CN112175809A - Integrated nucleic acid detection workstation - Google Patents
Integrated nucleic acid detection workstation Download PDFInfo
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- CN112175809A CN112175809A CN202011166955.6A CN202011166955A CN112175809A CN 112175809 A CN112175809 A CN 112175809A CN 202011166955 A CN202011166955 A CN 202011166955A CN 112175809 A CN112175809 A CN 112175809A
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- nucleic acid
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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
- C12Q1/6844—Nucleic acid amplification reactions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
Abstract
The invention discloses an integrated nucleic acid detection workstation, which comprises a shell and a workstation body arranged in the shell, wherein the workstation body comprises a reagent rack assembly, consumables, a mechanical arm assembly, a magnetic rack assembly and a fluorescence detection assembly; the consumable comprises a plurality of test tubes, and the test tubes are used as carriers for processing samples; the reagent rack assembly comprises a temperature control unit, and the temperature control unit provides required temperature for sample treatment; the mechanical arm assembly comprises a movably arranged liquid transferring head and a liquid transferring head which is used for sucking, blowing, beating and uniformly mixing liquid in the sample treatment process; the magnetic frame component is used for extracting and separating nucleic acid of a sample; the fluorescence detection component is used for carrying out fluorescence detection analysis on the amplified nucleic acid. The operator only needs to place the consumable and the nucleic acid amplification tube which are provided with the sample behind the reagent rack assembly, and the whole nucleic acid detection process is automatically carried out, so that the efficiency of nucleic acid detection is improved, the pollution risk is reduced, and the accuracy of nucleic acid detection is improved.
Description
Technical Field
The invention relates to the technical field of molecular diagnosis, in particular to an integrated nucleic acid detection workstation.
Background
The pretreatment of biological samples, nucleic acid extraction and purification, and nucleic acid amplification detection are important experimental means for research in molecular biology, and the traditional experimental methods are all performed manually by a user or performed by stages by a machine, including but not limited to sample pretreatment, nucleic acid extraction and purification, preparation and subpackaging of nucleic acid amplification reagents, sample adding of nucleic acid solutions, closing of amplification reagent tubes, detection in a fluorescence PCR instrument or other chemical reaction instruments which can be used for nucleic acid amplification, and result analysis and interpretation in other ways, so that the time consumption is long, biological sample pollution or amplification product pollution is easy to occur, and the experimental results are easily influenced by individual operation differences of experimenters, so that the experimental results are unreliable, and the actual application of the molecular detection technology is seriously influenced by the occurrence of false positive and false negative results.
At present, based on the traditional solution, the instruments for extracting, amplifying and detecting nucleic acid commercially used for diagnosis are all separated, such as a sample preparation instrument for extracting nucleic acid, a nucleic acid extractor, a quantitative PCR instrument for amplification detection, or other nucleic acid amplification detectors. The method comprises the specific processes of manually adding a sample into a full-automatic nucleic acid extractor, manually transferring the sample to a nucleic acid amplification detection instrument after the nucleic acid extraction and purification are carried out on the biological sample by using the full-automatic nucleic acid extractor, carrying out amplification detection on the extract by using the nucleic acid amplification detection instrument, and manually completing all the transfer operation links by experimenters. The key point of the current nucleic acid detection field is that the current physicochemical means and various automatic tools are utilized to realize the 'sample in and out' and the whole molecular diagnosis process in one instrument and equipment.
Disclosure of Invention
The invention aims to provide an integrated nucleic acid detection workstation, which can integrate sample pretreatment, nucleic acid extraction and purification, amplification and detection into a whole and realize automatic and efficient detection of nucleic acid.
In order to achieve the purpose, the invention adopts the following technical scheme:
the integrated nucleic acid detection workstation comprises a shell and a workstation body arranged in the shell, wherein the workstation body comprises a reagent rack assembly, consumables, a mechanical arm assembly, a magnetic rack assembly and a fluorescence detection assembly;
the reagent rack assembly comprises a consumable support for placing the consumable, the consumable comprises a plurality of test tubes, and the test tubes are used as carriers for processing samples;
the reagent rack assembly further comprises a temperature control unit, the temperature control unit is arranged below the consumable support and provides required temperature for sample treatment;
the mechanical arm assembly is arranged above the reagent rack assembly, and comprises a movably arranged liquid transferring head which is used for sucking, blowing, beating and uniformly mixing liquid in the sample processing process;
the magnetic frame assembly is used for extracting and separating nucleic acid of a sample;
the fluorescence detection assembly comprises a movably arranged detection unit, and the detection unit is used for carrying out fluorescence detection analysis on the amplified nucleic acid.
As a preferable scheme of the invention, the consumable further comprises a substrate, a plurality of suckers, a sucker placing groove and a waste liquid groove, wherein the plurality of test tubes comprise sample tubes for placing samples, reagent tubes for cracking the samples and reaction tubes for extracting and separating nucleic acids of the samples, the waste liquid groove is close to the reaction tubes and is arranged at intervals with the reaction tubes, hole sites for placing nucleic acid amplification tubes are arranged on the reagent frame assembly, and the nucleic acids of the samples are transferred into the nucleic acid amplification tubes from the reaction tubes for amplification.
In a preferred embodiment of the present invention, the consumable surface further comprises a sealing film that seals openings of the tip placement groove, the waste liquid tank, the sample tube, the reagent tube, and the reaction tube.
As a preferred embodiment of the present invention, the integrated nucleic acid detecting workstation further comprises an amplification tube cap assembly, the amplification tube cap assembly comprises a first motor, a rotating shaft, a first fixing seat, a second fixing seat and a plurality of pressing blocks, two ends of the rotating shaft are respectively connected to the first fixing seat and the second fixing seat, the first fixing seat and the second fixing seat are disposed on the reagent rack assembly, the pressing blocks are disposed on the rotating shaft, and the first motor is disposed at one end of the rotating shaft and can drive the rotating shaft to rotate, so that the pressing blocks can selectively seal the nucleic acid amplification tube.
As a preferable scheme of the present invention, the liquid-transferring head includes a fixed support, a guide sleeve, a second motor, a screw rod, a nut, and a plurality of head rods, the head rods are used for inserting and taking out the heads, the guide sleeve and the second motor are both disposed on the fixed support, the screw rod is connected to the second motor and can rotate under the driving of the second motor, the screw rod is vertically disposed, the nut is sleeved on the screw rod and can move along the screw rod, one end of the head rod is fixedly connected to the nut, a plurality of through holes for inserting the other end of the head rod and moving up and down are disposed in the guide sleeve, the through holes are stepped holes, and the through holes can limit the heads.
As a preferable mode of the present invention, the mechanical arm assembly further includes a syringe pump, a first driving member, and a second driving member, an end of the pipette tip rod away from the guide sleeve is connected to the syringe pump through a hose, the first driving member is connected to the pipette tip, the first driving member moves the pipette tip in a first direction within a horizontal plane, the first driving member is provided on the second driving member, the second driving member moves the first driving member in a second direction within the horizontal plane, and the first direction is perpendicular to the second direction.
As a preferable scheme of the present invention, the magnetic frame assembly includes a support plate, a lifting unit, a magnet fixing seat and a plurality of magnets, the lifting unit is disposed on the support plate, the magnet fixing seat is connected to the lifting unit, the plurality of magnets are disposed on the magnet fixing seat at intervals, and the lifting unit can drive the fixing seat to move in a vertical direction so as to enable the magnets to approach or leave the reaction tube.
As a preferable scheme of the invention, the reagent rack assembly further comprises a positioning plate, a plurality of fans and a fixing plate, the consumable support can be placed on and fixed on the positioning plate, the plurality of fans, the positioning plate and the temperature control unit are all connected with the fixing plate, the temperature control unit comprises a first temperature control part and a second temperature control part, the first temperature control part is used for heating the reaction tube, and the second temperature control part is used for heating or cooling the nucleic acid amplification tube.
As a preferable scheme of the invention, the fluorescence detection assembly further comprises a slide block, a guide rail and a driving unit, the detection unit is arranged on the slide block, and the driving unit can drive the slide block to do reciprocating linear motion along the guide rail so as to enable the detection unit to perform fluorescence detection on the nucleic acid amplification tube.
As a preferable scheme of the invention, an air draft filtering device and an ultraviolet lamp sterilizing device are further arranged inside the shell.
The invention has the beneficial effects that:
the integrated nucleic acid detection workstation of the invention is provided with a reagent rack assembly, a consumable, a mechanical arm assembly, a magnetic rack assembly and a fluorescence detection assembly in a shell, wherein the consumable comprises a plurality of test tubes which are used as carriers for processing samples, the processing process comprises the pretreatment of the samples and the extraction and purification of nucleic acids of the samples, the consumable can adopt a form without a nucleic acid amplification tube and directly uses the nucleic acid amplification tube configured in the detection kit, thereby saving resources, a temperature control unit in the reagent rack assembly provides required temperature for the nucleic acid extraction and purification of the samples and the amplification process of the nucleic acids, a pipetting head movably arranged in the mechanical arm assembly can perform pipetting and blowing operations on various types of test tubes to complete the processing process, and after the nucleic acid amplification is completed, the fluorescence detection assembly can perform fluorescence detection analysis, therefore, the operator only needs to place the consumptive material and the nucleic acid amplification tube that are equipped with the sample behind the reagent frame subassembly, and whole nucleic acid testing process will go on inside the shell automatically, need not to carry out artifical manual operation, has not only improved nucleic acid testing's efficiency greatly, has still reduced the pollution risk, has improved nucleic acid testing's accuracy.
Drawings
FIG. 1 is a schematic perspective view of an integrated nucleic acid detection workstation according to an embodiment of the present invention;
FIG. 2 is a schematic side view of a consumable according to an embodiment of the invention;
FIG. 3 is a perspective view of one embodiment of a workstation body;
FIG. 4 is a second perspective view of the workstation body according to the embodiment of the present invention;
FIG. 5 is an exploded view of the workstation body of FIG. 1;
FIG. 6 is a schematic perspective view of an embodiment of an amplification tube closure assembly;
FIG. 7 is a schematic perspective view of a pipetting head according to an embodiment of the invention;
FIG. 8 is a perspective view of the pipetting head body of FIG. 7;
FIG. 9 is a schematic cross-sectional view of a guide sleeve of an embodiment of the present invention;
FIG. 10 is a perspective view of a magnetic carrier assembly according to an embodiment of the present invention;
FIG. 11 is one of the schematic perspective views of a reagent rack assembly according to an embodiment of the present invention;
FIG. 12 is an exploded view of FIG. 11;
FIG. 13 is a second schematic perspective view of a reagent rack assembly according to an embodiment of the present invention;
FIG. 14 is a perspective view of a fluorescence detection assembly according to an embodiment of the invention.
In the figure:
1. a housing; 2. a reagent rack assembly; 21. a consumable support; 211. opening a hole; 22. positioning a plate; 23. a fan; 24. a fixing plate; 251. a first temperature control member; 252. a second temperature control member; 2521. a semiconductor refrigeration sheet; 2522. a heat sink; 26. fixing the rod; 3. consumable materials; 31. a sample tube; 32. a reagent tube; 33. a reaction tube; 34. a substrate; 35. a suction head; 36. a suction head placing groove; 37. a waste liquid tank; 38. a liquid washing pipe;
4. a mechanical arm assembly; 41. a pipetting head; 411. a protective shell; 412. a pipetting head body; 4121. a suction head rod; 4122. fixing a bracket; 4123. a guide sleeve; 41231. a through hole; 4124. a second motor; 4125. a screw rod; 4126. a nut; 42. an injection pump; 43. a first driving member; 44. a second driving member;
5. a magnetic frame assembly; 51. a support plate; 52. a lifting unit; 53. a magnet fixing seat; 54. a magnet; 6. a fluorescence detection component; 61. a detection unit; 62. a slider; 63. a guide rail; 64. a drive unit; 7. a workstation base; 71. a base plate; 72. a main frame; 8. a battery; 9. an amplification tube cap assembly; 91. a first motor; 92. a rotating shaft; 93. a first fixed seat; 94. a second fixed seat; 95. and (7) briquetting.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
In the present invention, unless otherwise expressly stated or limited, "above" or "below" a first feature means that the first and second features are in direct contact, and that the first and second features are not in direct contact but are in contact with each other via another feature therebetween. Also, the word "over" a first feature or feature in a second feature may include the word "over" or "over" the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature "under" a second feature may include a first feature that is directly under and obliquely under the second feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
Furthermore, in the description of the present invention, unless explicitly stated or limited otherwise, the terms "first", "second", and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicit indication of the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.
As shown in fig. 1 to 5, the integrated nucleic acid detecting workstation of the embodiment of the present invention includes a housing 1 and a workstation body disposed inside the housing 1, wherein the workstation body includes a reagent rack assembly 2, a consumable material 3, a mechanical arm assembly 4, a magnetic rack assembly 5 and a fluorescence detecting assembly 6;
the consumable 3 can be placed on the reagent rack assembly 2, and the consumable 3 comprises a plurality of test tubes which are used as carriers for processing samples;
the reagent rack assembly 2 comprises a temperature control unit which provides required temperature for sample treatment;
as shown in fig. 4, the robot arm assembly 4 includes a movably disposed pipetting head 41, and the pipetting head 41 is used for sucking, blowing, and mixing liquid in the sample processing process;
the magnetic frame component 5 is used for extracting and separating nucleic acid of a sample;
the fluorescence detection component 6 is used for carrying out fluorescence detection analysis on the amplified nucleic acid.
The integrated nucleic acid detection workstation of the invention is provided with a reagent rack assembly 2, a consumable 3, a mechanical arm assembly 4, a magnetic rack assembly 5 and a fluorescence detection assembly 6 in a shell 1, wherein the consumable 3 comprises a plurality of test tubes, the plurality of test tubes are used as carriers for processing samples, the processing process comprises the steps of sample pretreatment, nucleic acid extraction and purification of the samples, the consumable 3 can be in a form without a nucleic acid amplification tube, the nucleic acid amplification tube arranged in a detection kit is directly used for nucleic acid amplification, thereby saving resources, a temperature control unit in the reagent rack assembly 2 provides required temperature for the nucleic acid extraction and purification of the samples and the nucleic acid amplification process, a liquid transfer head 41 movably arranged in the mechanical arm assembly 4 can carry out liquid transfer and blow-blow operations on various types of test tubes to complete the processing process of the samples, after the nucleic acid amplification is completed, fluorescence detection subassembly 6 can carry out fluorescence detection analysis, consequently, operating personnel only need will be equipped with the consumptive material 3 and the nucleic acid amplification pipe of sample and place behind reagent frame subassembly 2, and whole nucleic acid detects the flow and will go on in that the shell 1 is inside automatically, need not to carry out artifical manual operation, has not only improved nucleic acid detection's efficiency greatly, has still reduced the pollution risk, has improved nucleic acid detection's accuracy.
In this embodiment, the workstation body further comprises a workstation base 7 and a battery 8, and the reagent rack assembly 2, the robot arm assembly 4, the magnetic rack assembly 5 and the fluorescence detection assembly 6 are all mounted on the workstation base 7. Preferably, the workstation base 7 comprises a bottom plate 71 and a main frame 72 disposed on the bottom plate 71, the reagent rack assembly 2 is disposed on the main frame 72 and is fixedly connected with the bottom plate 71, the robot arm assembly 4 is disposed on the main frame 72, and the magnetic rack assembly 5 and the fluorescence detection assembly 6 are disposed on the bottom plate 71. The battery 8 is fixed on the bottom plate 71, so that the integrated nucleic acid detecting workstation of the embodiment can be operated without plugging.
As shown in fig. 2, further, the plurality of test tubes include a sample tube 31, a reagent tube 32 and a reaction tube 33, the reagent tube 32 contains reagents such as proteinase K and magnetic beads, the sample is transferred from the sample tube 31 to the reagent tube 32 through a liquid transfer head 41 for lysis, then the mixed liquid containing proteinase K, magnetic beads and the sample after being mixed is transferred to the reaction tube 33 containing a magnetic bead binding liquid for binding the magnetic beads with free nucleic acids, the nucleic acids are purified and eluted, separated from the magnetic beads through a magnetic rack assembly 5 and transferred to a nucleic acid amplification tube for amplification, a temperature control unit in the reagent rack assembly 2 heats or cools the reaction tube 33 and the nucleic acid amplification tube to provide a temperature required for reaction, and the fluorescence detection assembly 6 performs fluorescence detection analysis on the nucleic acids amplified in the nucleic acid amplification tube. The whole treatment process of the sample is automatically completed in various test tubes of the consumable 3 placed on the reagent rack assembly 2 by means of the pipetting head 41, and the whole process is efficient, convenient and applicable to clinical detection.
As shown in fig. 6, in some embodiments, the integrated nucleic acid detecting station further includes an amplification tube cover assembly 9, the amplification tube cover assembly 9 includes a first motor 91, a rotating shaft 92, a first fixing seat 93, a second fixing seat 94 and a plurality of pressing blocks 95, two ends of the rotating shaft 92 are respectively connected to the first fixing seat 93 and the second fixing seat 94, the first fixing seat 93 and the second fixing seat 94 are disposed on the reagent rack assembly 2, the plurality of pressing blocks 95 are disposed on the rotating shaft 92, and the first motor 91 is disposed at one end of the rotating shaft 92 and can drive the rotating shaft 92 to rotate, so that the pressing blocks 95 selectively close the nucleic acid amplification tubes.
When liquid transfer operation or nucleic acid amplification reaction is carried out, along with air and liquid surface friction or liquid volatilization, nucleic acid aerosol can be generated, the nucleic acid aerosol is tiny water drops containing nucleic acid fragments with different lengths, and the tiny water drops are settled in other samples to cause aerosol pollution, so that the detection result is inaccurate. The amplification tube cover assembly 9 seals the nucleic acid amplification tube when the nucleic acid amplification tube does not need to be subjected to pipetting operation, so that the possibility of aerosol pollution can be greatly reduced, and the accuracy of a detection result is improved.
As shown in fig. 2, further, consumable 3 further includes a substrate 34 and a plurality of suction heads 35, the substrate 34 is provided with a plurality of suction head placement grooves 36, a waste liquid groove 37 and a washing liquid pipe 38, the plurality of suction heads 35 are placed in the suction head placement grooves 36, the waste liquid groove 37 is close to the reaction tube 33 and is spaced from the reaction tube 33, consumable 3 of the present embodiment adopts an integrated structure, that is, the sample tube 31, the reagent tube 32, the reaction tube 33, the substrate 34, suction head placement grooves 36, waste liquid groove 37 and washing liquid pipe 38 are integrally formed, all types of test tubes do not shake or fall, and daily use is facilitated.
As shown in FIG. 7, the pipette tip 41 includes a plurality of tip rods 4121 movable in the vertical direction, the tip rods 4121 are used for inserting the tips 35 in the tip placement grooves 36, and the wash solution tube 38 contains a wash solution or an eluent for purification of nucleic acids and separation of magnetic beads from nucleic acids. The waste liquid tank 37 is arranged close to the reaction tube 33, so that the moving distance of the liquid transfer head 41 when the waste liquid in the reaction tube 33 is discharged can be reduced, the magnetic frame assembly 5 needs to be frequently close to and far away from the reaction tube 33, therefore, the reaction tube 33 needs to be separated from the waste liquid tank 37 and other test tubes to provide enough moving space for the magnetic frame assembly 5, the suction head placing groove 36 enables the suction heads 35 to be placed neatly, the suction heads 35 can be conveniently inserted and taken by the suction head rod 4121, and when the suction heads 35 need to be discarded, the suction heads 35 needing to be discarded can be placed back to the original positions in the suction head placing groove 36. The substrate 34 may also be plastic-sealed with a sealing film, such as a plastic film, an aluminum film or a paper film, for sealing the consumable 3 for loading reagents and protecting the consumable during transportation.
As shown in fig. 7 to 9, further, the pipetting head 41 comprises a protective housing 411 and a pipetting head body 412, the pipetting head body 412 further comprises a fixed bracket 4122, a guide sleeve 4123, a second motor 4124, a screw rod 4125 and a nut 4126 besides the pipette tip rod 4121, the guide sleeve 4123 and the second motor 4124 are both arranged on the fixed bracket 4122, the screw rod 4125 is connected with the second motor 4124 and can rotate under the driving of the second motor 4124, the screw rod 4125 is vertically arranged, the nut 4126 is sleeved on the screw rod 4125 and can move along the screw rod 4125, one end of the pipette tip rod 4121 is fixedly connected with the nut 4126, a plurality of through holes 41231 for inserting and moving up and down the other end of the pipette tip rod 4121 are arranged in the guide sleeve 4123, the through holes 41231 are step holes, and the through holes 41231 can limit the pipette tip 35. The transmission mode of the ball screw has the characteristics of high precision and stability, so that the sucker 4121 can be accurately positioned. When the through hole 41231 formed in the guide sleeve 4123 is a stepped hole, and the suction head bar 4121 with the suction head 35 is lifted in the through hole 41231 when the suction head 35 needs to be discarded, the top of the suction head 35 is abutted against the stepped structure, so that the suction head 35 cannot continuously lift along with the suction head bar 4121, and the separation of the suction head 35 and the suction head bar 4121 is realized.
As shown in fig. 4 and 5, the robotic arm assembly 4 preferably further comprises a syringe pump 42, a first driving member 43 and a second driving member 44, wherein the ends of the plurality of pipette tip rods 4121 away from the guide sleeve 4123 are connected to the syringe pump 42 through a hose (not shown), the first driving member 43 is connected to the pipette tip 41, the first driving member 43 moves the pipette tip 41 in a first direction within the horizontal plane, the first driving member 43 is disposed on the second driving member 44, and the second driving member 44 moves the first driving member 43 in a second direction within the horizontal plane, the first direction being perpendicular to the second direction. Since the pipette tip 4121 can move up and down in the vertical direction, and the first driving member 43 and the second driving member 44 enable the pipetting head 41 to move freely in the horizontal plane, the pipette tip 4121 can have the ability to move and position precisely in three-dimensional space, and of course, in order to achieve this function, a plurality of sensors (not shown) are further provided on the robot arm assembly 4, so as to precisely monitor the distance and direction of the displacement, and ensure that the pipette tip 4121 is always in the correct position.
As shown in fig. 10, preferably, the magnetic frame assembly 5 includes a support plate 51, a lifting unit 52, a magnet holder 53 and a plurality of magnets 54, wherein the lifting unit 52 is disposed on the support plate 51, the magnet holder 53 is connected to the lifting unit 52, the plurality of magnets 54 are disposed on the magnet holder 53 at intervals, and the lifting unit 52 can drive the magnet holder 53 to move in a vertical direction so that the magnets 54 are close to or far from the reaction tube 33. The magnet fixing seat 53 is configured to be elongated, and can be smoothly inserted into the side surface of the reaction tube 33, so that the magnet 54 disposed on the magnet fixing seat 53 can adsorb the magnetic beads in the reaction tube 33, and since the reaction tubes 33 generally have a large number, the magnets 54 are disposed at intervals, and each magnet 54 corresponds to one reaction tube 33, thereby ensuring a good magnetic bead adsorption effect. The elevator unit 52 may also employ a ball screw drive pattern similar to that in the pipetting head 41 to achieve precise movement of the position of the magnet 54.
Preferably, as shown in fig. 11 to 13, the reagent rack assembly 2 further includes a plurality of consumable supports 21, a positioning plate 22, a plurality of fans 23 and a fixing plate 24, the consumable 3 is disposed on the consumable support 21, the consumable support 21 is disposed on the positioning plate 22, the plurality of fans 23, the positioning plate 22 and the temperature control unit are all connected to the fixing plate 24, the fixing plate 24 is fixed on the bottom plate 71 through a plurality of fixing rods 26, the temperature control unit includes a first temperature control 251 and a second temperature control 252, the first temperature control 251 is used for heating the reaction tube 33, the second temperature control 252 is used for heating or cooling the nucleic acid amplification tube, the second temperature control 252 includes a plurality of semiconductor cooling fins 2521 and a plurality of cooling fins 2522, and the plurality of fans 23 are used for assisting in cooling the second temperature control 252 and the nucleic acid amplification tube. Consumable support 21 can wholly take off from locating plate 22, and offers the trompil 211 of the staff gripping of being convenient for on the consumable support 21, makes things convenient for putting into and taking out of consumable support 21 and the consumptive material 3 of placing on consumable support 21. The nucleic acid amplification reaction needs to be heated and then cooled, and has certain requirements on the cooling rate, so that the nucleic acid amplification tube is heated or cooled by the semiconductor cooling sheet 2521 which can heat and cool, and is matched with the plurality of cooling fins 2522 and the plurality of fans 23 to rapidly cool.
As shown in FIG. 14, it is preferable that the fluorescence detection unit 6 includes two detection units 61, a slide block 62, a guide rail 63 and a driving unit 64, the detection unit 61 is disposed on the slide block 62, and the driving unit 64 drives the slide block 62 to reciprocate linearly along the guide rail 63, so that the detection unit 61 is used for emission of excitation light and collection of reflected light to perform fluorescence detection of the nucleic acid amplification tube. The number of the detecting units 61 is at least one, and the fluorescence detecting assembly 6 of the present embodiment includes two detecting units 61, so that two different nucleic acid amplification tubes can be detected at the same time, and when the number of the nucleic acid amplification tubes to be detected is large, the detection can be completed in a short time. The driving unit 64 may also adopt a transmission mode similar to a ball screw in the pipetting head 41 to achieve stabilization of the movement process of the detection unit 61.
Preferably, an air draft filtering device (not shown in the figure) and an ultraviolet lamp sterilizing device (not shown in the figure) are further arranged inside the casing 1. The air draft filtering device can play a role in heat dissipation and ventilation, and can prevent external pollutants from entering the shell 1, and the ultraviolet lamp sterilizing device can reduce or prevent biological sample pollution and/or nucleic acid pollution in the detection process.
The following is an embodiment of an integrated nucleic acid detection workstation of an embodiment to illustrate specific detection steps:
one, manual operation part
(1) Taking out the consumable support 21 from the integrated nucleic acid detection workstation, and placing the consumable 3 on the consumable support 21;
(2) adding a sample to be detected into the sample tube 31, inserting the nucleic acid amplification tube into the matched hole, putting the consumable support 21 and the consumable 3 into the integrated nucleic acid detection workstation, starting up, selecting a program, and starting to operate.
Secondly, automatic operation:
before the experiment begins, the amplification tube cover assembly 9 is started, the nucleic acid amplification tube is sealed by the pressing block 95, the air draft filtering device is started, and then nucleic acid extraction is carried out, wherein the method comprises the following steps:
(1) inserting the pipette tip 35 into the pipette tip 41, taking the sample liquid from the sample tube 31, adding the sample liquid into the reagent tube 32 containing the proteinase K and the magnetic beads, repeatedly blowing the liquid for at least one time to blow away the magnetic beads, and returning the pipette tip 41 to the original position;
(2) transferring the uniformly mixed protease K, magnetic beads and sample mixed solution to a reaction tube 33 containing a magnetic bead binding solution;
(3) heating the reaction tube 33, during which the liquid is repeatedly blown at least once;
(4) the magnet 54 in the magnetic frame assembly 5 moves to the middle part of the sample solution on the side of the reaction tube 33 for staying and attracting magnetism;
(5) completely sucking the whole liquid in the reaction tube 33 by at least one liquid suction of the liquid-transferring head 41 and injecting the liquid into the waste liquid tank 37;
(6) sucking the washing liquid from the washing liquid pipe 38 into the reaction tube 33, and slowly moving the magnet 54 away from the reaction tube 33;
(7) repeatedly pumping the liquid for at least one time to ensure that the magnetic beads are fully washed;
(8) the magnet 54 in the magnetic frame assembly 5 moves to the middle part of the sample solution on the side of the reaction tube 33 again for staying and magnetic absorption;
(9) the magnet 54 is far away from the reaction tube 33, and the temperature of the first temperature control 251 is increased to evaporate the liquid left in the reaction tube 33;
(10) inserting the pipette tip 41 into the pipette tip 35, sucking the eluent from the wash tube 38 into the reaction tube 33, repeatedly blowing the liquid at least once, and heating the first temperature control member 251 to elute the nucleic acid;
(11) moving the magnet 54 to the middle of the sample solution on the side of the reaction tube 33 for magnetic absorption, moving the magnet 54 further upward, and collecting all magnetic beads on the upper part of the tube wall and far away from the tube bottom;
(12) the press block 95 is opened, the nucleic acid amplification tube is opened, a certain amount of nucleic acid is taken out from the reaction tube 33 into the nucleic acid amplification tube, the blowing is repeated, and the nucleic acid amplification detection step is performed.
The nucleic acid amplification detection comprises the following steps:
firstly, the cover assembly 9 of the amplification tube is started to seal the nucleic acid amplification tube, the second temperature control element 252 starts to heat to provide the temperature required by the nucleic acid amplification reaction, and the nucleic acid reaction is ensured to be carried out;
then, in the detection stage, the detection unit 61 in the fluorescence detection assembly 6 sequentially detects each nucleic acid amplification tube, and obtains the detection result through the real-time fluorescence signal.
After the sterilization, the ultraviolet lamp is automatically turned on for ultraviolet sterilization.
This detection step simulates people manual operation process, will draw, amplify and detect and concentrate on an organic whole, and operating personnel only need carry out simple preparation action and can accomplish whole testing process, reachs the result, has that sample handling capacity is big, characteristics that whole process is high-efficient, convenient, is applicable to clinical testing.
Reference throughout this specification to the description of the terms "preferred," "further," or the like, as used in describing preferred embodiments of the present invention, means that a particular feature, structure, material, or characteristic described in connection with the example or illustration is included in at least one example or illustration of the present invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The above examples are only intended to illustrate the details of the invention, which is not limited to the above details, i.e. it is not intended that the invention must be implemented in such detail. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (10)
1. An integrated nucleic acid detection workstation is characterized by comprising a shell and a workstation body arranged in the shell, wherein the workstation body comprises a reagent rack assembly, consumables, a mechanical arm assembly, a magnetic rack assembly and a fluorescence detection assembly;
the reagent rack assembly comprises a consumable support for placing the consumable, the consumable comprises a plurality of test tubes, and the test tubes are used as carriers for processing samples;
the reagent rack assembly further comprises a temperature control unit, the temperature control unit is arranged below the consumable support and provides required temperature for sample treatment;
the mechanical arm assembly is arranged above the reagent rack assembly, and comprises a movably arranged liquid transferring head which is used for sucking, blowing, beating and uniformly mixing liquid in the sample processing process;
the magnetic frame assembly is used for extracting and separating nucleic acid of a sample;
the fluorescence detection assembly comprises a movably arranged detection unit, and the detection unit is used for carrying out fluorescence detection analysis on the amplified nucleic acid.
2. The integrated nucleic acid detection workstation according to claim 1, wherein the consumable further comprises a substrate, a plurality of suction heads, a suction head placement groove and a waste liquid groove, wherein the plurality of test tubes comprise sample tubes for placing samples, reagent tubes for cracking the samples and reaction tubes for extracting and separating nucleic acids of the samples, the waste liquid groove is arranged close to and spaced from the reaction tubes, hole sites for placing nucleic acid amplification tubes are arranged on the reagent frame assembly, and the nucleic acids of the samples are transferred from the reaction tubes to the nucleic acid amplification tubes for amplification.
3. The integrated nucleic acid detecting workstation according to claim 2, wherein the consumable surface is further provided with a sealing film that closes openings of the tip placement groove, the waste liquid groove, the sample tube, the reagent tube, and the reaction tube.
4. The integrated nucleic acid detecting workstation according to claim 2, further comprising an amplification tube cover assembly, wherein the amplification tube cover assembly comprises a first motor, a rotating shaft, a first fixing seat, a second fixing seat and a plurality of pressing blocks, two ends of the rotating shaft are respectively connected with the first fixing seat and the second fixing seat, the first fixing seat and the second fixing seat are disposed on the reagent rack assembly, the pressing blocks are disposed on the rotating shaft, and the first motor is disposed at one end of the rotating shaft and can drive the rotating shaft to rotate, so that the pressing blocks can selectively close the nucleic acid amplification tubes.
5. The integrated nucleic acid detecting workstation according to any one of claims 2 to 4, wherein the pipetting head comprises a fixed support, a guide sleeve, a second motor, a screw rod, a nut and a plurality of pipette head rods, the pipette head rods are used for inserting and taking the pipette heads, the guide sleeve and the second motor are both arranged on the fixed support, the screw rod is connected with the second motor and can rotate under the driving of the second motor, the screw rod is vertically arranged, the nut is sleeved on the screw rod and can move along the screw rod, one end of the pipette head rod is fixedly connected with the nut, a plurality of through holes for inserting the other end of the pipette head rod and moving up and down are arranged in the guide sleeve, the through holes are step holes, and the through holes can limit the pipette heads.
6. The integrated nucleic acid detecting workstation according to claim 5, wherein the mechanical arm assembly further comprises a syringe pump, a first driving member and a second driving member, an end of the pipette tip rod away from the guide sleeve is connected with the syringe pump through a hose, the first driving member is connected with the pipette tip, the first driving member moves the pipette tip in a first direction within a horizontal plane, the first driving member is disposed on the second driving member, the second driving member moves the first driving member in a second direction within the horizontal plane, and the first direction is perpendicular to the second direction.
7. The integrated nucleic acid detecting workstation according to any one of claims 2 to 4, wherein the magnetic frame assembly comprises a support plate, a lifting unit, a magnet fixing seat and a plurality of magnets, the lifting unit is disposed on the support plate, the magnet fixing seat is connected with the lifting unit, the plurality of magnets are disposed on the magnet fixing seat at intervals, and the lifting unit can drive the fixing seat to move in a vertical direction so as to enable the magnets to approach or depart from the reaction tubes.
8. The integrated nucleic acid detecting workstation according to any one of claims 2 to 4, wherein the reagent rack assembly further comprises a positioning plate, a plurality of fans and a fixing plate, the consumable support can be placed on and fixed on the positioning plate, the plurality of fans, the positioning plate and the temperature control unit are all connected with the fixing plate, the temperature control unit comprises a first temperature control part and a second temperature control part, the first temperature control part is used for heating the reaction tube, and the second temperature control part is used for heating or cooling the nucleic acid amplification tube.
9. The integrated nucleic acid detection workstation according to any one of claims 2 to 4, wherein the fluorescence detection assembly further comprises a slide block, a guide rail and a driving unit, the detection unit is disposed on the slide block, and the driving unit can drive the slide block to perform reciprocating linear motion along the guide rail, so that the detection unit can perform fluorescence detection on the nucleic acid amplification tube.
10. The integrated nucleic acid detection workstation of any one of claims 1 to 4, wherein an air draft filtering device and an ultraviolet lamp disinfection device are further arranged inside the housing.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011166955.6A CN112175809A (en) | 2020-10-27 | 2020-10-27 | Integrated nucleic acid detection workstation |
| PCT/CN2020/140654 WO2022088472A1 (en) | 2020-10-27 | 2020-12-29 | Integrated nucleic acid detection workstation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011166955.6A CN112175809A (en) | 2020-10-27 | 2020-10-27 | Integrated nucleic acid detection workstation |
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| CN112175809A true CN112175809A (en) | 2021-01-05 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202011166955.6A Pending CN112175809A (en) | 2020-10-27 | 2020-10-27 | Integrated nucleic acid detection workstation |
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| CN (1) | CN112175809A (en) |
| WO (1) | WO2022088472A1 (en) |
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| CN112300911A (en) * | 2020-10-27 | 2021-02-02 | 广州和实生物技术有限公司 | Nucleic acid detector and nucleic acid detection method |
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