CN116554996B - Portable full-automatic fluorescent nucleic acid detector and nucleic acid extraction method thereof - Google Patents
Portable full-automatic fluorescent nucleic acid detector and nucleic acid extraction method thereof Download PDFInfo
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- CN116554996B CN116554996B CN202310556541.1A CN202310556541A CN116554996B CN 116554996 B CN116554996 B CN 116554996B CN 202310556541 A CN202310556541 A CN 202310556541A CN 116554996 B CN116554996 B CN 116554996B
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- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 72
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 71
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 71
- 238000000605 extraction Methods 0.000 title claims abstract description 42
- 238000006243 chemical reaction Methods 0.000 claims abstract description 154
- 239000011324 bead Substances 0.000 claims abstract description 127
- 239000007788 liquid Substances 0.000 claims abstract description 83
- 238000005336 cracking Methods 0.000 claims abstract description 68
- 238000000197 pyrolysis Methods 0.000 claims abstract description 58
- 230000033001 locomotion Effects 0.000 claims abstract description 27
- 239000002699 waste material Substances 0.000 claims abstract description 21
- 230000010355 oscillation Effects 0.000 claims abstract description 10
- 230000005389 magnetism Effects 0.000 claims abstract description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 53
- 239000000523 sample Substances 0.000 claims description 39
- 238000007789 sealing Methods 0.000 claims description 31
- 238000005406 washing Methods 0.000 claims description 27
- 238000003776 cleavage reaction Methods 0.000 claims description 26
- 238000000034 method Methods 0.000 claims description 22
- 238000010828 elution Methods 0.000 claims description 18
- 238000012546 transfer Methods 0.000 claims description 17
- 239000012295 chemical reaction liquid Substances 0.000 claims description 15
- 239000006166 lysate Substances 0.000 claims description 13
- 230000002093 peripheral effect Effects 0.000 claims description 13
- 230000005540 biological transmission Effects 0.000 claims description 7
- 239000012488 sample solution Substances 0.000 claims description 7
- 230000007017 scission Effects 0.000 claims description 4
- 230000005611 electricity Effects 0.000 claims 1
- 239000000243 solution Substances 0.000 description 41
- 230000000694 effects Effects 0.000 description 11
- 230000001276 controlling effect Effects 0.000 description 7
- 238000002156 mixing Methods 0.000 description 7
- 238000001179 sorption measurement Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 239000012535 impurity Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000008961 swelling Effects 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000009514 concussion Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000010030 laminating Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 210000003437 trachea Anatomy 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000011344 liquid material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The application discloses a portable full-automatic fluorescent nucleic acid detector and a nucleic acid extraction method thereof, wherein the portable full-automatic fluorescent nucleic acid detector comprises a sample extraction box, a cracking reaction chamber is arranged in the sample extraction box, a cracking reaction chamber bag is arranged in the cracking reaction chamber, and the top of the cracking reaction chamber bag is connected with a cracking liquid storage module, a magnetic bead liquid storage module and a sample liquid storage module; the magnetic rod assembly comprises a moving slide plate, a magnetic rod arranged on the moving slide plate, a magnetic rod motion control module for driving the moving slide plate to move along the horizontal and vertical directions, and a magnetic rod oscillation module for driving the magnetic rod to rotate in a gap and reciprocate along the vertical direction; a pyrolysis waste liquid cavity bag; wherein, the magnetic rod can extrude the outer wall of schizolysis reaction chamber bag in order to make schizolysis reaction chamber bag deformation and make the outer wall parcel magnetic rod's of schizolysis reaction chamber bag periphery wall, and the magnetism of magnetic rod can be controlled according to switching on and off. The application has the function of improving the extraction precision of nucleic acid.
Description
Technical Field
The application relates to the technical field of nucleic acid detection, in particular to a portable full-automatic fluorescent nucleic acid detector and a nucleic acid extraction method thereof.
Background
Nucleic acid extraction by the magnetic bead method can be generally divided into four steps: cleavage-binding-washing-eluting.
Cracking: adding a lysate into a sample, uniformly mixing and fully reacting the reaction solution through mechanical movement and heating, and performing cell lysis to release nucleic acid. Combining: adding magnetic beads into the sample lysate, fully and uniformly mixing, adsorbing nucleic acid by utilizing the characteristic that the magnetic beads have strong affinity to nucleic acid under the conditions of high salt and low pH value, separating the magnetic beads from the solution under the action of an external magnetic field, removing the liquid by utilizing a suction head, and discarding the liquid to a waste liquid tank. Washing: removing the external magnetic field, replacing the new suction head with the washing buffer solution, fully and uniformly mixing, removing impurities, and removing the liquid under the action of the external magnetic field. Eluting: removing the external magnetic field, replacing the magnetic head with a new suction head, adding the eluting buffer solution, fully and uniformly mixing, and separating the combined nucleic acid from the magnetic beads to obtain the purified nucleic acid.
In the related art, a magnetic rod method is generally adopted to magnetically separate nucleic acid, a special magnetic rod sleeve stretches into a solution, magnetic beads are directionally agglomerated on the special magnetic rod sleeve on the surface of the magnetic rod, and the magnetic beads are precisely moved to realize the process of automatically extracting nucleic acid from sample lysate to washing solution and then to eluent, so that the automatic extraction process of nucleic acid detection is completed.
For the related technology, nucleic acid and impurities can be attached to the magnetic rod sleeve and transferred along with the magnetic rod, so that errors exist in subsequent reactions, and the test result is not accurate enough.
Disclosure of Invention
In order to help to improve the extraction precision of nucleic acid, the application provides a portable full-automatic fluorescent nucleic acid detector and a nucleic acid extraction method thereof.
The application provides a portable full-automatic fluorescent nucleic acid detector and a nucleic acid extraction method thereof, which adopts the following technical scheme:
first aspect
A portable fully automatic fluorescent nucleic acid detector comprising:
the sample extraction box is internally provided with a cracking reaction chamber, a cracking reaction chamber bag is arranged in the cracking reaction chamber, the top of the cracking reaction chamber bag is connected with a cracking liquid storage module, a magnetic bead liquid storage module and a sample liquid storage module, and the cracking liquid storage module, the magnetic bead liquid storage module and the sample liquid storage module can inject liquid into the cracking reaction chamber bag;
the magnetic rod assembly comprises a moving slide plate, a magnetic rod arranged on the moving slide plate, a magnetic rod movement control module for driving the moving slide plate to move along the horizontal and vertical directions, and a magnetic rod oscillation module for driving the magnetic rod to rotate in a clearance and reciprocate along the vertical direction;
a cavity bag domain-dividing sealing clamp is arranged between the pyrolysis waste liquid cavity bag and the pyrolysis reaction cavity bag;
the magnetic rod can extrude the outer wall of the cracking reaction cavity bag so as to deform the cracking reaction cavity bag and enable the outer wall of the cracking reaction cavity bag to wrap the outer peripheral wall of the magnetic rod, and the magnetism of the magnetic rod can be controlled according to the on-off power.
Through adopting above-mentioned technical scheme, at first pour into the pyrolysis reaction chamber bag with sample liquid and pyrolysis liquid into, sample liquid and pyrolysis liquid mix, in this in-process, bar magnet motion control module control bar magnet moves pyrolysis reaction chamber bag top, bar magnet motion control module control bar magnet moves down, the bar magnet is down-stamped, pyrolysis reaction chamber bag warp, then, bar magnet concussion module control bar magnet vertical repetitive motion uses the bar magnet to vibrate pyrolysis reaction chamber bag, the reaction liquid intensive mixing of pyrolysis reaction chamber bag makes the schizolysis effect of sample more thoroughly. Then, injecting magnetic bead liquid into the cracking reaction cavity bag, mixing the magnetic beads with the reaction liquid, adsorbing DNA by the magnetic beads, and enabling the magnetic rod to be not electrified, wherein the magnetic rod vibration module is used for controlling the magnetic rod to vertically and repeatedly move, the magnetic beads are dispersed in the reaction liquid and move in the reaction liquid, the collision probability of the magnetic beads and the reaction liquid is increased, and the DNA/RNA adsorbed by the magnetic beads is increased and the adsorption efficiency is increased. After a period of time, the magnetic rod is electrified, the magnetic beads are adsorbed on the inner wall of the cracking reaction cavity bag close to the magnetic rod, the magnetic rod oscillation module controls the magnetic rod to control the magnetic rod to intermittently rotate and vertically repeatedly move, the magnetic beads do rotary motion, under the action of centrifugal force, protein, polysaccharide and other impurities are far away from the center of the magnetic rod, meanwhile, the magnetic beads are far away from the magnetic rod, but due to the intermittent rotation of the magnetic rod, the magnetic beads have stronger adsorption force on the magnetic beads, the magnetic beads are close to the magnetic rod, and further, the magnetic beads are enabled to rotate in the cracking liquid to reciprocally approach or be far away from the magnetic rod, so that the collision probability of the magnetic beads and the cracking liquid is increased to the greatest extent, and the accuracy of adsorbing DNA/RNA by the magnetic beads is improved; finally, the magnetic rod stops rotating and vertically reciprocates, the cavity bag domain sealing pliers are opened, the pyrolysis waste liquid material of the pyrolysis reaction cavity bag is discharged into the pyrolysis waste liquid cavity bag, the magnetic rod moves upwards, and the magnetic rod adsorbs the magnetic beads to move the magnetic beads to the next nucleic acid extraction step.
Optionally, the magnetic rod includes the body and articulates the polylith magnetic sheet at body bottom week side, works as the outer wall parcel of schizolysis reaction chamber bag the outer peripheral wall of magnetic rod, the magnetic sheet laminating is in the outer peripheral wall of body, works as the bottom butt of body is in when the top of schizolysis reaction chamber bag, the magnetic sheet laminating is in the top of schizolysis reaction chamber bag.
By adopting the technical scheme, when the magnetic rod is poked into the cracking reaction cavity bag, the magnetic sheet is attached to the outer peripheral wall of the rod body, and the magnetic sheet can fully adsorb the magnetic beads in the cracking reaction cavity bag, so that the magnetic beads can be dispersed on the peripheral side of the magnetic sheet, and the magnetic beads can adsorb more DNA/RNA; meanwhile, when the magnetic beads need to be transferred, the magnetic sheets are opened, even if the amount of the magnetic beads is large, the magnetic beads can be stably adsorbed on the inner wall of the top of the cracking reaction cavity bag, and the magnetic beads can be driven to be transferred through the magnetic rods and the magnetic sheets. Through the technical scheme, the whole reaction process and the transfer process are carried out in the sealed environment, so that pollution caused in the reaction process and the transfer process can be effectively reduced, and the nucleic acid extraction effect is improved.
Optionally, a washing reaction chamber is further arranged in the sample extraction box, a washing reaction chamber bag is arranged in the washing reaction chamber, a connecting pipe is connected between the side wall of the washing reaction chamber bag close to the top and the side wall of the cracking reaction chamber bag close to the top, and a chamber bag split-domain sealing clamp is also arranged on the connecting pipe.
Through adopting above-mentioned technical scheme, the bar magnet removes along pyrolysis reaction chamber bag, connecting pipe and washing reaction chamber bag's top outer wall, can shift magnetic bead and adhesion material thereof in totally closed environment, and the furthest prevents the pollution from external environment, ensures the accuracy of extraction result.
Optionally, the bar magnet is along vertical and motion slide sliding connection, the bar magnet concussion module is including being used for driving the rotatory bar magnet rotary driving subassembly of bar magnet and being used for driving the reciprocal sloping cam plate control assembly who goes up and down of bar magnet.
Optionally, the magnetic rod rotation driving assembly includes:
the magnetic rod driving motor is arranged on the sport skateboard;
the driving shaft is fixed at the output end of the magnetic rod driving motor;
an incomplete driving wheel fixed on the periphery of the driving shaft;
the driven wheel is slidably arranged on the periphery of the magnetic rod, and a transmission belt is connected between the incomplete driving wheel and the driven wheel;
the swash plate control assembly includes:
a swash plate fixed to an outer circumference of the driving shaft;
and the inclined cutting shaft is fixed on the outer wall of the magnetic rod, and the peripheral surface of the inclined disc is in sliding contact with the end part of the inclined cutting shaft.
Through adopting above-mentioned technical scheme, bar magnet driving motor drives the drive shaft and rotates, utilizes incomplete action wheel and from the driving wheel to drive the bar magnet and rotate, simultaneously, the drive shaft drives the sloping cam plate and rotates, and sloping cam plate and oblique cutting axle cooperation effect drive bar magnet up-and-down reciprocating motion, and then can both drive the bar magnet through bar magnet driving motor and rotate, also drive bar magnet up-and-down reciprocating motion. Through the scheme, the cracking reaction cavity bag can be extruded to deform, so that the solution of the cracking reaction cavity bag can frequently move, and the magnetic beads and the reaction solution are fully mixed and stirred, so that good adhesion and combination are realized.
Optionally, the lysate storage module, the magnetic bead solution storage module, and the sample solution storage module all include:
the tube body is connected with the cracking reaction cavity bag;
the reagent tube is used for storing the lysate, the magnetic bead liquid or the sample liquid, and is arranged in the tube body in a vertically sliding and sealing manner, and a puncturable sealing film is arranged at the bottom of the reagent tube;
and the puncturing piece is arranged at the bottom of the inner side of the tube body, and can puncture the sealing film when the reagent tube moves downwards.
By adopting the technical scheme, when nucleic acid extraction is required, the reagent tube is pressed down, and the sealing film is punctured by the puncturing piece, so that the solution in the reagent tube can flow into the cracking reaction cavity bag, and the addition of a certain solution is realized; the reagent tube can independently make a sealed finished product, thereby being beneficial to improving the accuracy of nucleic acid extraction.
Optionally, the bottom of reagent pipe is provided with the tube housing, tube housing and the inner wall sliding connection of body, form the atmospheric pressure room between the outer wall of inner wall, body and the tube housing of body, install the trachea in the atmospheric pressure room.
By adopting the technical scheme, the position of the pipe body is controlled by utilizing air pressure, so that the operation and control are convenient, and the cost is low.
Optionally, the puncture piece includes the toper puncture arrow body, the fixture block is installed to reagent pipe inner wall, the fixture block can with the toper puncture arrow body joint is in order to prevent reagent pipe to move towards the direction of keeping away from the pyrolysis reaction chamber bag, the gas of trachea output can get into in the reagent pipe through the gap between tube cover and the body to in entering the pyrolysis reaction chamber bag through the reagent pipe.
Through adopting above-mentioned technical scheme, utilize atmospheric pressure to make the body move down and utilize the puncture piece to puncture the sealing membrane after, the toper punctures arrow body and fixture block joint, and the reaction chamber bag of pyrolysis can't keep away from to the reagent pipe this moment, continues to inject gas, and gas gets into in the reagent pipe through the gap between pipe cover and the body to can impress in the reaction chamber bag of pyrolysis with the solution in the reagent pipe, ensure that the solution in the reagent pipe can get into the reaction chamber bag of pyrolysis completely, and then ensure the accuracy of nucleic acid extraction. Meanwhile, gas can enter the cracking reaction cavity bag through the reagent pipe, so that the gas pressure in the cracking reaction cavity bag can be regulated, and a gas pressure environment for experimental reaction and a swelling space for movement of the magnetic beads are provided.
Optionally, an elution reaction chamber is further arranged in the sample extraction box, an elution reaction chamber bag is arranged in the elution reaction chamber, the elution reaction chamber bag is connected with the washing reaction chamber bag, and the magnetic rod can drive the magnetic beads to move and transfer in the cracking reaction chamber bag, the washing reaction chamber bag and the elution reaction chamber bag.
By adopting the technical scheme, the magnetic rod provided by the application can accelerate the movement of the solution, improve the cracking, washing and eluting efficiencies of nucleic acid, fully drive the magnetic beads to move, ensure the effective transfer of the magnetic beads and ensure the extraction effect of nucleic acid.
Second aspect
A method for extracting nucleic acid, comprising the steps of:
s1, adding corresponding solutions into a lysate storage module, a magnetic bead solution storage module and a sample solution storage module;
s2, injecting the sample liquid in the sample liquid storage module and the pyrolysis liquid in the pyrolysis liquid storage module into a pyrolysis reaction cavity bag, and oscillating the reaction liquid in the pyrolysis reaction cavity bag by using the magnetic rod assembly;
s3, injecting the magnetic bead liquid in the magnetic bead liquid storage module into the cracking reaction cavity bag, and controlling the positions of the magnetic beads in the cracking reaction cavity bag and adsorbing the magnetic beads by using the magnetic rod assembly;
s4, driving the magnetic beads to transfer by using the magnetic rod assembly.
By adopting the technical scheme, the magnetic rod assembly can accelerate the movement of the solution, so that the cracking speed of nucleic acid can be accelerated on one hand, and the magnetic beads and the solution can be promoted to be fully mixed and stirred on the other hand, and the adhesion and bonding effects of the magnetic beads are improved; meanwhile, the magnetic rod assembly can adsorb the magnetic beads to the greatest extent, so that the magnetic beads and adhesion substances thereof can be transferred to the adjacent chamber to the greatest extent, and the extraction purity of nucleic acid is improved.
In summary, the present application includes at least one of the following beneficial technical effects:
1. the magnetic rod assembly can accelerate the movement of the solution, so that the cracking speed of nucleic acid can be accelerated, and the magnetic beads and the solution can be fully mixed and stirred, so that the adhesion and bonding effects of the magnetic beads are improved; meanwhile, the magnetic rod assembly can adsorb the magnetic beads to the greatest extent, so that the magnetic beads and adhesion substances thereof can be transferred to the adjacent chamber to the greatest extent, and the extraction purity of nucleic acid is improved.
2. The gas output by the gas pipe can ensure that the solution in the reagent pipe completely enters the cracking reaction cavity bag, thereby ensuring the accuracy of nucleic acid extraction. Meanwhile, gas can enter the cracking reaction cavity bag through the reagent pipe, so that the gas pressure in the cracking reaction cavity bag can be regulated, and a gas pressure environment for experimental reaction and a swelling space for movement of the magnetic beads are provided.
3. The whole reaction process and the transfer process are carried out in an environment with a sealed environment, so that the pollution caused in the reaction process and the transfer process can be effectively reduced, and the nucleic acid extraction effect is improved.
Drawings
Fig. 1 is an exploded schematic view of the overall structure of an embodiment of the present application.
Fig. 2 is a schematic view of the embodiment of the present application mainly showing a state in which each of the reaction chambers are communicated with each other.
FIG. 3 is a schematic view of a memory module and a cavity bag according to an embodiment of the present application
Fig. 4 is an enlarged schematic view of the portion a in fig. 3.
Fig. 5 is a schematic diagram mainly showing a state that a magnetic rod assembly is not poked into a reaction chamber bag according to an embodiment of the application.
Fig. 6 is an enlarged schematic view of the portion B in fig. 5.
Fig. 7 is a schematic diagram mainly showing a state that a magnetic rod assembly is poked into a reaction chamber bag according to an embodiment of the application.
Reference numerals illustrate:
10. a housing; 11. a sample extraction cartridge; 12. a cleavage reaction chamber; 121. cracking reaction chamber bags; 122. a pyrolysis waste liquid cavity bag; 13. washing the reaction chamber; 131. washing the reaction chamber bag; 132. a connecting pipe; 14. eluting the reaction chamber; 141. eluting the reaction cavity bag; 142. a nucleic acid solution collection chamber bag; 15. a magnetic bead discarding chamber;
20. a lysate storage module; 21. a magnetic bead liquid storage module; 22. a sample fluid storage module;
30. a tube body; 301. sealing cover; 40. a reagent tube; 401. a sealing film; 402. a tube cover; 403. a clamping block; 50. a piercing member;
60. sealing pliers for cavity and bag division; 61. an air pressure chamber; 62. an air pipe; .
80. A sport skateboard; 81. a magnetic rod; 811. a rod body; 812. a magnetic sheet; 82. a magnetic rod motion control module; 821. a magnetic bar bracket; 822. a rack and pinion mechanism; 83. the magnetic rod oscillation module; 841. a magnetic bar driving motor; 842. a drive shaft; 843. an incomplete drive wheel; 844. driven wheel; 845. a transmission key; 846. a drive belt; 851. a swash plate; 852. and (3) chamfering the shaft.
Detailed Description
The application is described in further detail below with reference to fig. 1-7.
The embodiment of the application discloses a portable full-automatic fluorescent nucleic acid detector. Referring to fig. 1, the full-automatic fluorescent nucleic acid detector comprises a housing 10 and a sample extraction box 11 installed in the housing 10, wherein the sample extraction box 11 is used for carrying out splitting, combining, washing and eluting on nucleic acid so as to extract the nucleic acid, a nucleic acid PCR amplification module and a nucleic acid marker fluorescent detection module are also installed in the sample extraction box 11, and the nucleic acid PCR amplification module mainly has the functions of processing the nucleic acid extracted and purified by the sample extraction box 11 and then pumped into a PCR reaction chamber, adding a PCR reactant, and carrying out experimental process control on the nucleic acid, so as to complete the multiple cycle processes of denaturation, annealing and extension, and realize the amplification of nucleic acid fragments. The nucleic acid marker fluorescence detection module mainly realizes detection of the characteristic sequence, and the detection process can be carried out in each round of circulation process, so that real-time quantitative analysis is realized. Through the functional module, a simple, efficient, accurate, reliable, cost-effective and adaptive nucleic acid automation solution can be provided for a laboratory.
Referring to fig. 1, the sample extraction cartridge 11 is provided in a cylindrical shape, and a cleavage reaction chamber 12, a washing reaction chamber 13, an elution reaction chamber 14, and a magnetic bead discarding chamber 15 are sequentially provided in the sample extraction cartridge 11.
Referring to fig. 1 and 2, a cleavage reaction chamber bag 121 is installed in the cleavage reaction chamber 12, a cleavage liquid storage module 20, a magnetic bead liquid storage module 21 and a sample liquid storage module 22 are connected to the top of the cleavage reaction chamber bag 121, and the cleavage liquid storage module 20, the magnetic bead liquid storage module 21 and the sample liquid storage module 22 can inject liquid into the cleavage reaction chamber bag 121 according to requirements.
Referring to fig. 3 and 4, the lysate storage module 20, the bead solution storage module 21 and the sample solution storage module 22 each include a tube body 30, a reagent tube 40 and a piercing member 50, the tube body 30 is connected with a cleavage reaction chamber bag 121, a chamber bag domain sealing clamp 60 is disposed between the tube body 30 and the cleavage reaction chamber bag 121, and the chamber bag domain sealing clamp 60 is composed of a set of normally closed electric control valves.
The reagent tube 40 is used for storing solutions such as lysate, magnetic bead solution or sample solution, and the reagent tube 40 is mounted in the tube body 30 in a vertically sliding sealing manner, and in order to facilitate replacement of the reagent tube 40 and ensure that solution transfer is performed in a sealed environment, a sealing cover 301 is in threaded connection with the top of the tube body 30, and the reagent tube 40 penetrates through the center of the sealing cover 301 and is in sliding sealing with the sealing cover 301.
Meanwhile, a sealing film 401 capable of being pierced is arranged at the bottom of the reagent tube 40, the piercing member 50 is arranged at the inner bottom of the tube body 30 near the center, when the reagent tube 40 moves downwards, the piercing member 50 can pierce the sealing film 401, so that the solution in the reagent tube 40 can flow into the tube body 30, and when the cavity bag split-domain sealing clamp 60 is opened, the solution in the tube body 30 can enter the cleavage reaction cavity bag 121.
In order to make the solution in the tube body 30 fully enter the cracking reaction cavity bag 121, the bottom end of the reagent tube 40 is integrally formed with a tube cover 402, the peripheral side edge of the tube cover 402 is in sliding contact with the inner wall of the tube body 30, a pneumatic chamber 61 is formed among the inner wall of the tube body 30, the tube body 30 and the outer wall of the tube cover 402, a pneumatic tube 62 is installed in the pneumatic chamber 61, the pneumatic tube 62 is connected with a pneumatic pump and an electric control feedback pneumatic valve, and the pneumatic pressure in the cavity bag can be controlled through the structure. When the gas is outputted from the gas pipe 62, a part of the gas in the gas pressure chamber 61 can enter the reagent vessel 40 through the gap between the outer peripheral side of the pipe cover 402 and the inner wall of the pipe body 30.
Referring to fig. 4, the puncturing member 50 includes a tapered puncturing arrow body having elasticity, and a clamping block 403 is mounted on an inner wall of the reagent tube 40, and when the puncturing member 50 punctures the sealing membrane 401 and enters the reagent tube 40, the puncturing member 50 can be clamped with the clamping block 403 to prevent the reagent tube 40 from moving in a direction away from the cleavage reaction chamber bag 121. At this time, the gas output from the gas pipe 62 can enter the reagent pipe 40 through the gap between the pipe cover 402 and the pipe body 30, the reagent pipe 40 cannot be far away from the cleavage reaction chamber bag 121, and after the chamber bag split-domain sealing clamp 60 is opened, the gas pressure can press the solution in the reagent pipe 40 into the cleavage reaction chamber bag 121, so as to realize sufficient transfer of the solution in the reagent pipe 40.
Meanwhile, gas can enter the cracking reaction cavity bag 121 through the reagent tube 40, so that the gas pressure in the cracking reaction cavity bag 121 can be regulated, and a gas pressure environment for experimental reaction and a swelling space for movement of the magnetic beads are provided.
In addition, for the convenience of replacing the reagent vessel 40, the clamping block 403 may be configured to be broken after a certain force is applied, so that when the reagent vessel 40 is applied with air pressure, the clamping block 403 can prevent the reagent vessel 40 from moving reversely, and when the reagent vessel 40 is applied with a sufficient external force, the clamping block 403 is broken, and the reagent vessel 40 can be removed normally.
Referring to fig. 1, the washing reaction chamber 13 is provided with a rinsing liquid storage module, and the eluting reaction chamber 14 is also provided with an eluent storage module, and the structures and functions of the two modules are similar to those of the above modules, and are not described herein.
Referring to fig. 5 and 6, a portion of the memory module is hidden for ease of illustration. The full-automatic fluorescent nucleic acid detector further comprises a magnetic rod assembly arranged inside the housing 10, wherein the magnetic rod assembly comprises a moving slide plate 80, a magnetic rod 81 arranged on the moving slide plate 80, a magnetic rod motion control module 82 for driving the moving slide plate 80 to move along the horizontal and vertical directions, and a magnetic rod oscillation module 83 for driving the magnetic rod 81 to rotate in a clearance mode and reciprocate along the vertical direction.
Referring to fig. 6 and 7, the cleavage reaction chamber bag 121 is made of a transparent polymer material which is easily deformable, resistant to high temperature, pressure and corrosion, and has a fully closed structure. After the magnetic rod motion control module 82 controls the magnetic rod 81 to move downwards, the magnetic rod 81 can squeeze the outer wall of the cracking reaction cavity bag 121 to deform the cracking reaction cavity bag 121 and enable the outer wall of the cracking reaction cavity bag 121 to wrap the outer peripheral wall of the magnetic rod 81, and the magnetic rod 81 is magnetic after being electrified and can adsorb magnetic beads in reaction liquid so that the magnetic beads are adsorbed on the inner wall of the cracking reaction cavity bag 121, which is close to the magnetic rod 81.
Referring to fig. 7, the magnetic rod 81 includes a rod body 811 and a plurality of magnetic pieces 812 hinged to the bottom end circumference side of the rod body 811, and a torsion spring is provided between the magnetic pieces 812 and the rod body 811 so that the magnetic pieces 812 can be separated from the surface of the rod body 811 when no external force is applied. The number and shape of the magnetic sheets 812 can be designed according to specific requirements to satisfy the requirement of completely adsorbing the magnetic beads in the cleavage reaction chamber bag 121.
When the magnetic rod 81 moves downwards, the magnetic rod 81 extrudes the outer wall of the cracking reaction cavity bag 121, the outer wall of the cracking reaction cavity bag 121 extrudes the magnetic rod 81 as well, at the moment, the magnetic sheet 812 is folded and attached to the outer peripheral wall of the rod 811, and the magnetic sheet 812 and the rod 811 can absorb magnetic beads together; referring to fig. 6, when the magnetic rod 81 moves up and moves to the top of the cleavage reaction chamber pocket 121, the magnetic sheet 812 receives no other external force, the rod body 811 is perpendicular to the magnetic sheet 812, the plurality of magnetic sheets 812 are opened, and the ends of the rod body 811 and the magnetic sheet 812 adsorb the magnetic beads so that the magnetic beads are adsorbed near the top of the cleavage reaction chamber pocket 121 for facilitating the transfer of the subsequent magnetic beads. Therefore, even if the number of magnetic beads in the cleavage reaction chamber bag 121 is relatively large, the above-described structure can sufficiently adsorb and transfer the magnetic beads.
In this process, the reaction solution is not full of the whole cleavage reaction chamber bag 121, and when the magnetic beads are adsorbed and moved to the top of the cleavage reaction chamber bag 121, the magnetic beads are separated from the reaction solution, and meanwhile, the magnetic beads can throw off the waste liquid on the surface, so that the waste liquid is transferred into the next reaction chamber along with the magnetic beads, and the accuracy of nucleic acid extraction is improved.
Referring to fig. 5 and 6, the magnetic rod motion control module 82 includes a magnetic rod support 821, the magnetic rod support 821 is slidably connected with a top side inner wall of the housing 10 along a circumferential direction of the housing 10, the motion slide 80 is vertically slidably connected to the magnetic rod support 821, a rack and pinion mechanism 822 is disposed between the magnetic rod support 821 and the housing 10, and the magnetic rod 81 can be driven to move between each chamber by using the rack and pinion mechanism 822, so that the magnetic beads can be driven to move between each chamber.
Referring to fig. 6, a rack and pinion mechanism 822 is provided between the moving slide 80 and the bar magnet holder 821, and the bar magnet 81 is moved up or down by the rack and pinion mechanism 822. So that the magnetic rod 81 can be close to or far from the cleavage reaction chamber bag 121 to realize the adsorption and transfer of the magnetic beads.
Referring to fig. 6, a magnetic rod 81 is slidably connected with a moving slide 80 along a vertical direction, and a magnetic rod oscillating module 83 includes a magnetic rod rotation driving assembly for driving the magnetic rod 81 to rotate and a swash plate control assembly for driving the magnetic rod 81 to reciprocate.
The magnetic rod rotation driving assembly comprises a magnetic rod driving motor 841, a driving shaft 842, an incomplete driving wheel 843 and a driven wheel 844, wherein the magnetic rod driving motor 841 is installed on the movement sliding plate 80, the driving shaft 842 is fixed at the output end of the magnetic rod driving motor 841 and extends vertically, the incomplete driving wheel 843 is fixed at the periphery of the driving shaft 842, the incomplete driving wheel 843 can be 1/3 of full teeth, the driven wheel 844 is slidably installed at the periphery of the magnetic rod 81, a transmission key 845 is fixed at the periphery of the magnetic rod 81 along the circumferential direction, and the driven wheel 844 can drive the magnetic rod 81 to rotate and also enable the magnetic rod 81 to slide relative to the driven wheel 844 through the transmission key 845. Wherein, a transmission belt 846 is connected between the incomplete driving wheel 843 and the driven wheel 844. In addition, the structure for preventing axial movement of the drive belt 846 is conventional and will not be shown here too much.
Referring to fig. 6, the swash plate control assembly includes a swash plate 851 and a swash shaft 852, the swash plate 851 is fixed to the outer circumference of the drive shaft 842, the swash shaft 852 is fixed to the outer wall of the magnetic rod 81, and the circumferential surface of the swash plate 851 is in sliding contact with the end of the swash shaft 852. When the drive shaft 842 rotates the swash plate 851, the magnetic rod 81 reciprocates in the axial direction under the transmission of the bevel shaft 852.
The nucleic acid is cracked and the adsorption and transfer process of the magnetic beads is as follows: firstly, injecting sample liquid and pyrolysis liquid into the pyrolysis reaction chamber bag 121, mixing the sample liquid and the pyrolysis liquid, in the process, controlling the magnetic rod 81 to move to the upper part of the pyrolysis reaction chamber bag 121 by the magnetic rod motion control module 82, controlling the magnetic rod 81 to move downwards by the magnetic rod motion control module 82, stamping the magnetic rod 81 downwards, deforming the pyrolysis reaction chamber bag 121, then controlling the magnetic rod 81 to vertically and repeatedly move by the magnetic rod oscillation module 83, oscillating the pyrolysis reaction chamber bag 121 by using the magnetic rod 81, and fully stirring the reaction liquid of the pyrolysis reaction chamber bag 121, so that the pyrolysis effect of a sample is more thorough. Then, the magnetic bead liquid is injected into the cracking reaction cavity bag 121, the magnetic beads are combined with the reaction liquid, the magnetic beads adsorb DNA/RNA, at this time, the magnetic rod 81 is not electrified, firstly, the magnetic rod oscillation module 83 is used for controlling the magnetic rod 81 to vertically and repeatedly move, the magnetic beads are dispersed in the reaction liquid and move in the reaction liquid, the collision probability of the magnetic beads and the reaction liquid is increased, the DNA/RNA adsorbed by the magnetic beads is increased, and the adsorption efficiency is increased. After a period of time, the magnetic rod 81 is electrified, the magnetic beads are adsorbed onto the inner wall of the cracking reaction cavity bag 121 close to the magnetic rod 81, the magnetic rod oscillation module 83 controls the magnetic rod 81 to control the magnetic rod 81 to intermittently rotate and vertically repeatedly move, the magnetic beads are rotated, under the action of centrifugal force, impurities such as proteins and polysaccharides are far away from the center of the magnetic rod 81, meanwhile, the magnetic beads are also far away from the magnetic rod 81, but due to the fact that the magnetic rod 81 intermittently rotates, the magnetic beads have stronger adsorption force on the magnetic beads, the magnetic beads are close to the magnetic rod 81 in the intermittent process, and further the magnetic beads are rotated in the cracking liquid and are reciprocally close to or far away from the magnetic rod 81, so that the collision probability of the magnetic beads and the cracking liquid is increased to the greatest extent, and the accuracy of adsorbing DNA/RNA by the magnetic beads is improved.
Referring to fig. 4 and 7, a waste solution chamber 122 is further connected to the bottom of the reaction chamber 121, and a chamber-bag split-domain sealing clamp 60 is also provided between the waste solution chamber 122 and the reaction chamber 121. After the reaction is completed, the magnetic rod 81 stops rotating and vertically reciprocating, the cavity bag domain sealing clamp 60 is opened, the pyrolysis waste liquid of the pyrolysis reaction cavity bag 121 is discharged into the pyrolysis waste liquid cavity bag 122, and in the process, the pyrolysis waste liquid can be conveyed by utilizing the air pressure output by the air pipe 62. After that, the magnetic rod 81 moves upwards, the magnetic beads are adsorbed by the magnetic rod 81 to move upwards, and the magnetic beads are separated from the pyrolysis waste liquid, so that the gas can blow off the pyrolysis waste liquid remained on the surfaces of the magnetic beads to a certain extent in the process, and the pyrolysis waste liquid is furthest reduced from being transferred to the next nucleic acid extraction step.
Referring to fig. 1, a washing reaction chamber bag 131 is installed in the washing reaction chamber 13, a connection pipe 132 is connected between a side wall of the washing reaction chamber bag 131 near the top and a side wall of the cleavage reaction chamber bag 121 near the top, and a chamber bag split-domain sealing jaw 60 is also provided on the connection pipe 132. The connection pipe 132 is disposed near the top of the washing reaction chamber bag 131 and the cleavage reaction chamber bag 121, and can reduce the transfer of residual waste between the chamber bags.
Similarly, the elution reaction chamber 14 and the magnetic bead discarding chamber 15 are respectively provided with a corresponding chamber bag and a connecting pipe 132, for example, an elution reaction chamber bag 141 is provided in the elution reaction chamber 14, the elution reaction chamber bag 141 is connected with a nucleic acid solution collecting chamber bag 142, nucleic acid is eluted in the elution reaction chamber bag 141, the eluted nucleic acid solution enters the nucleic acid solution collecting chamber bag 142, and the magnetic beads can be transferred to the magnetic bead discarding chamber 15 for recycling through the magnetic rod assembly.
In addition, still be provided with the temperature control device of the chamber of branch territory in each chamber bag, divide the chamber temperature control device of territory to include semiconductor refrigeration heating module, temperature sensing module and MCU temperature control module, heat or refrigerate each chamber bag group according to the experiment demand, ensure to satisfy the experiment requirement.
The embodiment of the application relates to a portable full-automatic fluorescent nucleic acid detector, which comprises the following implementation principles: the application can accelerate the movement of the solution through the special magnetic rod assembly. On one hand, the method can accelerate the cracking speed of nucleic acid, on the other hand, the method can promote the magnetic beads and the solution to be fully mixed and stirred, and the adhesion and bonding effects of the magnetic beads are improved; meanwhile, the magnetic rod assembly can adsorb the magnetic beads to the greatest extent, so that the magnetic beads and adhesion substances thereof can be transferred to the adjacent chamber to the greatest extent, and the extraction purity of nucleic acid is improved.
The embodiment of the application also discloses a nucleic acid extraction method. The method comprises the following steps:
s1, adding corresponding reagent tubes 40 into the lysate storage module 20, the magnetic bead liquid storage module 21 and the sample liquid storage module 22, wherein corresponding solutions are pre-stored in the reagent tubes 40.
S2, injecting the sample liquid in the sample liquid storage module 22 and the pyrolysis liquid in the pyrolysis liquid storage module 20 into the pyrolysis reaction cavity bag 121, and vibrating the reaction liquid in the pyrolysis reaction cavity bag 121 by using the magnetic rod assembly when the sample liquid reacts with the pyrolysis liquid.
S3, injecting the magnetic bead liquid in the magnetic bead liquid storage module 21 into the cracking reaction cavity bag 121, controlling the positions of the magnetic beads in the cracking reaction cavity bag 121 by utilizing the magnetic rod assembly, improving the collision probability of the magnetic beads and the reaction liquid, adsorbing the magnetic beads after the magnetic beads are combined with DNA/RNA of the demand, and discharging waste liquid, wherein the magnetic beads are separated from waste materials.
S4, magnetic beads are driven to transfer by the magnetic rod assembly, and in each cavity bag, washing efficiency and elution efficiency are quickened by utilizing the vibration function of the magnetic bead assembly, and washing and elution effects are enhanced.
The above embodiments are not intended to limit the scope of the present application, so: all equivalent changes in structure, shape and principle of the application should be covered in the scope of protection of the application.
Claims (5)
1. A portable fully automatic fluorescent nucleic acid detector, comprising:
the sample extraction box (11), be equipped with pyrolysis reaction chamber (12) in sample extraction box (11), install pyrolysis reaction chamber bag (121) in pyrolysis reaction chamber (12), pyrolysis reaction chamber bag (121) top is connected with pyrolysis liquid storage module (20), magnetic bead liquid storage module (21) and sample liquid storage module (22), pyrolysis liquid storage module (20), magnetic bead liquid storage module (21) and sample liquid storage module (22) can annotate liquid to pyrolysis reaction chamber bag (121);
the magnetic rod assembly comprises a moving slide plate (80), a magnetic rod (81) arranged on the moving slide plate (80), a magnetic rod movement control module (82) for driving the moving slide plate (80) to move along the horizontal and vertical directions, and a magnetic rod oscillation module (83) for driving the magnetic rod (81) to rotate in a clearance and reciprocate along the vertical direction;
a pyrolysis waste liquid cavity bag (122), wherein a cavity bag domain sealing clamp (60) is arranged between the pyrolysis waste liquid cavity bag (122) and the bottom of the pyrolysis reaction cavity bag (121);
the magnetic rod (81) can squeeze the outer wall of the cracking reaction cavity bag (121) to deform the cracking reaction cavity bag (121) and enable the outer wall of the cracking reaction cavity bag (121) to wrap the outer peripheral wall of the magnetic rod (81), and the magnetism of the magnetic rod (81) can be controlled according to on-off electricity; the magnetic rod (81) comprises a rod body (811) and a plurality of magnetic sheets (812) hinged to the periphery side of the bottom end of the rod body (811), when the outer wall of the cracking reaction cavity bag (121) wraps the outer peripheral wall of the magnetic rod (81), the magnetic sheets (812) are attached to the outer peripheral wall of the rod body (811), and when the bottom end of the rod body (811) is abutted to the top of the cracking reaction cavity bag (121), the magnetic sheets (812) are attached to the top of the cracking reaction cavity bag (121);
a washing reaction chamber (13) is further arranged in the sample extraction box (11), a washing reaction chamber bag (131) is arranged in the washing reaction chamber (13), a connecting pipe (132) is connected between the side wall of the washing reaction chamber bag (131) close to the top and the side wall of the cracking reaction chamber bag (121) close to the top, and a chamber bag split-domain sealing clamp (60) is also arranged on the connecting pipe (132);
the lysate storage module (20), the bead solution storage module (21) and the sample solution storage module (22) each include:
the tube body (30), the said tube body (30) is connected with reaction chamber bag of cleavage (121);
the reagent tube (40) is used for storing a lysate, a magnetic bead solution or a sample solution, the reagent tube (40) is arranged in the tube body (30) in a vertically sliding and sealing manner, and a puncturable sealing film (401) is arranged at the bottom of the reagent tube (40);
a puncturing member (50) mounted on the inner bottom of the tube body (30), the puncturing member (50) being capable of puncturing the sealing film (401) when the reagent tube (40) moves downward;
the bottom end of the reagent tube (40) is provided with a tube cover (402), the tube cover (402) is in sliding connection with the inner wall of the tube body (30), an air pressure chamber (61) is formed among the inner wall of the tube body (30), the tube body (30) and the outer wall of the tube cover (402), and an air tube (62) is arranged in the air pressure chamber (61);
the sample extraction box (11) is internally provided with an elution reaction chamber (14), the elution reaction chamber (14) is internally provided with an elution reaction chamber bag (141), the elution reaction chamber bag (141) is connected with the washing reaction chamber bag (131), and the magnetic rod (81) can drive the magnetic beads to move and transfer in the cracking reaction chamber bag (121), the washing reaction chamber bag (131) and the elution reaction chamber bag (141).
2. The portable full-automatic fluorescent nucleic acid detector according to claim 1, wherein the magnetic rod (81) is slidably connected with the moving slide plate (80) along the vertical direction, and the magnetic rod oscillation module (83) comprises a magnetic rod rotation driving assembly for driving the magnetic rod (81) to rotate and a swash plate control assembly for driving the magnetic rod (81) to reciprocate.
3. The portable fully automatic fluorescent nucleic acid detector of claim 2, wherein: the magnetic rod rotation driving assembly includes:
a magnetic rod driving motor (841) mounted on the moving slide plate (80);
a drive shaft (842) fixed to the output end of the magnetic rod drive motor (841);
an incomplete drive wheel (843) fixed to the outer periphery of the drive shaft (842);
a driven wheel (844) which is slidably mounted on the periphery of the magnetic rod (81), and a transmission belt (846) is connected between the incomplete driving wheel (843) and the driven wheel (844);
the swash plate control assembly includes:
a swash plate (851) fixed to the outer periphery of the drive shaft (842);
and a bevel shaft (852) fixed to the outer wall of the magnetic rod (81), wherein the circumferential surface of the swash plate (851) is in sliding contact with the end of the bevel shaft (852).
4. The portable fully automatic fluorescent nucleic acid detector of claim 1, wherein: the piercing member (50) comprises a conical piercing arrow body, a clamping block (403) is arranged on the inner wall of the reagent tube (40), the clamping block (403) can be connected with the conical piercing arrow body in a clamping mode to prevent the reagent tube (40) from moving towards a direction away from the cracking reaction cavity bag (121), and gas output by the gas tube (62) can enter the reagent tube (40) through a gap between the tube cover (402) and the tube body (30) and enter the cracking reaction cavity bag (121) through the reagent tube (40).
5. A method of nucleic acid extraction based on the portable fully automatic fluorescent nucleic acid detector of any one of claims 1-4, comprising the steps of:
s1, adding corresponding solution into a lysate storage module (20), a magnetic bead solution storage module (21) and a sample solution storage module (22);
s2, injecting the sample liquid in the sample liquid storage module (22) and the pyrolysis liquid in the pyrolysis liquid storage module (20) into the pyrolysis reaction cavity bag (121), and oscillating the reaction liquid in the pyrolysis reaction cavity bag (121) by using the magnetic rod assembly;
s3, injecting the magnetic bead liquid in the magnetic bead liquid storage module (21) into the cracking reaction cavity bag (121), and controlling the positions of the magnetic beads in the cracking reaction cavity bag (121) and adsorbing the magnetic beads by using the magnetic rod assembly;
s4, driving the magnetic beads to transfer by using the magnetic rod assembly.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108410859A (en) * | 2018-04-28 | 2018-08-17 | 绍兴迅敏康生物科技有限公司 | Method for extracting nucleic acid |
| CN112048429A (en) * | 2020-09-25 | 2020-12-08 | 杭州迪安生物技术有限公司 | High-efficient of nucleic acid extraction appearance is drawed and is shaken module and nucleic acid extraction appearance |
| CN113234591A (en) * | 2021-05-28 | 2021-08-10 | 宁波康程德诺生物医药有限公司 | Integrated nucleic acid rapid-extraction test tube, rapid-extraction detection device and method |
| CN216764861U (en) * | 2021-11-03 | 2022-06-17 | 杭州比格飞序生物科技有限公司 | Nucleic acid extraction and amplification device |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108410859A (en) * | 2018-04-28 | 2018-08-17 | 绍兴迅敏康生物科技有限公司 | Method for extracting nucleic acid |
| CN112048429A (en) * | 2020-09-25 | 2020-12-08 | 杭州迪安生物技术有限公司 | High-efficient of nucleic acid extraction appearance is drawed and is shaken module and nucleic acid extraction appearance |
| CN113234591A (en) * | 2021-05-28 | 2021-08-10 | 宁波康程德诺生物医药有限公司 | Integrated nucleic acid rapid-extraction test tube, rapid-extraction detection device and method |
| CN216764861U (en) * | 2021-11-03 | 2022-06-17 | 杭州比格飞序生物科技有限公司 | Nucleic acid extraction and amplification device |
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