JP2017112842A - Heat cycle device and control method thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a heat cycle device capable of suppressing convection.SOLUTION: A heat cycle device comprises: an attachment part to which a substance amplification reaction container can be attached, the substance amplification reaction container stores a reaction liquid and a liquid having a specific gravity smaller than that of the reaction liquid, and not being mixed with the reaction liquid, and forms a channel in which a first channel and a second channel are connected through a bent part, and where the reaction liquid is moved; a first heating part for heating a first area on the channel to a first temperature; a second heating part for heating a second area on the channel to a second temperature higher than the first temperature; and a third heating part for heating a third area on the channel to a third temperature equal to the second temperature or higher than the second temperature, in which the first area is an end on the first channel side of the channel, the second area is an end on the second channel side of the channel and the third area is an area on the second channel different from the second area, or an area of the bent part.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a heat cycle device and a control method for the heat cycle device.

  At present, simple test kits for rapidly diagnosing infectious diseases typified by influenza using a sample such as a nasal wipe are widely used. Most of them use an immunochromatography method, but a polymerase chain reaction (PCR) method is effective for conducting a test with higher accuracy.

  For example, Patent Document 1 describes a PCR method in which a heating and cooling cycle is performed in a tube by moving droplets in oil having two levels of temperature with a specific gravity difference. The method described in Patent Document 1 can shorten the time required for the PCR reaction as compared with a method of repeating overheating cooling of the entire heat block or a method of moving a tube to a plurality of heat blocks.

JP 2012-115208 A

  However, in the method of Patent Document 1, when a droplet (reaction liquid) is held at the end of a high-temperature container during a thermal cycle, the low-temperature oil is positioned immediately above the high-temperature oil. Therefore, in the method of Patent Document 1, convection of oil occurs, the temperature in the container fluctuates, the PCR reaction becomes unstable, and the nucleic acid amplification amount may vary.

  One of the objects according to some aspects of the present invention is to provide a thermal cycler capable of suppressing convection. Another object of some aspects of the present invention is to provide a method for controlling a thermal cycler that can suppress convection.

The thermal cycle apparatus according to the present invention is
The reaction liquid and a liquid having a specific gravity smaller than that of the reaction liquid and immiscible with the reaction liquid are filled, and the first flow path and the second flow path are connected via a bent portion, so that the reaction liquid is A mounting portion on which a substance amplification reaction vessel forming a moving flow path can be mounted;
A first heating section capable of heating the first region of the flow path to a first temperature;
A second heating part capable of heating the second region of the flow path to a second temperature higher than the first temperature;
A third heating section capable of heating the third region of the flow path to a third temperature that is the same as or higher than the second temperature;
Including
The first region is an end of the channel on the first channel side,
The second region is an end of the channel on the second channel side,
The third region is a region of the second flow path different from the second region, or a region of the bent portion.

In such a heat cycle apparatus, while the reaction solution is held in the second region, the first region that is lower in temperature than the second region is located above the second region in the direction in which gravity acts (second A third region that is higher in temperature than the second region is positioned directly above the second region, not directly above the region.
Therefore, in such a heat cycle device, convection of the liquid can be suppressed.

In the thermal cycle apparatus according to the present invention,
The third heating unit may heat the third region of the second flow path.

  In such a heat cycle device, the distance between the first heating unit and the third heating unit can be increased as compared with the case where the third heating unit heats the bent portion. Thereby, in such a heat cycle device, interference of heat between the liquid in the first region and the liquid in the third region can be suppressed.

In the thermal cycle apparatus according to the present invention,
The angle formed by the central axis of the first flow path and the central axis of the second flow path may be 90 °.

  In such a heat cycle device, the first region can be more reliably not positioned immediately above the second region while the reaction solution is held in the second region.

In the thermal cycle apparatus according to the present invention,
A drive mechanism for switching the substance amplification reaction vessel between the first arrangement and the second arrangement;
A controller that controls the drive mechanism so that the substance amplification reaction container is maintained in the second arrangement for a predetermined time after being maintained in the first arrangement;
Including
The first arrangement is an arrangement in which the second region is lower than the first region and the third region in the direction in which gravity acts,
The second arrangement may be an arrangement in which the first area is lower than the second area and the third area in a direction in which gravity acts.

  In such a heat cycle apparatus, the substance amplification reaction vessel can be automatically switched between the first arrangement and the second arrangement.

In the thermal cycle apparatus according to the present invention,
The drive mechanism switches the substance amplification reaction vessel between a third arrangement and a fourth arrangement,
The first arrangement is an arrangement in which the second region is lower than the bent part in a direction in which gravity acts,
The second arrangement is an arrangement in which the first region is lower than the bent portion in the direction in which gravity acts,
The third arrangement is an arrangement in which the bent portion is below the second region in the direction in which gravity acts,
The fourth arrangement may be an arrangement in which the bent portion is below the first region in the direction in which gravity acts.

  In such a heat cycle device, liquid convection can be suppressed.

In the thermal cycle apparatus according to the present invention,
The first heating unit is provided with a first opening into which the first region can be inserted,
The second heating unit is provided with a second opening into which the second region can be inserted,
The third heating unit may be provided with a third opening into which the third region can be inserted.

  In such a heat cycle device, liquid convection can be suppressed.

In the thermal cycle apparatus according to the present invention,
The first opening, the second opening, and the third opening may be the mounting portion.

  In such a heat cycle device, convection can be suppressed.

In the thermal cycle apparatus according to the present invention,
The reaction solution may contain a nucleic acid.

  In such a heat cycle device, convection can be suppressed.

The control method of the heat cycle apparatus according to the present invention is as follows:
A control method for a thermal cycle apparatus that performs control for switching a substance amplification reaction vessel between a first arrangement and a second arrangement,
The substance amplification reaction vessel is filled with a reaction liquid and a liquid having a specific gravity smaller than that of the reaction liquid and immiscible with the reaction liquid, and the first flow path and the second flow path are connected via a bent portion. Forming a flow path through which the reaction solution moves,
The first region of the flow path is heated at a first temperature;
The second region of the flow path is heated at a second temperature higher than the first temperature;
The third region of the flow path is heated at a third temperature that is the same as or higher than the second temperature,
The first region is an end of the channel on the first channel side,
The second region is an end of the channel on the second channel side,
The third region is a region of the second flow path different from the second region, or a region of the bent portion,
The first arrangement is an arrangement in which the second region is lower than the first region and the third region in the direction in which gravity acts,
The second arrangement is an arrangement in which the first region is lower than the first region and the third region in the direction in which gravity acts,
Control is performed so that the second arrangement is maintained for a predetermined time after the first arrangement is maintained for a predetermined time.

  In such a control method of the heat cycle apparatus, convection can be suppressed.

The figure which shows typically the thermal cycle apparatus which concerns on this embodiment. The schematic diagram for demonstrating the control method of the heat cycle apparatus which concerns on this embodiment. The schematic diagram for demonstrating the control method of the heat cycle apparatus which concerns on this embodiment. The schematic diagram for demonstrating the control method of the heat cycle apparatus which concerns on this embodiment. The flowchart for demonstrating the process of the heat cycle apparatus which concerns on this embodiment. The figure which shows typically the heat cycle apparatus which concerns on the 1st modification of this embodiment. The figure which shows typically the heat cycle apparatus which concerns on the 2nd modification of this embodiment. The figure which shows typically the heat cycle apparatus which concerns on the 2nd modification of this embodiment. The figure which shows typically the heat cycle apparatus which concerns on the 3rd modification of this embodiment.

  DESCRIPTION OF EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings. The embodiments described below do not unduly limit the contents of the present invention described in the claims. In addition, not all of the configurations described below are essential constituent requirements of the present invention.

1. Thermal cycle apparatus 1.1. Configuration First, a thermal cycle apparatus according to the present embodiment will be described with reference to the drawings. FIG. 1 is a diagram schematically showing a heat cycle apparatus 100 according to the present embodiment. In FIG. 1, the direction in which gravity acts (hereinafter also referred to as “gravity acting direction”), and the downward direction is indicated by an arrow g.

  As shown in FIG. 1, the thermal cycle apparatus 100 includes heating units 20, 22, and 24 to which the substance amplification reaction vessel 10 can be attached, a drive mechanism 30, and a control unit 40. In FIG. 1, the substance amplification reaction vessel 10 is fixed to the heating units 20, 22, and 24.

  The substance amplification reaction vessel 10 is filled with the reaction solution 4 and the liquid 6. The reaction liquid 4 is held in the liquid 6 in the form of droplets. Since the specific gravity of the reaction liquid 4 is larger than that of the liquid 6, for example, the reaction liquid 4 is located in the lowermost region in the gravity acting direction of the substance amplification reaction vessel 10. The reaction solution 4 includes, for example, DNA (target nucleic acid) amplified by PCR, an enzyme such as DNA polymerase necessary for amplifying the DNA, a primer, a fluorescent probe for detecting a nucleic acid amplification product, and the like. . The liquid 6 is a liquid that is not miscible with the reaction liquid 4, that is, does not mix. The liquid 6 has a specific gravity smaller than that of the reaction liquid 4. The liquid 6 is, for example, dimethyl silicone oil or paraffin oil.

  The material of the substance amplification reaction vessel 10 is, for example, glass, polymer, metal or the like. The substance amplification reaction vessel 10 forms a flow path 12 through which the reaction solution 4 moves. The flow channel 12 includes a first flow channel 14, a second flow channel 16, and a bent portion 18.

  The first flow path 14 and the second flow path 16 are connected via a bent portion 18. That is, the channel 12 has a bent portion. The flow paths 14 and 16 extend linearly. In the illustrated example, the angle θ formed by the central axis α of the first flow path 14 and the central axis β of the second flow path 16 is 90 °. The substance amplification reaction vessel 10 has a lid (not shown) for injecting the reaction solution 4 and the liquid 6, and the lid is provided at a portion where the bent portion 18 of the substance amplification reaction vessel 10 is formed. May be.

  The first flow path 14 has a first region 12a. The first region 12a is an end of the flow channel 12 on the first flow channel 14 side. That is, the first region 12a is an end portion of the first flow path 14 on the side opposite to the bent portion 18 side. The second flow path 16 has a second region 12b and a third region 12c. The second region 12b is an end of the flow channel 12 on the second flow channel 16 side. That is, the second region 12b is an end portion of the second flow path 16 on the side opposite to the bent portion 18 side. The third region 12c is a region of the second flow path 16 that is different from the second region 12b. The third region 12c is provided closer to the bent portion 18 than the second region 12b.

  The first heating unit 20, the second heating unit 22, and the third heating unit 24 are members that transmit heat generated from a heater (not shown) to the substance amplification reaction vessel 10. The heating units 20, 22, and 24 are in contact with the substance amplification reaction vessel 10, for example. The heating units 20, 22, and 24 are, for example, aluminum heat blocks. By making the heat block made of aluminum, the substance amplification reaction vessel 10 can be efficiently heated. Moreover, the uniformity of temperature can be improved in the heat block, and a highly accurate thermal cycle can be realized. Moreover, since processing is easy, a heat block can be shape | molded with high precision.

The first heating unit 20 is provided with a first opening 21 into which the first region 12a can be inserted. In the illustrated example, the first opening 21 is a bottomed hole. The first heating unit 20 includes the first region 12a.
The liquid 6 can be heated to the first temperature. The first temperature is, for example, 55 ° C. or more and 72 ° C. or less, and preferably 60 ° C. The first temperature is a temperature suitable for annealing (reaction that binds to the single-stranded DNA of the primer) and extension reaction (reaction in which a complementary strand of DNA is formed starting from the primer) in the PCR reaction. Compared with the case where the 1st opening part 21 is a through-hole, the 1st heating part 20 heats the liquid 6 of the 1st area | region 12a more efficiently than the case where the 1st opening part 21 is a through-hole. Can do.

  The second heating unit 22 is provided with a second opening 23 into which the second region 12b can be inserted. In the illustrated example, the second opening 23 is a hole with a bottom. The second heating unit 22 can heat the liquid 6 in the second region 12b to the second temperature. The second temperature is higher than the first temperature, for example, 92 ° C. or more and 97 ° C. or less, and preferably 95 ° C. The second temperature is a temperature suitable for denaturation (dissociation of double-stranded DNA) in the PCR reaction. Compared to the case where the second opening 23 is a through-hole, the second heating unit 22 efficiently heats the liquid 6 in the second region 12b because the second opening 23 is a hole having a height. Can do.

  The third heating unit 24 is provided with a third opening 25 into which the third region 12c can be inserted. In the illustrated example, the third opening 25 is a through hole. The third heating unit 24 can heat the liquid 6 in the third region 12c to the third temperature. The third temperature is the same as or higher than the second temperature (the third temperature is equal to or higher than the second temperature), and is, for example, 92 ° C. or higher and 110 ° C. or lower, preferably 100 ° C. The third temperature is lower than the temperature at which the enzyme contained in the reaction solution 4 is deactivated.

  The temperatures of the heating units 20, 22, and 24 may be controlled by a temperature sensor and a control unit 40 (not shown). Openings 21, 23, and 25 provided in the heating units 20, 22, and 24 are mounting units to which the substance amplification reaction vessel 10 can be mounted.

  The drive mechanism 30 is a mechanism that switches the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 to the first arrangement, the second arrangement, the third arrangement, and the fourth arrangement. Although not shown, the drive mechanism 30 includes a support unit that supports the substance amplification reaction vessel 10 and the heating units 20, 22, and 24, and a motor that rotates the support unit. The rotation axis Q of the support portion is, for example, a direction orthogonal to the central axes α and β. In the illustrated example, the position of the rotation axis Q is centered on the imaginary straight line L connecting the end of the flow path 12 on the first flow path 14 side and the end of the flow path 12 on the first flow path 14 side. It is the position of the leg of the perpendicular drawn from the intersection P. Thereby, size reduction of the drive mechanism 30 can be achieved, and size reduction of the heat cycle apparatus 100 can be achieved. The drive mechanism 30 rotates the support portion around the rotation axis Q while maintaining the positional relationship between the substance amplification reaction vessel 10 and the heating portions 20, 22, and 24 by the operation of the motor.

  The control unit 40 controls the drive mechanism 30 so that the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 are maintained in the first arrangement for a predetermined time and then maintained in the second arrangement for a predetermined time. The control unit 40 includes, for example, a CPU (Central Processing Unit) and a storage unit (ROM (Read Only Memory), RAM (Random Access Memory)). For example, various programs and data for controlling the drive mechanism 30 are recorded in the storage unit. The storage unit temporarily records, for example, data during processing of various processes performed by the control unit 40, processing results, and the like.

1.2. Control Method FIGS. 2 to 4 are schematic diagrams for explaining a control method of the heat cycle apparatus 100. FIG. 5 is a flowchart for explaining the processing of the heat cycle apparatus 100. For convenience, illustration of the drive mechanism 30 and the control unit 40 is omitted in FIGS. Moreover, in FIGS. 2-4, it is a gravity action direction, Comprising: The downward direction is shown by the arrow g.

  First, the substance amplification reaction vessel 10 filled with the reaction solution 4 and the liquid 6 is fixed to the heating units 20, 22, and 24 (step S101). For example, the substance amplification reaction vessel 10 is made of a flexible resin, and the substance amplification reaction vessel 10 is inserted into the openings 21, 23, 25 while being bent, and fixed to the heating units 20, 22, 24.

  In step S101, the arrangement of the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 is the first arrangement (see FIG. 1). As shown in FIG. 1, the first arrangement is an arrangement in which the second region 12 b is below the first region 12 a, the third region 12 c, and the bent portion 18 in the direction of gravity action. In the first arrangement, for example, the second region 12b is located at the lowest part of the flow path 12 in the direction of gravity action. Therefore, the reaction liquid 4 having a larger specific gravity than the liquid 6 is located in the second region 12b. In the illustrated example, the second region 12 a is located directly below the bent portion 18.

  Next, the substance amplification reaction vessel 10 is heated by the heating units 20, 22, and 24 (step S102). Specifically, the first heating unit 20 heats the liquid 6 in the first region 12a to the first temperature. The second heating unit 22 heats the liquid 6 in the second region 12b to the second temperature. The third heating unit 24 heats the liquid 6 in the third region 12c to the third temperature.

  In step S102, since the arrangement of the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 is the first arrangement, the reaction solution 4 is heated to the second temperature. Therefore, in step S102, the reaction (denaturation) is performed on the reaction solution 4 at the second temperature.

  Next, the control unit 40 determines whether or not the first time has elapsed in the first arrangement (step S103). The first time is the time for maintaining the first arrangement, and is the time required for denaturation. Specifically, the control unit 40 includes a timer, and determines whether or not the preset time has elapsed. In this embodiment, when step S103 is performed following step S102, that is, when the first step S103 is performed, the control unit 40 is a temperature sensor (a temperature sensor connected to the second heating unit 22). It is determined whether or not the time elapsed since the temperature reached the predetermined temperature is the first time.

  When it is determined in step S103 that the first time has elapsed (in the case of Yes), the process proceeds to step S104. If it is determined in step S103 that the first time has not elapsed (in the case of No), step S103 is repeated.

  In the case of Yes in step S103, the control unit 40 controls the drive mechanism 30 to switch the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 from the first arrangement to the second arrangement (step S104). Thereby, the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 are changed to the state (second arrangement) shown in FIG. 3 through the state (third arrangement) shown in FIG. Specifically, from the state shown in FIG. 1, it is rotated counterclockwise by 180 ° around the rotation axis Q to be in the state shown in FIG. 2, and further from the state shown in FIG. 2 around the rotation axis Q. It is rotated by 90 ° counterclockwise to reach the state shown in FIG.

  The state shown in FIG. 2 (third arrangement) is an arrangement in which the bent portion 18 is below the second region 12b and the third region 12c in the direction of gravity action. Therefore, the reaction solution 4 reaches the bent portion 18 from the second region 12b through the third region 12c. In the illustrated example, the bent portion 18 is located immediately below the second region 12b.

The state (second arrangement) shown in FIG. 3 is an arrangement in which the first region 12a is below the second region 12b, the third region 12c, and the bent portion 18 in the direction of gravity action. In the second arrangement, for example, the second region 12b is located at the lowest part of the flow path 12 in the direction of gravity action. Therefore, the reaction solution 4 reaches the first region 12a from the bent portion 18. In the illustrated example, the first region 12 a is located directly below the bent portion 18.

  Next, the control unit 40 determines whether or not the second time has elapsed in the second arrangement (step S105). The second time is the time for maintaining the second configuration, and is the time required for the annealing and extension reaction. The control unit 40 determines whether or not the time after reaching the second arrangement has reached the second time. Specifically, the control unit 40 includes a timer, and determines whether or not the preset time has elapsed. In step S105, since the arrangement of the substance amplification reaction vessel 10 and the heating units 20, 22, 24 is the second arrangement, the reaction solution 4 is heated to the first temperature. Therefore, in step S105, the reaction at the first temperature (annealing and extension reaction) is performed on the reaction solution 4.

  When it is determined in step S105 that the second time has elapsed (in the case of Yes), the process proceeds to step S106. If it is determined in step S105 that the second time has not elapsed (in the case of No), step S105 is repeated.

  In the case of Yes in step S105, the control unit 40 determines whether or not the number of thermal cycles has reached a predetermined number of cycles (step S106). Specifically, the control unit 40 determines whether or not the procedure from step S103 to step S105 has been completed a predetermined number of times. In the present embodiment, the number of times the procedure from step S103 to step S105 is completed is determined by the number of times determined as “Yes” in steps S103 and S105. When Steps S103 to S105 are performed once (when “Yes” is determined once in Steps S103 and S105), the reaction solution 4 is subjected to one thermal cycle, so Steps S103 to S105 are performed. The number of times until the above procedure is completed (the number of thermal cycles) is one cycle.

  In step S106, when the control unit 40 determines that the predetermined number of thermal cycles has been performed (Yes), the control unit 40 completes the processing (END). When the control unit 40 determines that the predetermined number of thermal cycles have not been performed (No), the process proceeds to step S107.

  In the case of No in step S106, the control unit 40 controls the drive mechanism 30 to switch the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 from the second arrangement to the first arrangement (step S107). Thereby, the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 are changed to the state (first arrangement) shown in FIG. 1 through the state (fourth arrangement) shown in FIG. Specifically, the state shown in FIG. 3 is rotated counterclockwise by 180 ° around the rotation axis Q to be in the state shown in FIG. 4, and further, the state shown in FIG. It is rotated 90 ° clockwise to the state shown in FIG.

  The state shown in FIG. 4 (fourth arrangement) is an arrangement in which the bent portion 18 is below the first region 12a in the direction of gravity action. Accordingly, the reaction solution 4 reaches the bent portion 18 from the first region 12a. In the illustrated example, the bent portion 18 is located immediately below the first region 12a.

  The state (first arrangement) shown in FIG. 1 is an arrangement in which the second region 12b is below the first region 12a, the third region 12c, and the bent portion 18 in the direction of gravity action. Accordingly, the reaction solution 4 reaches the second region 12b from the bent portion 18. When the substance amplification reaction vessel 10 and the heating units 20, 22, and 24 reach the first arrangement, step S103 is started.

When step S103 is performed subsequent to step S107, that is, in the second and subsequent steps S103, the control unit 40 includes the substance amplification reaction vessel 10 and the heating unit 20,
It is determined whether or not the time since the arrival of the first arrangement 22 and 24 has reached the first time.

The first temperature, the second temperature, the third temperature, the first time, the second time, and the number of cycles are the types and amounts of the reaction solution 4 and the liquid 6, the size of the substance amplification reaction vessel, etc. Is determined as appropriate,
The thermal cycle apparatus 100 has the following characteristics, for example.

  The thermal cycle apparatus 100 includes a mounting portion to which the substance amplification reaction vessel 10 in which the first flow path 14 and the second flow path 16 are connected via the bent portion 18 to form the flow path 12 can be mounted. Further, in the heat cycle apparatus 100, the first heating unit 20 capable of heating the first region 12a at the first temperature and the second heating unit 22 capable of heating the second region 12b at the second temperature higher than the first temperature. And a third heating unit 24 capable of heating the third region 12c at a third temperature higher than the second temperature. Therefore, in the heat cycle apparatus 100, as shown in FIG. 1, while the reaction solution 4 is held in the second region 12b, the first region 12a, which is lower in temperature than the second region 12b, is converted into the second region 12b. On the other hand, the third region 12c that is higher in temperature than the second region 12b is positioned directly above the second region 12b, not positioned above the second region 12b in the direction of gravity action. Therefore, in the heat cycle apparatus 100, the convection of the liquid 6 can be suppressed, and the temperature gradient of the liquid 6 can be prevented from collapsing due to the convection. As a result, in the thermal cycle apparatus 100, the temperature in the substance amplification reaction vessel 10 can be stabilized and the PCR reaction can be performed stably, so that the uniformity of the nucleic acid amplification amount can be improved.

  In the heat cycle apparatus 100, the third heating unit 24 heats the third region 12 c of the second flow path 16. Therefore, in the heat cycle apparatus 100, the distance between the first heating unit 20 and the third heating unit 24 can be increased as compared with the case where the third heating unit 24 heats the bent portion 18. Thereby, in the heat cycle apparatus 100, the heat interference between the liquid 6 in the first region 12a and the liquid 6 in the third region 12c can be suppressed.

  Furthermore, when the 3rd heating part 24 heats the bending part 18, the 3rd heating part 24 consists of a heat block divided | segmented into 2 parts, for example, the bending part 18 is inserted | pinched with this heat block, and the substance amplification reaction container 10 is included. Is fixed. On the other hand, when the third heating unit 24 heats the third region 12c of the second flow path 16, the third heating unit 24 is a heat block provided with a through hole (opening 25), and the through hole The substance amplification reaction vessel 10 can be fixed by inserting the second flow channel 16 into the tube. Therefore, when the 3rd heating part 24 heats the 3rd field 12c of the 2nd channel 16, the 3rd heating part 24 can be manufactured easily, and material amplification reaction container 10 is fixed easily. be able to.

  In the heat cycle apparatus 100, the angle θ formed by the central axis α of the first flow path 14 and the central axis β of the second flow path 16 is 90 °. Therefore, in the heat cycle apparatus 100, the first region 12a can be more reliably not positioned immediately above the second region 12b while the reaction solution 4 is held in the second region 12b.

  In the thermal cycle apparatus 100, the drive mechanism 30 that switches the substance amplification reaction vessel 10 to the first arrangement and the second arrangement, and the substance amplification reaction container 10 is maintained in the first arrangement for a predetermined time, and then in the second arrangement for a predetermined time. And a control unit 40 that controls the drive mechanism 30 so as to be maintained. Therefore, in the thermal cycle apparatus 100, the substance amplification reaction vessel 10 can be automatically switched between the first arrangement and the second arrangement.

2. Modification of heat cycle apparatus 2.1. First Modification Next, a thermal cycler according to a first modification of the present embodiment will be described with reference to the drawings. FIG. 6 is a diagram schematically showing a heat cycle apparatus 200 according to a first modification of the present embodiment. In addition, in FIG. 6, it is a gravity action direction, Comprising: The downward direction is shown by the arrow g.

  Hereinafter, in the thermal cycle apparatus 200 according to the first modification of the present embodiment, members having the same functions as those of the structural members of the thermal cycle apparatus 100 according to the present embodiment are denoted by the same reference numerals, and detailed description thereof is given. Is omitted. The same applies to the thermal cyclers according to the second and third modifications of the present embodiment described below.

  In the thermal cycle apparatus 100 described above, as shown in FIG. 1, the third heating unit 24 can heat the third region 12 c of the second flow path 16. On the other hand, in the heat cycle apparatus 200, as shown in FIG. 6, the 3rd heating part 24 can heat the 3rd area | region 12c which is the area | region of the bending part 18. As shown in FIG.

  In the heat cycle apparatus 200, the convection of the liquid 6 can be suppressed as in the heat cycle apparatus 100.

2.2. Second Modification Example Next, a thermal cycler according to a second modification example of the present embodiment will be described with reference to the drawings. FIG. 7 is a diagram schematically showing a heat cycle apparatus 300 according to the second modification of the present embodiment. In FIG. 7, the gravity direction is indicated by the arrow g in the downward direction.

  In the heat cycle apparatus 100 described above, as shown in FIG. 1, the angle θ formed by the central axis α of the first flow path 14 and the central axis β of the second flow path 16 was 90 °. On the other hand, in the heat cycle apparatus 300, as shown in FIG. 7, the angle θ is not 90 °. The angle θ is, for example, not less than 60 ° and not more than 120 °. In the heat cycle apparatus 300, as shown in FIG. 8, the third heating unit 24 may be able to heat the third region 12 c that is the region of the bent portion 18.

  In the heat cycle apparatus 300, the convection of the liquid 6 can be suppressed as in the heat cycle apparatus 100.

2.3. Third Modification Next, a thermal cycler according to a third modification of the present embodiment will be described with reference to the drawings. FIG. 9 is a diagram schematically showing a heat cycle apparatus 400 according to a third modification of the present embodiment. In FIG. 9, the gravity direction is indicated by the arrow g in the downward direction.

  As shown in FIG. 9, the thermal cycle apparatus 400 is different from the thermal cycle apparatus 100 described above in that it includes a fluorescence detection unit 50.

  The fluorescence detection unit 50 irradiates the reaction solution 4 with light under the control of the control unit 40, for example, and detects fluorescence emitted from the reaction solution 4 (fluorescence of the reaction solution 4). The fluorescence detection unit 50 is provided at a position where the first region 12a can be irradiated with light. In the illustrated example, the first opening 21 provided in the first heating unit 20 is a through hole, and the fluorescence detection unit 50 reacts in the first region 12a through the first opening 21 in the third arrangement. The liquid 4 is irradiated with light. The fluorescence detection unit 50 may continue to emit light while performing PCR. The fluorescence detection unit 50 is fixed without being rotated by the drive mechanism 30.

The detection result of the fluorescence detection unit 50 may be stored in the storage unit of the control unit 40. The control unit 40 can acquire, for example, a PCR nucleic acid amplification curve based on the fluorescence brightness (fluorescence brightness) detected by the fluorescence detection unit 50 and determine the number of PCR temperature cycles. For example, in step S106 (see FIG. 5), the control unit 40 determines whether or not the detected fluorescence brightness is equal to or greater than a predetermined value stored in the storage unit in advance. When the detected fluorescence brightness is equal to or higher than a predetermined value, the control unit 40 determines that the predetermined number of cycles has been reached. On the other hand, when the detected fluorescence luminance is less than the predetermined value, the control unit 40 determines that the predetermined number of cycles has not been reached.

  The thermal cycle apparatus 400 includes a fluorescence detection unit 50. Therefore, the thermal cycle apparatus 400 can determine the number of PCR temperature cycles based on the fluorescence brightness detected by the fluorescence detection unit 50.

  In the heat cycle apparatus 400, the fluorescence detection unit 50 irradiates light to the reaction solution 4 in the first region 12a. Therefore, in the heat cycle apparatus 400, the fluorescence measurement can be accurately performed as compared with the case where the second region 12b, which is higher in temperature than the first region 12a, is irradiated with light. For example, when the fluorescence measurement is performed using an intercalator type dye, the fluorescence measurement cannot be performed in a high temperature region such as the second region 12b.

  Although not shown, the thermal cycle devices 200 and 300 described above may include the fluorescence detection unit 50 as in the thermal cycle device 400.

  In the present invention, a part of the configuration may be omitted within a range having the characteristics and effects described in the present application, or each embodiment or modification may be combined.

  The present invention includes configurations that are substantially the same as the configurations described in the embodiments (for example, configurations that have the same functions, methods, and results, or configurations that have the same objects and effects). In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that exhibits the same operational effects as the configuration described in the embodiment or a configuration that can achieve the same object. Further, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

4 ... reaction liquid, 6 ... liquid, 10 ... substance amplification reaction vessel, 12 ... channel, 12a ... first region, 12b ... second region, 12c ... third region, 14 ... first channel, 16 ... second Flow path, 18 ... bent portion, 20 ... first heating portion, 21 ... first opening, 22 ... second heating portion, 23 ... second opening, 24 ... third heating portion, 25 ... third opening, DESCRIPTION OF SYMBOLS 30 ... Drive mechanism, 40 ... Control part, 50 ... Fluorescence detection part, 100, 200, 300, 400 ... Thermal cycle apparatus

Claims (9)

The reaction liquid and a liquid having a specific gravity smaller than that of the reaction liquid and immiscible with the reaction liquid are filled, and the first flow path and the second flow path are connected via a bent portion, so that the reaction liquid is A mounting portion on which a substance amplification reaction vessel forming a moving flow path can be mounted;
A first heating section capable of heating the first region of the flow path to a first temperature;
A second heating part capable of heating the second region of the flow path to a second temperature higher than the first temperature;
A third heating section capable of heating the third region of the flow path to a third temperature that is the same as or higher than the second temperature;
Including
The first region is an end of the channel on the first channel side,
The second region is an end of the channel on the second channel side,
The thermal cycle apparatus, wherein the third region is a region of the second flow path different from the second region, or a region of the bent portion. In claim 1,
The third heating unit is a thermal cycle device that heats the third region of the second flow path. In claim 1 or 2,
The thermal cycle device, wherein an angle formed by the central axis of the first flow path and the central axis of the second flow path is 90 °. In any one of Claims 1 thru | or 3,
A drive mechanism for switching the substance amplification reaction vessel between the first arrangement and the second arrangement;
A controller that controls the drive mechanism so that the substance amplification reaction container is maintained in the second arrangement for a predetermined time after being maintained in the first arrangement;
Including
The first arrangement is an arrangement in which the second region is lower than the first region and the third region in the direction in which gravity acts,
The second arrangement is a thermal cycle apparatus in which the first area is arranged below in the direction in which gravity acts than the second area and the third area. In claim 4,
The drive mechanism switches the substance amplification reaction vessel between a third arrangement and a fourth arrangement,
The first arrangement is an arrangement in which the second region is lower than the bent part in a direction in which gravity acts,
The second arrangement is an arrangement in which the first region is lower than the bent portion in the direction in which gravity acts,
The third arrangement is an arrangement in which the bent portion is below the second region in the direction in which gravity acts,
Said 4th arrangement | positioning is a thermal cycle apparatus which is an arrangement | positioning in which the said bending part becomes lower than the said 1st area | region in the direction where gravity acts. In any one of Claims 1 thru | or 5,
The first heating unit is provided with a first opening into which the first region can be inserted,
The second heating unit is provided with a second opening into which the second region can be inserted,
The heat cycle apparatus, wherein the third heating unit is provided with a third opening into which the third region can be inserted. In claim 6,
The thermal opening device, wherein the first opening, the second opening, and the third opening are the mounting portions. In any one of Claims 1 thru | or 7,
The thermal reaction apparatus, wherein the reaction solution contains a nucleic acid. A control method for a thermal cycle apparatus that performs control for switching a substance amplification reaction vessel between a first arrangement and a second arrangement,
The substance amplification reaction vessel is filled with a reaction liquid and a liquid having a specific gravity smaller than that of the reaction liquid and immiscible with the reaction liquid, and the first flow path and the second flow path are connected via a bent portion. Forming a flow path through which the reaction solution moves,
The first region of the flow path is heated at a first temperature;
The second region of the flow path is heated at a second temperature higher than the first temperature;
The third region of the flow path is heated at a third temperature that is the same as or higher than the second temperature,
The first region is an end of the channel on the first channel side,
The second region is an end of the channel on the second channel side,
The third region is a region of the second flow path different from the second region, or a region of the bent portion,
The first arrangement is an arrangement in which the second region is lower than the first region and the third region in the direction in which gravity acts,
The second arrangement is an arrangement in which the first region is lower than the first region and the third region in the direction in which gravity acts,
A control method for a heat cycle apparatus, wherein control is performed so that the second arrangement is maintained for a predetermined time after being maintained for a predetermined time in the first arrangement.

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