JP2007189979A - Reaction vessel - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction vessel capable of avoiding losing a reaction reagent and of preventing the vessel itself from being stained by the reaction reagent. <P>SOLUTION: The reaction vessel includes a reaction section 4 having openings 15A and 15B formed on one side 2A of a base material 2 and a lid component 17 covering the openings 15A and 15B, wherein the lid component 17 has a port 22A communicating with the openings 15A and 15B and an element 23 functioning to open/close the port 22A in response to the movement, relative to the lid component 17, of a pipette chip butted on the lid component 17. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a reaction vessel used for, for example, a biochemical reaction.

2. Description of the Related Art Conventionally, for example, a reaction vessel that processes a small amount of reagent in a biochemical reaction or the like has been provided with a plurality of recesses or flow paths as a reaction field (see, for example, Patent Document 1). In such a reaction vessel, the reaction reagent supplied to each recess or channel is heated by a reaction device including a temperature control device including a Peltier element that can control the temperature state of each recess or channel.
Here, in the case of a reaction vessel having a flow channel-like reaction section, the reaction reagent supplied to the flow path is exposed to the outside only from the opening of the flow path, thereby reducing the evaporation of the reaction reagent during the reaction. be able to. However, due to the expansion of mixed bubbles, the surface condition of the flow path-like reaction part and the surface condition such as processing stripes, the reaction liquid moves due to the pinching condition of the temperature control device, etc. Furthermore, the reaction solution may reach the opening and evaporate. At this time, in order to further prevent evaporation of the reaction reagent, for example, a sealing liquid such as mineral oil is further supplied to the flow path to which the reaction reagent is supplied, so that the flow path is sealed with the sealing liquid. The reaction reagent may be blocked by the flow path and the sealing liquid.
Japanese Patent No. 2759071

  However, the following problems remain in the conventional reaction vessel. That is, when a reaction reagent or sealing liquid is supplied to the flow path, air may be mixed as bubbles in the flow path. The bubbles expand when the reaction reagent is heated, which may cause the sealing liquid to be pushed out of the opening and out of the flow path. For this reason, there exists a problem that the reaction reagent evaporates and the loss of the reaction reagent occurs. Further, there is a problem that contamination by the reaction reagent occurs due to the reaction reagent flowing out.

  The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a reaction vessel that can avoid loss of a reaction reagent and prevent contamination by the reaction reagent.

  The present invention employs the following configuration in order to solve the above problems. That is, the reaction container of the present invention includes a reaction part having an opening formed on one surface of a base material and a lid member covering the opening, and the communication port through which the lid member communicates with the opening And an opening / closing part that opens and closes the communication port based on the movement of the dispensing member brought into contact with the lid member with respect to the lid member.

  In this invention, even if the communicating port is opened by moving the tip of a dispensing member such as a pipette tip or an injection needle in contact with the lid member, the pipette tip or the like is separated. When the communication port is closed, the sealed state of the reaction part is maintained. Thereby, even if bubbles are mixed when a reaction reagent or the like is supplied into the reaction part, evaporation and outflow of the reagent accommodated in the reaction part and scattering to the outside of the reaction container are suppressed. Therefore, loss of the reaction reagent can be avoided and contamination by the reaction reagent can be prevented.

Further, in the reaction container of the present invention, the lid member has a fixing portion that is fixed to the base material and supports the opening / closing portion, and the opening / closing portion can be moved toward and away from each other, and the dispensing member abuts on the container. It is preferable to include a plurality of movable portions and a connection portion that connects the plurality of movable portions to the fixed portion in a state where the movable portions are biased so that the communication port is closed.
In the present invention, the communication port is opened by moving the tip of a pipette tip or the like in contact with the movable part. Here, since the movable part is urged by the connection part so that the communication port is closed, the communication port is closed when the tip part such as a pipette tip is separated.

Moreover, the reaction container of this invention is good also as the said connection part being provided with the rotation axis | shaft which rotates the said movable part around the axis along the said one surface.
In this invention, when the pipette tip or the like is in contact with the movable part and moved so as to be pushed toward the opening, the plurality of movable parts are inclined about the axis along one surface of the base material, and the communication port is in the open state. It becomes. When the tip of the pipette tip or the like is moved away from the movable part, the movable part is biased as described above, so that it moves around the axis opposite to the above, and the communication port is closed.

In the reaction container of the present invention, it is preferable that the connection portion includes a rotation shaft that rotates the movable portion around an axis perpendicular to the one surface.
In this invention, when the tip of a pipette tip or the like is moved so as to be pushed toward the opening in contact with the movable part, the plurality of movable parts are separated from each other around an axis perpendicular to one surface of the substrate. And the communication port is opened. Then, when the tip of the pipette tip or the like is separated from the movable part, the movable part is biased as described above, so that it rotates in the opposite direction to the above, and the communication port is closed.

Moreover, it is preferable that the reaction container of this invention is provided with the detection part which can be optically analyzed on the surface of the said base material.
According to the present invention, it is possible to continuously and efficiently execute at least a process for causing a desired reaction and a detection process on a single substrate.

Moreover, it is preferable that the reaction container of this invention is provided with the reagent storage part which accommodates the reaction reagent on the surface of the said base material.
In this invention, the process which accommodates at least a reaction reagent and the process which produces a desired reaction with respect to a single base material can be performed continuously and efficiently.

  According to the reaction container of the present invention, even when the communication port is in an open state, the communication port is closed when the pipette tip or the like is separated, so that the sealed state of the reaction unit is maintained. As a result, evaporation and outflow of the reagent and scattering to the outside of the reaction vessel are suppressed, and loss of the reaction reagent can be avoided and contamination by the reaction reagent can be prevented.

Hereinafter, an embodiment of a reaction container according to the present invention will be described with reference to the drawings.
For example, as shown in FIGS. 1A to 1C, the reaction container 1 according to the present embodiment includes a reagent container 3 provided on a single substantially rectangular plate-like substrate 2, a reaction unit 4, And a detector 5.

  The base material 2 is composed of, for example, each plastic such as PC (polycarbonate), PP (polypropylene), cycloolefin polymer, fluorine polymer, silicon resin, or an appropriate combination of these plastics. Is formed. Moreover, the base material 2 is excellent in heat resistance, chemical resistance, moldability, and the like.

The reagent storage unit 3 is provided, for example, at one end along the longitudinal direction of the base material 2, and has a recessed hole shape formed at a plurality of locations (four locations) on the surface (one surface) 2 </ b> A of the base material 2. It is comprised by the reagent accommodation recessed part 11 of this.
In the plurality of reagent storage recesses 11, various reagents such as reaction reagents used in various reaction processes such as PCR (Polymerase Chain Reaction), a diluted solution or a buffer solution are stored. Here, the magnitude | size of the reagent accommodation recessed part 11 is suitably set according to the quantity of the reagent to accommodate, for example, the opening diameter is 0.1 mm-10 mm, and the depth is 0.1 mm-10 mm.

The shape of the reagent containing recess 11 is not particularly limited, and may be any appropriate well shape such as a truncated cone shape, a truncated pyramid shape, a cone shape, a truncated pyramid shape, or a curved bottom shape. It is set appropriately depending on the injection property of the solution. Further, the inner surface of the reagent containing recess 11 may be subjected to a surface treatment such as hydrophilicity or water repellency.
The inner surface of the reagent containing recess 11 is formed of, for example, PC, PP, PS (polystyrene), PE (polyethylene), PET (polyethylene terephthalate), POM (polyacetal), PA (polyamide), PAN (polyacrylonitrile), PMMA (polyethylene). Methyl pentene films such as methyl methacrylate) and TPX films (registered trademark, manufactured by Mitsui Chemicals), cycloolefin films such as ZEONOR (registered trademark, manufactured by ZEON CORPORATION), silicon resin films, fluorine polymer films, etc. Each of these plastics or a film obtained by appropriately combining a plurality of these plastics may be used.

The reaction part 4 is provided in the center part along the longitudinal direction of the base material 2, for example, and is formed by the film 13 which covers the groove part 12 formed in the back surface 5B of the base material 2, and the opening end 12A of this groove part 12. A through-hole that communicates each of the openings 14A and 15B and the groove 12 provided on the surface 2A of the base 2 through the flow path 14 that is a space and the thickness of the base 2 A certain liquid injection part 16A, 16B is provided.
That is, the reaction unit 4 has a flow path shape, and the solution supplied into the reaction unit 4 from one opening 15A that opens on the surface 2A of the substrate 2 is sequentially supplied to one injection unit 16A. The flow path 14 formed by the groove part 12 and the film 13 and the other liquid injection part 16B can be circulated. Here, each opening 15A, 15B has a circular shape in plan view.
Moreover, the reaction part 4 is provided with the cover member 17 arrange | positioned so that attachment or detachment is possible so that each of each opening part 15A, 15B may be covered.

  The lid member 17 is made of, for example, each plastic such as PC, PP, cycloolefin polymer, fluorine polymer, silicon resin, or an appropriate combination of these plastics, and has a substantially bottomed cylindrical shape. Yes. And the cover member 17 is provided with the fixing | fixed part 22 and the opening-and-closing part 23 which were formed of the notch 21, as shown to Fig.1 (a)-(c) and Fig.2 (a).

  The fixing part 22 is formed integrally with the opening / closing part 23 and has a cylindrical shape surrounding the periphery of the opening 15 </ b> A, and is erected on the base material 2. A communication port 22 </ b> A that communicates with the opening 15 </ b> A is formed at the upper end of the fixed portion 22. The fixing portion 22 is formed with an engaging claw (not shown) that fixes the lid member 17 to the base material 2 by engaging with an engaging groove (not shown) formed on one surface of the base material 2. ing.

The opening / closing part 23 is provided on the fixed part 22, and includes two movable parts 25 and a connection part 26 that connects the movable part 25 and the fixed part 22.
The movable part 25 is formed at two places on the fixed part 22 by providing the notches 21 in the lid member 17. Further, the movable portion 25 is formed with an abutting recess 25A for abutting a pipette tip P described later when the opening / closing portion 23 is in a closed state.
Similarly to the movable portion 25, the connecting portion 26 is provided by providing a cut 21 in the lid member 17, and connects each movable portion 25 and the fixed portion 22. In addition, the connection portion 26 supports the movable portion 25 so as to be rotatable about the axis of the virtually formed rotation shaft 27 by forming the notch 21 deep and making the lid member 17 thin. Yes. Here, the rotation shaft 27 is provided so as to be substantially parallel to the one surface 2A of the substrate 2. Furthermore, each connection part 26 is urging | biasing the direction which makes the movable part 25 formed in two places approach mutually with the elastic force of the cover member 17. As shown in FIG. That is, each movable portion 25 is fixed to each fixed portion 22 in a state where it is urged to close the communication port 22A.

  Note that the film 13 is a methylpentene film such as PC, PP, PS, PE, PET, POM, PA, PAN, PMMA, TPX film (registered trademark, manufactured by Mitsui Chemicals), Zeonore (registered trademark, Nippon Zeon) (Made by Co., Ltd.) and other plastics such as cycloolefin films, silicon resin films, fluorine polymer films, etc., or a film having a single layer structure or multilayer structure in which a plurality of these plastics are appropriately combined, or, for example, aluminum, copper, gold Each of these metals or the like, or a film having a single layer structure or a multilayer structure in which a plurality of these metals are appropriately combined, and further a film having a multilayer structure by a combination of plastic and metal.

  And the thickness of the film 13 is 1-500 micrometers, for example, Preferably it is 1-100 micrometers, Comprising: It becomes more preferable as it becomes thin within this range. When the thickness is less than 1 μm, thermal deformation becomes excessively large and desired strength cannot be ensured. On the other hand, when the thickness of the film 13 is greater than 500 μm, the thermal conductivity is excessively reduced, and the temperature state of the solution in the reaction unit 4 is controlled uniformly from the outside when the temperature state of the solution in the reaction unit 4 is controlled from the outside. It becomes difficult to control, and the desired uniformity with respect to the reaction state cannot be ensured. Moreover, the film 13 made of metal preferably has a thickness of 1 to 50 μm.

The plastic film 13 preferably has a thermal conductivity of 0.1 kcal / mh ° C. or higher. For example, PP has a thermal conductivity of about 0.119 kcal / mh ° C., and PC has a thermal conductivity of about 0.1. It is about 166 kcal / mh ° C., and the thermal conductivity of PE is about 0.252 kcal / mh ° C.
The film 13 made of metal preferably has a thermal conductivity of 100 kcal / mh ° C. or higher, for example, aluminum has a thermal conductivity of about 177 kcal / mh ° C., and copper has a thermal conductivity of 324 kcal / mh ° C. The thermal conductivity of gold is about 254 kcal / mh ° C.

The single-layer film 13 made of plastic preferably has a thickness of about 10 μm to 100 μm.
In addition, the film 13 having a single layer structure made of metal preferably has a thickness of about 5 μm to 80 μm in the case of soft aluminum, for example, and preferably has a thickness of about 5 μm to 50 μm in the case of hard aluminum.

The multilayer film 13 made of plastic is formed of, for example, PET or OPP (stretched polypropylene), and preferably has a desired toughness and flexibility by setting the thickness to about 1 μm to 20 μm. Secured.
Further, the film 13 having a multilayer structure made of a combination of plastic and metal, for example, in the case of aluminum, preferably has a thickness of about 7 μm to 50 μm. Further, the base material 2 of the reaction vessel 1 is formed on the surface of the aluminum. A seal layer that can be attached to the surface by, for example, heat welding or pressure bonding is provided so as to be integrated with aluminum. This seal layer is formed by laminating a sealant in the form of a resin film such as nylon on the surface of aluminum, or by coating maleic acid-modified polypropylene or the like on the surface of aluminum. In the film 13, a film such as PET or OPP may be laminated on the aluminum layer side in order to further increase the strength.

The detection unit 5 is provided at, for example, the other end portion along the longitudinal direction of the base material 2, and is formed by a concave hole-shaped detection concave portion 29 formed at a plurality of locations (16 locations) on the surface of the base material 2. It is configured.
Here, the detection recess 29 is appropriately set according to the amount of reagent used for DNA analysis. However, since the amount of the reagent is very small, for example, the opening diameter is 0.01 mm to 5 mm, and the depth is 0.00. It is 01 mm to 5 mm.
The shape of the detection recess 29 is not particularly limited, as is the case with the reagent storage recess 11, and may be an appropriate well shape as described above, and may be appropriately set depending on molding processability, solution injection property, and the like. The The inner surface of the detection recess 29 may be subjected to a surface treatment such as hydrophilicity or water repellency.
Further, the inner surface of the detection recess 29 may be covered with a covering film in which each plastic or a plurality of these plastics is appropriately combined, as described above.

The reaction vessel 1 configured as described above is used for performing a biochemical reaction test using a biochemical reaction apparatus 30 as shown in FIG.
The biochemical reaction device 30 includes a reaction device 31 that generates a predetermined reaction such as PCR, which is an enzyme reaction, and a detection device 32 that detects a specimen such as DNA by optical analysis, for example.

The reaction device 31 includes a temperature control device 33 including a Peltier element that controls a temperature state of a reaction reagent described later. For example, as shown in FIG. 3, the temperature control device 33 is disposed so as to sandwich the reaction portion 7 of the reaction vessel 1 from both sides in the thickness direction (that is, the front surface side and the back surface side of the reaction vessel 1). Two Peltier element portions 34a and 34b are provided. Here, each Peltier element part 34a, 34b that contacts the surface of the reaction container 1 has a shape (for example, a concave shape) along the surface shape (for example, a convex shape) of the reaction part 7 of the reaction container 1. It is configured as follows.
The detection device 32 reacts a sample adjusted by a predetermined reaction such as PCR by the reaction device 31 with various reagents for detection in the detection unit 8 of the reaction container 1, and a label attached to the sample or nucleic acid probe in advance. Luminescence detection is performed to detect the presence or absence of a substance (for example, a fluorescent substance) from, for example, the back side of the detection unit 8 of the reaction container 1.
In the present embodiment, the biochemical reaction device 30 includes the reaction device 31 and the detection device 32, but it is sufficient that at least the reaction device 31 is provided.

Next, a biochemical reaction test by the biochemical reaction apparatus 30 using the reaction vessel 1 will be described.
First, for example, a reaction step that causes PCR or the like is performed. This consists of a reaction reagent supply step, a sealing step, and a reaction generation step.
First, a reaction reagent supply step for supplying a reaction reagent is performed. For example, the reaction reagent is supplied into the flow path 14 from the opening 15 </ b> A of the reaction unit 4. That is, firstly, the reaction reagent accommodated in the reagent accommodating recess 11 is filled into a pipette tip P which is a dispensing means described later. Then, as shown in FIG. 4A, the tip of the pipette tip P is brought into contact with the contact recess 25A of the lid member 17, and the pipette tip P is pushed toward the opening 15A. As a result, the two movable parts 25 rotate around the rotation axis 27 in the direction of the arrow R1 shown in FIG. 4A and are separated from each other, thereby opening the communication port 22A. In this way, the tip of the pipette tip P is made to enter the flow path 14, and the reaction reagent filled therein is supplied into the flow path 14.
Thereafter, when the pipette tip P is removed from the lid member 17, the two movable portions 25 are urged by the connecting portion 26 so as to rotate in directions approaching each other, so that the two movable portions 25 are elastically restored. The communication port 22A is closed by rotating in the direction of the arrow R2 shown in FIG. Thereby, the sealed state of the flow path 14 is maintained. In this way, the opening / closing part 23 opens and closes the communication port 22A based on the movement of the pipette tip P relative to the lid member 17.
Here, the reaction reagent is supplied so as to be stored in the flow path 14. As a reaction reagent for PCR, for example, DNA extracted from blood or a pre-purified template DNA, polymerase enzyme, dNTP (deoxynucleotide triphosphate) which is a material of each base, and pH and concentration adjustment Dilute solution or buffer solution.

  And the sealing process which supplies mineral oil as sealing liquid is performed. This is because mineral oil is supplied from the openings 15A and 15B to the inside of the flow path 14 storing the reaction reagent, and on the liquid level of the reaction reagent exposed to the atmosphere inside the flow path 14. Mineral oil is overlaid to seal the inside of the flow path 14. Note that bubbles may be mixed in the flow path 14 when supplying the reaction reagent or mineral oil.

Then, the reaction production | generation process which produces PCR is performed. This consists of a denaturation step, an annealing step and an extension reaction step.
First, a denaturation step is performed in which DNA in the reaction reagent is thermally denatured. This is because the temperature controller 33 controls the temperature state of the reaction unit 4 to be a predetermined temperature (for example, about 90 ° C. to 100 ° C.) over a predetermined time (for example, 5 seconds to 25 seconds). DNA is heat denatured.

  Next, an annealing step for binding (annealing) DNA is performed. This is because the temperature control device 33 controls the temperature state of the reaction unit 4 to be a predetermined temperature (for example, about 50 ° C. to 60 ° C.) over a predetermined time (for example, 15 seconds to 60 seconds). The DNA fragment is bound to the desired gene sequence.

Then, an extension reaction step for synthesizing complementary strands by DNA polymerase is performed. This is because the temperature control device 33 controls the temperature state of the reaction unit 4 to be a predetermined temperature (for example, about 65 ° C. to 75 ° C.) over a predetermined time (for example, 1 minute to 5 minutes). Perform complementary strand synthesis with polymerase. Here, even if the bubbles mixed in the flow path 14 are expanded by heating by heating the reaction section 4 in the denaturation process, annealing process, and extension reaction process, the flow path 14 is sealed by the lid member 17. Therefore, the reaction reagent is prevented from flowing out from the lid member 17, and loss due to evaporation of the reaction reagent is avoided.
Thereafter, it is determined whether or not to continue the series of processes. If the process is continued, the process returns to the denaturing process.

Next, the detection process using the specimen and various reagents for detection is performed. In this method, a specimen prepared by PCR in a reaction generation step and various detection reagents (for example, a nucleic acid probe) are reacted in the detection unit 5 of the reaction vessel 1 by hybridization or the like. That is, the tip of the pipette tip P is caused to enter the flow path 14 as in the above-described reaction reagent supply step. Then, the sample in the flow channel 14 is filled into the pipette tip P. Thereafter, the sample filled in the pipette chip P is supplied to the detection recess 29, and the sample and the detection reagent accommodated in the detection recess 29 are reacted by hybridization or the like.
Thereafter, luminescence detection is performed in which the presence or absence of a labeling substance (for example, a fluorescent substance) previously attached to the specimen or the nucleic acid probe is detected from, for example, the back side of the detection unit 5 of the reaction container 1.
As described above, the biochemical reaction test using the reaction vessel 1 is performed.

According to the reaction container 1 configured as described above, even when the communication port 22A is opened, the communication port 22A is closed when the pipette tip P is separated, whereby the sealed state of the flow path 14 is maintained. Therefore, evaporation and outflow of the reagent accommodated in the flow path 14 and scattering to the outside of the reaction container 1 are suppressed, so that loss of the reaction reagent can be avoided and contamination by the reaction reagent can be prevented.
Here, since the movable portion 25 is urged by the connecting portion 26 so that the communication port 22A is closed, the communication port 22A is closed when the tip of the pipette tip P is separated.
Moreover, since the reagent storage unit 3, the reaction unit 4, and the detection unit 5 are provided for the single base material 2, a series of reaction steps and detection steps can be executed continuously and efficiently.

In addition, this invention is not limited to the said embodiment, A various change can be added in the range which does not deviate from the meaning of this invention.
For example, in the above-described embodiment, the connecting portion 26 of each lid member 17 allows the movable portion 25 to rotate about an axis parallel to the one surface 2A of the base material 2. However, around the axis perpendicular to the one surface 2A of the base material 2 It is good also as rotation possible.
That is, the lid member 50 includes a fixing portion 52 and an opening / closing portion 53 formed by the cuts 51 as shown in FIGS. 5 and 6A and 6B.
The fixed portion 52 has a communication port 52 </ b> A, and the opening / closing portion 53 includes two movable portions 55 and a connection portion 56 that connects each movable portion 55 to the fixed portion 52. The movable portion 55 is formed with a contact recess 55A. And the connection part 56 is supporting each movable part 55 so that rotation around the axis | shaft of the rotating shaft 57 provided virtually is possible. Here, the rotation shaft 57 is provided so as to be substantially perpendicular to the one surface 2A of the substrate 2. Thereby, each movable part 55 is urged | biased by the connection part 56 so that it may rotate in the direction which mutually approaches.
As shown in FIG. 6A, the lid member 50 configured as described above rotates each movable portion 55 by bringing the pipette tip P into contact with the contact recess 55A and pushing it toward the opening. Each of the shafts 57 is rotated in the direction of arrow R3 around the axis 57, and the communication port 52A is opened. Then, as shown in FIG. 6B, by removing the pipette tip P, each movable portion 55 is elastically restored and rotated in the direction of the arrow R4, and the communication port 52A is closed.
Even the lid member 50 having such a configuration exhibits the same operations and effects as described above.

Further, the pipette tip P is used as a dispensing means, and the pipette tip P is brought into contact with the contact concave portion 25A of the opening / closing portion 23 and pushed toward the opening 15A, thereby opening and closing the opening / closing portion 23. Although the opening / closing part 23 is closed, the dispensing means may have an opening / closing jig for changing the opening / closing state of the opening / closing part, and the opening / closing state of the opening / closing part may be changed using this opening / closing jig.
That is, as shown in FIGS. 7 and 8A and 8B, the lid member 60 is a jig that is formed in the movable portion 25 and into which the tip end portion of the bar plate-shaped opening / closing jig P ′ is inserted. It has a recess 61 for use. Further, the pipetting tip P and the opening / closing jig P ′ shown in FIG. 8B constitute dispensing means.
Then, as shown in FIG. 8 (a), the lid member 60 is inserted approximately 90 times in the direction of the arrow R5 as shown in FIG. 8 (b) after inserting the opening / closing jig P ′ into the jig recess 61. By rotating it, each movable part 25 is rotated in the direction of arrow R6 around the axis of a rotation shaft 27 (not shown), and the communication port 22A is opened. After that, the lid member 60 causes the communication port 22A to be closed by separating the opening / closing jig P ′ from the jig recess 61.
Even the lid member 60 having such a configuration provides the same operations and effects as described above. Here, a jig recess 61 similar to the lid member 60 may be provided in the lid member 50 described above, and the communication port 52A may be opened and closed using the opening / closing jig P ′.

Moreover, although the lid member includes the fixing portion and the opening / closing portion, the fixing member may not be provided.
In addition, the lid member has a configuration in which the fixing portion and the opening / closing portion are integrally formed by providing a cut, but the fixing portion and the opening / closing portion may be separate members.
Moreover, although the lid member is detachable from the base material, the lid member and the base material may be integrally formed.
Moreover, although the lid member is detachable from the base material, the base material and the frame body may be integrally formed so as not to be detachable.
Moreover, although the container main body is provided with the reagent storage part, the reaction part, and the detection part, what is necessary is just to provide the reaction part at least.
In addition, the container body may be configured to include a plurality of reagent storage units, a plurality of reaction units, and a plurality of detection units, for example, depending on the type and number of reagents, the type and number of samples, and the like.
Further, the base material of the container main body is formed with the reagent containing concave portion, groove portion, and detection concave portion by an injection molding method, but the thickness of the base material may be appropriately changed according to the design, and a cutting method is used. Recesses and grooves may be formed. Furthermore, although plastic is used as the base material of the container body, other materials such as glass may be used as long as they have heat resistance, chemical resistance, moldability, and the like.
Further, the flow path of the reaction part is formed by covering the opening end of the groove part formed in the base material with a film, but the other part formed by using the same material as the base material for the opening part of the groove part. It may be formed by covering with a material or by providing a channel-shaped through-hole communicating with each opening in the substrate.
Further, the reaction unit is not limited to the flow channel-like reaction unit, and may be configured by a recess formed in the base material, similar to the reagent storage recess and the detection recess.
Moreover, although mineral oil is added to the reaction part as a sealing liquid, other solutions may be added as long as the specific gravity is lighter than the reaction reagent.
In addition, the sample DNA or antigen may be fixed in the reaction part or may be held without being fixed.
In addition, a reaction reagent or the like is stored in advance in each reagent storage recess of the reagent storage unit, and a probe nucleic acid is stored in the detection recess of the detection unit. The reagent storage unit is provided in the biochemical reaction apparatus, and this reagent storage unit The reagent reagent may be stored in the reagent container and the probe nucleic acid may be stored in the detector. Here, the reaction reagent or the like may be stored manually without using the reagent storage device.

Further, although the annealing step and the extension reaction step are sequentially executed, the annealing step and the extension reaction step may be executed simultaneously. At this time, the temperature state of the reaction part is controlled by the temperature control device so as to be a predetermined temperature (for example, about 50 ° C. to 70 ° C.) over a predetermined time (for example, 1 minute to 5 minutes). A primer (that is, a DNA fragment) is combined with a desired gene sequence, and complementary strand synthesis is performed with a DNA polymerase.
The PCR may be multiplex PCR. In this multiplex PCR, it is preferable to apply a hot start method in which the execution of the extension reaction step is started after the reaction reagent is at a relatively high temperature in order to suppress the occurrence of primer misannealing and oligomerization.

The biochemical reaction device can be used for various biochemical reactions such as detection of antigen-antibody reaction and DNA reaction.
In the case of antigen detection by antigen-antibody reaction, for example, the presence or absence of a reaction can be detected by adding a reagent containing an antigen in the reaction part in advance and attaching a labeling substance to the antigen or antibody. Here, as the labeling substance, a luminescent substance such as fluorescence is generally used.

In the case of detection of DNA, for example, a nucleic acid probe is prepared in advance in the detection unit, and then the sample DNA is supplied to the well-shaped detection unit and a DNA reaction is performed by a hybridization reaction between the nucleic acid probe and the sample DNA. Can be detected. Further, as the sample DNA, a DNA extracted from blood or the like prepared by the PCR method, the LAMP method or the like can be used. Further, by preparing a plurality of nucleic acids having different sequences as nucleic acid probes, it is possible to detect the sequence of the sample DNA.
Furthermore, the biochemical reaction apparatus can be used for analysis of SNP (Single Nucleotide Polymorphism). At this time, there may be a plurality of probe nucleic acids and substances used for detection thereof, and one of those substances only needs to be labeled.

  Further, as the labeling substance, a substance that specifically acts on the bound probe nucleic acid and sample DNA can be added after the reaction. Such a thing includes an intercalator. Further, the labeling substance here includes indirect substances. That is, a substance that binds to a fluorescent substance or the like may be bound to the sample DNA as a labeling substance, and the fluorescent substance may be added later.

Alternatively, SNP or DNA may be detected by performing a multistep reaction. For example, an invader assay method (Third Wave Technologies Inc. (Madison, Wis., USA)) may be used. This makes it possible to realize SNP analysis.
In this case, there may be a plurality of types of substances such as probe nucleic acids used for detecting the sample DNA, and at least one type of substance is previously placed in the detection unit, and then the sample DNA and other substances are injected simultaneously or sequentially, A reaction may be performed.

The reaction container in one Embodiment of this invention is shown, (a) is a top view, (b) is AA arrow sectional drawing of (a), (c) is BB arrow view of (a). It is sectional drawing. It is a perspective view which shows the cover member of FIG. It is a block diagram which shows the structure of a biochemical reaction apparatus. FIGS. 2A and 2B show an open / closed state of the lid member of FIG. 1, in which FIG. It is a perspective view which shows the other cover member which can apply this invention. FIGS. 6A and 6B are plan views showing an open / closed state of the lid member of FIG. 5, in which FIG. 5A is an open state and FIG. 5B is a closed state. Similarly, it is a perspective view which shows the other cover member which can apply this invention. FIGS. 8A and 8B show the open / closed state of the lid member in FIG. 7, where FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Reaction container 2 Base material 3 Reagent storage part 4 Reaction part 5 Detection part 15A, 15B Opening part 17,50,60 Lid member 22,52 Fixing part 22A, 52A Communication port 23,53 Opening / closing part 25,55 Movable part 26, 56 Connection 27, 57 Rotating shaft P Pipette tip (dispensing means)
P 'Open / close jig (dispensing means)

Claims (6)

A reaction portion having an opening formed on one surface of the substrate and a lid member covering the opening;
The lid member includes a communication port that communicates with the opening, and an opening / closing portion that opens and closes the communication port based on movement of the dispensing member that is in contact with the lid member with respect to the lid member. A reaction vessel. The lid member has a fixing portion that is fixed to the base material and supports the opening and closing portion;
A plurality of movable parts that can be moved toward and away from each other and that are in contact with the dispensing member, and a plurality of movable parts that are biased in a direction in which the communication port is closed to the fixed part The reaction container according to claim 1, further comprising a connection portion to be connected.   The reaction container according to claim 2, wherein the connection portion includes a rotation shaft that rotates the movable portion around an axis parallel to the one surface.   The reaction container according to claim 2, wherein the connection portion includes a rotation shaft that rotates the movable portion about an axis perpendicular to the one surface.   The reaction container according to any one of claims 1 to 4, wherein a detection unit capable of optical analysis is provided on a surface of the base material. The reaction container according to any one of claims 1 to 5, wherein a reagent storage section for storing a reaction reagent is provided on the surface of the base material.

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