JP2021132539A - Nucleic acid extraction device and nucleic acid extraction method - Google Patents

Abstract

To provide a nucleic acid extraction device for extracting simultaneously, a nucleic acid at a plurality of extraction containers, capable of preventing waste fluid discharged from a certain extraction container from adhering to other extraction container.SOLUTION: A nucleic acid extraction device 1 comprises: a plurality of extraction containers 11 for storing an object extraction solution for extracting a nucleic acid therefrom, and having a filter for extracting the nucleic acid included in the extraction solution; and a plurality of waste fluid collecting tools 12 each of which is coupled to one extraction container 11 out of the plurality of extraction containers 11, and collects the waste fluid which is left after extracting the nucleic acid by the filter 111 provided on each extraction container 11, in which, one extraction container 11 and one waste fluid collecting tool 12 for collecting the waste fluid from the extraction container 11 out of the plurality of waste fluid collecting tools 12, are tight closed and coupled.SELECTED DRAWING: Figure 2

Description

The present invention relates to a nucleic acid extractor for extracting nucleic acid from a solution containing nucleic acid.

Conventionally, a nucleic acid extraction device for extracting nucleic acid from a solution containing nucleic acid is known. Patent Document 1 discloses a nucleic acid extraction device including a plurality of extraction cartridges for extracting a plurality of solutions.

Japanese Unexamined Patent Publication No. 2005-58149

In the conventional nucleic acid extraction device, a waste liquid extraction container is provided for each of the plurality of extraction cartridges. However, since there is a space between the extraction cartridge and the waste liquid extraction container in which a plurality of extraction cartridges face in common, the waste liquid discharged from one extraction cartridge through the space is used for other extraction. There is a problem that contamination that is mixed in the cartridge occurs.

Therefore, the present invention has been made in view of these points, and in a nucleic acid extraction device that extracts nucleic acids simultaneously in a plurality of extraction containers, the waste liquid discharged from one extraction container is prevented from adhering to another extraction container. The purpose is to do.

The nucleic acid extraction device according to the first aspect of the present invention is a nucleic acid extraction device for simultaneously extracting a plurality of nucleic acids, and contains an extraction solution of a target for extracting nucleic acids, and is included in the contained extraction solution. Waste liquid that is bound to a plurality of extraction containers having filters for extracting the nucleic acid and one of the plurality of extraction containers, and remains after the nucleic acid is extracted by the filter provided in the extraction container. The one extraction container and one of the plurality of waste liquid collectors for collecting the waste liquid discharged from the extraction container are hermetically sealed. It is combined in the state of being.

The plurality of waste liquid collectors may be independently and detachably coupled to the one extraction container.

Each of the plurality of waste liquid collectors may have an aspirator that sucks the extract solution through the filter.

The nucleic acid extraction device further includes a control unit that controls the plurality of suction devices, and the control unit includes an operation reception unit that receives a suction operation for starting suction by the suction device, and the operation reception unit. It may have a drive unit that moves at least one of the plurality of suction devices according to the acceptance of the suction operation.

The nucleic acid extraction device further includes a plurality of operation units that receive inputs of the suction operation to each of the plurality of suction devices, and the operation reception unit is based on the suction operation performed in the plurality of operation units. The at least one aspirator may be moved.

The drive unit may move at least one of the plurality of aspirators at a different speed than the other aspirators.

The nucleic acid extraction device further includes a control unit that controls the plurality of aspirators, and the control unit identifies the state of the extraction solution contained in at least one extraction container among the plurality of extraction containers. It may have a state specifying unit and a driving unit that moves at least one of the plurality of aspirators based on the state of the extraction solution specified by the state specifying unit.

The state specifying unit identifies the state of each of the plurality of extraction solutions contained in each of the plurality of extraction containers, and the driving unit identifies each state of the plurality of extraction solutions specified by the state specifying unit. The plurality of aspirators may be moved independently based on the above.

In this case, the driving unit may move the plurality of aspirators at a speed determined based on each state of the plurality of extraction solutions specified by the state specifying unit.

The nucleic acid extraction method according to the second aspect of the present invention includes a plurality of extraction containers containing an extraction solution of a target for extracting nucleic acid and having a filter for extracting the nucleic acid contained in the contained extraction solution, and the plurality of extraction containers. It has a plurality of waste liquid collectors which are bound to one of the extraction containers of the above and collect the waste liquid remaining after the nucleic acid is extracted by the filter provided in the extraction container. Using a nucleic acid extraction device, one extraction container and one waste liquid recovery device that collects the waste liquid discharged from the extraction container among the plurality of waste liquid recovery devices are bound in a sealed state. A nucleic acid extraction method for simultaneously extracting a plurality of nucleic acids, in which a step of injecting an extraction solution of a target for extracting nucleic acid into a plurality of extraction containers of the nucleic acid extraction device and a step of injecting an extraction solution of a target to be extracted into each of the plurality of waste liquid collectors. A step of collecting the extract solution discharged through the filter as a waste liquid by moving an aspirator that sucks the extract solution through the filter, and a nucleic acid extracted by the filter. Has a step of recovering.

The recovery step includes a step of monitoring the state of the extraction solution in the plurality of extraction containers and a step of moving the aspirator when the state of the extraction solution reaches a predetermined state. You may.

According to the present invention, in a nucleic acid extraction device that simultaneously extracts nucleic acids in a plurality of extraction containers, it is possible to prevent the waste liquid discharged from one extraction container from adhering to another extraction container.

It is a perspective view of the nucleic acid extractor. It is a figure which shows typically the structure of the nucleic acid extractor. It is a figure which shows typically the structure of the nucleic acid extraction apparatus which concerns on a modification. It is a figure which shows the result of having performed the comparative experiment of the nucleic acid extraction amount.

[Structure of Nucleic Acid Extractor 1]
FIG. 1 is a perspective view of the nucleic acid extraction device 1. FIG. 2 is a diagram schematically showing the configuration of the nucleic acid extraction device 1. The nucleic acid extraction device 1 is a device for simultaneously extracting a plurality of nucleic acids from a plurality of extraction solutions containing nucleic acids. Hereinafter, the configuration of the nucleic acid extraction device 1 will be described with reference to FIGS. 1 and 2.

The nucleic acid extraction device 1 includes an extraction unit 10 and a control unit 20. The extraction unit 10 is a site for extracting nucleic acid from the extraction solution, and executes an operation for extracting nucleic acid based on the control of the control unit 20. The control unit 20 has, for example, a processor inside, and controls the extraction unit 10 based on the operation of the user of the nucleic acid extraction device 1, so that the extraction unit 10 extracts nucleic acid from the extraction solution.

[Structure of extraction unit 10]
The extraction unit 10 includes a plurality of extraction containers 11 (extraction containers 11a, 11b, 11c in FIG. 1), a plurality of waste liquid collectors 12 (waste liquid collectors 12a, 12b, 12c in FIG. 1), and a plurality of coupling portions 13. And have. The extraction container 11 is also referred to as a column, and the waste liquid collector 12 is also referred to as a syringe.

As shown in FIG. 2, the extraction container 11 contains an extraction solution of a target for extracting nucleic acid. The extraction container 11 has, for example, a cylindrical shape, and is provided with an opening at one end for a user to put an extraction solution. The other end of the extraction container 11 is detachably connected to the waste liquid collector 12 via the coupling portion 13. The extraction container 11 has a filter 111 for extracting nucleic acids contained in the extraction solution.

The filter 111 is made of a material that does not allow nucleic acids to pass through but allows solutions other than nucleic acids to pass through. The filter 111 is formed of, for example, silica gel, silica beads, silica membrane, or silica monolith. The filter 111 has a cylindrical shape and is fixed to the inside of the extraction container 11 on the side close to the other end of the extraction container 11. In the following description, the solution that has passed through the filter 111 among the extraction solutions is referred to as a waste solution.

The waste liquid collector 12 is bound to one of the plurality of extraction containers 11 via the binding portion 13, and the waste liquid remaining after the nucleic acid is extracted by the filter 111 provided in the extraction container 11. To collect. Each of the plurality of waste liquid collectors 12 has a piston 121 (pistons 121a, 121b, 121c in FIG. 2) that functions as an aspirator that sucks the extraction solution through the filter 111.

Specifically, the waste liquid recovery device 12 has, for example, a cylindrical shape, and a cylindrical piston 121 that moves in the longitudinal direction of the waste liquid recovery device 12 under the control of the control unit 20 is provided inside. The piston 121 is in close contact with the inner wall of the waste liquid collector 12, and moves away from the joint portion 13 to suck the waste liquid through the joint portion 13. When the piston 121 sucks the waste liquid, the waste liquid is collected in the space between the filter 111 and the piston 121 in the waste liquid recovery device 12.

When the waste liquid is accumulated in the waste liquid collector 12, the user is expected to remove the waste liquid collector 12 from the joint portion 13 and dispose of the waste liquid. By connecting the waste liquid collector 12 to the coupling portion 13 after the user disposes of the waste liquid, the piston 121 can suck the waste liquid again.

The connecting portion 13 is a tubular member that connects the extraction container 11 and the waste liquid collector 12. Although the connecting portion 13 is shown in a linear shape in FIG. 2, the connecting portion 13 may have an L-shape as shown in FIG. Further, the coupling portion 13 may be fixed to either the extraction container 11 or the waste liquid collector 12 and may be detachably configured to be detachable from the other. Further, each of the extraction container 11 and the waste liquid collector 12 has a protruding cylindrical portion that becomes a part of the coupling portion 13, and the coupling portion 13 may be formed by connecting the respective cylindrical portions. ..

Here, in the nucleic acid extraction device 1, one extraction container 11 and one waste liquid recovery device 12 (for example, one waste liquid recovery device 12 (for example) that collects the waste liquid discharged from the extraction container 11 (for example, the extraction container 11a) among the plurality of waste liquid recovery devices 12). For example, the waste liquid collector 12a) is bound in a sealed state. That is, the plurality of waste liquid collectors 12 are independently and detachably coupled to one extraction container 11. In other words, one set of the extraction container 11, the waste liquid collector 12, and the connecting portion 13 does not communicate with the other set and is independent of each other. Therefore, since the waste liquid collected in the waste liquid recovery device 12 other than the corresponding waste liquid recovery device 12 does not adhere to the extraction container 11, impurities are not mixed in the extracted nucleic acid.

[Structure of control unit 20]
The control unit 20 controls a plurality of pistons 121 to move the pistons 121 inside the waste liquid recovery device 12. The control unit 20 has at least an operation unit 201 (first operation unit 201a, second operation unit 201b, third operation unit 201c), an operation reception unit 202, and a drive unit 203.

The operation unit 201 is a device for the user to operate, for example, a push button switch or a touch panel. The first operation unit 201a receives an operation for moving the piston 121a away from the extraction container 11 to suck the waste liquid, and an operation for stopping the movement of the piston 121a to end the suction of the waste liquid. Similarly, the second operation unit 201b and the third operation unit 201c are operated to move the piston 121b and the piston 121c, respectively.

The operation unit 201 notifies the operation reception unit 202 of information indicating the contents of the operation. When the first operation unit 201a, the second operation unit 201b, and the third operation unit 201c are push button switches, the push button switch is pressed on the first operation unit 201a, the second operation unit 201b, and the third operation unit 201c. The operation reception unit 202 is notified of a signal indicating whether the switch is in the operating state or not pressed. The operation unit 201 may receive an operation for designating the speed at which each of the plurality of pistons 121 is moved, and may notify the operation reception unit 202 of information indicating the designated speed.

The operation receiving unit 202 is realized by, for example, a CPU (Central Processing Unit) that executes a program, and receives a suction operation for starting suction by the piston 121. The operation reception unit 202 moves at least one piston 121 based on the suction operation performed by the plurality of operation units 201. Specifically, the operation reception unit 202 determines whether or not to move the piston 121 based on the notification input from the operation unit 201, and if it is determined to move the piston 121, drives the drive unit 203. This moves the piston 121.

When the notification input from the operation unit 201 indicates that an operation for suction has been performed, the operation reception unit 202 causes the drive unit 203 to move the piston 121 in a direction away from the extraction container 11. Control. When the notification input from the operation unit 201 indicates that the operation for stopping the suction has been performed, the drive unit 203 is controlled so as to stop the movement of the piston 121. Further, when the notification input from the operation unit 201 indicates the speed at which the piston 121 is moved, the operation reception unit 202 controls the drive unit 203 so as to move the piston 121 at the speed indicated by the notification.

The drive unit 203 moves at least one piston 121 of the plurality of pistons 121 in response to the operation reception unit 202 receiving the suction operation. The drive unit 203 has a solenoid or a motor capable of moving an actuator coupled to the piston 121, for example, by flowing an electric current.

The drive unit 203 moves each of the plurality of pistons 121 independently, for example, based on an instruction from the operation reception unit 202. The drive unit 203 may move the two or more pistons 121 in a synchronized state based on the instruction of the operation reception unit 202. Further, the drive unit 203 may move at least one piston 121 of the plurality of pistons 121 at a speed different from that of the other pistons 121. By moving the plurality of pistons 121 at different speeds in this way, the drive unit 203 moves the plurality of pistons 121 at a speed suitable for each of the extraction solutions when different types of extraction solutions are filled in the plurality of extraction containers 11. Can be moved.

When the drive unit 203 moves the piston 121 away from the extraction container 11, the extraction solution contained in the extraction container 11 passes through the filter 111 and then is discharged to the waste liquid collector 12. Then, when the waste liquid is accumulated in the waste liquid recovery device 12, the user removes the waste liquid recovery device 12 from the joint portion 13 and disposes of the waste liquid. After that, when the user attaches the waste liquid collector 12 to the binding portion 13, the process of extracting the nucleic acid from the extraction solution can be continued again.

[Modification example]
In the example shown in FIG. 2, the drive unit 203 moves the piston 121 by the user's operation. On the other hand, the drive unit 203 may move the piston 121 without any operation by the user.

FIG. 3 is a diagram schematically showing the configuration of the nucleic acid extraction device 1a according to the modified example. The control unit 20 in the nucleic acid extraction device 1a shown in FIG. 3 is different from the nucleic acid extraction device 1 shown in FIG. 2 in that it further includes a state identification unit 204, and is the same in other respects.

The state specifying unit 204 identifies the state of the extraction solution contained in at least one of the plurality of extraction containers 11. The state of the extract solution is, for example, color, permeability or viscosity. The state specifying unit 204 has, for example, a color sensor, and identifies the color of the extraction solution contained in the extraction container 11. The state specifying unit 204 may specify the speed at which the extraction solution passes through the filter 111 (that is, the speed at which the liquid level of the extraction solution drops) as the state of the extraction solution. The driving unit 203 moves at least one of the plurality of pistons 121 based on the state of the extraction solution specified by the state specifying unit 204.

The drive unit 203 may move the piston 121 so that the speed at which the height of the liquid level of each of the extraction solutions of the plurality of extraction containers 11 changes is the same. The drive unit 203 is the liquid in the extraction container 11 (that is, the extraction container 11 in which the liquid level of the extraction solution drops the slowest) in which the height of the liquid level of each of the extraction solutions of the plurality of extraction containers 11 has the slowest decomposition rate. The plurality of pistons 121 may be moved so as to be the same as the rate of change in the height of the surface. By operating the drive unit 203 in this way, the extraction solution can be discharged according to the extraction container 11 having the slowest decomposition rate, so that nucleic acid can be efficiently extracted.

The state specifying unit 204 identifies the states of the plurality of extraction solutions contained in each of the plurality of extraction containers 11, and the driving unit 203 is based on the respective states of the plurality of extraction solutions specified by the state specifying unit 204. The plurality of pistons 121 may be moved independently. The driving unit 203 moves the plurality of pistons 121 at a speed determined based on the respective states of the plurality of extraction solutions specified by the state specifying unit 204, for example. By operating the drive unit 203 and the state specifying unit 204 in this way, when different types of extraction solutions are contained in a plurality of extraction containers 11, the corresponding piston 121 is moved at a speed suitable for each extraction solution. Can be moved.

[Nucleic acid extraction method by nucleic acid extraction device 1]
Nucleic acid can be extracted by the following procedure using the nucleic acid extraction device 1. First, a step of injecting an extraction solution of a target for extracting nucleic acid into a plurality of extraction containers 11 of the nucleic acid extraction device 1 is executed. Subsequently, by moving the piston 121 which is provided in each of the plurality of waste liquid collectors 12 and sucks the extract solution through the filter 111, the extract solution discharged through the filter 111 is collected as waste liquid. Perform the process. Then, the step of recovering the nucleic acid extracted by the filter 111 is executed.

In the step of collecting the extraction solution, the state of the extraction solution in the plurality of extraction containers 11 may be monitored, and the piston 121 may be moved when the state of the extraction solution reaches a predetermined state. For example, when the color, permeability or viscosity of the extraction solution reaches a predetermined state, the piston 121 is moved to discharge the extraction solution. By such a procedure, nucleic acid can be efficiently extracted.

[Experimental example]
In the nucleic acid extraction device 1, an experiment was conducted in which the state of the extraction solution contained in the extraction container 11 was monitored and the nucleic acid was extracted by discharging the extraction solution at an appropriate timing. FIG. 4 is a diagram showing the results of a comparative experiment on the amount of nucleic acid extracted.

In FIG. 4, when the existing centrifuge is used (broken line), when the extraction solution is discharged at high speed without monitoring the state of the extraction solution (■), extraction is performed at low speed while monitoring the state of the extraction solution. The case where the solution is discharged (●) is shown. The horizontal axis of FIG. 4 shows the number of base pairs constituting the nucleic acid, and the vertical axis shows the amount of nucleic acid recovered when the amount of nucleic acid recovered when the centrifuge is used is 1.0. ing.

From FIG. 4, when the extraction solution is discharged at a low speed while monitoring the state of the extraction solution, the amount of nucleic acid recovered can be increased as compared with the case where the extraction solution is discharged at a high speed without monitoring the state of the extraction solution. You can check. It was also confirmed that when the number of base pairs is 50 or more, the amount of nucleic acid recovered is increased by about 10% compared to the case of using a centrifuge by discharging the extraction solution at a low speed while monitoring the state of the extraction solution. did it.

[Effect of Nucleic Acid Extractor 1]
As described above, in the nucleic acid extraction device 1, the set of the extraction container 11 and the waste liquid collector 12 is sealed and does not communicate with other sets. Therefore, when the user disposes of the waste liquid, the waste liquid discharged from one extraction container 11 does not adhere to the other extraction container 11. As a result, the user can extract the nucleic acid with high purity in the extraction container 11.

Although the present invention has been described above using the embodiments, the technical scope of the present invention is not limited to the scope described in the above embodiments, and various modifications and changes can be made within the scope of the gist thereof. be. For example, all or a part of the device can be functionally or physically distributed / integrated in any unit. Also included in the embodiments of the present invention are new embodiments resulting from any combination of the plurality of embodiments. The effect of the new embodiment produced by the combination also has the effect of the original embodiment.

For example, in the above description, the case where the nucleic acid extraction device 1 has three extraction containers 11, a waste liquid recovery device 12, and a binding portion 13 has been illustrated, but the extraction container 11, the waste liquid recovery device 12, and the nucleic acid extraction device 1 have. The number of connecting portions 13 is not limited to this, and is arbitrary.

1 Nucleic acid extraction device 10 Extraction unit 11 Extraction container 12 Waste liquid collector 13 Binding unit 20 Control unit 111 Filter 121 Piston 201 Operation unit 202 Operation reception unit 203 Drive unit 204 State identification unit

Claims (11)

A nucleic acid extractor for extracting multiple nucleic acids at the same time.
A plurality of extraction containers containing a target extraction solution for extracting nucleic acid and having a filter for extracting nucleic acid contained in the contained extraction solution, and a plurality of extraction containers.
A plurality of waste liquid collectors which are bound to one of the plurality of extraction containers and collect the waste liquid remaining after nucleic acid is extracted by the filter provided in the extraction container.
Have,
A nucleic acid extraction device in which the one extraction container and one waste liquid recovery device that collects the waste liquid discharged from the extraction container among the plurality of waste liquid recovery devices are bound in a sealed state. The plurality of waste liquid collectors are independently and detachably coupled to the one extraction container.
The nucleic acid extraction device according to claim 1. Each of the plurality of waste liquid collectors has an aspirator that sucks the extract solution through the filter.
The nucleic acid extractor according to claim 1 or 2. Further having a control unit for controlling the plurality of aspirators,
The control unit
An operation reception unit that accepts a suction operation for starting suction by the suction device,
A drive unit that moves at least one of the plurality of suction devices in response to the operation reception unit receiving the suction operation, and a drive unit.
Have,
The nucleic acid extraction device according to claim 3. It further has a plurality of operating units that receive the input of the suction operation for each of the plurality of suction devices.
The operation receiving unit moves the at least one suction device based on the suction operation performed in the plurality of operation units.
The nucleic acid extraction device according to claim 4. The drive unit moves at least one of the plurality of aspirators at a speed different from that of the other aspirators.
The nucleic acid extractor according to claim 4 or 5. Further having a control unit for controlling the plurality of aspirators,
The control unit includes a state specifying unit that specifies the state of the extraction solution contained in at least one of the plurality of extraction containers.
A drive unit that moves at least one of the plurality of aspirators based on the state of the extraction solution specified by the state-specific unit, and a drive unit.
Have,
The nucleic acid extraction device according to claim 3. The state specifying unit identifies the states of the plurality of extraction solutions contained in each of the plurality of extraction containers.
The driving unit independently moves the plurality of aspirators based on the respective states of the plurality of extraction solutions specified by the state specifying unit.
The nucleic acid extraction device according to claim 7. The driving unit moves the plurality of aspirators at a speed determined based on each state of the plurality of extraction solutions specified by the state specifying unit.
The nucleic acid extractor according to claim 7 or 8. A plurality of extraction containers having a filter for extracting the nucleic acid contained in the contained extraction solution and a plurality of extraction containers for containing the extraction solution to be extracted with the nucleic acid, and one of the plurality of extraction containers are combined with each other. It has a plurality of waste liquid collectors for collecting the waste liquid remaining after the nucleic acid is extracted by the filter provided in the extraction container, and the one extraction container and the plurality of waste liquid collectors. It is a nucleic acid extraction method for simultaneously extracting a plurality of nucleic acids by using a nucleic acid extraction device in which one waste liquid collector for collecting the waste liquid discharged from the extraction container is bound in a sealed state. hand,
A step of injecting an extraction solution of a target for extracting nucleic acid into a plurality of extraction containers of the nucleic acid extraction device, and
A step of recovering the extract solution discharged through the filter as a waste liquid by moving an aspirator that is provided in each of the plurality of waste liquid collectors and sucks the extract solution through the filter. When,
The step of recovering the nucleic acid extracted by the filter and
Nucleic acid extraction method having. The recovery step is
A step of monitoring the state of the extraction solution in the plurality of extraction containers, and
The step of moving the aspirator when the state of the extraction solution reaches a predetermined state, and
Have,
The nucleic acid extraction method according to claim 10.

Images