JP2010057403A - Reaction vessel plate and method for reaction treatment - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide such a reaction vessel plate as to include a bellows for smoothly running a liquid such as a sample liquid in a closed system, and also include a passive valve on the inlet side of a reaction vessel, ensuring negative pressure generated by the bellows' restorative force not to be applied onto the passive valve. <P>SOLUTION: One end of a degassing part 19 is connected to each of reaction vessels 5, the other end of the degassing part 19 being connected to the degassing flow channel 21 of the reaction vessel. One end of the flow channel 21 communicates with the connective port of a changeover valve 63. One end of an atmosphere releasing flow channel 26 communicates with the connective port of the changeover valve 63, the other end communicating with the outside of the reaction vessel plate 1. The changeover valve 63 is designed to connect a syringe 51 and the main flow channel 21 to each other and also connect the degassing flow channel 21 and the atmosphere releasing flow channel 26 to each other. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  INDUSTRIAL APPLICABILITY The present invention relates to a reaction vessel plate suitable for performing various analyzes and analyzes in the field of medical analysis and chemistry in the field of biological analysis, biochemical analysis, or chemical analysis in general, and a method for processing the reaction vessel plate. The present invention relates to a reaction processing method.

A micro multi-chamber apparatus is used as a small reaction apparatus used for biochemical analysis or normal chemical analysis. As such an apparatus, for example, a microwell reaction vessel plate such as a microtiter plate in which a plurality of wells are formed on a flat substrate surface is used (see, for example, Patent Document 1).
Moreover, as a trace liquid weighing structure capable of quantitatively handling a trace amount of liquid, a first channel and a second channel, a third channel opening in the channel wall of the first channel, A structure having a fourth channel that opens to the channel walls of the two channels and connects the one end of the third channel and the second channel and has a property that the capillary attraction is less likely to work than the third channel. Some are provided (see, for example, Patent Documents 2 and 3). According to the trace liquid weighing structure, after the liquid introduced into the first flow path is drawn into the third flow path, the liquid remaining in the first flow path is removed, and the volume of the third flow path is increased. A corresponding volume of liquid can be weighed into the second flow path.
JP-A-2005-177749 JP 2004-163104 A JP 2005-114430 A Japanese Patent No. 3454717 WO2008 / 096492

When the conventional microwell reaction container plate is used, the upper surface of the reaction container plate is open to the atmosphere. Therefore, foreign matter may enter the sample from the outside, and conversely, the reaction product may contaminate the external environment.
Further, in the trace liquid weighing structure disclosed in Patent Documents 2 and 3, liquid introduction ports are formed at both ends of the first flow path and at both ends of the second flow path. It is possible that the reaction products are open and pollute the external environment through these ports.

  Therefore, the present inventors include a reaction vessel for causing a sample to react, a reaction vessel channel connected to the reaction vessel for circulating a sample, a reaction reagent, etc., and a sample, a reagent, etc. In addition to a sealing container, a sealing container flow path that can be connected to the sealing container, a switching valve for switching and connecting a reaction container flow path or a sealing container flow path to a syringe for sending a liquid or a syringe And the like, and a reaction vessel plate in which a reaction vessel, a reaction vessel channel, a sealed vessel, and a sealed vessel channel are used as a closed system has been proposed (see Patent Document 5). According to the proposed reaction vessel plate, it is possible to prevent foreign substances from entering from the outside of the reaction vessel plate and environmental pollution to the outside.

  In the above reaction container plate, the liquid is circulated in a closed system. For example, when a sample solution contained in a sealed container is sucked with a syringe and injected into a reaction container, a path for circulating air is necessary to relieve pressure changes in the sealed container and the reaction container. Become. In view of this, the present inventors provide an elastic capacity variable portion that allows air to flow between the sealed container and the reaction container, and allows liquid to flow between the sealed container and the syringe or between the syringe and the reaction container. It has been proposed that air can be circulated between the container and the capacity variable portion at the same time when it is circulated. In this case, a metering channel having a certain capacity is arranged at the inlet portion from the reaction vessel channel of the reaction vessel via a passive valve that does not allow liquid to pass until a certain level of pressure is applied. It has also been proposed to control the amount of liquid injected into the reaction vessel to a constant amount by applying a pressure above a certain level in a state filled with liquid.

When a liquid such as a sample solution is sucked from the sealing container with the syringe, air flows from the variable capacity part into the sealing container, and thereby the variable capacity part contracts. Since the contracted capacity variable portion tries to return by elastic force, if the reaction container is connected to the contracted capacity variable portion, a negative pressure is applied to the inside of the reaction container. If a passive valve is provided at the inlet portion of the reaction vessel from the reaction vessel flow path, the passive valve has a weak pressure resistance. The amount of liquid to be injected cannot be accurately controlled.
Therefore, the present invention provides a reaction vessel plate having a stretchable volume variable portion for smoothly flowing a liquid such as a sample solution in a closed system, and having a passive valve on the inlet side of the reaction vessel. The object is to prevent the negative pressure generated by the restoring force of the variable portion from being applied to the passive valve.

  The reaction container plate according to the present invention includes a sealed reaction container, a syringe for feeding a liquid, a switching valve for switching a flow path to be connected, and air and passively contains an internal volume. Changeable, elastic capacity variable part connected to syringe via switching valve, main flow path with one end connected to drain and other end connected to syringe via switching valve, branched from main flow path A metering channel with a predetermined capacity, one end connected to the metering channel and the other end connected to the reaction vessel, having a channel resistance higher than the metering channel, and when liquid is introduced into the main channel and the metering channel A passive valve that does not allow liquid to pass through in the liquid introduction pressure state and the purge pressure state when the liquid in the main channel is discharged to the drain, and allows the liquid to pass in a pressure state greater than the liquid introduction pressure and the purge pressure, and a reaction vessel An air vent part for venting air, and an open air channel with one end leading to the outside of the reaction vessel plate and the other end connected to the air vent part via the switch valve, At the same time as connecting the syringe and the main flow path, the air vent and the air release flow path can be connected.

The reaction processing method using the reaction container plate includes a step of filling the main flow path and the measurement flow path with the liquid to be injected into the reaction container, and the main flow path using the air of the variable volume portion while leaving the liquid in the measurement flow path. The process of purging the liquid in the chamber and the process of injecting the liquid in the measuring channel into the reaction vessel using the air of the variable volume unit are performed in that order with the air vent and the atmosphere open channel connected. It is.
Thus, in the present invention, the step of filling the main channel and the metering channel with the liquid to be injected into the reaction vessel, the liquid in the main channel using the air of the variable capacity portion while leaving the liquid in the metering channel. The step of purging and the step of injecting the liquid in the measuring channel into the reaction vessel using the air of the variable capacity portion can be performed with the air venting portion opened to the atmosphere.

In the reaction vessel plate according to the present invention, the contact angle of the passive valve with respect to the water droplets is preferably 90 degrees or more, and the area of the boundary between the passive valve and the metering channel is preferably 1 to 10000000 μm ² (square micrometer). By doing so, it is difficult for the liquid to enter the passive valve when the liquid is introduced into the main channel and the metering channel, and the introduction pressure when the liquid is introduced into the main channel and the metering channel can be increased.
If the passive valve is composed of multiple flow paths, the “area of the boundary between the passive valve and the metering flow path” is the area of the boundary between each of the multiple flow paths that make up the passive valve. means.

  Examples of the sealed container include a sample container for containing a sample liquid, a sample pretreatment liquid (a liquid for pretreating a sample) or a reagent, or a gene amplification reaction. Includes a gene amplification container.

  The gene amplification reaction includes a PCR method and a LAMP method. Focusing on the PCR method for amplifying DNA, a method of directly performing a PCR reaction from a sample such as blood without pretreatment has also been proposed. In the nucleic acid synthesis method for amplifying a target gene in a sample containing a gene, the gene inclusion body in the sample containing the gene or the sample containing the gene itself is added to the gene amplification reaction solution, The target gene in the sample containing the gene is amplified when the pH of the reaction solution is 8.5 to 9.5 (25 ° C.) (see Patent Document 4).

  The reaction vessel is preferably made of a light transmissive material that can be optically measured from the bottom or from above. Then, the liquid in the reaction vessel can be measured optically without moving to another vessel.

  The reaction container plate and the reaction processing method of the present invention include a step of filling the main channel and the metering channel with the liquid to be injected into the reaction container, the main channel using the air of the variable capacity portion while leaving the liquid in the metering channel. The step of purging the liquid inside and the step of injecting the liquid in the measuring channel into the reaction vessel using the air of the variable volume part are performed in an open state without connecting the air vent part to the variable volume part. Therefore, the negative pressure due to the restoring force of the variable capacity portion is not applied to the passive valve, and the passive valve can be prevented from being broken.

1A and 1B are diagrams showing an embodiment of a reaction vessel plate, where FIG. 1A is a schematic plan view, and FIG. 1B is a cross-sectional view at a position AA in FIG. , A schematic cross-sectional view of the passive valve 17, the air vents 19 and 21, the liquid drain space 29, the sample container 35, and the bellows 53, and FIG. FIG. FIG. 2 is an exploded sectional view showing this embodiment and a schematic exploded perspective view of the switching valve. FIG. 3 is a schematic view showing the vicinity of one reaction vessel of this embodiment, where (A) is a plan view, (B) is a perspective view, and (C) is a cross-sectional view. FIG. 4 is an enlarged view of the sample container housing part and the sample container, (A) is a plan view of the sample container housing part, (B) is a cross-sectional view at the BB position of (A), (C) is a plan view of the sample container, (D) is a cross-sectional view at the C-C position in (C), (E) is a cross-sectional view in which the sample container is arranged in the sample container accommodating portion at the first holding position, (F) ) Is a cross-sectional view in which the sample container is disposed in the sample container housing portion at the second holding position. FIG. 5 is an enlarged view of the reagent container housing part and the reagent container, (A) is a plan view of the reagent container housing part, (B) is a cross-sectional view at the DD position of (A), (C) is a plan view of the reagent container, (D) is a cross-sectional view at the EE position of (C), (E) is a cross-sectional view in which the reagent container is arranged in the reagent container housing portion at the first holding position, (F) ) Is a cross-sectional view in which the reagent container is arranged in the reagent container housing portion at the second holding position. FIG. 6 is an enlarged view of the air suction container accommodating portion and the air suction container, (A) is a plan view of the air suction container accommodating portion, and (B) is an FF position of (A). (C) is a plan view of the air suction container, (D) is a cross-sectional view at the GG position of (C), and (E) is the air suction container accommodating portion. Sectional drawing arrange | positioned in the 1st holding position, (F) is sectional drawing which has arrange | positioned the air suction container in the air suction container accommodating part in the 2nd holding position.
An embodiment of the reaction vessel plate will be described with reference to FIGS.

  The reaction vessel plate 1 includes a plurality of reaction vessels 5 having openings on one surface of the vessel base 3. In this embodiment, a set of 6 × 2 reaction vessels 5 arranged in a staggered pattern is provided for each of the two flow path units 6a and 6b. A reagent 7 and wax 9 are accommodated in the reaction vessel 5.

  The material of the container base 3 including the reaction container 5 is not particularly limited. However, when the reaction container plate 1 is used as disposable, it is preferable that there is a material available at a low cost. As such a material, for example, a resin material such as polypropylene and polycarbonate is preferable. When the substance in the reaction vessel 5 is detected by absorbance, fluorescence, chemiluminescence, bioluminescence or the like, it must be formed of a light transmissive resin so that optical detection can be performed from the bottom side. Is preferred. In particular, when fluorescence detection is performed, the container base 3 is made of a material such as a resin having a low autofluorescence property (a property of generating less fluorescence from itself) and a light transmitting resin, such as polycarbonate. Is preferred. The thickness of the container base 3 is 0.2 to 4.0 mm (millimeters), preferably 1.0 to 2.0 mm. From the viewpoint of low autofluorescence for fluorescence detection, the container base 3 is preferably thinner.

  Referring to FIGS. 1 and 3, the flow path base 11 is disposed on the container base 3 so as to cover the arrangement region of the reaction containers 5. The channel base 11 is made of, for example, PDMS (polydimethylsiloxane) or silicone rubber. The thickness of the channel base 11 is, for example, 1.0 to 5.0 mm. The flow path base 11 has a groove on the joint surface with the container base 3. A main channel 13, a metering channel 15, a passive valve 17, and air vents 19 and 21 are formed by the groove and the surface of the container base 3. The main channel 13, the metering channel 15 and the passive valve 17 constitute a reaction vessel channel. A concave portion 27 disposed on the reaction vessel 5 is also formed on the joint surface of the flow path base 11 with the vessel base 3. In FIG. 1A and FIGS. 3A and 3B, only the groove and the concave portion of the flow path base 11 are illustrated.

  The flow path base 11 may be formed by integral molding including the formation regions of the flow path units 6a and 6b, or may be molded for each flow path unit 6a and 6b. When the flow path base 11 is formed for each of the flow path units 6a and 6b, the planar size of the flow path base 11 is smaller than that in which the flow path base 11 is integrally formed including the formation region of the flow path units 6a and 6b. .

When the channel base 11 is formed of an elastic resin such as PDMS or silicone rubber, the adhesion between the channel base 11 and the container base 3 can be improved by reducing the planar size of the channel base 11. . Thereby, the manufacturing yield of the reaction vessel plate 1 can be improved, and the manufacturing cost can be reduced.
In addition, when the flow path base 11 is formed of a hard resin such as polypropylene or polycarbonate, by reducing the planar size of the flow path base 11, distortion and sink marks (undesired recesses) in the flow path base 11 are formed. Can be reduced. Thereby, the manufacturing yield of the flow path base 11 and by extension, the reaction vessel plate 1 can be improved, and the manufacturing cost can be reduced.

  The main channel 13 is provided for each channel unit 6a, 6b. The main flow path 13 is formed so as to pass through the vicinity of all the reaction vessels 5 in the flow path units 6a and 6b. One end of the main flow path 13 is connected to a flow path 14 formed of a through hole provided in the container base 3. The flow path 14 is connected to a port of a switching valve 63 described later. The other end of the main channel 13 is connected to a liquid drain space 29 formed in the container base 3. The liquid drain space 29 is provided for each flow path unit 6a, 6b. The dimensions of the grooves constituting the main flow path 13 are, for example, a depth of 400 μm (micrometer) and a width of 500 μm. In addition, the main flow path 13 has a predetermined length downstream of the position where the measurement flow path 15 is connected, for example, a 250 μm portion is formed to be narrower than other portions, for example, the width is 250 μm. It is.

  The measuring channel 15 is branched from the main channel 13 and provided for each reaction vessel 5. The end of the measuring channel 15 opposite to the main channel 13 is disposed in the vicinity of the reaction vessel 5. The depth of the groove constituting the measuring channel 15 is 400 μm, for example. The measuring channel 15 has an internal capacity of a predetermined capacity, for example, 2.5 μL (microliter). The width dimension of the part connected to the main flow path 13 of the measurement flow path 15 is larger than the narrow part of the main flow path 13 described above, for example, 500 μm. Thereby, the flow resistance of the main flow path 13 is larger than that of the measurement flow path 15 in the portion where the measurement flow path 15 is branched with respect to the liquid flowing from one end of the main flow path 13. The liquid flowing from one end of the main channel 13 first flows into the metering channel 15, and after the metering channel 15 is filled with the liquid, it flows downstream through the narrowed portion of the main channel 13. It has become.

A passive valve 17 is also provided for each reaction vessel 5. One end of the passive valve 17 is connected to the measuring channel 15. The other end of the passive valve 17 is connected to a recess 27 disposed on the reaction vessel 5 and led to the reaction vessel 5. The passive valve 17 is formed with a dimension that maintains liquid tightness in the reaction vessel 5 in a state where there is no pressure difference between the reaction vessel 5 and the passive valve 17. In this embodiment, the passive valve 17 is composed of a plurality of grooves, and the dimensions of the grooves are, for example, a depth of 10 μm, a width of 20 μm, a pitch of 20 μm, and 13 grooves formed in a width region of 500 μm. Has been. Here, the area of the boundary between the groove constituting the passive valve 17 and the metering flow path 15, that is, the cross-sectional area of the groove constituting the passive valve 17 is 200 μm ² . The recess 27 has a depth of, for example, 400 μm, and the planar shape is a circle smaller than the reaction vessel 5.

  An air vent 19 is provided for each reaction vessel 5. One end of the air vent 19 communicates with the recess 27 disposed on the reaction vessel 5, the other end communicates with the reaction vessel air vent channel 21, and there is no pressure difference between the reaction vessel 5 and the air vent 19. The flow path is formed so as to keep liquid tightness in the reaction vessel 5. In this embodiment, the air vent 19 is composed of a plurality of grooves, and the dimensions of the grooves are, for example, a depth of 10 μm, a width of 20 μm, a pitch of 20 μm, and 13 grooves in a width region of 500 μm. Is formed.

  In this embodiment, two reaction vessel air vent channels 21 are provided in each of the channel units 6a and 6b. A plurality of air vents 19 are connected to each reaction vessel air vent channel 21. The reaction container air vent channel 21 is connected to a channel 22 formed of a through hole provided in the container base 3. The channel 22 is provided for each channel unit 6a, 6b. The flow path 22 is connected to a port of a switching valve 63 described later. The dimensions of the grooves constituting the reaction vessel air vent channel 21 are, for example, a depth of 400 μm and a width of 500 μm.

  The drain space air vent channel 23 is for connecting the liquid drain space 29 to a port of a switching valve 63 described later. One end of the drain space air vent channel 23 is disposed on the liquid drain space 29. The other end of the drain space air vent channel 23 is connected to a channel 24 formed of a through hole provided in the container base 3. The flow path 24 is connected to a port of a switching valve 63 described later. The dimensions of the grooves constituting the drain space air vent channel 23 are, for example, a depth of 400 μm and a width of 500 μm.

  A flow path cover 33 (not shown in FIG. 1A) is disposed on the flow path base 11. The flow path cover 33 is for fixing the flow path base 11 to the container base 3. A through hole is formed in the flow path cover 33 at a position on the reaction vessel 5.

  Referring to FIG. 1 and FIGS. 4 to 6, the sample container storage unit 36, the reagent container storage unit 38, and the air suction container are placed in the container base 3 at a position different from the arrangement region of the reaction container 5 and the drain space 29. The accommodating part 40 is formed. A sample container 35 is accommodated in the sample container accommodating portion 36, a reagent container 37 is accommodated in the reagent container accommodating portion 38, and an air aspirating 39 is accommodated in the air aspirating container accommodating portion 40. The sample container 35, the reagent container 37, and the air suction container 39 constitute a sealed container for the reaction container plate of the present invention.

  As shown in FIG. 4, a sample channel 35 a penetrating from the bottom to the back surface of the sample container housing portion 36 and a sample container air vent penetrating from the front surface to the back surface are disposed in the container base 3 near the sample container housing portion 36. A flow path 35b is formed. Three locking claws 35 c for holding the sample container 35 are arranged on the surface of the container base 3 around the opening of the sample container housing portion 36.

  At the bottom of the sample container housing part 36, a projecting projection channel 35d is provided which projects toward the opening side of the sample container housing part 36. The base end side end of the projection channel 35d is connected to the sample channel 35a. The front end surface of the protruding channel 35d is inclined with respect to the protruding direction of the protruding channel 35d.

  On the surface of the container base 3 in the vicinity of the sample container accommodating portion 36, a protruding second protruding flow path 35e provided to protrude upward is formed. The end portion on the proximal end side of the second protruding flow path 35e is connected to the sample container air vent flow path 35b. The tip surface of the second protrusion channel 35e is inclined with respect to the protruding direction of the second protrusion channel 35e.

  An annular packing 35f is provided on the outer peripheral side surface of the base end portion of the projection flow path 35d and the outer peripheral side surface of the base end portion of the second projection flow path 35e. For example, it is formed of an elastic material such as silicone rubber or PDMS.

  The sample container 35 disposed in the sample container housing portion 36 includes a sample container main space 35g, a sample container air vent channel 35h, and a sample container air vent space 35i.

  The sample container main space 35g is provided so as to penetrate from the upper surface to the lower surface of the sample container main body formed of a resin material such as polypropylene or polycarbonate. The opening on the lower surface side of the sample container main space 35g is sealed with a film 35j (penetrable portion) made of, for example, aluminum and attached to the lower surface of the sample container main body. The opening on the upper surface side of the sample container main space 35g is sealed with a film 35k made of, for example, aluminum attached to the upper surface of the sample container main body.

  The sample container air vent channel 35h is formed by covering a groove formed on the upper surface of the sample container body with a film 35k. The groove for forming the sample container air vent channel 35h is formed by, for example, one or a plurality of pores having a width of 5 to 200 μm and a depth of 5 to 200 μm. This is for keeping the liquid tightness of the sample container main space 35g in a state where there is no pressure difference in the container air vent space 35i.

  The sample container air vent space 35i is formed in a protruding portion provided on the upper side surface of the sample container main body, and is provided so as to penetrate from the upper surface to the lower surface of the protruding portion. The opening on the lower surface side of the sample container air vent space 35i is sealed with a film 35l (second penetrable portion) made of, for example, aluminum, which is attached to the lower surface of the protruding portion. The opening on the upper surface side of the sample container air vent space 35i is sealed with a film 35k.

  A septum 41, which is an elastic member, is formed on the film 35k. The septum 41 is made of, for example, an elastic material such as silicone rubber or PDMS. The septum 41 can be penetrated by a dispensing device having a sharp tip, and the through-hole can be closed by elasticity when the dispensing device is pulled out after penetration. A septum stopper 43 for fixing the septum 41 is disposed on the septum 41. The septum stopper 43 has an opening on the sample container 35. In this embodiment, the reagent 45 is stored in advance in the sample container main space 35g. In FIG. 4, the septum stopper 43 is fixed to the sample container main body by a locking claw provided on the septum stopper 43, but the number of locking claw provided on the septum stopper 43 is arbitrary. The septum stopper 43 may be fixed to the sample container main body by any method. For example, the septum stopper 43 may be fixed to the sample container main body by an adhesive.

  Locking grooves 35m and 35n are formed on the side surface of the sample container main body. The locking grooves 35m and 35n are for holding the sample container 35 in the sample container housing portion 36 at the first holding position or the second holding position by the locking claw 35c. The locking groove 35m is formed below the locking groove 35n, and is for holding the sample container 35 at the first holding position (see (E)). The locking groove 35n is for holding the sample container 35 at the second holding position (see (F)). The locking claw 35c and the locking grooves 35m and 35n constitute a sealed container holding mechanism for the reaction container plate of the present invention. However, the sealing container holding mechanism is not limited to the one constituted by the locking claw 35c and the locking grooves 35m and 35n, and the sample container 35 (sealing container) is held at the first holding position and the second holding position. Any configuration may be used as long as it can be held.

  In the first holding position, as shown in (E), the film 35j and the protruding flow path 35d are arranged to face each other, and the film 35l and the second protruding flow path 35e are arranged to face each other. By pushing the sample container 35 toward the container base 3 from the state of the first holding position, the sample container 35 can be moved to the second holding position.

  At the second holding position, as shown in (F), the tip of the projection channel 35d penetrates the film 35j and is inserted into the sample container main space 35g, and the tip of the second projection channel 35e is the film 35l. And is inserted into the sample container air vent space 35i. In the state of the second holding position, the sample container main space 35g and the sample container channel 35a are connected via the projection channel 35d, and the sample container air vent space 35i and the sample container air vent channel 35b pass through the projection channel 35e. Connected through. At this time, the lower surface of the sample container main body of the sample container 35 is pressed against the packings 35f and 35f. Thereby, the sample container main space 35g and the sample container channel 35a, and the sample container air vent space 35i and the sample container air vent channel 35b are connected with high airtightness. Thereby, liquid leakage and air leakage can be prevented. However, the method for preventing liquid leakage and air leakage is not limited to the provision of the packings 35f and 35f, as long as the method can connect the sample container 35 and the flow paths 35a and 35b with airtightness. Any method may be used.

  If the sample container 35 is arranged at the first holding position, the reaction container plate can be stored in a state where the sample container 35 and the sample container flow path 35a are separated, so that the reagent 45 and the dilution 45 are diluted in the sample container main space 35g. Even if a liquid solid such as water or a powdered solid such as a reagent is enclosed and stored in advance, the liquid or solid does not enter the sample container channel 35a during storage.

  Furthermore, in the first holding position, the sample container main space 35g and the sample container air vent space 35i are sealed, so that liquid such as the reagent 45 and dilution water may be sealed in the sample container main space 35g in advance. The liquid can be prevented from evaporating.

Next, the reagent container 37 and the reagent container storage unit 38 will be described with reference to FIG.
The reagent container housing portion 38 has the same structure as the sample container housing portion 36 described with reference to FIG. That is, a reagent channel 37a, a reagent container air vent channel 37b, a locking claw 37c, a projection channel 37d, a second projection channel 37e, and packings 37f and 37f are provided.

  The reagent container 37 has the same structure as the sample container 35 described with reference to FIG. However, the reagent container 37 does not include the septum 41 as compared with the sample container 35, and includes a cover 47 instead of the septum stopper 43. That is, the reagent container 37 includes a reagent container main space 37g, a reagent container air vent channel 37h, a reagent container air vent space 37i, films 37j, 37k, and 37l, locking grooves 37m and 37n, and a cover 47. . The cover 47 is for preventing the film 37k affixed on the upper surface of the reagent container main body from being damaged. Dilution water 49 is accommodated in the reagent container main space 37g. In FIG. 5, the cover 47 is fixed to the reagent container main body by a locking claw provided on the cover 47, but the number of the locking claw provided on the cover 47 is arbitrary. Further, any method may be used for fixing the cover 47 to the reagent container main body. For example, the cover 47 may be fixed to the reagent container main body with an adhesive.

  Similarly to the sample container 35, the reagent container 37 is also arranged in the reagent container housing portion 38 at the first holding position (see (E)) and the second holding position (see (F)). The connection between the reagent container 37 and the reagent container channel 37a and the reagent container air vent channel 37b is the same as the connection between the sample container 35, the sample container channel 35a and the reagent container air vent channel 35b described with reference to FIG. It is.

  If the reagent container 37 is arranged at the first holding position, the reaction container plate can be stored in a state where the reagent container 37 and the reagent container flow path 37a are separated, so that the reagent or dilution water is stored in the reagent container main space 37g. Even if a liquid such as 49 or a powdery solid such as a reagent is enclosed and stored in advance, the liquid or solid does not enter the reagent container channel 37a during storage.

  Furthermore, in the first holding position, the reagent container main space 37g and the reagent container air vent space 37i are sealed, so that a liquid such as a reagent or dilution water 49 may be sealed in the reagent container main space 37g in advance. The liquid can be prevented from evaporating.

Next, the air suction container 39 and the air suction container housing 40 will be described with reference to FIG.
The air suction container container 40 has the same structure as the reagent container container 38 described with reference to FIG. That is, an air suction channel 39a, an air suction container air vent channel 39b, a locking claw 39c, a projection channel 39d, a second projection channel 39e, and packings 39f and 39f are provided.

  The air suction container 39 has the same structure as the reagent container 37 described with reference to FIG. That is, the air suction container 39 includes an air suction container main space 39g, an air suction container air vent channel 39h, an air suction container air vent space 39i, films 39j, 39k, 39l, and locking grooves 39m, 39n. , And a cover 47. Liquid and solid are not accommodated in the air suction container main space 39g and are filled with air.

  Similarly to the sample container 35 and the reagent container 37, the air suction container 39 is also arranged in the air suction container housing portion 40 at the first holding position (see (E)) and the second holding position (see (F)). The The connection between the air suction container 39, the air suction container channel 39a, and the air suction container air vent channel 39b is the same as that of the sample container 35, the sample container channel 35a, and the reagent container air vent flow described with reference to FIG. This is the same as the connection of the path 35b.

  1 and 2, the syringe 51 is provided on the surface of the container base 3 at a position different from the arrangement region of the reaction vessel 5, the drain space 29, and the container accommodating portions 36, 38, and 40. Yes. The syringe 51 is formed by a cylinder 51a formed in the container base 3, a plunger 51b disposed in the cylinder 51a, and a cover body 51d. A syringe channel 51c penetrating from the discharge port provided at the bottom of the cylinder 51a to the back surface is formed in the container base 3.

  The cover body 51d has flexibility in the sliding direction of the plunger 51b, and is connected to the cylinder 51a and the plunger 51b. The cover body 51d is for blocking the portion of the inner wall of the cylinder 51a that the plunger 51b contacts with the atmosphere outside the cylinder 51a while maintaining airtightness, and is surrounded by the cylinder 51a, the plunger 51b, and the cover body 51d. A sealing space 51e is formed. The end of the cover body 51d on the side connected to the cylinder 51a is fixed to the upper end of the cylinder 51a with a cylinder cap 51f while ensuring airtightness. The end of the cover body 51d on the side connected to the plunger 51b is connected to the upper surface of the plunger 51b with an adhesive while ensuring airtightness. However, the method and position of connecting the cover body 51d to the cylinder 51a and the plunger 51b are not limited to this.

  Thus, the cover body 51d is connected to the cylinder 51a and the plunger 51b to form a sealed space 51e surrounded by the cylinder 51a, the plunger 51b, and the cover body 51d, so that the space between the cylinder 51a and the plunger 51b is formed. Therefore, it is possible to prevent foreign substances from entering and environmental contamination of the liquid to the outside. Since the cover body 51d is flexible in the sliding direction of the plunger 51b, the sliding operation of the plunger 51b is possible.

  In this embodiment, the plunger 51b and the cover body 51d are formed by separate members, but the plunger and the cover body may be integrally formed. An example of the integrally formed plunger and cover material is silicone rubber.

  The container base 3 is also provided with a bellows (capacity variable portion) 53 at a position different from the arrangement region of the reaction container 5, the drain space 29, the containers 35, 37, 39 and the syringe 51. The bellows 53 is in close contact with the surface of the container bottom 55, and its internal capacity is passively variable by expanding and contracting. The bellows 53 is disposed, for example, in a through hole 53 a provided in the container base 3.

  A container bottom 55 is attached to the back surface of the container base 3 at a position different from the arrangement region of the reaction containers 5. The container bottom 55 is provided with a flow path for forming two open air flow paths 26 corresponding to the flow path units 6a and 6b. One end of each open air channel 26 communicates with a port of a switching valve 63, which will be described later, and the other end 26a faces the bonding surface of the container base 3 and the container bottom 55 or the bonding surface of the container base 3 and the flow channel base 11. ing. The portion of each atmosphere opening channel 26 facing the other end 26 a is opened to the atmosphere by providing a gap or the like that leads to the outside of the reaction vessel plate 1. The air release channel 26 is connected to the air vent channels 21 and 23 by a switching valve 63, and discharges the gas pushed out from these channels 21 and 23 to the outside of the reaction vessel plate 1. The switching valve 63 can also connect the atmosphere release flow path 26 to any flow path so that the reaction vessel plate 1 is completely sealed.

  In addition, an air vent channel 53b is provided in the container bottom 55 at a position where it communicates with the bellows 53, and further, an arc-shaped channel 55a (see FIG. 1 (A)) formed of an arc-shaped groove or a channel. 14, 22, 24, 35a, 35b, 37a, 37b, 39a, 39b, 51c, 53b are provided with channels for guiding them to a predetermined port of the switching valve 63.

The container base 3 container and the bottom 55 are provided with a syringe air vent channel 53 c having one end connected to the sealed space 51 e and the other end made a bellows 53. Illustration of the syringe air vent channel 53c in FIG. 1 (A) is omitted.
Thus, since the syringe air vent channel 53c having one end connected to the sealed space 51e and the other end bellows 53 is provided, the sealed space 51e is blocked from the atmosphere outside the reaction vessel plate 1. When the plunger 51b slides, the pressure change in the sealed space 51e accompanying the change in the internal capacity of the sealed space 51e can be alleviated, and the plunger 51b can be slid smoothly.

  A rotary switching valve 63 including a disc-shaped seal plate 57, a rotor upper 59, and a rotor base 61 is provided on the surface of the container bottom 55 opposite to the container base 3. The switching valve 63 is attached to the container bottom 55 by a lock 65.

The seal plate 57 is provided in the vicinity of the peripheral edge thereof, and the through hole 57a connected to any one of the flow paths 14, 35a, 37a, 39a and the through grooves 57b, 57d provided on the inner concentric circles. 57e and a through hole 57c provided in the center and connected to the syringe flow path 51c.
The rotor upper 59 has a through hole 59a provided at the same position as the through hole 57a of the seal plate 57 and grooves 59b, 59d, 59e provided on the surface corresponding to the through grooves 57b, 57d, 57e of the seal plate 57. And a through hole 59c provided in the center.
The rotor base 61 is provided with a groove 61a on its surface for connecting two peripheral holes 59a and 59c disposed at the center and the peripheral portion of the rotor upper 59.
For convenience, FIG. 1A also shows the grooves 59b, 59d, 59e, and 61a (see thick lines).

  In the position of the switching valve 63 shown in FIG. 1A, the syringe flow path 51c is not connected to any of the flow paths 14, 35a, 37a, 39a, and the air vent flow path 53b is also connected to the flow paths 22, 24. , 35b, 37b, 39b are not connected to the initial position. The switching valve 63 configured as described above can connect the syringe 51 and the main channel 21 and simultaneously connect the reaction vessel air vent channel 21, the drain space air vent channel 23, and the atmosphere release channel 26. it can.

  In the reaction vessel plate 1, the passive valve 17 is formed so as to maintain the liquid tightness of the reaction vessel 5 with no pressure difference between the reaction vessel 5 and the passive valve 17. The air vent 19 is also formed so as to maintain the liquid tightness of the reaction vessel 5 in the state where there is no pressure difference between the reaction vessel 5 and the air vent 19. The main flow path 13 of the reaction container flow path, the liquid drain space 29 to which the main flow path 13 is connected, and the drain space air vent flow path 23 can be sealed by switching the switching valve 63. The containers 35, 37, and 39 are sealed with a septum 41 or a film 47. The flow paths 35a, 35b, 37a, 37b, 39a, 39b connected to the containers 35, 37, 39 can be sealed by switching the switching valve 63. One end of the air vent channel 53b is connected to the bellows 53 and sealed. Thus, the container and flow path inside the reaction container plate 1 are formed in a closed system. Even when the air vent channel 53b is connected to the atmosphere outside the reaction vessel plate 1 without the bellows 53, the air vent channel 53b is switched to the reaction vessel by switching the switching valve 63. Since it can block | block with the flow paths other than the container inside the plate 1 and the air vent flow path 53b, the container and flow path in which a liquid is accommodated or a liquid flows can be made into a closed system.

FIG. 7 is a cross-sectional view showing a reaction processing apparatus for processing the reaction container plate 1 shown in FIG. Since the structure of the reaction vessel plate 1 is the same as that shown in FIG.
The reaction processing apparatus includes a temperature adjustment mechanism 67 for adjusting the temperature of the reaction vessel 5, a syringe drive unit 69 for driving the syringe 51, and a switching valve drive unit 71 for switching the switching valve 63.

  8 to 14 are plan views for explaining the operation of introducing the sample liquid from the sample container 35 into the reaction container 5. This operation will be described with reference to FIGS. 1 and 8 to 14.

  In a state where the sample container 35, the reagent container 37, and the air suction container 39 are held at the first holding position, a dispensing device having a sharp point (not shown) is used to penetrate the septum 41 on the sample container 35, for example. Dispense 5 μL of the sample solution into the sample container 35. After dispensing the sample solution, pull out the dispensing device. The through hole of the septum 41 when the dispensing instrument is pulled out is closed by the elasticity of the septum 41.

  The sample container 35, the reagent container 37, and the air suction container 39 held at the first holding position are pushed into the container base 3 and moved to the second holding position. Thereby, the sample container main space 35g and the sample container channel 35a, the sample container air vent space 35i and the sample container air vent channel 35b, the reagent container main space 37g and the reagent container channel 37a, the reagent container air vent space 37i and the reagent The container air vent channel 37b, the air suction container main space 39g, the air suction container channel 39a, the air suction container air vent space 39i, and the air suction container air vent channel 39b are connected with airtightness. Before dispensing the sample liquid into the sample container 35, the sample container 35, the reagent container 37, and the air suction container 39 are pushed into the container base 3 and moved to the second holding position, and then the sample container 35 is moved. The sample solution may be dispensed inside.

The syringe drive unit 69 is connected to the plunger 51 b of the syringe 51, and the switching valve drive unit 71 is connected to the switching valve 63.
As shown in FIG. 8, the switching valve 63 is rotated from the state of the switching valve 63 shown in FIG. 1A, and the sample channel 35a and the syringe channel 51c are connected by the groove 61a. At the same time, the sample container air vent channel 35b is connected to the air vent channel 53b via the channel 55a by the groove 59e. For example, 45 μL of the reagent 45 is accommodated in the sample container 35.

  The plunger 51b of the syringe 51 is slid to mix the sample liquid and the reagent 45 in the sample container 35. Thereafter, the mixed solution in the sample container 35 is sucked into the flow path in the switching valve 63, the syringe flow path 51c, and the syringe 51 by, for example, 10 μL. At this time, since the sample container 35 is connected to the bellows 53 via the air vent channels 35e, 35d, 35b, the switching valve 63 and the air vent channel 53b, the gas capacity in the sample container 35 is changed. The bellows 53 expands and contracts. Further, the sliding of the plunger 51b deforms the cover body 51d, and the internal capacity of the sealed space 51e (see FIG. 1C) changes. Since the sealed space 51e is connected to the bellows 53 via the syringe air vent channel 53c, the bellows 53 expands and contracts even when the internal capacity of the sealed space 51e changes. Even in the operation process described below, the bellows 53 expands and contracts with the change in the internal capacity of the sealed space 51e due to the sliding of the plunger 51b.

  As shown in FIG. 9, the switching valve 63 is rotated to connect the reagent channel 37a and the syringe channel 51c by the groove 61a. At the same time, the reagent container air vent channel 37b is connected to the air vent channel 53b via the channel 55a by the groove 59d. For example, 190 μL of dilution water 49 is accommodated in the reagent container 37. The mixed solution sucked into the flow path in the switching valve 63, the syringe flow path 51 c and the syringe 51 is injected into the reagent container 37, and the syringe 51 is slid to mix the mixed liquid and the dilution water 49. For example, the diluted mixed solution is sucked into the flow path in the switching valve 63, the syringe flow path 51c, and the syringe 51, for example, half, that is, 100 μL. At this time, since the reagent container 37 is connected to the bellows 53 via the air vent channels 37e, 37d, 37b, the switching valve 63 and the air vent channel 53b, the gas capacity in the reagent container 37 is changed. The bellows 53 expands and contracts.

  As shown in FIG. 10, the switching valve 63 is rotated to connect the flow path 14 connected to one end of the main flow path 13 of the flow path unit 6a and the syringe flow path 51c by the groove 61a. At the same time, the flow path 22 connected to the liquid drain space 29 and the flow path 24 connected to the reaction vessel air vent flow path 21 are connected to the atmosphere open flow path 26 by the groove 59b. The syringe 51 is driven in the extruding direction, and the diluted mixed solution sucked into the flow path in the switching valve 63, the syringe flow path 51c, and the syringe 51 is sent to the main flow path 13 of the flow path unit 6a. In the flow path unit 6 a, the diluted mixed liquid injected from the flow path 14 side to the main flow path 13 fills the measurement flow path 15 in order from the flow path 14 side and reaches the liquid drain space 29 as indicated by the embossments and arrows. To do. The passive valve 17 allows only a gas to pass therethrough and does not allow the diluted mixture to pass through in an introduction pressure state when the diluted mixture is introduced into the main channel 13 and the metering channel 15. The gas in the measurement flow path 15 moves into the reaction vessel 5 through the passive valve 17 as the measurement flow path 15 is filled with the diluted mixed liquid. By this operation, the gas pushed out from the reaction vessel 5, the main flow channel 21, and the liquid drain space 29 is discharged to the outside of the reaction vessel plate 1 through the atmosphere release flow channel 26 (see the white arrow).

  As shown in FIG. 11, the switching valve 63 is rotated to connect the air suction flow path 39a and the syringe flow path 51c by the groove 61a. At the same time, the air suction container air vent channel 39b is connected to the air vent channel 53b by the groove 59b. The syringe 51 is driven to the suction side, and the gas in the air suction container 39 is sucked into the flow path in the switching valve 63, the syringe flow path 51 c, and the syringe 51. At this time, since the air suction container 39 is connected to the bellows 53 via the air vent channels 39e, 39d, 39b, the switching valve 63 and the air vent channel 53b, the pressure inside the air suction container 39 is reduced. As a result, the bellows 53 contracts (see the white arrow).

  As shown in FIG. 12, the switching valve 63 is rotated to connect the flow path 14 and the syringe flow path 51c to the flow path unit 6a through the groove 61a and flow through the groove 59b as in the connection state of FIG. The paths 22 and 24 are connected to the atmosphere open flow path 26. The syringe 51 is driven in the extrusion direction, and the flow path in the switching valve 63, the syringe flow path 51c, and the gas in the syringe 51 are sent to the main flow path 13 of the flow path unit 6a to purge the diluted mixed liquid in the main flow path 13 (Refer to the white arrow). In this purge pressure state, the passive valve 17 does not pass the diluted mixed solution, so that the diluted mixed solution remains in the measuring flow path 15 (see embossing). The purged diluted liquid mixture is accommodated in the liquid drain space 29. The gas pushed out from the liquid drain space 29 by this operation is discharged to the outside of the reaction vessel plate 1 through the atmosphere opening flow path 26 (see the white arrow).

  As shown in FIG. 13, the switching valve 63 is rotated to connect the air suction flow path 39a and the syringe flow path 51c by the groove 61a, and the air suction container is vented by the groove 59b, as in the connection state of FIG. The flow path 39b and the air vent flow path 53b are connected. The syringe 51 is driven to the suction side, and the gas in the air suction container 39 is sucked into the flow path in the switching valve 63, the syringe flow path 51 c, and the syringe 51. At this time, the bellows 53 contracts as described with reference to FIG. 11 (see the white arrow).

  As shown in FIG. 14, the switching valve 63 is rotated to connect the flow path 14 and the syringe flow path 51c to the flow path unit 6a through the groove 61a. At the same time, the flow path 24 and the air release flow path 26 are connected by the groove 59b. This connection state is different from the connection state shown in FIGS. 10 and 12 in that the liquid drain space 29 to which the downstream end of the main flow path 13 of the flow path unit 6a is connected is not connected to the air release flow path 26. Different. The syringe 51 is driven in the pushing direction. Since the downstream end of the main flow path 13 is closed without being connected to the atmosphere open flow path 26, the inside of the main flow path 13 is pressurized to be larger than the liquid introduction pressure and the purge introduction pressure. As a result, the diluted mixed solution in the metering channel 15 is injected into the reaction vessel 5 through the passive valve 17. After the diluted mixed liquid is injected into the reaction vessel 5, a part of the gas in the main channel 13 flows into the reaction vessel 5 through the metering channel 15 and the passive valve 17 and is pushed out of the reaction vessel 5. Is discharged to the outside of the reaction vessel plate 1 through the atmosphere opening channel 26 (see white arrow). Thereby, the operation | movement which introduce | transduces a diluted liquid mixture into the reaction container 5 with respect to the flow-path unit 6a is completed.

Subsequently, an operation of introducing the diluted mixed solution into the reaction vessel 5 with respect to the flow path unit 6b will be described.
As shown in FIG. 15, the switching valve 63 is rotated to connect the reagent flow path 37a and the syringe flow path 51c by the groove 61a, and the reagent container air vent flow path 37b by the groove 59d, as in the connection state of FIG. The air vent channel 53b is connected. For example, 100 μL of all of the diluted mixed solution remaining in the reagent container 37 is sucked into the flow path in the switching valve 63, the syringe flow path 51 c, and the syringe 51. At this time, since the reagent container 37 is connected to the bellows 53 in the same manner as described in the connection state of FIG. 9, the bellows 53 expands and contracts as the gas capacity in the reagent container 37 changes.

  As shown in FIG. 16, the switching valve 63 is rotated, and the flow path 14 connected to one end of the main flow path 13 of the flow path unit 6b and the syringe flow path 51c are connected by the groove 61a. At the same time, the flow path 22 connected to the liquid drain space 29 and the flow path 24 connected to the reaction vessel air vent flow path 21 are connected to the atmosphere open flow path 26 by the groove 59b. The syringe 51 is driven in the extrusion direction, and the diluted mixed solution sucked into the flow path in the switching valve 63, the syringe flow path 51c, and the syringe 51 is sent to the main flow path 13 of the flow path unit 6b. Similar to the description of the flow path unit 6a in the connected state of FIG. 10, in the flow path unit 6b, the diluted mixed liquid injected from the flow path 14 side to the main flow path 13 flows as shown by the embossments and arrows. The metering flow path 15 is filled in order from the path 14 side and reaches the liquid drain space 29. The gas in the main channel 13, the metering channel 15, and the liquid drain space 29 is pushed out by the diluted mixed solution, and is discharged to the outside of the reaction vessel plate 1 through the atmosphere release channel 26 (see the white arrow).

  As shown in FIG. 17, the switching valve 63 is rotated to connect the air suction flow path 39a and the syringe flow path 51c by the groove 61a, and the air suction container is vented by the groove 59b, as in the connection state of FIG. The flow path 39b and the air vent flow path 53b are connected. The syringe 51 is driven to the suction side, and the gas in the air suction container 39 is sucked into the flow path in the switching valve 63, the syringe flow path 51 c, and the syringe 51. At this time, the bellows 53 contracts as described with reference to FIG. 11 (see the white arrow).

  As shown in FIG. 18, the switching valve 63 is rotated to connect the flow path 14 and the syringe flow path 51c to the flow path unit 6b through the groove 61a and flow through the groove 59b as in the connection state of FIG. The paths 22 and 24 are connected to the atmosphere open flow path 26. The syringe 51 is driven in the pushing direction, and the flow path in the switching valve 63, the syringe flow path 51c, and the gas in the syringe 51 are sent to the main flow path 13 of the flow path unit 6b to purge the diluted mixed liquid in the main flow path 13 (Refer to the white arrow). In this purge pressure state, the passive valve 17 does not pass the diluted mixed solution, so that the diluted mixed solution remains in the measuring flow path 15 (see embossing). The purged diluted liquid mixture is accommodated in the liquid drain space 29. Further, when the diluted mixed liquid is injected into the liquid drain space 29, the gas in the channel from the liquid drain space 29 to the atmosphere opening channel 26 sequentially moves to the atmosphere opening channel 26 side (see white arrows). ).

  As shown in FIG. 19, the switching valve 63 is rotated to connect the air suction flow path 39a and the syringe flow path 51c by the groove 61a, and the air suction container is vented by the groove 59b, as in the connection state of FIG. The flow path 39b and the air vent flow path 53b are connected. The syringe 51 is driven to the suction side, and the gas in the air suction container 39 is sucked into the flow path in the switching valve 63, the syringe flow path 51 c, and the syringe 51. At this time, the bellows 53 contracts as described with reference to FIG. 11 (see the white arrow).

As shown in FIG. 20, the switching valve 63 is rotated to connect the flow path 14 and the syringe flow path 51c to the flow path unit 6b through the groove 61a. At the same time, the flow path 24 and the air release flow path 26 are connected by the groove 59b. This connection state is different from the connection state shown in FIGS. 16 and 18 in that the liquid drain space 29 to which the downstream end of the main flow path 13 of the flow path unit 6b is connected is not connected to the air release flow path 26. Different. The syringe 51 is driven in the pushing direction. Since the downstream end of the main flow path 13 is not connected to the atmosphere open flow path 26, the inside of the main flow path 13 is pressurized to be larger than the liquid introduction pressure and the purge introduction pressure. As a result, the diluted mixed solution in the metering channel 15 is injected into the reaction vessel 5 through the passive valve 17. After the diluted mixed solution is injected into the reaction vessel 5, a part of the gas in the main channel 13 flows into the reaction vessel 5 through the metering channel 15 and the passive valve 17. The gas pushed out of the reaction vessel 5 is discharged to the outside of the reaction vessel plate 1 through the atmosphere opening channel 26 (see the white arrow).
Thereby, the operation | movement which introduce | transduces a diluted liquid mixture into the reaction container 5 with respect to the flow-path unit 6b is completed.

After the switching valve 63 is connected as shown in FIG. 1 and the container, flow path and drain space inside the reaction container plate 1 are sealed, the reaction container 5 is heated by the temperature control mechanism 67 to melt the wax 9. Thereby, the diluted mixed solution injected into the reaction vessel 5 enters under the wax 9, and the diluted mixed solution and the reagent 7 are mixed and reacted. Thus, according to the reaction container plate 1, the reaction process can be performed in a closed system.
Before injecting the diluted mixed solution into the reaction vessel 5, the reaction vessel 5 is heated by the temperature control mechanism 67 to melt the wax 9, and the wax 9 is injected when the diluted mixed solution is injected into the reaction vessel 5. May be melted. In this case, the diluted mixed solution injected into the reaction vessel 5 immediately enters under the wax 9, and the diluted mixed solution and the reagent 7 are mixed and reacted. Even if the connection state of the switching valve 63 is the state shown in FIG. 14, the sealed system is secured by the bellows 53. If the switching valve 63 is brought into the connection state shown in FIG. 1 after the diluted mixed solution is injected, the container, flow path and drain space inside the reaction container plate 1 can be sealed. Here, the timing for switching the switching valve 63 to the connected state in FIG. 1 may be any timing from immediately after the injection of the diluted mixed solution to the end of the reaction of the diluted mixed solution and the reagent 7, or the diluted mixed solution and the reagent 7. It may be after the completion of the reaction.
Thus, according to the reaction vessel plate 1, the reaction process can be performed in a closed system, and a closed system can be formed before and after the reaction process.

In this embodiment, the diluted mixed solution is introduced into both the flow path units 6a and 6b, but the diluted mixed liquid may be introduced into only one of the flow path units 6a and 6b.
Further, the number of flow path units may be three or more.
Moreover, you may vary the sample liquid density | concentration of the dilution liquid mixture inject | poured by the flow-path units 6a and 6b. For example, in the above-described operation described with reference to FIGS. 8 to 20, the operation described with reference to FIG. 14 (operation for injecting the diluted mixed solution into the reaction vessel 5 of the flow path unit 6a) and FIG. 9 and the operation described above (operation for sucking the diluted mixed solution to be injected into the flow path unit 6b into the syringe 51), the sample liquid and the reagent mixed solution are placed in the syringe 51 in the connected state shown in FIG. An operation of sucking only a fixed amount and an operation of mixing the mixed liquid in the syringe 51 and the diluted mixed liquid in the reagent container 37 in the connected state of FIG. 14 may be added. Thereby, the diluted liquid mixture whose sample liquid concentration is higher than the diluted liquid mixture injected into the reaction container 5 of the flow path unit 6a can be injected into the reaction container 5 of the flow path unit 6b.

  Further, in this embodiment, the grooves for forming the flow paths 13, 15, 17, 19, 21, 23 are formed in the flow path base 11, but the present invention is not limited to these, A groove for forming all or part of the flow path may be formed on the surface of the container base 3.

  FIG. 21 is a schematic cross-sectional view showing, in an enlarged manner, the vicinity of the reaction vessel of another embodiment of the reaction vessel plate. This embodiment is the same as the above-described embodiment described with reference to FIGS. 1 to 6 except that a flow path spacer is disposed between the container base and the flow path base.

  A flow path spacer 73 is disposed on the container base 3 so as to cover the arrangement region of the reaction containers 5, and a flow path base 11 and a flow path cover 33 are further disposed in that order. The channel spacer 73 is made of, for example, PDMS or silicone rubber. The thickness of the channel spacer 73 is, for example, 0.5 to 5.0 mm. The flow path spacer 73 includes a convex portion 75 protruding into the reaction container 5 for each reaction container 5. The convex portion 75 has a substantially trapezoidal cross section. For example, the base end has a width of 1.0 to 2.8 mm, the tip has a width of 0.2 to 0.5 mm, and the tip has a base end. It is thinner than Further, the surface of the convex portion 75 is subjected to super water repellent treatment. However, the surface of the convex portion 75 may not necessarily be subjected to the water repellent treatment.

Further, the flow path spacer 73 is provided with a passive valve 77 formed of a through hole penetrating from the tip end portion of the convex portion 75 to the opposite surface for each position where the convex portion 75 is formed. The inner diameter of the passive valve 77 is, for example, 500 μm. The opening on the flow path base 11 side of the passive valve 77 is connected to the passive valve 17 of the flow path base 11. In this embodiment, the channel base 11 is not provided with the recess 27 as compared with the above-described embodiment described with reference to FIGS.
Further, the flow path spacer 73 is also provided with a reaction container air vent flow path 79 including a through hole for communicating the air vent portion 19 of the flow path base 11 and the reaction container 5.

  Although not shown, the channel spacer 73 includes both end portions of the main channel 13, end portions of the reaction vessel air vent channel 21 on the air drain space 31 side, and both ends of the drain space air vent channels 23 and 25. The part is provided with a through hole, and the flow paths 13, 21, 23, 25 are connected to the containers 29, 31 or the flow paths 24, 25 b provided in the container base 3.

  In this embodiment, the end of the passive valve 77 opposite to the passive valve 15 (the other end of the passive valve) is disposed at the tip of a convex portion 75 formed to protrude from the inner upper surface of the reaction vessel 5. Then, the liquid injected into the reaction vessel 5 through the passive valves 15 and 77 can be easily dropped into the reaction vessel 5.

  Further, when the liquid is discharged from the tip of the convex portion 75 through the passive valve 77, the tip of the convex portion 75 is reacted so that the droplet formed at the tip of the convex portion 75 contacts the side wall of the reaction vessel 5. If it arrange | positions in the side wall vicinity of the container 5, a liquid can be inject | poured in the reaction container 5 along the side wall of the reaction container 5, and a liquid can be inject | poured in the reaction container 5 more reliably. However, the formation position of the convex portion 75 may be a position where a droplet formed at the tip of the convex portion 75 does not contact the side wall of the reaction vessel 5.

FIG. 22 is a schematic cross-sectional view showing, in an enlarged manner, the vicinity of the reaction vessel of another embodiment of the reaction vessel plate.
This embodiment further includes a protrusion 81 inside the reaction vessel 5 as compared to the embodiment described with reference to FIG. The tip of the protrusion 81 is disposed below the tip of the protrusion 75. Thereby, it becomes easy to guide the droplet formed at the tip of the convex portion 75 into the reaction vessel 5. In particular, it is particularly effective if a hydrophilic treatment is applied to at least the tip surface of the protrusion 81.

FIG. 23 is a schematic cross-sectional view showing, in an enlarged manner, the vicinity of a reaction vessel of another embodiment of the reaction vessel plate.
In this embodiment, compared with the embodiment described with reference to FIG. 22, the stepped portion 83 formed on the side wall of the reaction vessel 5 and the upper surface of the reaction vessel 5 are formed on the upper surface of the stepped portion 83 with a gap. Further provided is a convex ridge 85. The step part 83 and the protruding line part 85 are formed in an annular shape when viewed from above. The tip of the ridge 85 is disposed at a distance from the side wall of the reaction vessel 5.
Since the tip of the ridge 85 is spaced from the upper surface and side surface of the reaction vessel 5, the liquid contained in the reaction vessel 5 reaches the upper surface of the reaction vessel 5 along the side wall of the reaction vessel 5. Can be prevented. This effect is particularly effective when a water-repellent treatment is performed on at least the tip of the ridge 85.

The configuration provided with the stepped portion 83 and the protruding strip portion 85 shown in FIG. 23 can also be applied to the embodiment shown in FIG.
Further, in each of the embodiments described with reference to FIG. 21, FIG. 22 or FIG. 23, grooves for forming the flow paths 13, 15, 17, 19, 21, 23 are formed in the flow path base 11. However, the present invention is not limited to this, and the grooves for forming all or part of the flow paths are the surface of the flow path spacer 73 on the flow path base 11 side, the container base 3 of the flow path spacer 73. It may be formed on either the side surface or the surface of the container base 3.

  The embodiments of the present invention have been described above, but the present invention is not limited to these, and the shape, material, arrangement, number, dimensions, flow path configuration, etc. are examples, and are described in the claims. Various modifications are possible within the scope of the present invention.

For example, the bellows 53 connected to the air vent channel 53b may have another structure as long as the capacity is a variable capacity member whose internal capacity is passively variable. Examples of such a structure include a bag-like material made of a flexible material and a syringe-like material.
Further, the capacity variable member such as the bellows 53 may not necessarily be provided.
In addition, if the containers 35, 37, and 39 do not contain a liquid such as a reagent in advance, it is not always necessary to provide the flow paths 35e, 37e, and 39e made of pores in a part of the air vent flow path.

Further, in the above embodiment, the air vent channels 35b, 37b, 39b provided in communication with the containers 35, 37, 39 as sealing containers are connected to the air vent channel 53b via the switching valve 63. However, the air vent channel provided in communication with the sealing container may be directly connected to the outside of the reaction container plate or a capacity variable part such as the bellows 53.
Moreover, you may use the cap which can be opened and closed as a sealing method of the containers 35, 37, and 39.

Moreover, in the said Example, although the container base 3 is formed with one component, the container base may be formed with several components.
The reagent in the reaction vessel 5 may be a dry reagent.
In addition, the reagent may not be stored in advance in the sample container 35 or the reaction container 5.
In the above embodiment, the dilution water 49 is stored in the reagent container 37, but the reagent may be stored instead of the dilution water 49.

  Further, the container base 3 may be provided with a gene amplification container for performing a gene amplification reaction. For example, if the reagent container 37 is emptied, it can be used as a gene amplification container.

In addition, if a reagent for performing a gene amplification reaction is accommodated in the reaction container 5, the gene amplification reaction can be performed in the reaction container 5.
Further, when a gene is contained in the liquid introduced into the main flow path 13, a probe that reacts with the gene may be provided in the reaction vessel 5.

Moreover, in the said Example, although the syringe 51 is arrange | positioned on the switching valve 63, the position which arrange | positions the syringe 51 is not limited on the switching valve 63, and may be anywhere.
In the above embodiment, the rotary switching valve 63 is used as the switching valve. However, the switching valve is not limited to this, and various flow path switching valves can be used. A plurality of switching valves may be provided.

  In the above embodiment, when the liquid filled in the metering channel 15 is injected into the reaction vessel 5 through the passive valve 17, the inside of the main channel 13 after the air purge is pressurized so that the liquid is put into the reaction vessel 5. However, the reaction treatment method of the present invention is not limited to this. For example, by changing the flow path configuration so that the inside of the reaction vessel air vent channel 21 can be made negative pressure using the syringe 51, and by making the inside of the reaction vessel air vent channel 21 and thus the reaction vessel 5 inside negative pressure. You may make it inject | pour into the reaction container 5 through the passive valve 17 the liquid with which the measurement flow path 15 was filled. Alternatively, a separate syringe may be prepared so that the liquid is injected into the reaction vessel 5 with a positive pressure in the main channel 13 and a negative pressure in the reaction vessel 5.

  In the above embodiment, the penetrable portion and the second penetrable portion of the sealed container are formed by, for example, films 35j, 35l, 37j, 37l, 39j, and 39l made of aluminum. The 2 penetrable portion is not limited to this, and may be a film of another material or may be formed of the same material as the container body. For example, when the container body is formed of a resin material such as polypropylene or polycarbonate, the thickness of the resin material in the penetrable portion and the second penetrable portion is set to a thickness that can penetrate through the protruding flow channel and the second protruding flow channel. Then, the penetrable part and the second penetrable part can be formed by integral molding with the container body. Here, the thicknesses of the penetrable part and the second penetrable part when the penetrable part and the second penetrable part are formed integrally with the container body are, for example, 0.01 to 0.5 mm.

  Moreover, in the said Example, the sealing containers 35, 37, 39 currently hold | maintained at the 1st holding position are pushed in to the container base 3 side, and are moved to the 2nd holding position, and a sealing container and a sealing container flow path are carried out. However, the present invention is not limited to this. The reaction container plate of the present invention may have a structure in which a sealing container and a sealing container channel are connected in advance.

  Moreover, in the said Example, although the one main flow path 13 was provided and all the measurement flow paths 15 were connected to the main flow path 13, a flow path structure is not limited to this. For example, a plurality of main channels may be provided, and one or a plurality of metering channels may be connected to each main channel.

In the reaction vessel plate of the present invention, the main channel can be sealed, but an example can be given in which the main channel can be sealed by opening and closing both ends of the main channel. Here, “the both ends of the main channel can be opened and closed” means that other space is connected to the end of the main channel, and the end of the other space opposite to the main channel can be opened and closed This includes cases where For example, in the above embodiment, the flow path 14, the liquid drain space 29, the drain space air vent flow path 23, and the flow path 22 correspond to the other spaces.
In the reaction vessel plate of the present invention, the reaction vessel air vent channel can be sealed, but the end of the reaction vessel air vent channel opposite to the reaction vessel can be opened and closed. An example in which the reaction vessel air vent channel can be hermetically sealed can be given. Here, “the end of the reaction vessel air vent channel opposite to the reaction vessel can be opened and closed” means that the other end of the reaction vessel air vent channel opposite to the reaction vessel This includes a case where a space is connected and the end of the other space opposite to the reaction vessel air vent channel can be opened and closed. For example, in the above embodiment, the flow path 24 corresponds to the other space.
In such an aspect, after the liquid is introduced into the main channel and the metering channel, the liquid in the main channel is then purged, and the liquid remaining in the metering channel is injected into the reaction vessel, Both ends of the main channel and the end of the reaction vessel air vent channel opposite to the reaction vessel are closed to seal the main channel and the reaction vessel air vent channel.

  The present invention can be used for measurement of various chemical reactions and biochemical reactions.

It is a figure which shows one Example of reaction container plate, (A) is a schematic top view, (B) is a cross section in the AA position of (A), reaction container, bellows, drain space, and measurement flow path FIG. 2 is a schematic cross-sectional view including a cross section of a passive valve, a sample container, and a sample container air vent channel, and FIG. It is sectional drawing which decomposes | disassembles and shows the Example, and a schematic exploded perspective view of a switching valve. It is the schematic which shows the one reaction container vicinity of the Example, (A) is a top view, (B) is a perspective view, (C) is sectional drawing. It is the figure which expanded and showed the sample container accommodating part and sample container of the Example, (A) is a top view of a sample container accommodating part, (B) is sectional drawing in the BB position of (A), (C) is a plan view of the sample container, (D) is a cross-sectional view at the C-C position of (C), (E) is a cross-sectional view in which the sample container is disposed in the sample container housing portion at the first holding position, F) is a cross-sectional view in which the sample container is arranged at the second holding position in the sample container accommodating portion. It is the figure which expanded and showed the reagent container accommodating part and reagent container of the Example, (A) is a top view of a reagent container accommodating part, (B) is sectional drawing in the DD position of (A), (C) is a plan view of the reagent container, (D) is a cross-sectional view at the EE position of (C), (E) is a cross-sectional view in which the reagent container is arranged in the reagent container housing portion at the first holding position, F) is a cross-sectional view in which the reagent container is disposed in the reagent container housing portion at the second holding position. It is the figure which expanded and showed the container container for air suction and the container for air suction of the Example, (A) is a top view of the container container for air suction, (B) is FF of (A). (C) is a plan view of the air suction container, (D) is a cross sectional view at the GG position of (C), and (E) is an air suction container housing portion. Sectional drawing arrange | positioned in the 1st holding position, (F) is sectional drawing which has arrange | positioned the air suction container to the air suction container accommodating part in the 2nd holding position. It is the schematic sectional drawing which showed the reaction processing apparatus for processing a reaction container plate with the reaction container plate. It is a top view for demonstrating the operation | movement which introduce | transduces a sample liquid into a reaction container from a sample container. It is a top view for demonstrating the operation | movement following FIG. FIG. 10 is a plan view for explaining the operation following FIG. 9. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. FIG. 16 is a plan view for explaining the operation following FIG. 15. FIG. 17 is a plan view for explaining the operation following FIG. 16. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. It is a top view for demonstrating the operation | movement following FIG. It is a schematic sectional drawing which expands and shows the reaction container vicinity of the other Example of the reaction container plate. It is a schematic sectional drawing which expands and shows the reaction container vicinity of the further another Example of the reaction container plate. It is a schematic sectional drawing which expands and shows the reaction container vicinity of the further another Example of the reaction container plate.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Reaction container plate 3 Container base 5 Reaction container 6a, 6b Flow path unit 11 Flow path base 13 Main flow path 15 Metering flow path 17 Passive valve 19, 21 Reaction container air vent flow path 35 Sample container (sealing container)
35a Sample container flow path (sealing container flow path)
35b Sample container air vent channel 337 Reagent container (sealed container)
37a Reagent container flow path (sealing container flow path)
37b Reagent container air vent channel 39 Air suction container (sealing container)
39a Air suction container channel (sealing container channel)
39b Air suction container air vent channel 41 Septum (elastic member)
51 Syringe 63 switching valve

Claims (7)

A sealed reaction vessel;
A syringe for feeding liquid;
A switching valve for switching the flow path to be connected;
An internal capacity is passively changed by containing air, and an elastic capacity variable section connected to the syringe via the switching valve;
A main flow path having one end connected to the drain and the other end connected to the syringe via the switching valve;
A metering channel of a predetermined capacity branched from the main channel;
Liquid introduction pressure when one end is connected to the metering channel and the other end is connected to the reaction vessel, has a channel resistance higher than the metering channel, and liquid is introduced into the main channel and the metering channel A passive valve that does not pass the liquid in the state and purge pressure state when the liquid in the main flow path is discharged to the drain, and passes the liquid in a pressure state greater than the liquid introduction pressure and the purge pressure;
An air vent for venting the reaction vessel;
An atmosphere open flow path having one end connected to the outside of the reaction vessel plate and the other end connected to the air vent via the switching valve;
The switching valve is a reaction vessel plate configured to connect the syringe and the main flow path and simultaneously connect the air vent and the air release flow path. ^2. The reaction vessel plate according to claim 1, wherein a contact angle of the passive valve with respect to water droplets is 90 ° or more, and an area of a boundary between the passive valve and the metering channel is 1 to 10000000 μm ² .   The reaction container plate according to claim 1, wherein the sealing container includes a sample container for containing a sample liquid.   The reaction container plate according to any one of claims 1 to 3, wherein the sealed container includes a sample pretreatment liquid or a reagent stored in advance.   The reaction container plate according to any one of claims 1 to 4, wherein the sealing container also includes a gene amplification container for performing a gene amplification reaction.   The reaction vessel plate according to any one of claims 1 to 5, wherein the reaction vessel is made of a light-transmitting material that can be measured optically from the bottom or from above. A reaction treatment method using the reaction container plate according to any one of claims 1 to 6,
Filling the main channel and the metering channel with the liquid to be injected into the reaction vessel; purging the liquid in the main channel using the air of the variable capacity portion while leaving the liquid in the metering channel And the step of injecting the liquid in the measuring channel into the reaction vessel using the air of the capacity variable unit in that order in a state where the air vent and the open air channel are connected. The reaction processing method characterized by this.

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