CN115433667A - Biochemical reaction device and nucleic acid sequencing equipment - Google Patents
Biochemical reaction device and nucleic acid sequencing equipment Download PDFInfo
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- CN115433667A CN115433667A CN202210927075.9A CN202210927075A CN115433667A CN 115433667 A CN115433667 A CN 115433667A CN 202210927075 A CN202210927075 A CN 202210927075A CN 115433667 A CN115433667 A CN 115433667A
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- 238000005842 biochemical reaction Methods 0.000 title claims abstract description 69
- 238000012163 sequencing technique Methods 0.000 title claims abstract description 33
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 30
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 28
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 181
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 162
- 238000006243 chemical reaction Methods 0.000 claims abstract description 116
- 230000007246 mechanism Effects 0.000 claims abstract description 72
- 238000007599 discharging Methods 0.000 claims description 22
- 239000002699 waste material Substances 0.000 claims description 18
- 238000002791 soaking Methods 0.000 claims description 16
- 230000003287 optical effect Effects 0.000 claims description 10
- 238000004891 communication Methods 0.000 claims description 5
- 230000000712 assembly Effects 0.000 claims description 4
- 238000000429 assembly Methods 0.000 claims description 4
- 238000012840 feeding operation Methods 0.000 claims 1
- 239000012530 fluid Substances 0.000 abstract description 10
- 238000001802 infusion Methods 0.000 abstract description 5
- 238000010586 diagram Methods 0.000 description 11
- 238000000034 method Methods 0.000 description 10
- 238000001514 detection method Methods 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 238000009434 installation Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005574 cross-species transmission Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000003203 nucleic acid sequencing method Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
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- 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
- C12Q1/6869—Methods for sequencing
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Abstract
The application relates to the technical field of nucleic acid sequencing, and discloses a biochemical reaction device and nucleic acid sequencing equipment, wherein the biochemical reaction device comprises a reaction container, a liquid inlet mechanism and a liquid discharge mechanism, the reaction container is provided with a plurality of reagent grooves with openings, and each reagent groove is respectively communicated with a liquid inlet hole and a liquid outlet hole; feed liquor mechanism includes feed liquor subassembly and inlet line way, drain mechanism includes flowing back subassembly and drain line way, can realize holding of multiple reagent through a plurality of reagent grooves, can be convenient for the chip soaks different reagents in different reagent grooves, and, can carry out feed liquor and flowing back to the reagent groove through feed liquor mechanism and drain mechanism, thereby be convenient for fill of reagent, fluid infusion and update, so that the chip soaks continuously of operation many times and carries out, help improving reaction efficiency, and then improve the efficiency of nucleic acid sequencing. The nucleic acid sequencing apparatus having the biochemical reaction device also has the above-described advantages.
Description
Technical Field
The application relates to the technical field of nucleic acid sequencing, in particular to a biochemical reaction device and nucleic acid sequencing equipment.
Background
In the current second generation sequencing technology, a biochemical reaction part is usually realized by adopting a microfluidic system, and the biochemical reaction is realized by controllably flowing a reagent box fluid through a fluid channel of a flow cell and contacting a nucleic acid chain to be sequenced, which is connected on a flow cell substrate, mainly through the cooperation of a power source, a selection valve, a pipeline and the like.
In the related art, an open nucleic acid sequencing method is provided, which allows a chip carrying nucleic acid molecules to be soaked in different reaction reagents according to a set sequence to realize nucleic acid sequencing, and can avoid the problems of high-precision temperature control requirement and reagent waste caused by a closed fluid system. However, the conventional apparatus for performing open sequencing has problems that the arrangement of reaction vessels is not reasonable, and the addition and discharge of reaction reagents are difficult. Because the chip needs to be soaked in different reagents for many times by adopting an open sequencing mode, the control is complex when the existing equipment is adopted for nucleic acid sequencing, and the detection efficiency is influenced.
Disclosure of Invention
The present application is directed to solving at least one of the problems in the prior art. For this reason, this application provides a biochemical reaction device, through the soaking of reagent groove support chip to can realize the feed liquor and the flowing back in reagent groove, make filling, fluid infusion or the renewal of reagent all comparatively convenient, help improving detection efficiency.
The application also provides a nucleic acid sequencing device with the biochemical reaction device.
The biochemical reaction device comprises a reaction container, a liquid inlet mechanism and a liquid discharge mechanism, wherein the reaction container is provided with a plurality of reagent grooves with openings, the reagent grooves are used for containing reaction reagents and are suitable for containing chips to be tested, and each reagent groove is respectively communicated with a liquid inlet hole and a liquid outlet hole; the liquid inlet mechanism comprises a liquid inlet component and a liquid inlet pipeline, each liquid inlet hole is connected with the liquid inlet pipeline, and the liquid inlet component is used for moving a set reagent and feeding the liquid into the reagent tank through the set liquid inlet pipeline; the liquid discharging mechanism comprises a liquid discharging assembly and liquid discharging pipelines, each liquid outlet hole is connected with the liquid discharging pipelines respectively, and the liquid discharging assembly is used for discharging the reagent in the reagent groove through the liquid discharging pipelines.
The reaction vessel of the embodiment of the first aspect of the present application has at least the following beneficial effects: the holding of multiple reagent can be realized through a plurality of reagent grooves in the reaction vessel, the different reagents of chip soaking in different reagent grooves of can being convenient for to, can carry out feed liquor and flowing back to the reagent groove through feed liquor mechanism and flowing back mechanism, thereby be convenient for fill, fluid infusion and the renewal of reagent, so that the chip soaks the continuation of operation many times and goes on, help improving reaction efficiency, and then improve the efficiency of nucleic acid sequencing.
According to the biochemical reaction device of some embodiments of this application, the feed liquor subassembly includes feed liquor pump, first control valve and second control valve, first control valve is suitable for to pass through the reagent of settlement is introduced into to the feed liquor pipeline, the second control valve passes through the liquid inlet pipe coupling in the reagent groove the feed liquor hole, the feed liquor pump passes through the feed liquor pipe coupling in first control valve with between the second control valve for pass through required reagent the liquid inlet pipeline first control valve with the second control valve lets in the reagent groove, first control valve is used for selecting to let in and sets for reagent, the second control valve is used for selecting to set for the reagent groove.
According to the biochemical reaction device of some embodiments of this application, the second control valve includes a plurality of second valves, the second valve is used for connecting the feed liquor hole of reagent groove, feed liquor mechanism includes the multiunit feed liquor subassembly, and is a plurality of the second control valve the quantity sum of the second valve is not less than the quantity of reagent groove, each the reagent groove passes through the feed liquor hole with it is connected with each the second valve one-to-one to advance the liquid pipeline.
According to the biochemical reaction device of some embodiments of the present application, the liquid discharge assembly includes a liquid discharge pump and a third control valve, the third control valve is connected to the liquid outlet hole of the reagent tank through the liquid discharge pipeline, the liquid discharge pump is connected to the third control valve through the liquid discharge pipeline, and is used for discharging the reagent in the reagent tank with setting, and the third control valve is used for selecting the reagent tank with setting.
According to biochemical reaction device of some embodiments of this application, third control valve includes a plurality of third valves, the third valve is used for connecting the reagent groove go out the liquid hole, the feed liquor mechanism includes the multiunit drainage subassembly, and is a plurality of the third control valve the sum of the quantity of third valve is not less than the quantity of reagent groove, each the reagent groove passes through go out the liquid hole with drainage pipeline and each third valve one-to-one is connected.
The biochemical reaction device according to some embodiments of the present application, further comprising a control box communicatively connected to the liquid inlet assembly for controlling a liquid inlet operation of the liquid inlet assembly; the control box is also in communication connection with the liquid drainage assembly and is used for controlling the liquid drainage operation of the liquid drainage assembly.
According to the biochemical reaction device of some embodiments of this application, reaction vessel includes supporting piece and a plurality of reaction tank, and is a plurality of the reaction tank is arranged according to the settlement mode to connect in supporting piece, each the reaction tank is inside to be equipped with the reagent groove, the reagent groove link up the top surface of reaction tank forms the opening, with the confession the chip business turn over, the feed liquor hole with go out the liquid hole set up in on the wall body of reaction tank and communicate in the reagent groove.
According to the biochemical reaction device of some embodiments of the present application, a temperature control module is further disposed on at least a portion of the reaction tank, and is configured to adjust a temperature of the reagent in the reagent tank.
According to biochemical reaction device of some embodiments of this application, temperature control module includes temperature sensor and temperature protection switch, at least part still establish a plurality of installation departments on the wall body of reaction tank, temperature sensor and/or temperature protection switch connect in the installation department.
According to the biochemical reaction device of some embodiments of this application, the holding piece includes tray and lateral wall, the tray connect in the top of lateral wall, the tray is provided with a plurality of installing ports that link up, the reaction tank set up in the installing port and support hold in the top of tray.
According to the biochemical reaction device of some embodiments of this application, the top of tray is provided with the collecting tank, a plurality of waste discharge holes have still been seted up to the tray, waste discharge hole connect in the collecting tank, waste discharge hole still is connected with the drainage pipeline, the flowing back subassembly is used for passing through the drainage pipeline discharges liquid in the collecting tank.
The biochemical reaction device according to some embodiments of the present application further includes a moving mechanism and a controller, wherein the moving mechanism includes a moving component and a clamping component, the clamping component is adapted to hold and take the chip, and the moving mechanism is connected to the clamping component and is configured to drive the clamping component to move relative to the reaction container so as to be placed into the set reagent tank for soaking; the controller is in communication connection with the moving mechanism and is used for controlling the moving operation of the moving assembly on the clamping assembly and controlling the picking and placing operation of the clamping assembly on the chip.
According to some embodiments of the biochemical reaction apparatus of the present application, in the reaction container, a plurality of reagent grooves are distributed in a predetermined manner to form a plurality of reaction regions, each of the reaction regions has a plurality of reagent grooves, and the moving mechanisms are disposed at positions corresponding to the reaction regions, and each of the moving mechanisms is configured to pick and place and move the chip in the corresponding reaction region.
According to the biochemical reaction device of some embodiments of this application, in the reaction container, still include the buffer, the buffer has a plurality of the reagent groove, the buffer is located adjacent between the reaction area, adjacent moving mechanism can get in the buffer and put and remove the chip.
The nucleic acid sequencing equipment according to the second aspect of the embodiment of the application comprises an optical device and the biochemical reaction device of the first aspect of the embodiment, wherein the optical device is used for acquiring images of the chip after the soaking reaction of the biochemical reaction device is completed.
The nucleic acid sequencing device of the embodiment of the second aspect of the application has at least the following advantages: the biochemical reaction device adopting the embodiment can carry out biochemical reaction by soaking, realizes an open reaction mode, and compared with closed sequencing adopting a microfluidic mode, the reagent can be recycled, thereby reducing waste and saving reagent cost to a certain extent. Chip images can be obtained through an optical device after biochemical reaction, so that nucleic acid information can be analyzed, a plurality of reagent grooves, a liquid inlet mechanism and a liquid discharging mechanism are matched to facilitate filling, liquid supplementing and updating of various reagents, the reaction efficiency can be improved, and the efficiency of nucleic acid sequencing can be effectively improved.
Additional aspects and advantages of the present application will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the present application.
Drawings
FIG. 1 is a schematic structural diagram of a biochemical reaction apparatus according to an embodiment of the present application;
FIG. 2 is a schematic structural view of a reaction vessel in an example of the present application;
FIG. 3 is a top view of a reaction vessel according to an embodiment of the present application;
FIG. 4 isbase:Sub>A cross-sectional view of section A-A of FIG. 3;
FIG. 5 is a cross-sectional view of section B-B of FIG. 3;
FIG. 6 is an exploded view of a reaction vessel according to an embodiment of the present disclosure;
FIG. 7 is a schematic view of the connection between the liquid inlet mechanism and the liquid discharge mechanism in the embodiment of the present application;
FIG. 8 is a schematic view of a portion of a liquid inlet mechanism according to an embodiment of the present application;
FIG. 9 is a schematic view of a portion of a drainage mechanism according to an embodiment of the present application;
FIG. 10 is a schematic view of another portion of the drainage mechanism of the embodiment of the present application;
FIG. 11 is a simplified diagram of a reaction vessel in an example of the present application;
FIG. 12 is a schematic structural diagram of a nucleic acid sequencing apparatus according to an embodiment of the present application.
Reference numerals:
the liquid inlet mechanism 200, the liquid inlet component 201, the liquid inlet pipeline 202, the liquid inlet pump 203, the first control valve 204 and the second control valve 205;
a drain mechanism 300, a drain assembly 301, a drain line 302, a drain pump 303, a third control valve 304, a waste liquid container 305, a waste liquid pump 306, a waste liquid line 307;
a control box 400;
a moving mechanism 500, a moving assembly 501, a clamping assembly 502;
a controller 600;
an optical device 700.
Detailed Description
The conception and the resulting technical effects of the present application will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, and not all embodiments, and other embodiments obtained by those skilled in the art without inventive efforts based on the embodiments of the present application belong to the protection scope of the present application.
In the description of the embodiments of the present application, if an orientation description is referred to, for example, the orientations or positional relationships indicated by "upper", "lower", "front", "rear", "left", "right", etc. are based on the orientations or positional relationships shown in the drawings, only for convenience of description and simplification of description, but not for indicating or implying that the referred device or apparatus must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present application.
In the description of the embodiments of the present application, if a feature is referred to as being "disposed", "fixed", "connected", or "mounted" to another feature, it can be directly disposed, fixed, or connected to the other feature or indirectly disposed, fixed, connected, or mounted to the other feature. In the description of the embodiments of the present application, if "a number" is referred to, it means one or more, if "a plurality" is referred to, it means two or more, if "greater than", "less than" or "more than" is referred to, it is understood that the number is not included, and if "greater than", "lower" or "inner" is referred to, it is understood that the number is included. If reference is made to "first" or "second", this should be understood to distinguish between features and not to indicate or imply relative importance or to implicitly indicate the number of indicated features or to implicitly indicate the precedence of the indicated features.
The method is characterized in that an open biochemical reaction is adopted in nucleic acid sequencing, and the method is different from the method of realizing the biochemical reaction by adopting a microfluidic system to enable a kit fluid to flow through a nucleic acid chain to be sequenced in the current second-generation sequencing technology. The embodiment of the first aspect of the application provides a biochemical reaction device, realizes the open soaking reaction of chip through a plurality of reagent grooves among the reaction vessel to the convenience of the feed liquor in feed liquor mechanism and flowing back mechanism improvement reagent groove and flowing back, thereby be convenient for fill, fluid infusion and the renewal of reagent.
Fig. 1 is a schematic structural diagram of a biochemical reaction apparatus according to an embodiment of the present application, fig. 2 is a schematic structural diagram of a reaction container according to an embodiment of the present application, fig. 3 is a top view of a reaction container according to an embodiment of the present application, and referring to fig. 1 to fig. 3, an embodiment of a first aspect of the present application provides a biochemical reaction apparatus, including a reaction container 100, a liquid inlet mechanism 200, and a liquid outlet mechanism 300, where the reaction container 100 is provided with a plurality of reagent grooves 101 having openings, the reagent grooves 101 are used for containing reaction reagents, and the reagent grooves 101 are adapted to accommodate chips to be tested, so that the chips can be conveniently immersed in different reagent grooves 101.
Wherein, each reagent groove 101 is respectively communicated with a liquid inlet hole 102 and a liquid outlet hole 103; the liquid inlet mechanism 200 comprises a liquid inlet assembly 201 and a liquid inlet pipeline 202, wherein each liquid inlet hole 102 is respectively connected with the liquid inlet pipeline 202, and the liquid inlet assembly 201 is used for moving a set reagent and feeding liquid into the reagent tank 101 through the set liquid inlet pipeline 202. The liquid discharge mechanism 300 includes a liquid discharge unit 301 and a liquid discharge pipe 302, each liquid outlet hole 103 is connected to the liquid discharge pipe 302, and the liquid discharge unit 301 is used for discharging the reagent in the reagent tank 101 through the liquid discharge pipe 302. From this, can carry out feed liquor and flowing back to reagent groove 101 through feed liquor hole 102, play liquid hole 103, feed liquor mechanism 200 and drainage mechanism 300 to be convenient for fill, fluid infusion and the renewal of reagent, so that the continuation of the many times soaking operation of chip is gone on, help improving reaction efficiency, and then improve the efficiency of nucleic acid sequencing.
Compared with the existing second-generation sequencing method, the use of a closed fluid system is avoided, and a plurality of chips can be soaked in the same reagent tank 101 in sequence, so that reagent waste is avoided. Moreover, the biochemical reaction system based on the biochemical reaction device has certain flexibility, the beat of chip feeding soaking and blanking after soaking is finished can be adjusted according to needs, working beats of front and rear procedures are configured according to the beat, adjustment is convenient, integral upgrading is not needed, and upgrading cost is saved.
The reaction vessel 100 may be an integrated structure or a split structure formed by assembling a plurality of structural members, and the reagent tank 101, the liquid inlet hole 102 and the liquid outlet hole 103 may be implemented in various ways. For example: referring to fig. 2 to 4, in some embodiments of the biochemical reaction apparatus, the reaction container 100 includes a support 104 and a plurality of reaction cells 107, the plurality of reaction cells 107 are arranged in a predetermined manner and connected to the support 104, a reagent tank 101 is disposed inside each reaction cell 107, the reagent tank 101 penetrates through a top surface of the reaction cell 107 to form an opening for the chip to enter and exit, and the liquid inlet hole 102 and the liquid outlet hole 103 are disposed on a wall of the reaction cell 107 and connected to the reagent tank 101. Accordingly, the support member 104 and the plurality of reaction cells 107 can be individually processed, the reagent vessel 101 has a predetermined depth so as to accommodate a chip, the size of the reaction cell 107 can be configured according to the size required for the reagent vessel 101, and the support member 104 only needs to be capable of connecting and supporting the reaction cell 107, thereby reducing the overall weight and material consumption of the reaction vessel 100.
In some embodiments, at least a portion of the reaction cells 107 may further include a temperature control module for adjusting the temperature of the reagent in the reagent tank 101, so as to facilitate a reaction with a certain temperature requirement. Wherein, temperature control module can include temperature controller, heater, temperature sensor and temperature protection switch, and the heater can be connected in the outer wall of reaction tank 107, through the temperature regulation to reaction tank 107 heating realization reagent in the reagent groove 101, and the heating subsides of paster form can be chooseed for use to the heater, can attach to the outer wall of reaction tank 107, also can form on the wall body of reaction tank 107 through embedded mode. The temperature sensor and the temperature protection switch are in communication connection with the temperature controller, the temperature sensor can sense the temperature of the reaction tank 107 to form a temperature signal, the temperature controller receives the temperature signal and indicates the heater to work, and the temperature protection switch can be used for switching the start and stop of the heater when the temperature exceeds a set value so as to stop heating. Compared with the existing second-generation sequencing method, the whole biochemical reaction device is not required to be provided with an accurate temperature control system, the cost can be effectively reduced, the temperature of part of the reagent tank 101 can be controlled in a targeted manner, the management of the reaction temperature can be facilitated, and the phenomenon of reaction imbalance can be avoided.
Fig. 4 isbase:Sub>A cross-sectional view of the sectionbase:Sub>A-base:Sub>A in fig. 3, referring to fig. 4, in some embodiments of the biochemical reaction apparatus, at leastbase:Sub>A portion of the wall of the reaction cell 107 is further provided withbase:Sub>A plurality of mounting portions 108, the temperature sensor and/or the temperature protection switch is connected to the mounting portions 108, the portion of the reaction cell 107 can be used for performingbase:Sub>A soaking reaction withbase:Sub>A set temperature requirement, the mounting portions 108 can facilitate the mounting of the temperature sensor and/or the temperature protection switch, the mounting portions 108 can bebase:Sub>A hole-shaped or groove-shaped structure disposed on the outer wall of the reaction cell 107, and the temperature sensor and/or the temperature protection switch can be disposed inside the hole-shaped or groove-shaped structure, so as to avoid protruding from the outer wall of the reaction cell 107 or reduce the portion protruding from the outer wall of the reaction cell 107, thereby avoiding collision damage.
Fig. 5 is a sectional view of a section B-B in fig. 3, fig. 6 is an exploded view of the reaction vessel 100 according to an embodiment of the present invention, and referring to fig. 2 to 6, in some embodiments of the biochemical reaction apparatus, the supporting member 104 includes a tray 105 and a sidewall 106, the tray 105 is connected to a top of the sidewall 106, the tray 105 is provided with a plurality of through mounting holes 111, and the reaction wells 107 are disposed in the mounting holes 111 and abut against a top of the tray 105, thereby supporting the reaction wells 107. In addition, the side wall 106 of the supporting member 104 is supported on the bottom of the tray 105, so that the tray 105 can be lifted, and the positioning and fixing of the reaction vessel 100 can be realized by the installation of the side wall 106 and the workbench, so as to facilitate the chip taking and placing and transferring by a robot. Wherein, tray 105's top can also be provided with collecting tank 109 for accept the dropping liquid of collecting the chip transfer in-process, and tray 105 has still seted up a plurality of rows of useless holes 110, and row useless hole 110 is connected in collecting tank 109, and row useless hole 110 still is connected with drainage pipeline 302, and drainage component 301 is used for through the liquid of drainage pipeline 302 discharge in the collecting tank 109, avoids liquid to spill over and pollutes mesa or other electrical parts.
In some embodiments, the reaction container 100 may also be of an integrated structure, for example, the reaction container 100 includes a body, the body is provided with reagent wells 101 at a plurality of positions, the reagent wells 101 penetrate through the top of the body to form openings for the chip to enter and exit, and the bottom of the body closes the reagent wells 101, so that reagents can be contained therein; the side part or the bottom of the body is provided with a liquid inlet hole 102 and a liquid outlet hole 103 which are communicated with the reagent groove 101, so that the requirements of liquid inlet and liquid discharge are met. The body can be arranged into an integrated structure, and the assembly can be simplified.
Fig. 7 is a schematic diagram showing a connection between a liquid feeding mechanism 200 and a liquid discharging mechanism 300 according to an embodiment of the present invention, and fig. 8 is a schematic diagram showing a connection between a portion of the liquid feeding mechanism 200 according to an embodiment of the present invention, and referring to fig. 7 and 8, in a biochemical reaction apparatus according to some embodiments, a liquid feeding module 201 includes a liquid feeding pump 203, a first control valve 204, and a second control valve 205, the first control valve 204 is adapted to introduce a set reagent through a liquid feeding line 202, the second control valve 205 is connected to a liquid feeding hole 102 of a reagent tank 101 through the liquid feeding line 202, the liquid feeding pump 203 is connected between the first control valve 204 and the second control valve 205 through the liquid feeding line 202, the first control valve 204, and the second control valve 205, and is adapted to introduce a desired reagent into the reagent tank 101 through the liquid feeding line 202, the first control valve 204 is adapted to selectively introduce the set reagent, and the second control valve 205 is adapted to select the set reagent tank 101. The liquid inlet pump 203 can be an injection pump or a plunger pump to pump the reagent, and the control valve can be a rotary valve or a three-way valve to switch the on and off of the corresponding liquid inlet pipeline 202. Fig. 7 and 8 illustrate the connection relationship of the first control valve 204 and the second control valve 205 by using a rotary valve as an example, and the embodiment of the present application is not limited to the connection manner illustrated in the drawing.
Referring to fig. 7 and 8, in the biochemical reaction apparatus according to some embodiments, the first control valve 204 includes a plurality of first valves for connecting the containers for storing the reagents through the inlet line 202, so that the desired reagent can be selected by turning on or off the plurality of first valves of the plurality of first control valves 204. The second control valve 205 includes a plurality of second valves for connecting the liquid inlet holes 102 of the reagent vessels 101, so that the reagent introduced from the first control valve 204 can be injected into the reagent vessel 101 through which the opening/closing of the plurality of second valves of the second control valve 205 is selectively set.
Referring to fig. 7, the liquid inlet mechanism 200 includes a plurality of sets of liquid inlet assemblies 201, the sum of the number of the second valves of the plurality of second control valves 205 is not less than the number of the reagent tanks 101, and each reagent tank 101 is connected with each second valve in a one-to-one correspondence manner through the liquid inlet hole 102 and the liquid inlet pipeline 202, so that each reagent tank 101 can select the type of the reagent to be injected through the first control valve 204 and the second control valve 205.
FIG. 9 is a schematic diagram showing a part of a connection of a liquid discharge mechanism 300 in an embodiment of the present application, and referring to FIGS. 7 and 9, in a biochemical reaction device according to some embodiments, a liquid discharge assembly 301 includes a liquid discharge pump 303 and a third control valve 304, the third control valve 304 is connected to the liquid outlet hole 103 of the reagent tank 101 through a liquid discharge pipe 302, the liquid discharge pump 303 is connected to the third control valve 304 through the liquid discharge pipe 302 for discharging a reagent in a set reagent tank 101, and the third control valve 304 is used for selecting a set reagent tank 101. The drainage mechanism 300 may also include a waste container 305 for collecting liquid drained by the drainage mechanism 300. In fig. 7 and 9, the third control valve 304 is a rotary valve as an example, and the connection relationship is illustrated in fig. 7 and 9, where the third control valve 304 is a rotary valve.
Referring to fig. 7 and 9, in the biochemical reaction apparatus according to some embodiments, the third control valve 304 includes a plurality of third valves, the third valves are used to connect to the outlet holes 103 of the reagent tanks 101, the liquid inlet mechanism 200 includes a plurality of sets of liquid discharging assemblies 301, the sum of the number of the third valves of the plurality of third control valves 304 is not less than the number of the reagent tanks 101, and each reagent tank 101 is connected to each third valve through the outlet hole 103 and the liquid discharging pipeline in a one-to-one correspondence manner. So that each reagent vessel 101 can be drained by selecting the reagent vessel 101 to be drained through the third control valve 304.
Referring to fig. 10, another connection diagram of a portion of a liquid discharge mechanism 300 according to an embodiment of the present invention is shown, and referring to fig. 10, for a reaction vessel 100 having a liquid collection tank 109 on a tray 105, the liquid discharge mechanism 300 may further include a waste liquid pump 306 and a waste liquid pipe 307 connected to a waste discharge hole 110 on the tray 105 to discharge liquid in the liquid collection tank 109, thereby removing drips generated during a chip transfer process.
Referring to fig. 1, in the above embodiment, the biochemical reaction apparatus further includes a control box 400, the control box 400 is communicatively connected to the liquid inlet module 201, and is used for controlling the liquid inlet operation of the liquid inlet module 201; the control box 400 is also communicatively connected to the drain assembly 301 for controlling the draining operation of the drain assembly 301, such as controlling the control valves to select the on/off of the valves, and controlling the intake pump 203 and the drain pump 303. The control of the pump valve by the control box 400 is a mature technology in the field, and the principle and the control method thereof are not described in detail herein.
Referring to fig. 1, in some embodiments, the biochemical reaction apparatus further comprises a moving mechanism 500 and a controller 600, wherein the moving mechanism 500 comprises a moving assembly 501 and a holding assembly 502, the holding assembly 502 is adapted to hold a chip, and the moving mechanism 500 is connected to the holding assembly 502 for driving the holding assembly 502 to move relative to the reaction container 100 to be placed in a set reagent tank 101 for soaking; the controller 600 is communicatively connected to the moving mechanism 500, and is used for controlling the moving operation of the moving assembly 501 to the clamping assembly 502 and controlling the chip pick-and-place operation of the clamping assembly 502. The movement of the chip between different reagent tanks 101 is realized through the moving mechanism 500, so that the continuous operation of multiple soaking operations of the chip is facilitated, the reaction efficiency is improved, and the efficiency of nucleic acid sequencing is improved.
The moving assembly 501 can be realized by a multi-axis manipulator commonly used in mechanical equipment, the clamping assembly 502 can be a gripper driven by an air cylinder, the gripper comprises two fingers and the air cylinder, the air cylinder drives the fingers to move towards or away from each other so as to be closed or opened, and the chip can be grabbed and released. The clamp assembly 502 may also be provided with a plurality of sensors, including a grip sensor, a release sensor, and a chip presence detection sensor. The grab sensor is used to sense whether the two fingers are correctly closed, which can be achieved by a distance sensor or by providing a photo sensor in the closed position. The release sensor is used to sense whether the two fingers are correctly opened, which detection can be achieved by a distance sensor or by providing a photo sensor in the open position. The chip presence or absence detection sensor is used for sensing whether a chip is present on the corresponding position of the gripper when the gripper starts to grip, sensing whether the chip is held on the gripper in the transferring process, and sensing whether the chip is present on the gripper when the chip is released, so that the chip can be smoothly gripped before the gripper is ensured, the chip can be identified to fall off when the gripper catches, and the chip can be ensured to be smoothly released after the gripper is released. The detection of the chip can be realized by a contact type or non-contact type sensor. The multiple sensors realize the relevant detection of chip taking and placing and transferring operation processes, so that the operation safety is improved, the abnormal conditions are conveniently identified, and the chip grabbing and transferring efficiency is improved.
The controller 600 is a feedback loop component widely used in industrial control applications, such as a programmable memory, in which instructions for performing operations such as logic operations, sequence control, timing, counting, and arithmetic operations are stored, and controls various types of machinery or manufacturing processes through digital or analog input and output. Therefore, in the embodiment of the present application, the moving operation of the moving assembly 501 on the clamping assembly 502 and the chip pick-and-place operation of the clamping assembly 502 on the chip are realized by a preset scheme, which can be easily realized by those skilled in the art according to the functions of the existing controller 600, and the principle and the control method thereof are not described herein again.
Fig. 11 is a simplified diagram of a reaction container 100 in an embodiment of the present application, referring to fig. 1 to 3, and fig. 11, in a biochemical reaction apparatus in some embodiments, in the reaction container 100, a plurality of reagent vessels 101 are distributed in a set manner to form a plurality of reaction regions 112, each reaction region 112 has a plurality of reagent vessels 101, and a moving mechanism 500 is disposed at a position corresponding to each reaction region 112, each moving mechanism 500 is used for picking and placing and moving a chip in the corresponding reaction region 112. In addition, the reaction vessel 100 may further include a buffer area 113 (shown by a dashed line frame), the buffer area 113 has a plurality of reagent wells 101, the buffer area 113 is located between adjacent reaction areas 112, and the adjacent moving mechanism 500 can pick and place and move the chip in the buffer area 113. Therefore, the cooperation of the adjacent moving mechanisms 500 can be realized, when the chip is transferred between the adjacent reaction areas 112, the chip can be placed in the reagent tank 101 of the buffer area 113 through the moving mechanism 500 of one reaction area 112, and then the chip in the buffer area 113 can be grabbed by the moving mechanism 500 of the other reaction area 112 to be transferred to the other reaction area 112 for soaking, so that the interference caused by space-time intersection can be avoided. The buffer 113 may also be used to temporarily store the chips that have completed the reaction and are waiting to be transferred to the optical device 700 for image acquisition.
In a second aspect of the present application, a nucleic acid sequencing apparatus is provided, and fig. 12 is a schematic structural diagram of the nucleic acid sequencing apparatus according to the embodiment of the present application, and with reference to fig. 12 and in combination with the above embodiments, the nucleic acid sequencing apparatus according to the embodiment of the present application includes an optical device 700 and the biochemical reaction device according to the embodiment of the first aspect, where the optical device 700 is configured to obtain an image of a chip after a soaking reaction in the biochemical reaction device is completed. The biochemical reaction device adopting the embodiment can realize an open biochemical reaction mode, and compared with a closed sequencing scheme adopting a microfluidic mode, the reagent can be recycled, so that the waste is reduced, and the reagent cost can be saved to a certain extent. After biochemical reaction, chip images can be acquired through the optical device 700 so as to analyze nucleic acid information, and the multiple reagent grooves 101, the liquid inlet mechanism 200 and the liquid discharge mechanism 300 are matched to facilitate the filling, liquid supplementing and updating of multiple reagents, so that the reaction efficiency can be improved, and the efficiency of nucleic acid sequencing can be effectively improved.
The embodiments of the present application have been described in detail with reference to the drawings, but the present application is not limited to the embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present application. Furthermore, the embodiments and features of the embodiments of the present application may be combined with each other without conflict.
Claims (15)
1. A biochemical reaction device for use in nucleic acid sequencing, comprising:
the reaction container is provided with a plurality of reagent grooves with openings, the reagent grooves are used for containing reaction reagents and are suitable for containing chips to be detected, and each reagent groove is respectively communicated with a liquid inlet hole and a liquid outlet hole;
the liquid inlet mechanism comprises a liquid inlet assembly and a liquid inlet pipeline, each liquid inlet hole is connected with the liquid inlet pipeline, the liquid inlet assembly is used for moving a set reagent and feeding the liquid into the reagent tank through the set liquid inlet pipeline;
the liquid discharging mechanism comprises a liquid discharging assembly and liquid discharging pipelines, each liquid outlet hole is connected with the liquid discharging pipelines respectively, and the liquid discharging assembly is used for discharging the reagent in the reagent groove through the liquid discharging pipelines.
2. The biochemical reaction apparatus according to claim 1, wherein the liquid inlet assembly comprises a liquid inlet pump, a first control valve and a second control valve, the first control valve is adapted to introduce a set reagent through the liquid inlet pipeline, the second control valve is connected to the liquid inlet hole of the reagent tank through the liquid inlet pipeline, the liquid inlet pump is connected between the first control valve and the second control valve through the liquid inlet pipeline for introducing a desired reagent into the reagent tank through the liquid inlet pipeline, the first control valve and the second control valve, the first control valve is used for selectively introducing a set reagent, and the second control valve is used for selectively introducing a set reagent into the reagent tank.
3. The biochemical reaction apparatus according to claim 2, wherein the second control valve includes a plurality of second valves for connecting the liquid inlet holes of the reagent tanks, the liquid inlet mechanism includes a plurality of sets of the liquid inlet assemblies, a sum of numbers of the second valves of the plurality of second control valves is not less than a number of the reagent tanks, and each of the reagent tanks is connected to each of the second valves in a one-to-one correspondence manner through the liquid inlet holes and the liquid inlet pipes.
4. The biochemical reaction apparatus according to claim 1, wherein the drain assembly includes a drain pump and a third control valve, the third control valve is connected to the outlet hole of the reagent tank through the drain line, the drain pump is connected to the third control valve through the drain line for discharging the set reagent in the reagent tank, and the third control valve is used for selecting the set reagent tank.
5. The biochemical reaction device according to claim 4, wherein the third control valve includes a plurality of third valves for connecting the liquid outlet holes of the reagent tanks, the liquid inlet mechanism includes a plurality of sets of the liquid discharge assemblies, the sum of the numbers of the third valves of the plurality of third control valves is not less than the number of the reagent tanks, and each of the reagent tanks is connected to each of the third valves in a one-to-one correspondence via the liquid outlet holes and the liquid discharge lines.
6. The biochemical reaction device according to claim 1, further comprising a control box communicatively connected to the liquid feeding assembly for controlling a liquid feeding operation of the liquid feeding assembly; the control box is also in communication connection with the liquid drainage assembly and is used for controlling the liquid drainage operation of the liquid drainage assembly.
7. The biochemical reaction apparatus according to any one of claims 1 to 6, wherein the reaction container includes a support member and a plurality of reaction cells, the plurality of reaction cells are arranged in a predetermined manner and connected to the support member, the reagent tank is disposed inside each reaction cell, the reagent tank penetrates through a top surface of the reaction cell to form the opening for the chip to enter and exit, and the liquid inlet hole and the liquid outlet hole are disposed on a wall body of the reaction cell and communicated with the reagent tank.
8. The biochemical reaction device according to claim 7, wherein a temperature control module is further disposed on at least a portion of the reaction chamber for adjusting the temperature of the reagent in the reagent tank.
9. The biochemical reaction device according to claim 8, wherein the temperature control module comprises a temperature sensor and a temperature protection switch, wherein at least some of the walls of the reaction chamber are further provided with a plurality of mounting portions, and the temperature sensor and/or the temperature protection switch are connected to the mounting portions.
10. The biochemical reaction device according to claim 7, wherein the supporting member comprises a tray and a side wall, the tray is connected to the top of the side wall, the tray is provided with a plurality of through mounting holes, and the reaction chamber is disposed in the mounting holes and abuts against the top of the tray.
11. The biochemical reaction device according to claim 10, wherein a liquid collecting tank is disposed on the top of the tray, the tray further defines a plurality of waste discharge holes, the waste discharge holes are connected to the liquid collecting tank, the waste discharge holes are further connected to a liquid discharge pipeline, and the liquid discharge assembly is configured to discharge liquid in the liquid collecting tank through the liquid discharge pipeline.
12. The biochemical reaction device according to claim 1, further comprising:
the moving mechanism comprises a moving assembly and a clamping assembly, the clamping assembly is suitable for holding the chip, and the moving mechanism is connected with the clamping assembly and used for driving the clamping assembly to move relative to the reaction container so as to be placed in the set reagent groove for soaking;
and the controller is in communication connection with the moving mechanism and is used for controlling the moving operation of the moving assembly on the clamping assembly and controlling the picking and placing operation of the clamping assembly on the chip.
13. The biochemical reaction apparatus according to claim 12, wherein a plurality of the reagent vessels are distributed in a predetermined manner to form a plurality of reaction regions in the reaction container, each of the reaction regions has a plurality of the reagent vessels, and the moving mechanism is disposed at a position corresponding to each of the reaction regions, and each of the moving mechanisms is configured to pick and place and move the chip in the corresponding reaction region.
14. The biochemical reaction device according to claim 13, further comprising a buffer area having a plurality of reagent wells, wherein the buffer area is located between adjacent reaction areas, and the adjacent moving mechanism is capable of picking and moving the chip in the buffer area.
15. Nucleic acid sequencing equipment, characterized in that it comprises an optical device and a biochemical reaction device according to any one of claims 1 to 14, the optical device is used for acquiring images of the chip after the completion of the soaking reaction of the biochemical reaction device.
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| CN202210927075.9A CN115433667B (en) | 2022-08-03 | 2022-08-03 | Biochemical reaction device and nucleic acid sequencing equipment |
| PCT/CN2023/075018 WO2024027121A1 (en) | 2022-08-03 | 2023-02-08 | Biochemical reaction apparatus and nucleic acid sequencing device |
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| CN202210927075.9A CN115433667B (en) | 2022-08-03 | 2022-08-03 | Biochemical reaction device and nucleic acid sequencing equipment |
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| CN115433667A true CN115433667A (en) | 2022-12-06 |
| CN115433667B CN115433667B (en) | 2024-02-23 |
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| WO2024027121A1 (en) * | 2022-08-03 | 2024-02-08 | 深圳赛陆医疗科技有限公司 | Biochemical reaction apparatus and nucleic acid sequencing device |
| WO2024027119A1 (en) * | 2022-08-03 | 2024-02-08 | 深圳赛陆医疗科技有限公司 | Nucleic acid sequencing system and sequencing control method |
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| WO2024027121A1 (en) | 2024-02-08 |
| CN115433667B (en) | 2024-02-23 |
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