig. 5-6: in an embodiment of the present application, the multiple door module 6 includes a first door assembly 61 and a second door assembly 62;
the first lid opening assembly 61 includes: a first slide rail 611, on which first slide rail 611 a movable sample clamping portion 612 is provided, which is adapted to clamp a sample tube;
the first lifting platform 613 is arranged above the first sliding rail, and a first rotary clamping part 615 capable of lifting is arranged on the first lifting platform, and the first rotary clamping part 615 is suitable for opening or closing a tube cover of a sample test tube.
The second door 62 assembly includes:
a second sliding rail 621, wherein a movable centrifugal test tube clamping part 622 is arranged on the second sliding rail 621, and the centrifugal test tube clamping part 622 is suitable for clamping a tube body of a centrifugal test tube;
the second lifting platform 623, the second lifting platform 623 is disposed above the second sliding rail 621, and the second lifting platform 623 is provided with a liquid adding needle 624 and a liftable second rotary clamping portion 625, and the second rotary clamping portion is suitable for opening or closing a tube cover of the centrifugal test tube. The centrifuge tube may be a 2ml tube and the sample tube may be a 50ml fecal sample tube, the sample grip 612 being sized larger than the second sample grip 622 and the first rotational grip 615 being sized larger than the second rotational grip 625.
As shown in fig. 3 and 7: in an embodiment of the present application, the dual-robot module 7 includes: the first mechanical arm 71 and the second mechanical arm 72, the first mechanical arm 71 is provided with a first clamping part 710, the first mechanical arm 71 is located above the first transferring active area 100, the first clamping part 710 transfers a sample test tube or a centrifugal test tube in the first transferring active area, the first mechanical arm is an mechanical arm with XYZ axes moving freely in three dimensions, the first mechanical arm 71 further comprises a first X-axis arm 711, a first Y-axis arm 712 and a first Z-axis arm 713, the first clamping part 710 moves in the left-right direction on the first X-axis arm 711, the first clamping part 710 moves in the front-back direction on the first Y-axis arm 712, and the first clamping part 710 moves in the up-down direction on the first Y-axis arm 713;
the first X-axis arm 711 is mounted on the first Y-axis arm 713, the first Y-axis arm 713 may drive the first X-axis arm 711 to move back and forth, the first Z-axis arm 713 is mounted on the first X-axis arm 711, the first X-axis arm 711 may drive the first Z-axis arm 713 to move left and right, the first Z-axis arm 713 is connected with the first clamping part 710, and the first Z-axis arm 713 may drive the first clamping part 710 to move up and down, thereby realizing the transfer of the first clamping part 710 into the sample tube and the centrifuge tube in the first transfer active area.
A second clamping part 720 and a pipetting part 721 are arranged on the second mechanical arm 72, the second mechanical arm 72 is positioned in a second transferring active area 200, the second clamping part 72 transfers test tubes in the second transferring active area 200, and the pipetting part 721 transfers solution in the second transferring active area through a TIP gun head on the TIP frame 3; the second robot arm 72 may be a three-axis robot arm, and the second robot 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 extending in the left-right direction, the second Y-axis arm 723 moving in the front-rear direction, and the second Y-axis arm 723 moving in the up-down direction. The second X-axis arm 722 may be connected to the frame 1, the second Y-axis arm 723 is mounted on the second X-axis arm 722, the second X-axis arm 722 may drive the second Y-axis arm 723 to move left and right, the second Z-axis arm 724 is mounted on the second Y-axis arm 723, the second Y-axis arm 723 may drive the second Z-axis arm 724 to move back and forth, the second Z-axis arm 724 includes two Z-axis lifting arms, one of the Z-axis lifting arms is provided with the second clamping portion 720, the other Z-axis lifting arm is provided with the pipetting portion 721, and the second Z-axis arm 724 may drive the second clamping portion 720 and the pipetting portion 721 to move up and down independently, so as to realize that the second clamping portion 721 transfers the test tube in the second transferring active area, and the pipetting portion 722 transfers the solution through the TIP in the second transferring active area.
The display control module 10 is used for respectively controlling the operation of the double centrifugal module 5, the double mixing module 9, the multiple cover opening module 6, the double mechanical arm module 7 and the step-by-step nucleic acid extraction module 8 so as to process nucleic acid samples.
When carrying out nucleic acid sample pretreatment, arm module 7 shifts to the multiple sample module 6 that uncaps with the nucleic acid sample test tube by sample module 2, uncaps the nucleic acid sample test tube through multiple uncap module 6, and arm module 7 rethread TIP rifle head shifts the solution in the nucleic acid sample test tube to nucleic acid extraction module 8 to extract the nucleic acid product through the nucleic acid extraction module, from this, realized the full-flow automation of the pretreatment of sample, and then be favorable to reducing experimenter intensity of labour, promote experimental result stability and uniformity.
As shown in fig. 2: in an embodiment of the application, the nucleic acid extraction and transformation integrated machine further includes:
the liquid adding module 11 is arranged on the rack and is used for storing reaction additive reagents; in an embodiment of the present application, the liquid adding module 11 includes a first liquid adding device 111 and a second liquid adding device 112, where the first liquid adding device 111 is disposed in the first transferring active area 100 and connected to the multiple cover opening modules 6 through a pipeline, and the first liquid adding device 111 is located near the multiple cover opening modules 6, and specifically, the first liquid adding device 111 provides liquid adding power for the liquid adding needle 624;
the second liquid adding device 112 is positioned in the second transferring active area 200 and is connected with the step-by-step nucleic acid extraction module 8 through a pipeline.
The temperature incubation module 12 is in the second transfer active zone 200, and is used for providing a constant temperature incubation environment for the test tube;
the display control module 10 is also used for controlling the operation of the first liquid adding device 111, the second liquid adding device 112 and the temperature incubation module 12 for nucleic acid sample treatment.
The nucleic acid extraction and conversion integrated machine further comprises an anti-pollution sterilizing device, wherein the anti-pollution sterilizing device comprises a UV lamp 13, an activated carbon filter 14 and a directional ventilation device 15 which are arranged at the top of the shell.
In an embodiment of the present application, the dual centrifugation module 5 is located in the second transfer activity zone 200, and includes a low-speed centrifugation device 51 and a high-speed centrifugation device 52, wherein the centrifugation speed of the low-speed centrifugation device 51 may be 8000rpm or less for the centrifugation of the sample tube, the centrifugation speed of the high-speed centrifugation device 52 may be 15000rpm to 30000rpm, and the centrifugation speed of the high-speed centrifugation device is used for the centrifugation delamination of the sample tube.
As shown in fig. 2: in one embodiment of the present application, the dual mixing module 9 comprises
The oscillation device 91 is arranged in the second transferring active area and is used for evenly mixing the sample test tubes in an oscillating way;
the mixing device 92 is reversed: the inversion mixing device is arranged in the first transfer active area and is used for inversion mixing of the centrifugal test tube.
As shown in fig. 8-9: in one embodiment of the present application, the reverse blending device 92 includes: a fixing bracket 921;
the rotating bracket 922 is arranged above the fixed bracket 921, a plurality of centrifugal test tube holes 920 are formed in the rotating bracket 922, the number of the centrifugal test tube holes is 4*4 square, 16 in total, a pair of elastic plates 925 for fixing the centrifugal test tubes are arranged at the edges of the centrifugal test tube holes, and two ends of the rotating bracket 922 penetrate through the fixed bracket 921 and are in rotary connection with the fixed bracket 921;
and the mixing motor 923, the mixing motor 923 is arranged below the fixed support 921, the mixing motor 923 is connected with the end part of the rotating support 922 through a conveying belt 924, and the mixing motor 923 drives the rotating support 922 to rotate through the conveying belt 924 so as to invert the mixing centrifugal test tube.
As shown in fig. 10: the application relates to a sample module 2, wherein the sample module 2 comprises a sample frame 21 and a sample support 22, the sample support 22 is clamped on a clamping groove 20 on the sample frame 22, the sample support 22 in the embodiment is a support for a fecal sample test tube, and of course, the sample support using other items, such as a nucleic acid sample and a body fluid sample, can be replaced, and different processing flows are designed according to different samples, so that different modules are called.
FIG. 11 is a comprehensive flow chart of the method of the application, a method for operating an integrated nucleic acid extraction and conversion machine capable of editing a flow,
example 1: when the sample to be detected is a fecal sample, the method comprises the steps of:
s1 sample feeding: installing a sample support carrying a plurality of fecal sample tubes, which are 50ml tubes, on a sample frame, clamping and moving the sample tubes to the sample clamping parts by a first clamping part on a first mechanical arm, clamping and fixing the sample tubes by the sample clamping parts, and moving the sample tubes to a second transfer activity area along a first sliding rail;
s2, oscillating low-speed centrifugation: the second mechanical arm moves the sample test tube to the vibration device for vibration, uniformly mixes the vibration of the sample test tube, and then moves the sample test tube to the low-speed centrifugal device for centrifugation;
s3, transferring centrifugate: after the centrifugation of the sample tube is completed, the second mechanical arm transfers the sample tube to the sample clamping part, the sample clamping part moves to the position right below the first rotary clamping part, the first rotary clamping part opens the tube cover of the sample tube, the first mechanical arm clamps the first centrifugal tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotary clamping part opens the tube cover of the first centrifugal tube, and the pipetting part on the second mechanical arm transfers the supernatant of the sample tube to the first centrifugal tube through the tip gun head;
s4, adding a reaction reagent: the liquid adding needle adds reagent into the first centrifugal test tube, the second rotary clamping part closes the centrifugal test tube cover, and the first rotary clamping part closes the sample test tube cover;
s5, reversely mixing and high-speed centrifuging: the first mechanical arm moves the first centrifugal test tube to the reversing and mixing device, after full mixing, the first mechanical arm moves the first centrifugal test tube to the centrifugal test tube clamping part, the centrifugal test tube clamping part moves to the second transferring movable area along the second sliding rail, and the centrifugal test tube is moved to the high-speed centrifugal device through the second mechanical arm for high-speed centrifugation;
s6, transferring centrifugate: the first mechanical arm clamps a second centrifugal test tube on the centrifugal tube rack to a centrifugal test tube clamping part, the second rotary clamping part opens a second centrifugal test tube cover, and the upper pipetting part of the second mechanical arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through a tip gun head;
s7, transferring a second centrifugal test tube to a temperature incubation module by a second mechanical arm;
s8, after incubation timing in one of the centrifugal test tubes is completed, carrying out an extraction step: the second mechanical 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.
According to the detection flow of different cancer types, the display control module deletes or repeatedly repeats at least one step of S2, S3, S4, S5 or S6; or, the execution sequence of the above S2, S3, S4, S5, S6 is readjusted according to the detection flow of different cancer species.
Example 1: and when the detection sample is a bladder cancer urine sample, the steps S2 and S3 are canceled, and the steps S4, S5 and S6 are repeated for 2 times.
Sample feeding: a sample bracket carrying a plurality of bladder cancer urine sample test tubes is arranged on a sample frame, the bladder cancer urine sample test tubes are 2ml test tubes, a first clamping part on a first mechanical arm clamps and moves the bladder cancer urine sample test tubes to a sample clamping part, and the sample clamping part clamps and fixes the sample test tubes and moves to a second transfer activity area along a first sliding rail;
adding a reaction reagent: the liquid adding needle adds a reagent into the bladder cancer urine sample tube, and the second rotary clamping part closes the tube cover of the bladder cancer urine sample tube;
mixing and high-speed centrifuging in a reversed way: the first mechanical arm moves the bladder cancer urine sample tube to the reversing and mixing device, after full mixing, the first mechanical arm moves the bladder cancer urine sample tube to the centrifugal tube clamping part, the centrifugal tube clamping part moves to the second transferring activity area along the second sliding rail, and the bladder cancer urine sample tube is moved to the high-speed centrifugal device for high-speed centrifugation through the second mechanical arm;
transfer centrifugate: the first mechanical arm clamps a first centrifugal test tube on the centrifugal tube rack to a centrifugal test tube clamping part, the second rotary clamping part opens a first centrifugal test tube cover, and the upper pipetting part of the second mechanical arm transfers the supernatant of the bladder cancer urine sample test tube to the first centrifugal test tube through a tip gun head;
the reaction reagent is added again: the liquid adding needle adds a reagent into the first centrifugal test tube, the second rotary clamping part closes the tube cover of the first centrifugal test tube, and the first rotary clamping part closes the tube cover of the bladder cancer urine sample test tube;
mixing and high-speed centrifuging in a reversed way: the first mechanical arm moves the first centrifugal test tube to the reversing and mixing device, after full mixing, the first mechanical arm moves the first centrifugal test tube to the centrifugal test tube clamping part, the centrifugal test tube clamping part moves to the second transferring movable area along the second sliding rail, and the centrifugal test tube is moved to the high-speed centrifugal device through the second mechanical arm for high-speed centrifugation;
transfer centrifugate: the first mechanical arm clamps a second centrifugal test tube on the centrifugal tube rack to a centrifugal test tube clamping part, the second rotary clamping part opens a second centrifugal test tube cover, and the upper pipetting part of the second mechanical arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through a tip gun head;
transferring the second centrifugal test tube to a temperature incubation module by a second mechanical arm;
after incubation in one of the centrifuge tubes was timed, the extraction step was performed: the second mechanical 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.
Meanwhile, according to the resource requirement condition of the current task flow, under the condition that the current task execution step flow is not influenced, the next new parallel step is started at the same time, the resources of all functional modules of the system are utilized to the maximum extent, parallel processing of multiple batches of samples can be realized, the detection productivity of the system is greatly improved, the detection labor cost is effectively reduced, the sample detection time is shortened, and the method has positive significance in promoting popularization of early detection of tumors.
The fecal sample pretreatment method comprises the following steps: a first centrifugation step: after the first reagent and the second reagent are added into the fecal sample tube, the fecal sample tube is oscillated and centrifuged to obtain a centrifuged first supernatant. Wherein 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. And a second centrifugation step: and adding a third reagent into the first supernatant, and oscillating and centrifuging the first supernatant liquid added with the third reagent to obtain a centrifuged second supernatant. Wherein the second reagent may be an inhibitor removing solution for removing PCR inhibitors such as polysaccharides and phenols. And a third centrifugation step: and adding a fourth reagent into the second supernatant, and oscillating, incubating at a controlled temperature and centrifuging the second supernatant added with the fourth reagent. Wherein the fourth reagent may be isopropanol which forms a flocculent precipitate of the nucleic acid; discarding the supernatant after centrifugation in the third centrifugation step, adding a fifth reagent into the residual liquid, oscillating the residual liquid added with the fifth reagent, and extracting a nucleic acid product from the oscillated residual liquid. Wherein the fifth reagent may be a lysis binding solution providing conditions for adsorbing nucleic acids to the magnetic beads.
In some embodiments of the present application, in the first centrifugation step, the first supernatant after centrifugation is obtained by centrifugation and supernatant extraction is repeated a plurality of times, and the number of times of repetition may be two times, so as to ensure the centrifugation effect in the first centrifugation step. In some embodiments of the present application, in the second centrifugation step, the second supernatant after centrifugation is obtained by centrifugation and supernatant extraction is repeated a plurality of times, and the number of times of repetition may be two, so as to ensure the centrifugation effect in the second centrifugation step.
When the pretreatment of the fecal sample is carried out, the nucleic acid product of the fecal sample is obtained through the first centrifugation step, the second centrifugation step, the third centrifugation step and the extraction step in sequence, so that the full-flow automation of the pretreatment of the fecal sample is realized, the labor intensity of experimental personnel is reduced, and the stability and consistency of experimental results are improved.
In addition, according to the fecal sample pretreatment method provided by the embodiment of the application, the nucleic acid extraction and conversion device can realize one-key intelligent operation through the display screen, the display screen can be provided with an intelligent operation interface, the digital storage record of the whole-flow experimental information can be realized, and the automation, informatization and intelligent construction of a laboratory are improved.
The application has the advantages of modular design and flexible configuration, capability of increasing or decreasing corresponding modules as required, higher compatibility, capability of being compatible with various sample test tubes and centrifugal test tubes by replacing different sample modules and clamps. The application also has good safety and stability, station integrated material in-place detection function, and can identify that consumable materials are placed in place and accurately, and the tube cover is opened and closed correctly. Meanwhile, according to the fecal sample pretreatment method provided by the embodiment of the application, the test is accurate and efficient, the operation can be carried out continuously for 24 hours, and one batch can be completed every 1.5 hours.
The speed of detecting 16 samples by the traditional manual efficiency is 6 hours, the device is simple and convenient to operate, and 5 devices can be simultaneously operated and observed by a single person, so that the detection speed is greatly improved in efficiency, and the device is more suitable for the requirements of large batch and high detection timeliness.
The technical features of the above embodiments may be combined arbitrarily, and the steps of the method are not limited to the execution sequence, so that all possible combinations of the technical features in the above embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description of the present specification.
The foregoing examples illustrate only a few embodiments of the application and are described in detail herein without thereby limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of the application should be assessed as that of the appended claims.
Claims (10)
1. The nucleic acid extraction conversion all-in-one machine capable of editing a flow is characterized by comprising: the machine comprises a rack, wherein a shell is arranged on the rack, and the shell surrounds an accommodating space; the sample module is arranged on the rack and used for placing a sample test tube;
the TIP frame is arranged on the rack and used for placing the TIP gun head;
the centrifuge tube rack is arranged on the rack and used for placing the centrifuge test tubes;
the double-centrifugal module is arranged on the rack and is used for centrifuging the test tube;
the multiple cover opening modules are arranged on the rack and used for opening and closing tube covers of the test tubes; the double mechanical arm module is arranged on the rack and used for transferring test tubes and transferring solutions through the TIP gun head;
the step-by-step nucleic acid extraction module is arranged on the rack and is used for extracting nucleic acid products of a sample;
the double mixing module is arranged on the rack and used for mixing the test tubes uniformly;
a display control module; the display control module is used for respectively controlling the double centrifugal modules, the double mixing module, the multiple uncovering module, the double mechanical arm module and the step-by-step nucleic acid extraction module to operate so as to process nucleic acid samples.
2. The flow-editable nucleic acid extraction conversion all-in-one machine of claim 1, wherein the multiple lid opening module comprises a first lid opening assembly and a second lid opening assembly;
the first door assembly includes: the sample clamping device comprises a first sliding rail, a second sliding rail and a first sliding rail, wherein a movable sample clamping part is arranged on the first sliding rail and is suitable for clamping a sample test tube;
the first lifting platform is arranged above the first sliding rail, and is provided with a first rotary clamping part capable of lifting, and the first rotary clamping part is suitable for opening or closing a tube cover of the sample test tube;
the second door assembly includes:
the second slide rail is provided with a movable centrifugal test tube clamping part, and the centrifugal test tube clamping part is suitable for clamping a tube body of a centrifugal test tube;
the second lifting platform is arranged above the second sliding rail, and is provided with a liquid adding needle head and a liftable second rotary clamping part, and the second rotary clamping part is suitable for opening or closing a tube cover of the centrifugal test tube.
3. The integrated nucleic acid extraction and transformation machine capable of editing a process according to claim 1, wherein the double mixing module comprises
The vibration device is arranged on the rack and is used for uniformly vibrating and mixing the sample test tubes;
reversing and evenly mixing device: the reversing and evenly mixing device is arranged on the frame and is used for reversing and evenly mixing the centrifugal test tube.
4. The integrated machine for extracting and converting nucleic acid capable of editing a process according to claim 3, wherein said reverse mixing device comprises: a fixed bracket;
the rotating bracket is arranged above the fixed bracket, a plurality of centrifugal test tube holes are formed in the rotating bracket, a pair of elastic plates used for fixing the centrifugal test tubes are arranged at the edges of the centrifugal test tube holes, and two ends of the rotating bracket penetrate through the fixed bracket and are in rotary connection with the fixed bracket;
the mixing motor is arranged below the fixed support and is connected with the end part of the rotating support through a conveying belt.
5. The integrated nucleic acid extraction and transformation machine capable of editing a process of claim 1, wherein the double-mechanical arm module comprises: the device comprises a first mechanical arm and a second mechanical arm, wherein a first clamping part is arranged on the first mechanical arm, a second clamping part and a liquid transferring part are arranged on the second mechanical arm, the second mechanical arm is provided with a second transferring active area, the second clamping part transfers test tubes in the second transferring active area, and the liquid transferring part transfers solution in the second transferring active area through a TIP gun head on a TIP frame.
6. The flow-editable nucleic acid extraction and conversion all-in-one machine of claim 1, further comprising an anti-fouling and disinfecting device comprising a UV lamp, an activated carbon filter, and a directional ventilation device disposed at the top of the housing.
7. The integrated nucleic acid extraction and conversion machine capable of editing a process according to claim 1, wherein 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 bar assembly, and the second nucleic acid extraction device comprises a second detection kit and a second magnetic bar assembly.
8. The integrated nucleic acid extraction and transformation machine capable of performing an editable process according to claim 7, further comprising:
the liquid adding module comprises a first liquid adding device and a second liquid adding device, the first liquid adding device is arranged on the rack and is connected with the multiple cover opening modules through pipelines, and the second liquid adding device is arranged on the rack and is connected with the step-by-step nucleic acid extraction modules through pipelines;
the temperature incubation module is arranged on the rack and used for controlling the temperature of the test tube for preservation;
the display control module is also used for controlling the operation of the liquid adding module and the temperature incubation module so as to process the nucleic acid sample.
9. The method of an integrated nucleic acid extraction and transformation machine capable of performing an editable procedure according to any one of claims 1 to 8, wherein the method comprises the steps of:
s1 sample feeding: mounting a sample support carrying a plurality of sample tubes on a sample frame, clamping and moving the sample tubes to the sample clamping parts by a first clamping part on a first mechanical arm, clamping and fixing the sample tubes by the sample clamping parts, and moving the sample tubes to a second transferring active area along a first sliding rail;
s2, oscillating low-speed centrifugation: the second mechanical arm moves the sample test tube to the vibration device for vibration, uniformly mixes the vibration of the sample test tube, and then moves the sample test tube to the low-speed centrifugal device for centrifugation;
s3, transferring centrifugate: after the centrifugation of the sample tube is completed, the second mechanical arm transfers the sample tube to the sample clamping part, the sample clamping part moves to the position right below the first rotary clamping part, the first rotary clamping part opens the tube cover of the sample tube, the first mechanical arm clamps the first centrifugal tube on the centrifuge tube rack to the centrifuge tube clamping part, the second rotary clamping part opens the tube cover of the first centrifugal tube, and the pipetting part on the second mechanical arm transfers the supernatant of the sample tube to the first centrifugal tube through the tip gun head;
s4, adding a reaction reagent: the liquid adding needle adds reagent into the first centrifugal test tube, the second rotary clamping part closes the centrifugal test tube cover, and the first rotary clamping part closes the sample test tube cover;
s5, reversely mixing and high-speed centrifuging: the first mechanical arm moves the first centrifugal test tube to the reversing and mixing device, after full mixing, the first mechanical arm moves the first centrifugal test tube to the centrifugal test tube clamping part, the centrifugal test tube clamping part moves to the second transferring movable area along the second sliding rail, and the centrifugal test tube is moved to the high-speed centrifugal device through the second mechanical arm for high-speed centrifugation;
s6, transferring centrifugate: the first mechanical arm clamps a second centrifugal test tube on the centrifugal tube rack to a centrifugal test tube clamping part, the second rotary clamping part opens a second centrifugal test tube cover, and the upper pipetting part of the second mechanical arm transfers the supernatant of the first centrifugal test tube to the second centrifugal test tube through a tip gun head;
s7, transferring a second centrifugal test tube to a temperature incubation module by a second mechanical arm;
s8, after incubation timing in one of the centrifugal test tubes is completed, carrying out an extraction step: the second mechanical 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 claim 9, wherein the display control module deletes or repeats at least one of steps S2, S3, S4, S5, or S6 multiple times according to the detection procedure of different cancer species; or, the execution sequence of the above S2, S3, S4, S5, S6 is readjusted according to the detection flow of different cancer species.
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| CN202310792849.6A CN117106570A (en) | 2023-06-30 | 2023-06-30 | Nucleic acid extraction and transformation integrated machine capable of editing flow and method thereof |
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