JP2018029514A - Material amplification equipment - Google Patents

Abstract

Provided is a substance amplifying device capable of rapidly changing a temperature of a reaction solution from a temperature at which thermal denaturation is performed to a temperature at which an annealing / extension reaction is performed. The substance amplifying apparatus 1 is a substance amplifying apparatus for amplifying a substance by subjecting a reaction liquid 150 to a heat cycle, and a reaction vessel 100 having a flow path 160 for moving the reaction liquid 150 in a longitudinal direction can be mounted. A mounting portion, a first region 170 disposed at one end of the flow channel 160 of the reaction vessel 100, a third region 190 disposed at the other end in the direction opposite to the one end of the flow channel 160, A second region 180 disposed in a portion of the path 160 between the first region 170 and the third region 190, a first heating unit 210 that controls the first region 170 to a first temperature, and a second region 180 A second heating unit 220 that controls the second temperature lower than the first temperature; and a drive mechanism that moves the reaction liquid 150 between the first region 170, the second region 180, and the third region 190. [Selection] Figure 2A

Description

  The present invention relates to a substance amplification device.

  Conventionally, a PCR (Polymerase Chain Reaction) method has been widely used as a method for amplifying nucleic acids at high speed, and the application range of PCR-related technology is expected to continue to expand in the future. The PCR method is a technique for amplifying a target nucleic acid by subjecting a solution (reaction solution) containing a nucleic acid (target nucleic acid) to be amplified and a reagent to thermal cycling. In the PCR method, it is currently required to efficiently amplify a target nucleic acid by shortening the time required for a thermal cycle between each step of thermal denaturation, annealing and extension.

  Patent Document 1 discloses a thermal cycle apparatus that performs PCR by moving droplets of a reaction solution between a first region (high temperature / about 95 ° C.) and a second region (low temperature / about 66 ° C.). Yes.

JP 2012-115208 A

In the thermal cycle apparatus described in Patent Document 1, when the reaction liquid is moved from the first area to the second area, the temperature of the surrounding liquid (oil) is relatively high. When a heat cycle is applied at a high speed, the temperature of the reaction solution may not be quickly taken away. Therefore, there is a problem that it is difficult to improve the speed of the heat cycle.
Accordingly, there has been a demand for a substance amplifying apparatus that can rapidly change the temperature of the reaction solution when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following forms or application examples.

  [Application Example 1] A substance amplification apparatus according to this application example is a substance amplification apparatus that amplifies a substance by subjecting a reaction liquid to a heat cycle, and is equipped with a reaction vessel having a flow path for moving the reaction liquid in the longitudinal direction. A possible mounting portion, a first region disposed at one end of the flow path of the reaction vessel, a third region disposed at the other end in the direction opposite to the one end of the flow path, and a first of the flow path A second region disposed in a region between the region and the third region, a first heating unit that controls the first region to a first temperature, and a second region that is controlled to a second temperature lower than the first temperature. A second heating unit and a drive mechanism that moves the reaction liquid between the first region, the second region, and the third region are provided.

According to the substance amplifying device of this application example, the first region is disposed at one end of the flow path, the third region is disposed at the other end, and the second region is disposed between the first region and the third region. An area is placed. And the 1st field is controlled by the 1st heating part to the 1st temperature (for example, temperature in which heat denaturation is performed). Further, the second heating unit controls the second region to a second temperature lower than the first temperature (for example, a temperature at which the annealing / elongation reaction is performed). The third region is in contact with, for example, the atmosphere (room temperature). Then, the reaction solution is moved between the first region, the second region, and the third region by the driving mechanism.
With this configuration, the reaction solution moves from the first region to the third region through the second region. Further, the reaction solution that has moved to the third region passes through the second region and moves to the first region. For this reason, the reaction liquid heated to a temperature close to the first temperature in the first region passes through the second region and moves to a third region that is lower than the second temperature, for example, in contact with room temperature. Can be reduced. Then, the reaction liquid whose temperature has been lowered moves from the third region to the second region, and moves from the second region to the first region, thereby being heated from the second temperature to the first temperature.
Thereby, for example, the reaction solution heated to a temperature close to the first temperature is thermally denatured in the first region. The reaction solution undergoes an annealing / extension reaction while moving from the second region to the first region.
Therefore, when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed (first temperature in the first region) to the temperature at which annealing / extension reaction is performed (second temperature in the second region), the temperature change Can be realized. Further, the heat cycle speed of the substance amplifying apparatus can be improved.

  [Application Example 2] A substance amplification apparatus according to this application example is a substance amplification apparatus that amplifies a substance by subjecting a reaction liquid to a heat cycle, and is equipped with a reaction vessel having a flow path for moving the reaction liquid in the longitudinal direction. A possible mounting portion, a first region disposed at one end of the flow path of the reaction vessel, a second region disposed at the other end in the direction opposite to the one end of the flow path, and a first of the flow path. A third region disposed in a region between the region and the second region, a first heating unit that controls the first region to a first temperature, and a second region that is controlled to a second temperature lower than the first temperature. A second heating unit and a drive mechanism that moves the reaction liquid between the first region, the second region, and the third region are provided.

According to the substance amplifying device of this application example, the first region is disposed at one end of the flow path, the second region is disposed at the other end, and the third region is disposed between the first region and the second region. An area is placed. And the 1st field is controlled by the 1st heating part to the 1st temperature (for example, temperature in which heat denaturation is performed). Further, the second heating unit controls the second region to a second temperature lower than the first temperature (for example, a temperature at which the annealing / elongation reaction is performed). The third region is in contact with, for example, the atmosphere (room temperature). Then, the reaction solution is moved between the first region, the second region, and the third region by the driving mechanism.
With this configuration, the reaction solution moves from the first region to the second region through the third region. Further, the reaction solution that has moved to the second region passes through the third region and moves to the first region. For this reason, the reaction liquid heated to the temperature close to the first temperature in the first region can be quickly lowered by moving the third region that is lower than the second temperature, for example, in contact with room temperature. Then, the reaction liquid whose temperature has been lowered moves from the third region to the second region and is heated at the second temperature.
Thereby, for example, the reaction solution heated to a temperature close to the first temperature is thermally denatured in the first region. In addition, the temperature of the reaction solution is decreased in the third region, and the reaction solution is heated at the second temperature in the second region to be annealed. In addition, an extension reaction is performed when moving from the second region to the first region through the third region.
Therefore, when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, a substance amplifying device capable of rapidly changing the temperature can be realized. Further, the heat cycle speed of the substance amplifying apparatus can be improved.

  [Application Example 3] In the substance amplification device according to this application example, a reaction vessel in which a reaction solution moves in a flow path formed by connecting a first flow path and a second flow path via a bent portion. A mounting portion that can be mounted, a first region that is an end region on the first channel side of the channel, a second region that is an end region on the second channel side of the channel, A third region that is a region of the bent portion; a first heating unit that controls the first region to a first temperature; a second heating unit that controls a second region to a second temperature lower than the first temperature; And a drive mechanism for moving the reaction solution between the region, the second region, and the third region.

According to the substance amplifying device of this application example, the flow path is formed by connecting the first flow path and the second flow path via the bent portion. The first region is disposed at the end of the flow channel on the first flow channel side, the second region is disposed at the end of the second flow channel side, and the third region is disposed at the bent portion. And the 1st field is controlled by the 1st heating part to the 1st temperature (for example, temperature in which heat denaturation is performed). Further, the second heating unit controls the second region to a second temperature lower than the first temperature (for example, a temperature at which the annealing / elongation reaction is performed). The third region is in contact with, for example, the atmosphere (room temperature). Then, the reaction solution is moved between the first region, the second region, and the third region by the driving mechanism.
With this configuration, the reaction solution is temporarily stopped from the first region to the third region disposed in the bent portion, and then moved to the second region. In addition, the reaction solution that has moved to the second region is temporarily stopped by the third region and then moved to the first region. For this reason, the reaction liquid that has been heated to a temperature close to the first temperature in the first region moves to a third region that is lower than the second temperature, for example, in contact with room temperature, and is temporarily stopped. The temperature can be lowered. Then, the reaction liquid whose temperature has been lowered moves from the third region to the second region and is heated at the second temperature.
Thereby, for example, the reaction solution heated to a temperature close to the first temperature is thermally denatured in the first region. In addition, the temperature of the reaction solution is decreased in the third region, and the reaction solution is heated at the second temperature in the second region to be annealed. In addition, an extension reaction is performed when moving from the second region to the first region through the third region.
Therefore, when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, a substance amplifying device capable of rapidly changing the temperature can be realized. Further, the heat cycle speed of the substance amplifying apparatus can be improved.

  Application Example 4 In the substance amplifying device described in the above application example, it is preferable to include a third heating unit that controls the third region to a third temperature lower than the second temperature.

  According to the substance amplifying device of this application example, the temperature of the reaction solution is annealed from the temperature at which heat denaturation is performed by including the third heating unit that controls the third region to a third temperature lower than the second temperature. When changing to the temperature at which the extension reaction takes place, the temperature change can be made quickly and accurately.

  Application Example 5 In the substance amplification device according to the application example described above, the substance amplification device includes a blocking unit that is arranged along the outer wall of the third region and blocks heat from the first heating unit and / or the second heating unit. preferable.

  According to the substance amplifying device of this application example, by including a blocking unit disposed along the outer wall of the third region, by blocking heat from the first heating unit and / or the second heating unit to the third region The heat of the first heating unit and the second heating unit can be prevented from affecting the third region. Accordingly, when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, the temperature change can be performed quickly.

  [Application Example 6] In the substance amplifying device described in the above application example, the reaction container has opposing inner walls, and the distance between the opposing inner walls is equal to both of the opposing inner walls when the reaction solution is arranged in the reaction container. It is preferable that it is the distance which a reaction liquid contacts.

  According to the substance amplifying apparatus of this application example, when the reaction solution is arranged in the reaction vessel, the heat of each heating unit is set to the distance that the reaction solution contacts both of the opposing inner walls. The heat of the reaction liquid is easily transmitted to each heating unit, and the reaction liquid can be efficiently changed to a predetermined temperature.

  Application Example 7 In the substance amplification device according to the application example described above, the drive mechanism rotates the mounting unit, the first heating unit, and the second heating unit when the reaction container is mounted on the mounting unit. Switching between the first arrangement and the second arrangement, the first arrangement is an arrangement in which the first region is below the second region in the direction of gravity, and the second arrangement is the second region in the direction of gravity. The arrangement is preferably lower than the first region.

  According to the substance amplifying device of this application example, the driving mechanism causes the first arrangement in which the first region is lower than the second region in the gravity direction, and the second arrangement in which the second region is lower than the first region in the gravity direction. Switch between 2 arrangements. By utilizing gravity in this way, for example, the time required for heat denaturation, annealing and elongation for amplifying nucleic acids can be shortened.

  Application Example 8 In the substance amplification device according to the application example described above, the drive mechanism includes a pressing unit that presses the reaction container and closely contacts the inner wall of the reaction container when the reaction container is mounted on the mounting unit. Is preferred.

  According to the substance amplifying device of this application example, the reaction liquid is moved between the first region, the second region, and the third region by providing the pressing unit and pressing the reaction vessel to closely contact the inner wall. Can do. As a result, when the temperature of the reaction solution is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, a substance amplifying device capable of rapidly changing the temperature can be realized. Further, heat denaturation, annealing / extension reaction can be performed regardless of the posture of the reaction vessel.

The side sectional view of the reaction container concerning a 1st embodiment. The plane sectional view of a reaction vessel. Sectional drawing which shows the state of the reaction container in 1st arrangement | positioning. Sectional drawing which shows the state of the reaction container in the middle toward 2nd arrangement | positioning from 1st arrangement | positioning. Sectional drawing which shows the state of the reaction container in 2nd arrangement | positioning. Sectional drawing which shows the state of the reaction container in the middle of going to 1st arrangement | positioning from 2nd arrangement | positioning. Sectional drawing which shows the state of the reaction container in the 1st arrangement | positioning which concerns on 2nd Embodiment. Sectional drawing which shows the state of the reaction container in the middle toward 2nd arrangement | positioning from 1st arrangement | positioning. Sectional drawing which shows the state of the reaction container in 2nd arrangement | positioning. Sectional drawing which shows the state of the reaction container in the middle of going to 1st arrangement | positioning from 2nd arrangement | positioning. The perspective view of the reaction container which concerns on 3rd Embodiment. The top view which shows a reaction container and a heating part. FIG. 3 is a schematic cross-sectional view of a reaction vessel and a heating unit. Sectional drawing which shows the state of the reaction container in 1st arrangement | positioning. Sectional drawing which shows the state which rotated the reaction container 135 degrees clockwise. Sectional drawing which shows the state which rotated the reaction container 45 degrees counterclockwise. Sectional drawing which shows the state which rotated the reaction container 135 degrees clockwise. Sectional drawing which shows the state which turned reaction container 45 degrees counterclockwise and was set as the 2nd arrangement | positioning. Sectional drawing which shows the state which rotated the reaction container 135 degrees counterclockwise. Sectional drawing which shows the state which rotated the reaction container 45 degrees clockwise. Sectional drawing which shows the state which rotated the reaction container 135 degrees counterclockwise. Sectional drawing which shows the state which rotated the reaction container 45 degree clockwise and returned to the 1st arrangement | positioning. Sectional drawing which shows the reaction container and heating part which concern on 4th Embodiment. The sectional side view which shows the interruption | blocking part which concerns on 5th Embodiment. The top view which looked at the interruption | blocking part from the 1st wall side. The figure which shows the state which the reaction liquid moved to the 1st area | region which concerns on 6th Embodiment. The figure which shows the state which moves the reaction liquid of a 1st area | region to a 3rd area | region. The figure which shows the state which moves the reaction liquid of a 3rd area | region to a 2nd area | region. The figure which shows the state which moves the reaction liquid of a 2nd area | region to a 1st area | region.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the following drawings, the scale of each member is illustrated differently from the actual scale for convenience of explanation.

[First Embodiment]
The substance amplifying apparatus 1 of this embodiment is configured as an apparatus that amplifies nucleic acids by subjecting the reaction solution 150 to a heat cycle by using a PCR (Polymerase Chain Reaction) method.

  FIG. 1A is a side sectional view of a reaction vessel 100 according to the present embodiment. Specifically, it is a cross-sectional view of the reaction vessel 100 from the third wall 130 side. FIG. 1B is a plan sectional view of the reaction vessel 100. Specifically, it is a cross-sectional view of the reaction vessel 100 from the first wall 110 side.

  As shown in FIGS. 1A and 1B, the reaction vessel 100 has a substantially rectangular parallelepiped shape, and includes a first wall 110, a second wall 120 facing the first wall 110, a first wall 110, and a second wall. It is comprised by the 3rd wall 130 which connects 120. FIG.

  A reaction solution 150 is sealed inside the reaction vessel 100 surrounded by the first wall 110, the second wall 120, and the third wall 130. The reaction liquid 150 according to the present embodiment is located in the lowermost region in the gravity direction of the reaction vessel 100, and the first wall 110, the second wall 120, and the third wall 130 are located at the lowermost region. Are in contact with the respective inner walls (first inner wall 110a, second inner wall 120a, and third inner wall 130a). In other words, the distance between the inner walls of the reaction vessel 100 is the distance at which the reaction solution 150 contacts both the first inner wall 110a and the second inner wall 120a facing each other when the reaction solution 150 is disposed in the reaction vessel 100. ing. With the above-described configuration, the reaction vessel 100 forms a flow channel 160 that is a path through which the reaction solution 150 moves in the longitudinal direction. The reaction solution 150 can move between a first region 170, a second region 180, and a third region 190, which will be described later, by the flow channel 160.

  The substance amplifying apparatus 1 of the present embodiment uses, as a reaction solution 150, a DNA sample (target nucleic acid) containing a DNA (deoxyribonucleic acid) sequence to be amplified by PCR, a DNA polymerase necessary for amplifying DNA, and primers (20 bases) An aqueous solution containing a certain degree of oligonucleotide, etc.).

  2A to 2D are diagrams schematically showing an operation when PCR is performed using the substance amplifying apparatus 1. 2A to 2D are schematic cross-sectional views showing the positional relationship between the reaction vessel 100 and the heating unit 200 in the substance amplifying apparatus 1, and are cross-sectional views seen from the short side of the reaction vessel 100.

  The heating unit 200 includes a first heating unit 210 and a second heating unit 220, as shown in FIG. 2A. The first heating unit 210 and the second heating unit 220 are members that transmit heat generated from respective heaters (not shown) to the reaction vessel 100. The first heating unit 210 includes a pair of first heat application units 211. Similarly, the second heating unit 220 includes a pair of second heat application units 221. In addition, the 1st heat application part 211 is holding temperature so that it may become 1st temperature. Moreover, the 2nd heat application part 221 is maintaining temperature so that it may become 2nd temperature.

  In the present embodiment, the first heat application unit 211 and the second heat application unit 221 are configured as aluminum blocks, so that the reaction vessel 100 can be efficiently heated. Moreover, since the heat conductivity is high, uneven heating is hardly generated in the block, and a highly accurate heat cycle is realized. Further, since the processing is easy, the block can be accurately molded, and the heating accuracy can be improved. Therefore, a more accurate thermal cycle can be realized.

  As shown in FIG. 2A, in the reaction vessel 100, a region disposed at one end of the flow channel 160 is referred to as a first region 170. In the present embodiment, when the reaction container 100 is attached to an attachment part (not shown), the first heating part 210 is disposed opposite to the first region 170. Specifically, when the reaction container 100 is attached to the attachment portion, the pair of first heat application portions 211 are disposed so as to abut on the first wall 110 and the second wall 120 of the first region 170, respectively. During the heat cycle operation, the first heat application unit 211 heats the first wall 110 and the second wall 120 at the first temperature.

  As shown in FIG. 2A, a region disposed at the other end portion in the direction opposite to the one end portion of the reaction vessel 100 (channel 160) is referred to as a third region 190. In the present embodiment, the heating unit 200 is not disposed at a position facing the third region 190. In other words, the third region 190 is in contact with the atmosphere and is at room temperature or the internal temperature of the substance amplifying apparatus 1.

  As shown in FIG. 2A, a region disposed at a portion between the first region 170 and the third region 190 is referred to as a second region 180. In the present embodiment, when the reaction vessel 100 is attached to an attachment part (not shown), the second heating part 220 is disposed opposite to the second region 180. Specifically, when the reaction container 100 is attached to the attachment portion, the pair of second heat application portions 221 are disposed so as to contact the first wall 110 and the second wall 120 of the second region 180, respectively. During the heat cycle operation, the second heat application unit 221 heats the first wall 110 and the second wall 120 at the second temperature.

  The substance amplifying apparatus 1 of this embodiment is provided with a drive mechanism (not shown). The drive mechanism is a mechanism that moves the reaction liquid 150 between the first region 170, the second region 180, and the third region 190. The drive mechanism is a mechanism that switches the mounting portion (not shown) on which the reaction vessel 100 is mounted and the heating unit 200 (the first heating unit 210 and the second heating unit 220) between the first arrangement and the second arrangement. .

  The drive mechanism includes a motor, a drive shaft, a mounting part (all not shown), and the like. The drive shaft is configured with reference to a rotation axis A that is set perpendicular to the approximate center of the first wall 110. The holding unit holds the reaction container 100 without being buffered with the heating unit 200. Then, by operating the motor, the mounting portion and the heating portion 200 rotate while maintaining the positional relationship around the drive shaft (rotation axis A), whereby the reaction vessel 100 rotates.

  In the present embodiment, the first arrangement is an arrangement in which the first region 170 is below the second region 180 in the direction of gravity (the direction in which gravity acts). In the first arrangement, the reaction solution 150 is located in the first region 170.

  In the present embodiment, the second arrangement is an arrangement in which the second region 180 is below the first region 170 in the direction of gravity (the direction in which gravity acts). In the present embodiment, the second arrangement is an arrangement in which the third region 190 is located at the lowermost part of the flow path 160 in the gravity direction. In the second arrangement, the reaction solution 150 is located in the third region 190. When the reaction vessel 100 is attached to the attachment portion, the drive mechanism rotates the reaction vessel 100 and switches the first region 170 and the second region 180 so as to be alternately up and down in the direction of gravity.

2A to 2D are diagrams schematically showing an operation in the case where PCR is performed using the substance amplifying apparatus 1 as described above. Specifically, FIG. 2A is a cross-sectional view showing a state of the reaction vessel 100 in the first arrangement. FIG. 2B is a cross-sectional view showing a state of the reaction vessel 100 on the way from the first arrangement to the second arrangement. FIG. 2C is a cross-sectional view showing a state of the reaction vessel 100 in the second arrangement. FIG. 2D is a cross-sectional view showing a state of the reaction vessel 100 on the way from the second arrangement toward the first arrangement. 2A to 2D, the direction of the arrow g (downward direction in the figure) indicates the direction of gravity.
With reference to FIG. 2A-FIG. 2D, operation | movement in the case of performing PCR using the substance amplification apparatus 1 is demonstrated.

  In the present embodiment, PCR is a technique for amplifying nucleic acid in the reaction solution 150 by repeatedly performing two-stage temperature treatment on the reaction solution 150. In high-temperature treatment, heat denaturation (denaturation (separation) of double-stranded DNA into single strands) is performed. In the low-temperature treatment, annealing (reaction in which the primer binds to single-stranded DNA) and extension reaction (reaction in which a DNA polymerase is reacted to form a complementary strand of DNA) are performed.

  In PCR, a high temperature (first temperature) at which heat denaturation is performed is set to a temperature between about 92 ° C. and 97 ° C., for example, and a low temperature (second temperature) is set to a temperature between about 65 ° C. and 72 ° C. for example. Is set. In the present embodiment, the first temperature is, for example, 95 ° C., and the second temperature is, for example, 70 ° C.

  The temperatures of the first heating unit 210 and the second heating unit 220 are controlled by a temperature sensor and a control unit (both not shown). The control unit controls the operation of the drive mechanism. The control unit controls the drive mechanism to repeat the first arrangement and the second arrangement at a predetermined cycle time.

  The control unit includes a CPU (Central Processing Unit) that performs various arithmetic processes as a processor. The control unit includes a storage device such as a ROM (Read Only Memory) and a RAM (Random Access Memory). In the storage device, a storage area for storing various programs and data for controlling each of the above operations is set. In addition, the storage device is set with a work area for temporarily storing data being processed and various processing results for the CPU, a storage area that functions as a temporary file, and other various storage areas. Yes.

The operation when performing PCR will be specifically described.
First, the reaction container 100 in which the reaction solution 150 is sealed is attached and fixed to the attachment portion. As a result, as shown in FIG. 2A, the first heating unit 210 is in contact with the reaction vessel 100 (the first wall 110 and the second wall 120) at a position facing the first region 170. The second heating unit 220 is in contact with the reaction vessel 100 (the first wall 110 and the second wall 120) at a position facing the second region 180. The state shown in FIG. 2A is the first arrangement.

  As shown in FIG. 2A, in the first arrangement, the reaction liquid 150 is in a state located in the first region 170. In a state where the reaction liquid 150 is positioned in the first region 170, the first heating unit 210 and the second heating unit 220 are heated to a predetermined temperature. Specifically, the first heating unit 210 is heated to the first temperature, and the second heating unit 220 is heated to the second temperature. In the present embodiment, the first temperature corresponds to a high temperature, and the second temperature corresponds to a low temperature.

  In this state, the first heat application unit 211 heats the first region 170 of the reaction vessel 100 to the first temperature. In the present embodiment, the reaction liquid 150 is in contact with both the first inner wall 110a and the second inner wall 120a facing each other of the reaction vessel 100. Therefore, in the first arrangement, the first temperature is transmitted to the reaction solution 150 at a high speed. Therefore, heat denaturation is performed on the reaction solution 150 to be a reaction at the first temperature.

  At this time, in the second region 180, the second heat application unit 221 heats the reaction vessel 100 to the second temperature. Moreover, in the 3rd area | region 190, it is the internal temperature of the substance amplification apparatus 1 (it is temporarily set as atmospheric temperature in this embodiment). Thereby, a temperature gradient in which the temperature gradually changes between the first temperature, the second temperature, and the atmospheric temperature is generated between the first region 170 and the second region 180 and between the second region 180 and the third region 190. It is formed.

  Next, the arrangement of the reaction vessel 100 and the heating unit 200 is switched from the first arrangement to the second arrangement by the drive mechanism. Specifically, when switching from the first arrangement to the second arrangement, the rotation is performed by 180 °. Note that when switching from the first arrangement to the second arrangement, the positional relationship between the first heat application unit 211 and the second heat application unit 221 in the first arrangement is maintained.

  As shown in FIG. 2B, the drive mechanism rotates about the rotation axis A. As shown in FIGS. 2A to 2B, when the reaction vessel 100 rotates, the reaction solution 150 passes from the first region 170 through the second region 180 toward the third region 190 by gravity.

  At this time, the reaction solution 150 in the first region 170 is close to the first temperature at which heat denaturation is performed. Then, by rotating, the reaction solution 150 passes through the second region 180 and is affected by the second temperature to lower the temperature. Then, as shown in FIG. 2C, when the second arrangement is adopted, the reaction solution 150 is located in the third region 190, and rapidly decreases the temperature due to the influence of the atmospheric temperature.

  Next, the drive mechanism switches from the second arrangement to the first arrangement (FIG. 2A). Specifically, when switching from the second arrangement to the first arrangement, it is performed by rotating 180 ° in the direction opposite to the rotation direction switched from the first arrangement to the second arrangement. As shown in FIGS. 2C to 2D, when the reaction vessel 100 rotates, the reaction solution 150 passes from the third region 190 through the second region 180 toward the first region 170 by gravity.

At this time, the temperature of the reaction liquid 150 is decreased in the third region 190, and the reaction liquid 150 is affected by the second temperature in the second region 180 due to the rotation of the reaction vessel 100. The temperature is close to. And when it becomes 1st arrangement | positioning (FIG. 2A), the reaction liquid 150 is heated from 2nd temperature to 1st temperature. In this case, during the movement from the second region 180 to the first region 170, the reaction solution 150 undergoes an annealing / elongation reaction that is a reaction at the second temperature.
In addition, after the case where it becomes the 1st arrangement | positioning, the operation | movement mentioned above is repeated.

  Note that the annealing / elongation reaction is also performed in the flow path 160 other than in the course of moving from the second region 180 to the first region 170 when the reaction solution 150 reaches a temperature suitable for the annealing and extension reaction.

  The substance amplifying apparatus 1 of the present embodiment is switched from the first arrangement to the second arrangement, and is performed in about 2 seconds from the second arrangement to the first arrangement in about 2 seconds, and the necessary number of cycles (for example, 40 cycles). Repeat until

According to the embodiment described above, the following effects can be obtained.
(1) According to the substance amplifying device 1 of the present embodiment, the first region 170 is disposed at one end of the flow channel 160, the third region 190 is disposed at the other end, and the first region 170 and the third region The second region 180 is disposed at a position between the first region 190 and the second region 180. Then, the first heating unit 210 controls the first region 170 to the first temperature (temperature at which heat denaturation is performed). Further, the second heating unit 220 controls the second region 180 to a second temperature lower than the first temperature (a temperature at which the annealing / elongation reaction is performed). Note that the third region 190 is in contact with the atmospheric temperature. Then, the reaction solution 150 is moved between the first region 170, the second region 180, and the third region 190 by the driving mechanism.
With this configuration, the reaction solution 150 moves from the first region 170 through the second region 180 to the third region 190. In addition, the reaction liquid 150 that has moved to the third region 190 passes through the second region 180 and moves to the first region 170. For this reason, the reaction liquid 150 heated to a temperature close to the first temperature in the first region 170 passes through the second region 180 and moves to the third region 190 that is in contact with the atmospheric temperature lower than the second temperature. Can quickly reduce the temperature. Then, the reaction liquid 150 whose temperature has been lowered moves from the third region 190 to the second region 180, and moves from the second region 180 to the first region 170, thereby being heated from the second temperature to the first temperature. .
Thus, the reaction solution 150 that has been heated to a temperature close to the first temperature is thermally denatured in the first region 170. The reaction solution 150 undergoes an annealing / elongation reaction while moving from the second region 180 to the first region 170.
Therefore, when the temperature of the reaction solution 150 is changed from the temperature at which thermal denaturation is performed (first temperature in the first region 170) to the temperature at which annealing / extension reaction is performed (second temperature in the second region 180). Thus, the substance amplifying apparatus 1 capable of rapidly changing the temperature can be realized. Moreover, the speed of the heat cycle of the substance amplifying apparatus 1 can be improved.

  (2) According to the substance amplifying apparatus 1 of the present embodiment, when the reaction solution 150 is disposed in the reaction vessel 100, both the inner walls (the first inner wall 110a and the second inner wall 120a) with which the reaction solution 150 is opposed are disposed. By setting the contact distance, the heat of the first heating unit 210 and the second heating unit 220 is easily transmitted to the reaction liquid 150 through the reaction vessel 100. Further, the heat of the reaction liquid 150 is easily transmitted to the first heating unit 210 and the second heating unit 220. Thereby, the reaction liquid 150 can be efficiently changed to a predetermined temperature.

  (3) According to the substance amplifying device 1 of the present embodiment, the driving mechanism causes the first arrangement in which the first region 170 is below the second region 180 in the direction of gravity and the second region 180 in the direction of gravity to The second arrangement below the first area 170 is switched. By using gravity in this way, the time required for heat denaturation, annealing and extension for amplifying nucleic acids can be shortened.

[Second Embodiment]
The substance amplifying device 2 of the present embodiment is different from the first embodiment in the arrangement of the first region 170, the second region 180, and the third region 190 in the reaction vessel 100 (channel 160). Other configurations are the same as those in the first embodiment. Similar components are denoted by similar reference numerals.

3A to 3D are diagrams schematically showing an operation when performing PCR using the substance amplifying apparatus 2 according to the present embodiment. Specifically, FIG. 3A is a cross-sectional view showing the state of the reaction vessel 100 in the first arrangement. FIG. 3B is a cross-sectional view showing the state of the reaction vessel 100 on the way from the first arrangement to the second arrangement. FIG. 3C is a cross-sectional view showing a state of the reaction vessel 100 in the second arrangement. FIG. 3D is a cross-sectional view showing a state of the reaction vessel 100 on the way from the second arrangement toward the first arrangement. 3A to 3D, the direction of the arrow g (downward direction in the figure) indicates the direction of gravity.
With reference to FIG. 3A-FIG. 3D, the positional relationship of the reaction container 100 and the heating part 200 of this embodiment is demonstrated.

  As shown in FIG. 3A, the reaction vessel 100 is the same as the reaction vessel 100 of the first embodiment. The difference is the positions of the second region 180 and the third region 190. In the reaction vessel 100, a region disposed at one end of the flow channel 160 is referred to as a first region 170. As in the first embodiment, when the reaction vessel 100 is attached to an attachment part (not shown), the first heating part 210 is disposed relative to the first region 170. Specifically, when the reaction container 100 is attached to the attachment portion, the pair of first heat application portions 211 are disposed so as to abut on the first wall 110 and the second wall 120 of the first region 170, respectively. During the heat cycle operation, the first heat application unit 211 heats the first wall 110 and the second wall 120 at the first temperature.

  As shown in FIG. 3A, in the present embodiment, a region disposed at the other end portion in the direction opposite to the one end portion of the reaction vessel 100 (flow channel 160) is the second region 180. Similar to the first embodiment, when the reaction vessel 100 is attached to an attachment part (not shown), the second heating part 220 is disposed relative to the second region 180. Specifically, when the reaction container 100 is attached to the attachment portion, the pair of second heat application portions 221 are disposed so as to contact the first wall 110 and the second wall 120 of the second region 180, respectively. During the heat cycle operation, the second heat application unit 221 heats the first wall 110 and the second wall 120 at the second temperature.

  As shown in FIG. 3A, in the present embodiment, a region arranged between the first region 170 and the second region 180 is the third region 190. In addition, the heating part 200 is not arrange | positioned in the position facing the 3rd area | region 190 similarly to 1st Embodiment. In other words, the third region 190 is in contact with the atmosphere and is at room temperature or the internal temperature of the substance amplifying device 2.

  The material amplification device 2 is provided with a drive mechanism (not shown) as in the first embodiment. The drive mechanism is a mechanism that moves the reaction liquid 150 between the first region 170, the second region 180, and the third region 190. Further, the drive mechanism switches the mounting unit and the heating unit 200 (the first heating unit 210 and the second heating unit 220) between the first arrangement and the second arrangement, whereby the reaction vessel 100 is arranged in the first arrangement and the second arrangement. It is a mechanism that switches between arrangements. A description of the general configuration of the drive mechanism is omitted.

  The first arrangement is the same as in the first embodiment, and the first area 170 is lower than the second area 180 in the direction of gravity (the direction in which gravity acts). In the first arrangement, the reaction solution 150 is located in the first region 170.

  In the present embodiment, the second arrangement is an arrangement in which the second region 180 is lower than the first region 170 in the direction of gravity (the direction in which gravity acts). In the second arrangement, the reaction solution 150 is located in the second region 180. When the reaction vessel 100 is attached to the attachment portion, the drive mechanism rotates the reaction vessel 100 and switches the first region 170 and the second region 180 so as to be alternately up and down in the direction of gravity.

  With reference to FIG. 3A-FIG. 3D, operation | movement in the case of performing PCR using the substance amplification apparatus 2 is demonstrated. In PCR, as in the first embodiment, the first temperature is, for example, 95 ° C., and the second temperature is, for example, 70 ° C.

  First, the reaction container 100 in which the reaction solution 150 is sealed is attached and fixed to the attachment portion. As a result, as shown in FIG. 3A, the first heating unit 210 is in contact with the reaction vessel 100 (the first wall 110 and the second wall 120) at a position facing the first region 170. The second heating unit 220 is in contact with the reaction vessel 100 (the first wall 110 and the second wall 120) at a position facing the second region 180. The state shown in FIG. 3A is the first arrangement.

  As shown in FIG. 3A, in the first arrangement, the reaction liquid 150 is in a state located in the first region 170. In a state where the reaction liquid 150 is positioned in the first region 170, the first heating unit 210 and the second heating unit 220 are heated to a predetermined temperature. Specifically, the first heating unit 210 is heated to the first temperature, and the second heating unit 220 is heated to the second temperature. In the present embodiment, the first temperature corresponds to a high temperature, and the second temperature corresponds to a low temperature.

  In this state, the first heat application unit 211 heats the first region 170 of the reaction vessel 100 to the first temperature. In the present embodiment, the reaction liquid 150 is in contact with both the first inner wall 110a and the second inner wall 120a facing each other of the reaction vessel 100. Therefore, in the first arrangement, the first temperature is transmitted to the reaction solution 150 at a high speed. Therefore, heat denaturation is performed on the reaction solution 150 to be a reaction at the first temperature.

  At this time, in the second region 180, the second heat application unit 221 heats the reaction vessel 100 to the second temperature. Moreover, in the 3rd area | region 190, it is the internal temperature of the substance amplification apparatus 2 (it is temporarily set as atmospheric temperature in this embodiment). As a result, a temperature gradient in which the temperature gradually changes between the first temperature, the atmospheric temperature, and the second temperature is generated between the first region 170 and the third region 190 and between the third region 190 and the second region 180. It is formed.

  Next, the arrangement of the reaction vessel 100 and the heating unit 200 is switched from the first arrangement to the second arrangement by the drive mechanism. Specifically, when switching from the first arrangement to the second arrangement, the rotation is performed by 180 °. Note that when switching from the first arrangement to the second arrangement, the positional relationship between the first heat application unit 211 and the second heat application unit 221 in the first arrangement is maintained.

  As shown in FIG. 3B, the drive mechanism rotates about the rotation axis A. As shown in FIGS. 3A to 3B, when the reaction vessel 100 rotates, the reaction solution 150 passes from the first region 170 through the third region 190 toward the second region 180 by gravity.

  At this time, the reaction solution 150 in the first region 170 is close to the first temperature at which heat denaturation is performed. And by rotating, the reaction liquid 150 passes through the 3rd area | region 190, receives the influence of atmospheric temperature, and falls temperature rapidly. And as shown to FIG. 3C, when it becomes 2nd arrangement | positioning, the reaction liquid 150 is located in the 2nd area | region 180, and becomes the temperature close | similar to 2nd temperature by receiving to the influence of 2nd temperature. Then, annealing that is a reaction at the second temperature is performed on the reaction solution 150.

  Next, the drive mechanism switches from the second arrangement to the first arrangement (FIG. 3A). Specifically, when switching from the second arrangement to the first arrangement, it is performed by rotating 180 ° in the direction opposite to the rotation direction switched from the first arrangement to the second arrangement. As shown in FIGS. 3C to 3D, when the reaction vessel 100 rotates, the reaction solution 150 passes from the second region 180 through the third region 190 to the first region 170 by gravity.

At this time, the temperature of the reaction solution 150 is decreased in the third region 190. Then, when the reaction container 100 is rotated to be in the first arrangement (FIG. 3A), the reaction solution 150 is heated from the lowered temperature to the first temperature. In this case, during the movement from the third region 190 to the first region 170, an extension reaction that is a reaction at the second temperature is performed on the reaction solution 150.
In addition, after the case where it becomes the 1st arrangement | positioning, the operation | movement mentioned above is repeated.

  In the annealing / elongation reaction, when the reaction solution 150 reaches a temperature suitable for the annealing and extension reaction, the flow path is not in the middle of moving to the second region 180 or the third region 190 to the first region 170. 160 is also performed.

According to the embodiment described above, the following effects can be obtained.
(1) According to the substance amplifying device 2 of the present embodiment, the first region 170 is disposed at one end of the flow channel 160, the second region 180 is disposed at the other end, and the first region 170 and the second region The third region 190 is disposed at a position between the first region 190 and the third region 190. Then, the first heating unit 210 controls the first region 170 to the first temperature (temperature at which heat denaturation is performed). Further, the second heating unit 220 controls the second region 180 to a second temperature lower than the first temperature (a temperature at which the annealing / elongation reaction is performed). Note that the third region 190 is in contact with the atmospheric temperature. Then, the reaction solution 150 is moved between the first region 170, the second region 180, and the third region 190 by the driving mechanism.
With this configuration, the reaction solution 150 moves from the first region 170 through the third region 190 to the second region 180. In addition, the reaction liquid 150 that has moved to the second region 180 passes through the third region 190 and moves to the first region 170. For this reason, the reaction liquid 150 that has been heated to a temperature close to the first temperature in the first region 170 passes through the third region 190 that is in contact with the atmospheric temperature lower than the second temperature, thereby quickly reducing the temperature. Can do. Then, the reaction liquid 150 whose temperature has been lowered moves from the third region 190 to the second region 180 and is heated at the second temperature.
Thus, the reaction solution 150 that has been heated to a temperature close to the first temperature is thermally denatured in the first region 170. In addition, the temperature of the reaction solution 150 is decreased in the third region 190 and heated at the second temperature in the second region 180 to perform an annealing / elongation reaction. The annealing / elongation reaction is also performed when moving from the second region 180 to the first region 170 through the third region 190.
Therefore, when the temperature of the reaction solution 150 is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, the substance amplifying device 2 capable of quickly changing the temperature can be realized. Moreover, the speed of the heat cycle of the substance amplifying apparatus 2 can be improved.

[Third Embodiment]
In the substance amplifying apparatus 3 of the present embodiment, the shape of the reaction vessel 100A is different from the reaction vessel 100 of the first embodiment. The arrangement of the first region 170, the second region 180, and the third region 190 in the reaction vessel 100A (channel 160A) of the present embodiment is different from that of the first embodiment, but is substantially the same as the second embodiment. It becomes. Other configurations are the same as those in the first embodiment.

  4A to 4C are diagrams showing the shape and positional relationship between the reaction vessel 100A and the heating unit 200 of the substance amplification device 3 according to this embodiment. Specifically, FIG. 4A is a perspective view of the reaction vessel 100A. FIG. 4B is a plan view showing the reaction vessel 100 </ b> A and the heating unit 200. 4B is shown as a perspective view. FIG. 4C is a schematic cross-sectional view of the reaction vessel 100 </ b> A and the heating unit 200. FIG. 4C is a schematic cross-sectional view of the reaction vessel 100A cut into an L shape and the bent portion 140 viewed from the short direction of the reaction vessel 100A. With reference to FIG. 4A-FIG. 4C, the shape and positional relationship of 100 A of reaction containers and the heating part 200 of this embodiment are demonstrated.

  The reaction vessel 100A is formed in a generally L-shaped flat bag shape. The reaction vessel 100A includes an L-shaped first wall 110A, a second wall 120A facing the first wall 110A, and a third wall 130A connecting the first wall 110A and the second wall 120A with a predetermined thickness. It consists of and. In the reaction vessel 100A, the first flow path 161 and the second flow path 162 are connected via the bent portion 140 to form the flow path 160A. The reaction liquid 150 sealed in the reaction vessel 100A moves through the flow path 160A.

  As shown in FIGS. 4A to 4C, in the reaction vessel 100 </ b> A, a region disposed at the end of the flow channel 160 </ b> A (end on the first flow channel 161 side) is defined as a first region 170. As in the first embodiment, when the reaction vessel 100 is attached to an attachment part (not shown), the first heating part 210 is disposed relative to the first region 170. Specifically, when the reaction container 100A is attached to the attachment portion, the pair of first heat application portions 211 are arranged so as to contact the first wall 110A and the second wall 120A of the first region 170, respectively. During the heat cycle operation, the first heat application unit 211 heats the first wall 110A and the second wall 120A at the first temperature.

  As shown in FIGS. 4A to 4C, in the reaction vessel 100 </ b> A, a region disposed at the end of the flow channel 160 </ b> A (end on the second flow channel 162 side) is defined as a second region 180. Similar to the first embodiment, when the reaction vessel 100 </ b> A is attached to an attachment part (not shown), the second heating part 220 is disposed opposite to the second region 180. Specifically, when the reaction vessel 100A is attached to the attachment portion, the pair of second heat application portions 221 are disposed so as to contact the first wall 110A and the second wall 120A of the second region 180, respectively. During the heat cycle operation, the second heat application unit 221 heats the first wall 110A and the second wall 120A at the second temperature.

  As shown in FIGS. 4A to 4C, a region disposed in the bent portion 140 of the flow path 160 </ b> A is a third region 190. As in the first embodiment, when the reaction vessel 100A is mounted on a mounting portion (not shown), the heating unit 200 is not disposed at a position facing the third region 190. In other words, the third region 190 is in contact with the atmosphere and is at room temperature or the internal temperature of the substance amplifying device 3.

  The substance amplifying device 3 is provided with a drive mechanism (not shown) as in the first embodiment. The drive mechanism is a mechanism that moves the reaction liquid 150 between the first region 170, the second region 180, and the third region 190. The drive mechanism is a mechanism that switches the reaction vessel 100A and the heating unit 200 (the first heating unit 210 and the second heating unit 220) between the first arrangement and the second arrangement. A description of the general configuration of the drive mechanism is omitted.

  The first arrangement is an arrangement in which the first area 170 is lower in the direction of gravity than the second area 180 and the third area 190. In the first arrangement, the reaction solution 150 is located in the first region 170.

  In the present embodiment, the second arrangement is an arrangement in which the second region 180 is lower in the direction of gravity than the first region 170 and the third region 190. In the second arrangement, the reaction solution 150 is located in the second region 180. When the reaction vessel 100 is attached to the attachment portion, the drive mechanism rotates the reaction vessel 100A and switches the first region 170 and the second region 180 so as to alternately rise and fall in the direction of gravity.

  FIG. 5A to FIG. 5I are diagrams schematically showing an operation when performing PCR using the substance amplifying device 3. Specifically, FIG. 5A is a cross-sectional view showing the state of the reaction vessel 100A in the first arrangement. FIG. 5B is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 135 ° clockwise. FIG. 5C is a cross-sectional view illustrating a state in which the reaction vessel 100A is rotated 45 ° counterclockwise. FIG. 5D is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 135 ° clockwise. FIG. 5E is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 45 ° counterclockwise to be in the second arrangement. FIG. 5F is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 135 ° counterclockwise. FIG. 5G is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 45 ° clockwise. FIG. 5H is a cross-sectional view showing a state in which the reaction vessel 100A is rotated 135 ° counterclockwise. FIG. 5I is a cross-sectional view (similar to FIG. 5A) showing a state in which the reaction vessel 100A is rotated clockwise by 45 ° and returned to the first arrangement.

  With reference to FIG. 5A to FIG. 5I, an operation when performing PCR using the substance amplifying device 3 will be described. In PCR, as in the first embodiment, the first temperature is, for example, 95 ° C., and the second temperature is, for example, 70 ° C.

  First, the reaction container 100A in which the reaction solution 150 is sealed is attached and fixed to the attachment portion. As a result, as shown in FIG. 5A, the first heating unit 210 is in contact with the reaction vessel 100A (the first wall 110A and the second wall 120A) at a position facing the first region 170. The second heating unit 220 is in contact with the reaction vessel 100A (first wall 110A, second wall 120A) at a position facing the second region 180.

  The state shown in FIG. 5A is the first arrangement. The reaction solution 150 is located in the first region 170. In a state where the reaction liquid 150 is positioned in the first region 170, the first heating unit 210 and the second heating unit 220 are heated to a predetermined temperature. Specifically, the first heating unit 210 is heated to the first temperature, and the second heating unit 220 is heated to the second temperature. In the present embodiment, the first temperature corresponds to a high temperature, and the second temperature corresponds to a low temperature.

  In this state, the first heat application unit 211 heats the first region 170 of the reaction vessel 100A to the first temperature. In the present embodiment, the reaction solution 150 is in contact with both the first inner wall 110 </ b> Aa and the second inner wall 120 </ b> Aa facing the reaction vessel 100. Therefore, in the first arrangement, the first temperature is transmitted to the reaction solution 150 at a high speed. Therefore, heat denaturation is performed on the reaction solution 150 to be a reaction at the first temperature.

  At this time, in the second region 180, the second heat application unit 221 heats the reaction vessel 100 to the second temperature. Moreover, in the 3rd area | region 190, it is the internal temperature of the material amplification apparatus 3 (it is temporarily set as atmospheric temperature in this embodiment). As a result, a temperature gradient in which the temperature gradually changes between the first temperature, the atmospheric temperature, and the second temperature is generated between the first region 170 and the third region 190 and between the third region 190 and the second region 180. It is formed.

  Next, the arrangement of the reaction vessel 100 and the heating unit 200 is switched from the first arrangement to the second arrangement by the drive mechanism. Specifically, first, the state of the first arrangement shown in FIG. 5A is set to 0 °, and the drive mechanism is rotated 135 ° clockwise around the rotation axis B to obtain the state shown in FIG. 5B. . In this state, the reaction solution 150 is located in the first region 170.

  Next, the state shown in FIG. 5B is rotated 45 degrees counterclockwise about the rotation axis B from the state shown in FIG. In the state of FIG. 5C, the third region 190 is in the lowest position in the direction of gravity, and the reaction solution 150 moves from the first region 170 to the third region 190. As a result, the reaction liquid 150 heated in the first region 170 is positioned in the third region 190, so that the temperature is rapidly lowered under the influence of the atmospheric temperature.

  Next, the state shown in FIG. 5C is rotated by 135 ° clockwise around the rotation axis B from the state shown in FIG. 5C. In this state, the reaction liquid 150 is located in the third region 190. Then, the state shown in FIG. 5D is turned 45 ° counterclockwise around the turning axis B to obtain the state shown in FIG. 5E. In the state of FIG. 5E, the second region 180 is in the lowest position in the direction of gravity, and is in the second arrangement. The reaction solution 150 moves from the third region 190 to the second region 180. Thereby, the reaction liquid 150 is affected by the second temperature and becomes a temperature close to the second temperature. Then, an annealing / elongation reaction that is a reaction at the second temperature is performed on the reaction solution 150.

  Next, the arrangement of the reaction vessel 100 and the heating unit 200 is switched from the second arrangement to the first arrangement by the drive mechanism. Specifically, first, the state shown in FIG. 5F is turned from the state shown in FIG. 5E by 135 ° counterclockwise around the turning axis B. In this state, the reaction liquid 150 is located in the second region 180.

  Next, the state shown in FIG. 5F is rotated 45 degrees clockwise around the rotation axis B from the state shown in FIG. 5F. In the state of FIG. 5G, the third region 190 is in the lowest position in the direction of gravity, and the reaction solution 150 moves from the second region 180 to the third region 190. As a result, the reaction liquid 150 heated in the second region 180 is positioned in the third region 190, so that the temperature is rapidly lowered under the influence of the atmospheric temperature.

Next, from the state shown in FIG. 5G, it is rotated 135 ° counterclockwise around the rotation axis B to obtain the state shown in FIG. 5H. In this state, the reaction liquid 150 is located in the third region 190. Next, the state shown in FIG. 5I is rotated 45 degrees clockwise around the rotation axis B from the state shown in FIG. 5H. In the state of FIG. 5I, the first region 170 is in the lowest position in the direction of gravity and is in the first arrangement. The diagram shown in FIG. 5I is the same as the diagram shown in FIG. 5A. The reaction solution 150 moves from the third region 190 to the first region 170. As a result, the reaction liquid 150 is affected by the first temperature and becomes a temperature close to the first temperature.
In addition, after the case where it becomes the 1st arrangement | positioning, the operation | movement mentioned above is repeated.

  The annealing / elongation reaction is also performed in the flow path 160 other than the second region 180 when the reaction solution 150 reaches a temperature suitable for the annealing and extension reaction.

According to the embodiment described above, the following effects can be obtained.
(1) According to the substance amplifying device 3 of the present embodiment, the flow path 160A is formed by connecting the first flow path 161 and the second flow path 162 via the bent portion 140. The first region 170 is disposed at the end on the first flow path 161 side, the second region 180 is disposed at the end on the second flow path 162 side, and the third region 190 is disposed at the bent portion 140. . Then, the first heating unit 210 controls the first region 170 to the first temperature (temperature at which heat denaturation is performed). Further, the second heating unit 220 controls the second region 180 to a second temperature lower than the first temperature (a temperature at which the annealing / elongation reaction is performed). Note that the third region 190 is in contact with the atmospheric temperature. Then, the reaction solution 150 is moved between the first region 170, the second region 180, and the third region 190 by the driving mechanism.
With this configuration, the reaction solution 150 moves from the first region 170 to the second region 180 after being temporarily stopped by the third region 190 disposed in the bent portion 140. Further, the reaction liquid 150 that has moved to the second region 180 is temporarily stopped by the third region 190 and then moved to the first region 170. For this reason, the reaction liquid 150 that has been heated to a temperature close to the first temperature in the first region 170 moves to the third region 190 in contact with the atmospheric temperature lower than the second temperature and is temporarily stopped from moving. Can quickly lower the temperature. Then, the reaction liquid 150 whose temperature has been lowered moves from the third region 190 to the second region 180 and is heated at the second temperature.
Thereby, for example, the reaction solution 150 that has been heated to a temperature close to the first temperature is thermally denatured in the first region 170. In addition, the temperature of the reaction solution 150 is decreased in the third region 190 and heated at the second temperature in the second region 180 to perform an annealing / elongation reaction. The annealing / elongation reaction is also performed when moving from the second region 180 to the first region 170 through the third region 190.
Therefore, when the temperature of the reaction solution 150 is changed from the temperature at which the thermal denaturation is performed to the temperature at which the annealing / extension reaction is performed, it is possible to realize the substance amplifying device 3 that can quickly change the temperature. Moreover, the speed of the heat cycle of the substance amplifying apparatus 3 can be improved.

[Fourth Embodiment]
The substance amplifying device 4 of the present embodiment is different from the first embodiment in that a third heating unit 230 for newly heating the third region 190 is added to the heating unit 200 of the first embodiment.
FIG. 6 is a cross-sectional view showing the reaction vessel 100 and the heating unit 200 according to the present embodiment.

  In the substance amplifying device 1 of the first embodiment, the third region 190 is in contact with the atmosphere and is at room temperature or the internal temperature of the substance amplifying device 1. However, in the substance amplifying device 4 of the present embodiment, the third heating unit 230 that heats the third region 190 is disposed. Similar to the first heating unit 210 and the second heating unit 220, the third heating unit 230 includes a pair of third heat application units 231.

  A pair of 3rd heat application parts 231 hold | maintain temperature so that it may become 3rd temperature. When the reaction container 100 is attached to the attachment portion, the pair of third heat application portions 231 are disposed so as to contact the first wall 110 and the second wall 120 of the third region 190, respectively. During the heat cycle operation, the third heat application unit 231 heats the first wall 110 and the second wall 120 at the third temperature. The third temperature is set lower than the second temperature. Specifically, the third temperature is the atmospheric temperature (25 ° C.) in the present embodiment.

  In addition, since the operation | movement of the substance amplification apparatus 4 at the time of using the 3rd heating part 230 is the same as that of 1st Embodiment, description is abbreviate | omitted.

According to the above-described embodiment, the following effects can be obtained in addition to the same effects as the first embodiment.
(1) According to the substance amplification device 4 of the present embodiment, the temperature of the reaction solution 150 is thermally denatured by including the third heating unit 230 that controls the third region 190 to a third temperature lower than the second temperature. When the temperature is changed from the temperature at which the annealing is performed to the temperature at which the annealing / extension reaction is performed, the temperature change can be performed quickly and accurately.

[Fifth Embodiment]
The substance amplification device 5 of the present embodiment is different from the substance amplification device 1 of the first embodiment in that a blocking unit 300 is newly provided.

FIG. 7A is a side cross-sectional view showing the blocking unit 300 according to the present embodiment. FIG. 7B is a plan view of the blocking unit 300 as viewed from the first wall 110 side.
As shown in FIG. 7A and FIG. 7B, a pair of blocking portions 300 are arranged along the outer walls (first wall 110 and second wall 120) of the third region 190. In this embodiment, the blocking unit 300 is configured by a heat insulating member that blocks heat from the second heating unit 220 (prevents heat from the second heating unit 220 from being transmitted to the third region 190). ing. The blocking unit 300 is provided on the side facing the second heating unit 220 and both ends in the short direction, and the end of the reaction vessel 100 on the third region 190 side is opened to surround the third region 190. Is arranged in. When the reaction container 100 is attached to the attachment part, the blocking part 300 comes into contact with the first wall 110 and the second wall 120 of the reaction container 100, respectively.

  Since the operation of the substance amplifying device 5 is the same as that of the substance amplifying device 1 of the first embodiment, the description thereof is omitted.

According to the above-described embodiment, the following effects can be obtained in addition to the same effects as the first embodiment.
(1) According to the substance amplifying device 5 of the present embodiment, the cutoff device 300 is disposed along the outer wall (first wall 110, second wall 120) of the third region 190, and the second heating unit 220 By blocking the heat to the third region 190, it is possible to prevent the heat of the second heating unit 220 from affecting the third region 190. Thereby, in this embodiment, since the temperature of the 3rd field 190 can be maintained at atmospheric temperature, the temperature of reaction liquid 150 is changed from the temperature in which heat denaturation is performed to the temperature in which annealing and extension reaction are performed. In this case, the temperature can be changed quickly.

[Sixth Embodiment]
The substance amplifying device 6 of the present embodiment is different from the substance amplifying apparatus 4 of the fourth embodiment in that three pressing portions 400 are newly provided. Further, the reaction vessel 100B of the present embodiment is different from the reaction vessel 100 of the fourth embodiment. Further, the substance amplifying device 6 of this embodiment does not include a drive mechanism.

  8A to 8D are cross-sectional views illustrating the operations of the reaction vessel 100B, the heating unit 200, and the pressing unit 400 of the substance amplifying device 6 according to the present embodiment. Specifically, FIG. 8A is a diagram illustrating a state where the reaction liquid 150 has moved to the first region 170. FIG. 8B is a diagram illustrating a state in which the reaction solution 150 in the first region 170 is moved to the third region 190. FIG. 8C is a diagram illustrating a state in which the reaction solution 150 in the third region 190 is moved to the second region 180. FIG. 8D is a diagram illustrating a state in which the reaction solution 150 in the second region 180 is moved to the first region 170.

  The reaction vessel 100B of the present embodiment is composed of a first wall 110B and a second wall 120B. The reaction vessel 100B is formed in a bag shape having the function of the third wall 130 of the reaction vessel 100 described above as a first wall 110B and sealing with the second wall 120B. In addition, the reaction vessel 100B of this embodiment has flexibility. Note that the reaction vessel 100B of the present embodiment is configured in substantially the same manner as the above-described reaction vessel 100 in appearance.

  As shown in FIG. 8A, the heating unit 200 of the present embodiment includes a first heating unit 210, a second heating unit 220, and a third heating unit 230. However, in the present embodiment, unlike the heating unit 200 of the fourth embodiment, one heat application unit (the first heat application unit 211, the second heat application unit 221, and the third heat application unit 231) is used instead of a pair. Configured and in contact with the second wall 120B.

  The pressing portion 400 is disposed on the first wall 110B, and corresponds to the first region 170, corresponds to the first pressing portion 410, and corresponds to the second region 180, and corresponds to the second pressing portion 420 and the third region 190. Three pressing parts 430 are arranged.

  The substance amplifying device 6 includes a moving mechanism (not shown) that moves the pressing unit 400 to an initial position and a pressing position. Under the control of the control unit, the moving mechanism is configured so that the first pressing unit 410, the second pressing unit 420, and the third pressing unit 430 are in contact with the outer wall of the first wall 110B as an initial position, and the first wall as a pressing position. 110B is pressed to move the first inner wall 110Ba to two positions, the position where the first inner wall 110Ba is brought into close contact with the second inner wall 120Ba.

The operation when performing PCR using the substance amplifying device 6 will be described.
Initially, when the reaction container 100 is mounted on the mounting portion, all of the pressing portions 400 are located at the initial position (similar to the state shown in FIG. 8B). The moving mechanism moves the third pressing portion 430 and the second pressing portion 420 to the pressing positions in the order (similar to the state shown in FIGS. 8C and 8D), so that the reaction liquid positioned in the third region 190 is reached. 150 is moved to the position of the first area 170 as shown in FIG. 8A.

  Next, the heating unit 200 heats the first heating unit 210 to the first temperature, the second heating unit 220 to the second temperature, and the third heating unit 230 to the third temperature. Accordingly, the reaction solution 150 located in the first region 170 is heat-denatured by being heated at the first temperature.

  Next, as shown in FIG. 8B, the moving mechanism moves the second pressing portion 420 and the third pressing portion 430 from the pressing position to the initial position substantially simultaneously. Thereby, the reaction liquid 150 also moves under the influence of gravity and moves from the first area 170 to the third area 190 through the second area 180. The reaction solution 150 rapidly decreases in temperature by changing from a state heated near the first temperature to a state heated by the third temperature.

  Next, as shown in FIG. 8C, the moving mechanism moves the third pressing portion 430 from the initial position to the pressing position. As a result, the reaction solution 150 moves from the third region 190 to the second region 180 and is heated by the second temperature. In this state, when the reaction solution 150 has a temperature necessary for annealing, annealing is performed. Next, as illustrated in FIG. 8D, the moving mechanism moves the second pressing portion 420 from the initial position to the pressing position. As a result, the reaction solution 150 moves from the second region 180 to the first region 170 and is heated by the first temperature. When moving from the second region 180 to the first region 170, an annealing / elongation reaction is performed.

According to the above-described embodiment, in addition to the same effects as the fourth embodiment, the following effects can be obtained.
(1) According to the substance amplifying device 6 of the present embodiment, the first pressing unit 400 (the first pressing unit 410, the second pressing unit 420, and the third pressing unit 430) includes the pressing unit 400 and presses the reaction vessel 100B. By bringing the inner wall 110Ba into close contact with the second inner wall 120Ba, the reaction solution 150 can be moved between the first region 170, the second region 180, and the third region 190. As a result, when the temperature of the reaction solution 150 is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, the substance amplifying device 6 that can quickly change the temperature can be realized. . Further, it is not necessary for the driving mechanism to rotate the reaction vessel 100B to switch between the first arrangement and the second arrangement, and heat denaturation, annealing / extension reaction can be performed.

  Note that the present invention is not limited to the above-described embodiment, and various modifications and improvements can be added to the above-described embodiment. A modification will be described below.

  (1) In the fourth embodiment, the heating unit 200 of the first embodiment includes a new third heating unit 230 and is disposed in the third region 190. However, this configuration may be applied as the heating unit 200 of the second embodiment or the third heating unit 230 of the heating unit 200 of the third embodiment.

  (2) In the said 6th Embodiment, the press part 400 is arrange | positioned facing the heating part 200 of 4th Embodiment. However, the present invention is not limited thereto, and the third heating unit 230 may be added to the heating unit 200 of the second embodiment, and the pressing unit 400 may be disposed with the same configuration as that of the present embodiment relative to the heating unit 200. .

  (3) In the fifth embodiment, the blocking unit 300 is arranged so that the heat of the second heating unit 220 does not affect the third region 190. However, the blocking unit 300 may be arranged in the third region 190 in the second embodiment or the third embodiment. In this configuration, the heat of the first heating unit 210 and the second heating unit 220 can be prevented from affecting the third region 190. As a result, when the temperature of the reaction solution 150 is changed from the temperature at which heat denaturation is performed to the temperature at which annealing / extension reaction is performed, the temperature change can be performed more rapidly.

  (4) In the sixth embodiment, the pressing part 400 (the first pressing part 410, the first pressing part 410, the first heat applying part 211, the second heat applying part 221, the third heat applying part 231) is made to be opposed to the heat applying part (first heat applying part 211, second heat applying part 221, third heat applying part 231). 2 pressing part 420 and 3rd pressing part 430) are arranged. The pressing unit 400 does not have a function as the heating unit 200. Therefore, the pressing unit 400 may also have a function as the heating unit 200. In other words, one of the pair of the first heat application unit 211, the second heat application unit 221, and the third heat application unit 231 can be moved between the initial position and the pressing position, and functions as the pressing unit 400. It may be allowed.

  (5) In the fourth embodiment, the third heating unit 230 (third heat application unit 231) is disposed in the third region 190. The third heat application unit 231 is a member that is heated by heat generated by the heater. However, the temperature of the reaction liquid 150 may be cooled by improving the heat dissipation performance using a heat sink as the third heating unit 230. Alternatively, the temperature of the reaction liquid 150 may be cooled using a Peltier element as the third heating unit 230. Further, this configuration may be applied as the heating unit 200 of the second embodiment or the third heating unit 230 of the heating unit 200 of the third embodiment.

  (6) In the first embodiment, the example in which the material of the first heating unit 210 and the second heating unit 220 is aluminum is shown. However, the material of the heating unit 200 is thermal conductivity, heat retention, and ease of processing. It can be selected in consideration of such conditions. For example, a copper alloy may be used and a plurality of materials may be combined. The first heating unit 210 and the second heating unit 220 may be made of different materials. The same applies to the second to sixth embodiments.

  (7) In the first embodiment, the reaction vessel 100 includes the first wall 110, the second wall 120, and the third wall 130 as shown in FIGS. 1A and 1B, and the third wall 130 is thin. It has a rectangular parallelepiped shape. However, it is not limited to this shape. For example, the third wall that is the side surface in the longitudinal direction is eliminated, and the third wall that forms the surface in the vertical direction is configured with one wall that is continuous so that the longitudinal direction connects the outer circumferences of the upper and lower third walls. It is good also as a container. In this case, the side surface in the longitudinal direction is formed by one wall, but in the positional relationship described in the first embodiment, the opposing wall is the first wall, the second wall, or the opposing inner wall is the first inner wall. The second inner wall may be used.

  1-6 ... Material amplification device, 100, 100A, 100B ... Reaction vessel, 110, 110A, 100B ... First wall, 110a, 110Aa, 110Ba ... First inner wall, 120, 120A, 120B ... Second wall, 120a, 120Aa , 120Ba ... second inner wall, 130, 130A ... third wall, 140 ... bent portion, 150 ... reaction solution, 160, 160A ... flow path, 161 ... first flow path, 162 ... second flow path, 170 ... first. Area, 180 ... second area, 200 ... heating part, 210 ... first heating part, 220 ... second heating part, 230 ... third heating part, 211 ... first heat application part, 221 ... second heat application part, 231 ... third heat applying unit, 300 ... blocking unit, 400 ... pressing unit, 410 ... first pressing unit, 420 ... second pressing unit, 430 ... third pressing unit, A ... rotating shaft.

Claims (8)

A substance amplifying apparatus for amplifying a substance by subjecting a reaction solution to a heat cycle,
A mounting portion on which a reaction container having a flow path for moving the reaction solution in the longitudinal direction can be mounted;
A first region disposed at one end of the flow path of the reaction vessel;
A third region disposed at the other end in the direction opposite to the one end of the flow path;
A second region disposed at a site between the first region and the third region of the flow path;
A first heating unit that controls the first region to a first temperature;
A second heating unit for controlling the second region to a second temperature lower than the first temperature;
A drive mechanism for moving the reaction solution between the first region, the second region, and the third region;
A substance amplifying apparatus comprising: A substance amplifying apparatus for amplifying a substance by subjecting a reaction solution to a heat cycle,
A mounting portion on which a reaction container having a flow path for moving the reaction solution in the longitudinal direction can be mounted;
A first region disposed at one end of the flow path of the reaction vessel;
A second region disposed at the other end in the direction opposite to the one end of the flow path;
A third region disposed at a site between the first region and the second region of the flow path;
A first heating unit that controls the first region to a first temperature;
A second heating unit for controlling the second region to a second temperature lower than the first temperature;
A drive mechanism for moving the reaction solution between the first region, the second region, and the third region;
A substance amplifying apparatus comprising: A mounting portion capable of mounting a reaction vessel in which a reaction solution moves in a flow channel formed by connecting the first flow channel and the second flow channel via a bent portion;
A first region which is a region of an end of the flow channel on the first flow channel side;
A second region that is an end region of the flow channel on the second flow channel side;
A third region which is a region of the bent portion of the flow path;
A first heating unit that controls the first region to a first temperature;
A second heating unit for controlling the second region to a second temperature lower than the first temperature;
A drive mechanism for moving the reaction solution between the first region, the second region, and the third region;
A substance amplifying apparatus comprising: The substance amplifying device according to any one of claims 1 to 3,
A substance amplifying apparatus comprising: a third heating unit that controls the third region to a third temperature lower than the second temperature. The substance amplifying device according to any one of claims 1 to 3,
A substance amplifying apparatus, comprising: a blocking unit that is disposed along an outer wall of the third region and blocks heat from the first heating unit and / or the second heating unit. The substance amplifying device according to any one of claims 1 to 5,
The reaction vessel has opposing inner walls;
The distance between the opposing inner walls is a distance at which the reaction solution contacts both of the opposing inner walls when the reaction solution is disposed in the reaction vessel. The substance amplifying device according to any one of claims 1 to 6,
When the reaction container is mounted on the mounting portion, the driving mechanism rotates the mounting portion, the first heating portion, and the second heating portion to rotate the first arrangement and the second arrangement. Switch
The first arrangement is an arrangement in which the first area is lower than the second area in the direction of gravity.
The substance amplifying apparatus according to claim 2, wherein the second arrangement is an arrangement in which the second region is below the first region in the direction of gravity. The substance amplifying device according to any one of claims 1 to 6,
The substance amplifying apparatus according to claim 1, wherein the drive mechanism includes a pressing unit that presses the reaction container to closely contact an inner wall of the reaction container when the reaction container is mounted on the mounting unit.

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