JP2018196365A - Nucleic acid amplification equipment - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a nucleic acid amplification device capable of easily fixing a microchannel device in close contact with a heating portion and easily aligning and attaching / detaching the microchannel device to the heating portion. .. SOLUTION: A movable mounting portion 10 on which a microchannel chip 100 having a microchannel 130 is mounted, a heating portion 20 for heating the microchannel chip 100, and a microchannel chip 100 are heating portions. A fixing portion 30 for fixing the microchannel tip 100 mounted on the mounting portion 10 is provided at a position in contact with the 20, and when the microchannel chip 100 is in contact with the heating portion 20, the fixing portion is provided. The 30 and the mounting portion 10 are arranged in this order along the direction from the microchannel chip 100 toward the heating portion 20. [Selection diagram] Fig. 2

Description

  The present invention relates to a nucleic acid amplification apparatus for amplifying nucleic acids (deoxyribonucleic acid, ribonucleic acid) contained in a specimen.

  In the inspection of microorganisms such as bacteria contained in beverages or foods, a PCR inspection method using a polymerase chain reaction (PCR) is known. The PCR inspection method has an advantage that the inspection process can be greatly speeded up and simplified as compared with the culture method.

  The PCR test is performed using, for example, a nucleic acid amplification device. As this type of nucleic acid amplification device, a microchannel chip having a microchannel is known. When performing a PCR test using a microchannel chip, a reaction solution containing a specimen (specimen stock solution) containing a microorganism to be tested and a reaction reagent (PCR reagent, etc.) is allowed to flow through the microchannel of the microchannel chip. , Subject the reaction solution to an elevated temperature cycle. As a result, the target nucleic acid contained in the specimen can be amplified exponentially at high speed, so that the specimen can be examined quickly.

  An example of a microchannel chip used for PCR inspection is disclosed in Patent Document 1. Patent Document 1 discloses a microchannel chip disposed on a heating unit that forms different temperature regions. The microchannel chip has a microchannel formed by meandering so as to pass through different temperature regions of the heating unit. Thereby, the raising / lowering temperature cycle can be given to a reaction solution only by sending a reaction solution to a microchannel.

International Publication No. 2015/019522

  In order to efficiently perform PCR by heating the microchannel chip with the heating unit, it is necessary to accurately form a desired temperature region in the microchannel of the microchannel chip.

  In the configuration disclosed in Patent Document 1, a desired temperature region is formed in the microchannel by placing the microchannel chip on the heating unit, and nucleic acid amplification is performed.

  Here, in order to efficiently perform nucleic acid amplification, it is required to form a desired temperature region in the microchannel with high accuracy by closely contacting the microchannel chip and the heating unit.

  In addition, in the method of manually placing the microchannel chip on the heating unit, it is difficult to accurately align the microchannel and the heating unit, and the position of the microchannel and the heating unit is accurate. Skillful operation is necessary to match well.

  In addition, the microchannel chip for which the PCR test has been completed is removed from the heating unit, and the next microchannel chip is set in the heating unit. Therefore, it is desirable that the microchannel chip can be easily attached to and detached from the heating unit.

  The present invention has been made to solve such a problem, and can easily fix the microchannel device in close contact with the heating unit, and align the microchannel device with respect to the heating unit. An object of the present invention is to provide a nucleic acid amplification device that can be easily attached and detached.

  In order to achieve the above object, one embodiment of the nucleic acid amplification device according to the present invention heats a movable mounting portion on which a microchannel chip having a microchannel is mounted, and the microchannel chip A heating unit; and a fixing unit that fixes the microchannel chip placed on the mounting unit at a position where the microchannel chip is in contact with the heating unit. When contacting the part, the fixing part and the mounting part are arranged in this order along the direction from the microchannel chip toward the heating part.

  According to the present invention, the microchannel device can be easily fixed in close contact with the heating unit, and the microchannel device can be easily aligned and detached from the heating unit.

It is a figure which shows an example of the microchannel chip | tip set to the nucleic acid amplifier which concerns on embodiment. It is sectional drawing of the nucleic acid amplifier which concerns on embodiment. It is a top view of the nucleic acid amplification device according to the embodiment. It is a figure for demonstrating the nucleic acid amplification method which concerns on embodiment. It is sectional drawing of the nucleic acid amplifier which concerns on the modification 1 of embodiment. It is a top view of the nucleic acid amplification device according to the first modification of the embodiment. It is sectional drawing of the nucleic acid amplifier which concerns on the modification 2 of embodiment.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. Note that each of the embodiments described below shows a preferred specific example of the present invention. Therefore, numerical values, shapes, materials, components, arrangement positions and connection forms of components, steps (steps) and order of steps, and the like shown in the following embodiments are examples and limit the present invention. It is not the purpose to do. Therefore, among the constituent elements in the following embodiments, constituent elements that are not described in the independent claims showing the highest concept of the present invention are described as optional constituent elements.

  Each figure is a schematic diagram and is not necessarily shown strictly. Accordingly, the scales and the like are not necessarily the same in each drawing, and the devices and components shown in each drawing may be omitted. In each figure, substantially the same configuration is denoted by the same reference numeral, and redundant description is omitted or simplified.

(Embodiment)
First, the configuration of the microchannel chip 100 set in the nucleic acid amplification device 1 will be described with reference to FIG. FIG. 1 is a diagram illustrating an example of a microchannel chip 100 set in the nucleic acid amplification device 1 according to the embodiment.

  As shown in FIG. 1, the microchannel chip 100 is a microfluidic chip through which a reaction solution that is a microfluid flows, and includes a first opening 110, a second opening 120, and a microchannel 130. The first opening 110 and the second opening 120 are connected by a microchannel 130.

  In the present embodiment, the microchannel chip 100 is a nucleic acid amplification device for amplifying a target nucleic acid of a sample contained in a reaction solution introduced into the microchannel chip 100.

  The reaction solution introduced into the microchannel chip 100 is, for example, an aqueous solution containing a specimen containing the target nucleic acid and a reaction reagent that reacts with the target nucleic acid. The sample is, for example, a sample stock solution containing a target nucleic acid previously extracted from a microorganism to be examined (bacteria, virus, tissue cell, or the like) with a nucleic acid extraction reagent. The reaction reagent is, for example, a reaction reagent solution such as a PCR reagent containing a fluorescent substance.

  The microchannel chip 100 includes a first substrate 101 that is a lower substrate and a second substrate 102 that is an upper substrate. As the first substrate 101 and the second substrate 102, for example, a resin substrate such as PET (polyethylene terephthalate), PC (polycarbonate), acrylic, a glass substrate, a silicon substrate, or the like can be used.

  The first opening 110 is provided at one end of the microchannel 130 and is the starting point of the microchannel 130. The first opening 110 is an inlet (inlet) of the microchannel 130, and is an inlet through which a reaction solution is introduced. That is, the reaction solution is introduced into the microchannel 130 via the first opening 110. Although the 1st opening part 110 is comprised by the recessed part provided in the 1st board | substrate 101, and the circular through-hole provided in the 2nd board | substrate 102, for example, it is not restricted to this.

  The second opening 120 is provided at the other end of the microchannel 130 and is the end point of the microchannel 130. That is, the 2nd opening part 120 becomes an end point of liquid feeding of a reaction solution. The second opening 120 is an outlet (outlet) of the microchannel 130. Similarly to the first opening 110, the second opening 120 includes, for example, a recess provided in the first substrate 101 and a circular through hole provided in the second substrate 102, but is not limited thereto. Absent. The second opening 120 is a discharge port that can discharge a part or all of the reaction solution flowing through the microchannel 130, but the reaction solution may not be discharged from the second opening 120. .

  The micro flow path 130 is a flow path for sending the reaction solution introduced from the first opening 110 to the second opening 120. The microchannel 130 is a microchannel whose channel width and channel depth are micro-order sizes. In the present embodiment, one micro flow path 130 is configured, and the reaction solution flows one-way through the micro flow path 130. Specifically, the reaction solution flows through the microchannel 130 in the direction from the first opening 110 to the second opening 120. In the present embodiment, the reaction solution is fed through the microchannel 130 by capillary force (capillary force). For example, the reaction solution can be fed by capillary force by coating the inner surface of the microchannel 130 with a surfactant or the like to make the surface hydrophilic.

  The microchannel 130 is a groove formed in the first substrate 101, for example. Note that the microchannel 130 may be formed on the second substrate 102. For example, the grooves constituting the microchannel 130 have a rectangular cross-sectional shape, and the channel width (groove width) and depth are constant throughout the microchannel 130. As an example, the grooves constituting the microchannel 130 have a channel width of 20 to 300 μm and a depth of 50 to 150 μm.

  The microchannel chip 100 includes a nucleic acid amplification unit 100a (nucleic acid amplification region) for amplifying a target nucleic acid contained in a reaction solution flowing through the microchannel 130. The micro flow path 130 in the nucleic acid amplification unit 100a is configured to pass through a temperature region in which the temperature is controlled by the heating unit 20 of the nucleic acid amplification device 1. Therefore, the reaction solution introduced into the microchannel chip 100 flows through the microchannel 130 and is heated by the heating unit 20 in the nucleic acid amplification unit 100a, whereby the target nucleic acid contained in the reaction solution is amplified.

  In the present embodiment, the microchannel 130 in the nucleic acid amplification unit 100a is a serpentine channel formed to meander, but is not limited thereto. Further, the meandering number of the meandering channel is about 20 to 70 cycles, for example, but only about 10 cycles are shown in FIG.

  Next, the configuration of the nucleic acid amplification device 1 according to the embodiment will be described with reference to FIGS. FIG. 2 is a cross-sectional view of the nucleic acid amplification device 1 according to the embodiment. FIG. 3 is a top view of the nucleic acid amplification device 1. 2 is a cross-sectional view taken along line II-II in FIG.

  The nucleic acid amplification device 1 is a device for heating and amplifying a target nucleic acid of a sample contained in a reaction solution introduced into the microchannel chip 100. As shown in FIG. 2 and FIG. The heating unit 20, the fixing unit 30, and the main body unit 40 are provided. The main body 40 is a base serving as a base of the nucleic acid amplification device 1 and is made of, for example, a metal material or a resin material.

  The mounting unit 10 is a mounting table on which the microchannel chip 100 is mounted. In the present embodiment, one microchannel chip 100 is placed on the placement unit 10. The placement unit 10 is, for example, a cassette, and the microchannel chip 100 placed on the placement unit 10 is held by the placement unit 10 so as not to fall.

  In this Embodiment, the mounting part 10 is comprised by the pair of 1st mounting bar 11 and 2nd mounting bar 12 which were isolate | separated. A pair of 1st mounting bar 11 and 2nd mounting bar are arrange | positioned at predetermined intervals in the horizontal direction. Therefore, the mounting part 10 has the opening part 10a comprised by the space space between the 1st mounting bar 11 and the 2nd mounting bar 12. FIG. Thereby, when the microchannel chip 100 is mounted on the mounting unit 10, the back surface of the microchannel chip 100 is exposed from the opening 10a.

  The first placement bar 11 includes a first support portion 11 a that supports the back surface of one end portion in the width direction of the microchannel chip 100, and a first surface that opposes one side surface in the width direction of the microchannel chip 100. Wall part 11b. Similarly, the second placement bar 12 faces the second support portion 12a that supports the back surface of the other end portion in the width direction of the microchannel chip 100 and the other side surface in the width direction of the microchannel chip 100. And a second wall portion 12b. Each of the first placement bar 11 and the second placement bar is, for example, a rod-shaped member having an L-shaped cross section. In addition, each of the 1st mounting bar 11 and the 2nd mounting bar 12 is comprised by the metal material or the resin material, for example. With the mounting unit 10 configured as described above, the micro-channel chip 100 mounted on the mounting unit 10 is restricted from vertical movement and horizontal movement. Thereby, the microchannel chip 100 is held by the mounting unit 10 without falling from the mounting unit 10.

  The placement unit 10 is positioned on the fixed unit 30 when the microchannel chip 100 is placed on the placement unit 10. That is, the mounting unit 10 is positioned on the fixed unit 30 in a state where the microchannel chip 100 is not mounted on the mounting unit 10.

  Further, when the microchannel chip 100 is mounted on the mounting unit 10, the mounting unit 10 is configured such that a gap exists between the inner surface of the mounting unit 10 and the side surface of the microchannel chip 100. Has been. Specifically, before the microchannel chip 100 is placed on the placement part 10, the inner surface of the first wall part 11 b of the first placement bar 11 and the second wall part 12 b of the second placement bar 12. The distance from the inner surface (the distance between the inner surfaces in the width direction of the mounting portion 10) is longer than the width of the microchannel chip 100. Thereby, before the microchannel chip 100 is mounted on the mounting unit 10, the inner surface of the first wall portion 11 b of the first mounting bar 11 and one side surface in the width direction of the microchannel chip 100. A gap can exist between them, and a gap can exist between the inner surface of the second wall portion 12 b of the second mounting bar 12 and the other side surface in the width direction of the microchannel chip 100.

  The mounting unit 10 is a movable mounting table and can move. The placement unit 10 can move in a direction that approaches or moves away from the heating unit 20. Specifically, the placement unit 10 can move in the vertical direction, and moves up and down in the vertical direction in the recess 41 provided in the main body unit 40. In addition, although the raising / lowering operation | movement of the mounting part 10 can be performed by an actuator, for example, you may carry out manually.

  In the present embodiment, the mounting unit 10 on which the microchannel chip 100 is mounted is lowered until the microchannel chip 100 is disposed on the heating unit 20. At this time, since there is an opening 10a that exposes the back surface of the microchannel chip 100 in the mounting unit 10, the microchannel can be lowered by lowering the mounting unit 10 on which the microchannel chip 100 is mounted. The back surface of the chip 100 is in contact with the upper surface of the heating unit 20 through the opening 10a. That is, the microchannel chip 100 is placed on the heating unit 20 with the back surface in contact with the upper surface of the heating unit 20 through the opening 10a.

  The placement unit 10 can be moved in the horizontal direction by the elastic member 13. The elastic member 13 is, for example, a spring. In the present embodiment, the elastic member 13 is a metal coil spring, and is attached to the mounting portion 10 so as to be expandable and contractable in the horizontal direction. Specifically, the end of the elastic member 13 in the longitudinal direction is fixed to the second placement bar 12 of the placement unit 10.

  Further, the end of the elastic member 13 in the longitudinal direction is fixed to the slide member 14. Thereby, the mounting part 10 is held by the main body part 40 by the elastic member 13 and the slide member 14. The slide member 14 is a slide bar made of, for example, metal or resin, and has a slide surface 14 a that contacts the inclined surface 40 a of the main body 40. In the present embodiment, the inclination angle of the slide surface 14a of the slide member 14 and the inclination angle of the inclined surface 40a of the main body 40 are the same.

  Thus, since the slide member 14 is fixed to the mounting part 10 via the elastic member 13, when the mounting part 10 moves up and down, the slide member 14 also moves up and down in conjunction with the lifting and lowering of the mounting part 10. . At this time, the slide member 14 moves up and down so that the slide surface 14 a is along the inclined surface 40 a of the main body 40. Thereby, the elastic member 13 is elastically deformed by extending or contracting as the slide member 14 moves up and down. For example, when the placement unit 10 is lowered in the vertical direction toward the heating unit 20, the slide member 14 is lowered and the elastic member 13 is contracted in conjunction with the lowering of the placement unit 10. Thus, the mounting part 10 can move in the horizontal direction within a range in which the elastic member 13 is elastically deformed.

  The heating unit 20 is a heating device (heater) that heats the microchannel chip 100. Specifically, the heating unit 20 heats the reaction solution introduced into the microchannel chip 100. As described above, when the mounting unit 10 on which the microchannel chip 100 is mounted is lowered, the microchannel chip 100 comes into contact with the heating unit 20. Thereby, the microchannel chip 100 can be directly heated by the heating unit 20.

  The heating unit 20 heats the reaction solution located in the nucleic acid amplification unit 100a of the microchannel chip 100. Therefore, the heating unit 20 only needs to be disposed so as to correspond to at least the nucleic acid amplification unit 100a of the microchannel chip 100. With this configuration, the heating unit 20 applies a predetermined temperature to the reaction solution sent to the microchannel 130 of the nucleic acid amplification unit 100a. Thereby, the target nucleic acid contained in the reaction solution can be amplified.

  The heating unit 20 is disposed below the mounting unit 10 and the fixed unit 30. In the present embodiment, the heating unit 20 is disposed on the bottom surface of the recess 41 of the main body 40.

  The heating unit 20 is disposed with a gap between the heating unit 20 and the side surface of the recess 41 of the main body unit 40. The placement unit 10 is accommodated in the gap when the placement unit 10 is lowered. Specifically, the first mounting bar 11 of the mounting unit 10 is accommodated in the gap between one side surface of the concave portion 41 of the main body 40 and one side surface of the heating unit 20, and the concave portion of the main body 40. In the gap between the other side surface of 41 and the other side surface of the heating unit 20, the second placement bar 12 of the placement unit 10 is accommodated. As described above, the gap between the heating unit 20 and the side surface of the concave portion 41 of the main body 40 is large enough to accommodate at least the placement unit 10.

  The heating unit 20 is, for example, a heater block. Specifically, the heating unit 20 is a metal block made of a metal such as a rectangular parallelepiped aluminum or stainless steel. In this case, the heating unit 20 is connected to a temperature control unit (not shown), and the temperature of the heating unit 20 is controlled by the temperature control unit.

  The fixing unit 30 is a fixing member for holding and fixing the microchannel chip 100 mounted on the mounting unit 10 at a predetermined position. Specifically, the fixing unit 30 fixes the microchannel chip 100 mounted on the mounting unit 10 at a position where the microchannel chip 100 contacts the heating unit 20. Thereby, the microchannel chip 100 can be adhered to the heating unit 20.

  In the present embodiment, the fixing portion 30 is constituted by a pair of separated first fixing body 31 and second fixing body 32. The pair of first fixed body 31 and second fixed body 32 are arranged at positions facing each other with a predetermined interval in the horizontal direction.

  The first fixed body 31 is provided on one side surface of the main body 40. Specifically, the first fixed body 31 is fixed to a first elastic member 33 housed in a recess 42 (first recess) provided on one side surface of the main body 40. The recess 42 is formed so as to extend in the horizontal direction. Accordingly, the first fixed body 31 moves in the horizontal direction as the first elastic member 33 expands and contracts. In the present embodiment, the first elastic member 33 is contracted so that the entire first fixed body 31 can be stored in the recess 42.

  On the other hand, the second fixed body 32 is provided on the other side surface of the main body 40. Specifically, the second fixed body 32 is fixed to the second elastic member 34 housed in the concave portion 43 (second concave portion) provided on the inclined surface 40 a of one side surface of the main body portion 40. . The recess 43 is formed so as to extend in the horizontal direction. Therefore, the second fixed body 32 moves in the horizontal direction as the second elastic member 34 expands and contracts. In the present embodiment, the second elastic member 34 is contracted so that the entire second fixed body 32 can be stored in the recess 43.

  Each of the first fixed body 31 and the second fixed body 32 is, for example, a member having a triangular cross section. Accordingly, the first fixed body 31 has a first inclined surface 31a that is a triangular inclined surface, and the second fixed body 32 has a second inclined surface 32a that is a triangular inclined surface. In addition, each of the 1st fixing body 31 and the 2nd fixing body 32 is comprised by the metal material or the resin material, for example.

  The fixing unit 30 is provided in the main body 40 at a position below the mounting unit 10 and above the heating unit 20 when the microchannel chip 100 is mounted on the mounting unit 10. It has been. That is, the fixing unit 30 is positioned below the mounting unit 10 and above the heating unit 20 in a state where the microchannel chip 100 is not mounted on the mounting unit 10.

  The fixing unit 30 is configured so that the mounting unit 10 can move below the fixing unit 30 when the mounting unit 10 on which the micro-channel chip 100 is mounted moves vertically downward. Move away from 100. In the present embodiment, the first fixed body 31 and the second fixed body 32 that are the fixed portions 30 are movable in the horizontal direction by the first elastic member 33 and the second elastic member 34, respectively.

  In the present embodiment, the fixing unit 30 moves in the horizontal direction by moving the mounting unit 10 vertically downward while contacting the fixing unit 30. Specifically, from the state where the first placement bar 11 and the second placement bar 12 are located above the first fixed body 31 and the second fixed body 32, the first placement bar 11 and the second placement bar 11 are arranged. When the mounting bar 12 is lowered, the back surface of the first mounting bar 11 comes into contact with the first inclined surface 31a of the first fixed body 31, so that the first fixed body 31 has the micro flow path. The second fixed body 32 moves in the horizontal direction so as to move away from the chip 100, and the second fixed body 32 is in contact with the second inclined surface 32 a of the second fixed body 32 by abutting the slide surface 14 a (inclined surface) of the slide member 14. The second fixed body 32 moves in the horizontal direction so that the fixed body 32 moves away from the microchannel chip 100.

  Next, the nucleic acid amplification method according to the embodiment will be described with reference to FIG. FIG. 4 is a diagram for explaining the nucleic acid amplification method according to the embodiment. 4 is a diagram corresponding to FIG. 2, and the right diagram in FIG. 4 is a diagram corresponding to FIG.

  The nucleic acid amplification method according to the present embodiment is performed using the nucleic acid amplification apparatus 1 in the above embodiment. In the nucleic acid amplification device 1, when the microchannel chip 100 is not placed on the placement unit 10 (that is, the state before the placement unit 10 is still lowered), the placement unit 10 and the fixing unit 30 are The placement unit 10 and the fixing unit 30 are arranged in this order along the direction from the microchannel chip 100 toward the heating unit 20.

  In the nucleic acid amplification method, using this state as an initial state, the microchannel chip 100 is placed on the placement unit 10 of the nucleic acid amplification device 1 as shown in FIG. At this time, the placement unit 10 is located above the fixed unit 30. That is, the first placement bar 11 and the second placement bar 12 are located above the first fixed body 31 and the second fixed body 32.

  In the present embodiment, in a state before the microchannel chip 100 is placed on the placement unit 10, the inner surface of the first wall 11 b of the first placement bar 11 and the second of the second placement bar 12. The distance from the inner surface of the wall 12 b is longer than the width of the microchannel chip 100. That is, when the microchannel chip 100 is mounted on the mounting unit 10, there is a gap between the inner surface of the mounting unit 10 and the side surface of the microchannel chip 100. By providing such a gap, the microchannel chip 100 can be easily placed on the placement portion 10.

  Next, as shown in FIG. 4B, the placement unit 10 on which the microchannel chip 100 is placed is moved vertically downward from the state where the placement unit 10 is located above the fixed unit 30. To lower. Specifically, from the state where the first placement bar 11 and the second placement bar 12 are located above the first fixed body 31 and the second fixed body 32, the first placement bar 11 and the second placement bar 11 are arranged. The mounting bar 12 is lowered vertically downward.

  Thereby, the back surface of the first mounting bar 11 abuts on the first inclined surface 31a of the first fixed body 31, and the sliding surface of the sliding member 14 connected to the second mounting bar 12 via the elastic member 13 is used. 14 a contacts the second inclined surface 32 a of the second fixed body 32. When the first placement bar 11 and the second placement bar 12 are further lowered from this state, the slide member 14 is lowered along the second inclined surface 32 a of the second fixed body 32.

  At this time, since the slide member 14 moves horizontally in a direction approaching the second placement bar 12, the elastic member 13 is elastically deformed so as to gradually contract. Thereby, the restoring force (elastic force) of the elastic member 13 is gradually increased, and the interval between the first placement bar 11 and the second placement bar 12 is gradually narrowed by the pressing by the restoring force. . That is, the gap between the inner surface of the placement unit 10 and the side surface of the microchannel chip 100 gradually disappears. As a result, finally, the side surface of the microchannel chip 100 comes into contact with the first mounting bar 11 and the second mounting bar 12, and the microchannel chip 100 is pressed by the elastic member 13 to be in contact with the first mounting bar. 11 and the second mounting bar 12. Therefore, the microchannel chip 100 is reliably held on the placement unit 10.

  Further, when the back surface of the first placement bar 11 comes into contact with the first inclined surface 31a of the first fixed body 31 and the first placement bar 11 is further lowered, as shown in FIG. The first fixed body 31 is moved in the horizontal direction by the pressing of the mounting bar 11 and stored in the recess 42. At this time, the first elastic member 33 is elastically deformed to be contracted by the pressing of the first fixed body 31.

  On the other hand, when the slide surface 14a of the slide member 14 comes into contact with the second inclined surface 32a of the second fixed body 32 and the slide member 14 is lowered together with the second mounting bar 12, as shown in FIG. The second fixed body 32 is moved in the horizontal direction by being pressed, and is stored in the recess 43. At this time, the second elastic member 34 is elastically deformed so as to be contracted by the pressing of the second fixed body 32.

  Next, as shown in (c) of FIG. 4, from the state where the first fixed body 31 and the second fixed body 32 are housed in the recesses 42 and 43, the mounting on which the microchannel chip 100 is mounted. The placement unit 10 (the first placement bar 11 and the second placement bar 12) is further lowered vertically.

  When the placement unit 10 passes the height position of the fixed unit 30, that is, when the first placement bar 11 and the second placement bar 12 pass the first fixed body 31 and the second fixed body 32, the same. As shown in the figure, the first fixed body 31 is pressed by the restoring force of the elastically deformed first elastic member 33 and jumps out of the recess 42, and the second fixed body 32 is elastically deformed by the second elastic member 34. It is pressed out by the restoring force of, and jumps out of the recess 43. That is, the first fixed body 31 and the second fixed body 32 jump out onto the microchannel chip 100 placed on the placement unit 10. Thereby, the microchannel chip 100 placed on the placement part 10 is locked and fixed by the protruding fixing part 30 (the first fixing body 31 and the second fixing body 32).

  At this time, the fixing unit 30 fixes the microchannel chip 100 by positioning a part of the microchannel chip 100 between the mounting unit 10 and the fixing unit 30. Specifically, the fixing unit 30 fixes the microchannel chip 100 by pressing the upper surface of the microchannel chip 100. Thereby, since a part of upper surface of the microchannel chip 100 is locked by the fixing portion 30, it is possible to prevent the microchannel chip 100 from jumping upward.

  In addition, in the present embodiment, when the placement unit 10 descends and passes through the height position of the fixed unit 30, the microchannel chip 100 placed on the placement unit 10 becomes an opening of the placement unit 10. It contacts the heating unit 20 via 10a.

  As described above, when the placement unit 10 passes the height position of the fixed unit 30, the microchannel chip 100 placed on the placement unit 10 is heated via the opening 10 a of the placement unit 10. And is pressed and fixed to the heating unit 20 by the fixing unit 30.

  Thus, the microchannel chip 100 can be fixed in close contact with the heating unit 20 by the fixing unit 30, and the microchannel chip 100 can be disposed at a predetermined position with respect to the heating unit 20.

  As described above, in the nucleic acid amplification device 1 according to the present embodiment, when the microchannel chip 100 is in contact (contact) with the heating unit 20, the placement unit 10 and the fixing unit 30 are configured as a microchannel chip. Along the direction from 100 toward the heating unit 20, the fixing unit 30 and the mounting unit 10 are arranged in this order.

  When the mounting unit 10 descends and passes through the height position of the fixed unit 30, the first mounting bar 11 is formed between the one side surface of the heating unit 20 and the one side surface of the recess 41 of the main body unit 40. It is stored in the gap between them. Similarly, when the mounting unit 10 descends and passes through the height position of the fixing unit 30, the second mounting bar 12 includes the other side surface of the heating unit 20 and the other side surface of the recess 41 of the main body unit 40. It is stored in the gap between.

  After the microchannel chip 100 is brought into close contact with the heating unit 20, the reaction solution is introduced into the microchannel chip 100. For example, the reaction solution is introduced into the first opening 110 of the microchannel chip 100 by dropping a reaction solution, which is prepared by previously mixing a sample containing the target nucleic acid and the reaction reagent, and weighing in advance with a pipette. The sample containing the target nucleic acid and the reaction reagent are not mixed in advance, but the sample containing the target nucleic acid and the reaction reagent are sequentially introduced into the first opening 110 and mixed in the vicinity of the first opening 110. The reaction solution may be introduced into the microchannel chip 100 by generating the reaction solution in the microchannel chip 100.

  The reaction solution introduced into the microchannel chip 100 advances in the microchannel 130 by capillary force and is heated by the heating unit 20 in the nucleic acid amplification unit 100a. Thereby, the target nucleic acid contained in the reaction solution can be amplified.

  After the reaction solution has spread over the entire microchannel 130, the amplification amount of the target nucleic acid contained in the reaction solution is detected using an optical detection device. For example, reflected light (fluorescence) is received while scanning laser light (excitation light) in a direction crossing the microchannel 130, and the target nucleic acid contained in the reaction solution is detected based on the received reflected light amount (fluorescence amount). Calculate the amount of amplification. Thereby, the inspection of the microchannel chip 100 is completed.

  After completion of the inspection, although not shown, the fixed state of the microchannel chip 100 by the fixing unit 30 is released, the mounting unit 10 is raised, and the microchannel chip 100 is separated from the heating unit 20. After that, the placement unit 10 is raised to the initial state (chip set) position (position (a) in FIG. 4), and the inspected microchannel chip 100 is taken out of the placement unit 10.

  Thereafter, the next microchannel chip 100 can be inspected in the same manner as described above by placing the microchannel chip 100 to be inspected next on the mounting portion 10.

  As described above, the nucleic acid amplification device 1 according to the present embodiment includes the movable mounting unit 10 on which the microchannel chip 100 is mounted, the heating unit 20 that heats the microchannel chip 100, and the microchannel chip 100. Is provided with a fixing unit 30 that fixes the microchannel chip 100 mounted on the mounting unit 10 at a position in contact with the heating unit 20. When the microchannel chip 100 is in contact with the heating unit 20, the fixing unit 30 and the mounting unit 10 are arranged in this order along the direction from the microchannel chip 100 toward the heating unit 20. Yes.

  With this configuration, the microchannel chip 100 mounted on the mounting unit 10 is brought into close contact with the heating unit 20 to be fixed or heated only by moving the mounting unit 10 on which the microchannel chip 100 is mounted. It can be separated from the part 20. In addition, the positioning of the microchannel chip 100 with respect to the heating unit 20 can be performed with high accuracy simply by moving the mounting unit 10 on which the microchannel chip 100 is mounted.

  As described above, according to the nucleic acid amplification device 1 in the present embodiment, the microchannel chip 100 can be easily adhered and fixed to the heating unit 20, and the microchannel chip 100 with respect to the heating unit 20 can be fixed. Positioning and attaching / detaching can be easily performed. Thereby, highly accurate nucleic acid amplification can be performed with respect to the target nucleic acid contained in the specimen of the reaction solution. That is, a highly accurate inspection can be performed.

  Further, in the nucleic acid amplification device 1 according to the present embodiment, the fixing unit 30 is configured so that the mounting unit 10 is more than the fixing unit 30 when the mounting unit 10 on which the microchannel chip 100 is mounted moves vertically downward. Also, it is configured to move in a direction away from the microchannel chip 100 so that it can move downward.

  With this configuration, immediately after the microchannel chip 100 is placed on the placement unit 10, even when the placement unit 10 is located above the fixing unit 30 protruding in the horizontal direction, the placement unit 10. Can be moved downward from the height position of the fixed portion 30.

  Further, in the nucleic acid amplification device 1 according to the present embodiment, the fixing unit 30 is movable in the horizontal direction by the elastic members (the first elastic member 33 and the second elastic member 34). Further, the fixing unit 30 moves in the horizontal direction by moving the mounting unit 10 vertically downward while contacting the fixing unit 30.

  With this configuration, by moving the mounting unit 10 down, the fixing unit 30 can be easily moved away from the microchannel chip 100, and the mounting unit 10 can be moved below the fixing unit 30. In addition, since the elastic member can be elastically deformed by the horizontal movement of the fixing portion 30, when the placement portion 10 passes the position of the fixing portion 30, the fixing portion 30 is restored by the restoring force of the elastic member that is elastically deformed. It automatically returns to the position of. Thereby, the microchannel chip 100 can be easily fixed by the fixing portion 30.

  Further, in the nucleic acid amplification device 1 according to the present embodiment, when the microchannel chip 100 is mounted on the mounting unit 10, the mounting unit 10 includes the inner surface of the mounting unit 10 and the side surface of the microchannel chip 100. It is comprised so that a clearance gap may exist between.

  With this configuration, when the microchannel chip 100 is set on the mounting unit 10 at the start of inspection, the microchannel chip 100 can be easily mounted on the mounting unit 10.

  Further, in the nucleic acid amplification device 1 according to the present embodiment, the placement unit 10 is movable in the horizontal direction by the elastic member 13.

  With this configuration, even when there is a gap between the inner surface of the mounting unit 10 and the side surface of the microchannel chip 100 when the microchannel chip 100 is set on the mounting unit 10, the elasticity of the elastic member 13. The gap can be eliminated by using force. Thereby, when the microchannel chip 100 mounted on the mounting unit 10 is moved, the microchannel chip 100 can be reliably held on the mounting unit 10. Therefore, when the microchannel chip 100 comes into contact with the heating unit 20 even when the microchannel chip 100 is not aligned with the heating unit 20 when the microchannel chip 100 is mounted on the mounting unit 10. The microchannel chip 100 is aligned with the heating unit 20.

  Moreover, in the nucleic acid amplification device 1 according to the present embodiment, the fixing unit 30 positions the microchannel chip 100 by placing a part of the microchannel chip 100 between the mounting unit 10 and the fixing unit 30. It is fixed.

  With this configuration, a part of the microchannel chip 100 can be sandwiched between the mounting unit 10 and the fixing unit 30, so that the microchannel chip 100 placed on the mounting unit 10 is placed in the heating unit 20. Even if they are brought into close contact with each other, the microchannel chip 100 can be prevented from jumping out of the mounting portion 10.

  In the nucleic acid amplification device 1 according to the present embodiment, the fixing unit 30 fixes the microchannel chip 100 by pressing the upper surface of the microchannel chip 100.

  With this configuration, the microchannel chip 100 placed on the placement unit 10 can be more closely attached to the heating unit 20 while being fixed by the fixing unit 30.

(Modification 1)
Next, a nucleic acid amplification device 1A according to Modification 1 of the embodiment will be described with reference to FIGS. FIG. 5 is a cross-sectional view of a nucleic acid amplification apparatus 1A according to Modification 1 of the embodiment. FIG. 6 is a top view of the nucleic acid amplification device 1B. 5 is a cross-sectional view taken along line VV in FIG.

  In the nucleic acid amplification device 1 </ b> A according to this modification, the heating unit 20 </ b> A is configured such that at least two temperature regions are formed in the microchannel chip 100. Specifically, the heating unit 20A includes a first heater 21 and a second heater 22 that are set to predetermined different temperatures. Thereby, in the nucleic acid amplification unit 100 a of the microchannel chip 100, two temperature regions set at different temperatures are formed by the two heaters of the first heater 21 and the second heater 22.

  For example, the temperature of the first heater 21 is set to be higher than the temperature of the second heater 22. In this case, the region corresponding to the first heater 21 is a high temperature region, and the region corresponding to the second heater 22 is a low temperature region. As a result, in the microchannel chip 100, a high temperature region is formed in the region on the first heater 21, and a low temperature region is formed in the region on the second heater 22.

  The set temperature of the first heater 21 for forming the high temperature region is, for example, 90 ° C. to 98 ° C., and in this embodiment, it is about 95 ° C., which is the denaturation reaction temperature of the nucleic acid amplification reaction. On the other hand, the set temperature of the second heater 22 for forming the low temperature region is, for example, 50 ° C. to 75 ° C., and in this embodiment, it is about 60 ° C., which is the annealing / elongation reaction temperature.

  The first heater 21 and the second heater 22 are arranged with a predetermined gap. The microchannel chip 100 is placed on the first heater 21 and the second heater 22 so that the nucleic acid amplification unit 100 a straddles the gap between the first heater 21 and the second heater 22. As a result, the micro flow path 130 in the nucleic acid amplification unit 100a is configured to reciprocate in two cycles between the two temperature regions of the first heater 21 and the second heater 22. With this configuration, a temperature increasing / decreasing temperature cycle (heat cycle) is given to the reaction solution flowing through the microchannel 130.

  The first heater 21 and the second heater 22 are, for example, heater blocks. Specifically, the first heater 21 and the second heater 22 are metal blocks made of a rectangular parallelepiped metal such as aluminum or stainless steel. In this case, the first heater 21 and the second heater 22 are connected to a temperature control unit (not shown), and each temperature of the first heater 21 and the second heater 22 is controlled by the temperature control unit.

  The nucleic acid amplification method in this modification can be performed in the same manner as the nucleic acid amplification method in the above-described embodiment shown in FIG. 4 using the nucleic acid amplification apparatus 1A in this modification.

  That is, also in this modification, the microchannel chip 100 can be closely attached and fixed to the heating unit 20A by the fixing unit 30 by lowering the mounting unit 10 on which the microchannel chip 100 is mounted.

  In this case, in the nucleic acid amplification device 1 </ b> A according to this modification, the heating unit 20 </ b> A includes the first heater 21 and the second heater 22. Further, the microchannel chip 100 is configured to reciprocate or periodically pass through a low temperature region and a high temperature region whose temperature is controlled by the heating unit 20A. Specifically, the microchannel 130 in the nucleic acid amplification unit 100a is a serpentine channel formed so as to meander, and in a plurality of cycles so as to reciprocate between a high temperature region and a low temperature region formed by the heating unit 20. Wrapped.

  As a result, the reaction solution introduced into the microchannel chip 100 passes through the high temperature region and the low temperature region by the meandering channel of the microchannel 130, and thus gives a periodic temperature change (temperature cycle) to the reaction solution. be able to.

  Specifically, the reaction solution introduced into the microchannel chip 100 passes through the microchannel 130 so as to reciprocate between the first heater 21 and the second heater 22 repeatedly. That is, the reaction solution is fed so as to alternately and repeatedly pass through the two temperature regions of the high temperature region (first heater 21) and the low temperature region (second heater 22). And cooling are repeatedly applied alternately. As a result, a heat cycle can be imparted to the reaction solution, so that the target nucleic acid contained in the reaction solution is amplified by repeating the denaturation reaction in the high temperature region and the annealing / extension reaction in the low temperature region. Become.

  As described above, since the reaction solution can be raised and lowered while the solution is fed, a very high-speed flow PCR can be realized. Therefore, the target nucleic acid contained in the reaction solution can be amplified at high speed.

  After the reaction solution has spread over the entire microchannel 130, the amount of amplification of the target nucleic acid contained in the reaction solution can be calculated in the same manner as in the above embodiment. For example, reflected light (fluorescence) is received while scanning laser light (excitation light) in a direction intersecting the microchannel 130, and each cycle of the meandering channel (temperature) based on the received reflected light amount (fluorescence amount). The amount of nucleic acid amplification in each cycle) is detected as an amplification curve. At this time, an amplification curve is obtained in which the nucleic acid amplification amount increases as the PCR cycle increases. Thereby, the amplification amount of the target nucleic acid contained in the reaction solution can be calculated.

(Modification 2)
Next, a nucleic acid amplification device 1B according to Modification 2 of the embodiment will be described with reference to FIG. FIG. 7 is a cross-sectional view of a nucleic acid amplification device 1B according to Modification 2 of the embodiment.

  The nucleic acid amplification device 1 </ b> B according to the present modification is configured to further include a pressing unit 50 that presses the heating unit 20 against the microchannel chip 100 with respect to the nucleic acid amplification device 1 according to the above embodiment. In this modification, the pressing unit 50 includes an elastic member 51 and a support member 52.

  The elastic member 51 is, for example, a spring. Specifically, the elastic member 51 is a metal coil spring, and is provided between the heating unit 20 and the support member 52 so as to expand and contract in the vertical direction. The support member 52 is a metal plate, for example, and supports the elastic member 51.

  The nucleic acid amplification method in this modification can be performed in the same manner as the nucleic acid amplification method in the above-described embodiment shown in FIG. 4 using the nucleic acid amplification device 1B in this modification.

  That is, also in this modification, the microchannel chip 100 can be closely attached and fixed to the heating unit 20 by the fixing unit 30 by lowering the mounting unit 10 on which the microchannel chip 100 is mounted.

  In this case, the nucleic acid amplification device 1 </ b> B according to this modification includes a pressing unit 50 that presses the heating unit 20 against the microchannel chip 100. With this configuration, when the placement unit 10 is lowered and the microchannel chip 100 is fixed to the heating unit 20 by the fixing unit 30, the heating unit 20 applies pressure upwardly by the pressing unit 50. The road chip 100 is sandwiched between the fixing unit 30 and the heating unit 20. Thereby, compared with the nucleic acid amplification device 1 in the above embodiment, the microchannel chip 100 can be more closely attached to the heating unit 20. Thereby, a more accurate inspection can be performed.

(Other variations)
As described above, the nucleic acid amplification device and the nucleic acid amplification method according to the present invention have been described based on the embodiments and the modifications. However, the present invention is not limited to the above embodiments.

  For example, in the above embodiment, a high thermal conductive sheet may be disposed as a part of the heating unit 20 between the heater block constituting the heating unit 20 and the micro-channel chip 100. That is, the microchannel chip 100 may be placed on the heating unit 20 via the high thermal conductive sheet. In this case, the microchannel chip 100 and the heating unit 20 are in contact with the high thermal conductive sheet, respectively. As the high thermal conductive sheet, for example, an elastic sheet made of a resin having high thermal conductivity can be used.

  Thus, by inserting the high heat conductive sheet between the microchannel chip 100 and the heating unit 20, the microchannel chip 100 and the high heat conductive sheet can be brought into close contact with each other and the heating unit 20 and the high heat conductive sheet can be adhered. Can be brought into close contact with each other. Thereby, an air layer is not interposed between the microchannel chip 100 and the heating unit 20, and the adhesion at the interface between the microchannel chip 100 and the heating unit 20 can be improved. Therefore, the microchannel chip 100 can be efficiently heated by the heating unit 20.

  Note that the insertion of the high thermal conductive sheet between the heater block and the microchannel chip 100 may be applied to the first and second modifications.

  Moreover, in the said embodiment, although the number of the microchannel chips 100 which can be set to the nucleic acid amplification apparatus 1 was set to one, it is not restricted to this. For example, the number of microchannel chips 100 that can be set in the nucleic acid amplification device 1 may be plural. This may be applied to the first and second modifications.

  In addition, a form obtained by making various modifications conceived by those skilled in the art with respect to each embodiment and modification, and any combination of components and functions in the embodiment without departing from the spirit of the present invention Forms to be made are also included in the present invention.

DESCRIPTION OF SYMBOLS 1, 1A, 1B Nucleic acid amplifier 10 Placement part 13 Elastic member 20, 20A Heating part 30 Fixing part 50 Pressing part 100 Microchannel chip 110 1st opening part 120 2nd opening part 130 Microchannel

Claims (10)

A movable mounting portion on which a microchannel chip having a microchannel is mounted;
A heating section for heating the microchannel chip;
A fixing portion for fixing the microchannel chip placed on the placement unit at a position where the microchannel chip is in contact with the heating unit;
When the microchannel chip is in contact with the heating unit, the fixing unit and the mounting unit are arranged in this order along the direction from the microchannel chip to the heating unit.
Nucleic acid amplification device. The mounting unit can move in a vertical direction, and is positioned on the fixed unit when the microchannel chip is mounted on the mounting unit,
The fixing portion is separated from the microchannel chip so that when the mounting portion on which the microchannel chip is mounted moves vertically downward, the mounting portion can move below the fixing portion. Move away,
The nucleic acid amplification device according to claim 1. The fixed portion is movable in the horizontal direction by an elastic member.
The nucleic acid amplification device according to claim 2. The fixed part moves in the horizontal direction by moving vertically downward while the mounting part is in contact with the fixed part.
The nucleic acid amplification device according to claim 2 or 3. When the microchannel chip is mounted on the mounting section, the mounting section is configured such that a gap exists between the inner surface of the mounting section and the side surface of the microchannel chip. Yes,
The nucleic acid amplification device according to any one of claims 1 to 4. The mounting portion is movable in the horizontal direction by an elastic member.
The nucleic acid amplification device according to claim 5. The fixing unit fixes the microchannel chip by positioning a part of the microchannel chip between the mounting unit and the fixing unit.
The nucleic acid amplification device according to any one of claims 1 to 6. The fixing portion fixes the microchannel chip by pressing the upper surface of the microchannel chip.
The nucleic acid amplification device according to claim 7. The heating unit is configured such that at least two temperature regions are formed in the microchannel chip.
The nucleic acid amplification device according to any one of claims 1 to 8. A pressing unit that presses the heating unit against the microchannel chip;
The nucleic acid amplification device according to any one of claims 1 to 9.

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