JP2016151444A - Nucleic acid amplification reaction cartridge set - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a nucleic acid amplification reaction cartridge set by which a plurality of vessels are assembled when a user uses it.SOLUTION: By a nucleic acid amplification reaction cartridge set 500 concerning the present invention, a plurality of vessels are assembled to be attached to a nucleic acid amplification reaction device. The nucleic acid amplification reaction cartridge set 500 comprises: an elution vessel 300 having a first flow channel 2a in which first fluid 32 is encapsulated and accommodated; and a reaction vessel 400 having a second flow channel 2b which is joined to the elution vessel 300, in communication with the first flow channel 2a, and in which second fluid 26 is encapsulated and accommodated. The elution vessel 300 has an elution insertion part 302 which is inserted into one end of the second flow channel 2b. The reaction vessel 400 has: a reaction acceptance part 404 which accepts the elution insertion part 302 in the second flow channel 2b; and a transfer restriction part 408 which restricts transfer of the elution insertion part 302 in an insertion direction S by exceeding a predetermined position.SELECTED DRAWING: Figure 13

Description

  The present invention relates to a nucleic acid amplification reaction cartridge set.

  In the field of biochemistry, a technique of PCR (Polymerase Chain Reaction) has been established. Recently, the accuracy and detection sensitivity of amplification in the PCR method have improved, and it has become possible to amplify, detect, and analyze a very small amount of sample (DNA, etc.). PCR is a technique for amplifying a target nucleic acid by subjecting a solution (reaction solution) containing a nucleic acid (target nucleic acid) to be amplified and a reagent to thermal cycling. As a thermal cycle of PCR, a method of performing a thermal cycle at two or three stages of temperatures is common.

  On the other hand, the diagnosis of infectious diseases such as influenza in the medical field is currently mainly performed using a simple test kit such as immunochromatography. However, in such a simple test, the accuracy may be insufficient, and it is desired to apply PCR that can be expected to have a higher test accuracy to the diagnosis of infectious diseases.

  In recent years, as a device used in the PCR method or the like, by laminating an aqueous liquid layer and a water-insoluble gel layer alternately in a capillary (in a cartridge) and passing a magnetic particle to which a nucleic acid is attached, A device for purifying a nucleic acid has been proposed (see Patent Document 1). Patent Document 1 describes that a nucleic acid amplification reaction solution is accommodated in the lowermost layer of a cartridge and a target nucleic acid is amplified in the nucleic acid amplification reaction solution.

International Publication No. 2012/086243

  However, the device as described above is composed of an integral container from the sample supply unit to the nucleic acid amplification reaction solution. For example, when stored in such a device for a long period of time, water contained in a washing solution or an eluate may diffuse and come into contact with a reagent for performing a nucleic acid amplification reaction, thereby inhibiting PCR.

  One of the objects according to some embodiments of the present invention is to provide a nucleic acid amplification reaction cartridge set in which a plurality of containers can be assembled when used by a user.

[Application Example 1]
The nucleic acid amplification reaction cartridge set according to the present invention includes:
A nucleic acid amplification reaction cartridge set comprising a first container having a first flow path in which a first fluid is sealed and housed, and a second container having a second flow path in which a second fluid is sealed and housed. ,
The first container has an insertion portion that is inserted into one end of the second flow path,
The second container includes a receiving portion that receives the insertion portion in the second flow path, and a movement restriction portion that restricts the insertion portion from moving beyond a predetermined position in the insertion direction.

According to the nucleic acid amplification reaction cartridge set according to this application example, the first fluid and the second fluid are sealed and housed in separate containers, thereby suppressing fluid movement between the containers until the containers are joined to each other. be able to. In particular, according to the nucleic acid amplification reaction cartridge set according to this application example, when the first container is joined to the second container, the insertion restriction part is inserted too much into the second flow path than the predetermined position by the movement restriction part. This can be prevented.

[Application Example 2]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The movement restriction unit may restrict movement of the insertion unit in the insertion direction by contacting the first container.

  According to the nucleic acid amplification reaction cartridge set according to this application example, the movement control unit can reliably limit the movement of the insertion unit.

[Application Example 3]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The first container is an elution container that stores an eluate as the first fluid,
The second container may be a reaction container including a reagent that reacts with the nucleic acid eluted in the eluate in the second channel.

  According to the nucleic acid amplification reaction cartridge set according to this application example, the elution container and the reaction container can be joined at a predetermined position.

[Application Example 4]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The movement restriction part is formed around the second flow path,
The first container has an abutment portion that is restricted from moving in the insertion direction of the insertion portion by contacting the movement restriction portion,
A plurality of the contact parts are arranged around the second flow path, and contact the movement restriction part in a direction parallel to the insertion direction when the first container and the second container are joined. Or may be arranged opposite each other with a predetermined interval.

  According to the nucleic acid amplification reaction cartridge set according to this application example, the insertion portion can be reliably prevented from being inserted beyond the predetermined position with respect to the second flow path.

[Application Example 5]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The movement restricting portion may be circular when viewed from the insertion direction.

  According to the nucleic acid amplification reaction cartridge set according to this application example, the circular movement restriction portion can reliably arrange that the insertion portion is inserted beyond the predetermined position with respect to the second flow path.

[Application Example 6]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The movement restriction part is formed around the second flow path,
The first container has an abutment portion that is restricted from moving in the insertion direction of the insertion portion by contacting the movement restriction portion,
The contact portion is formed at an opening end of a cylindrical portion formed around the first flow path,
The contact portion is a communication hole that connects the inside and the outside of the cylindrical portion in a state where the first container and the second container are joined and the contact portion is in contact with the movement restricting portion. You may have a notch which forms.

  According to the nucleic acid amplification reaction cartridge set according to this application example, when the first container and the second container are joined, the fluid flowing out from each container can pass through the communication hole. Therefore, when the fluid inside the cylindrical portion joins the first container and the second container, resistance does not occur, and the user can easily assemble the containers.

[Application Example 7]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The second container has a cylindrical body part that forms the second flow path, and a reservoir part that is opened on the receiving part side and formed around the body part,
The reservoir portion includes a cylindrical tube portion, a connection portion that connects the tube portion and the body portion, and functions as the movement restriction portion,
The first container and the second container may have a capacity for receiving fluid flowing out of the first container and the second container between the reservoir section and the body section when the first container and the second container are joined. .

  According to the nucleic acid amplification reaction cartridge set according to this application example, the fluid flowing out from each container when the first container and the second container are joined can be received by the reservoir unit.

[Application Example 8]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The first container may include a sealing portion that contacts the cylindrical portion and seals the opening of the cylindrical portion when the first container and the second container are joined.

May be.

  In the nucleic acid amplification reaction cartridge set according to this application example, when the first container and the second container are joined, the fluid flowing out from the first container and the second container can be reliably held in the reservoir portion.

[Application Example 9]
In the nucleic acid amplification reaction cartridge set according to the present invention,
The cylindrical portion has a through hole penetrating in a direction intersecting the insertion direction,
The first container has a protrusion protruding in a direction intersecting the insertion direction,
When the first container and the second container are joined, the protrusion may be fitted into the through hole, and the insertion part may be restricted from moving away from the receiving part.

  According to the nucleic acid amplification reaction cartridge set according to this application example, the joined state between the first container and the second container can be reliably maintained.

It is a front view of the container assembly 1 which concerns on embodiment. It is a side view of the container assembly 1 which concerns on embodiment. It is a top view of container assembly 1 concerning an embodiment. It is a perspective view of container assembly 1 concerning an embodiment. It is AA sectional drawing in FIG. 3 of the container assembly 1 which concerns on embodiment. It is CC sectional drawing in FIG. 3 of the container assembly 1 which concerns on embodiment. It is a schematic diagram explaining operation of the container assembly 1 which concerns on embodiment. It is a schematic diagram explaining operation of the container assembly 1 which concerns on embodiment. 1 is a schematic configuration diagram of a PCR device 50. FIG. 2 is a block diagram of a PCR device 50. FIG. 2 is a perspective view of an elution container 300. FIG. 4 is a perspective view of a reaction vessel 400. FIG. It is a BB sectional view of nucleic acid amplification reaction cartridge set 500 concerning an embodiment. It is a BB sectional view of nucleic acid amplification reaction cartridge 502 concerning an embodiment.

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

  The nucleic acid amplification reaction cartridge set according to this embodiment includes a first container having a first channel in which a first fluid is sealed and housed, a second container having a second channel in which a second fluid is sealed and housed, The first container has an insertion part that is inserted into one end of the second flow path, and the second container has the insertion part connected to the second flow path. It has a receiving part received in a path, and a movement restricting part which restricts that the insertion part moves beyond a predetermined position in the insertion direction.

  The nucleic acid amplification reaction cartridge set according to the present invention is a set for assembling a cartridge for performing a nucleic acid amplification reaction. That is, when the nucleic acid amplification reaction cartridge set according to the present invention is assembled, a cartridge for performing a nucleic acid amplification reaction can be obtained. In the following, the cartridge (container assembly) will be described first, and then the nucleic acid amplification reaction cartridge set will be described.

1. Outline of Container Assembly First, an outline of the container assembly 1 according to the present embodiment will be described with reference to FIGS. FIG. 1 is a front view of a container assembly 1 (hereinafter sometimes referred to as a cartridge) according to an embodiment. FIG. 2 is a side view of the container assembly 1 according to the embodiment. FIG. 3 is a plan view of the container assembly 1 according to the embodiment. FIG. 4 is a perspective view of the container assembly 1 according to the embodiment. In addition, the state of the container assembly 1 in FIGS. 1 to 3 will be described as an upright state.

  The container assembly 1 includes an adsorption container 100, a cleaning container 200, an elution container 300, and a reaction container 400. The container assembly 1 is a container that forms a flow path (not shown) that communicates from the adsorption container 100 to the reaction container 400. The flow path of the container assembly 1 is closed at one end by the cap 110 and at the other end by the bottom 402.

  The container assembly 1 binds a nucleic acid to magnetic beads (not shown) in the adsorption container 100, purifies the magnetic beads while moving in the washing container 200, and extracts the nucleic acid in an eluate droplet (not shown) in the elution container 300. This is a container that performs pre-treatment for elution and thermal cycle processing of polymerase reaction for the droplet of eluate containing nucleic acid in the reaction container 400.

Although the material of the container assembly 1 is not specifically limited, For example, glass, a polymer | macromolecule, a metal etc. can be used. It is more preferable to select a material having transparency in visible light such as glass or polymer as the material of the container assembly 1 because the inside (inside the cavity) can be observed from the outside of the container assembly 1. In addition, when a material that transmits magnetic force or a non-magnetic material is selected as the material of the container assembly 1, a magnetic force is applied from the outside of the container assembly 1 when a magnetic bead (not shown) is passed through the container assembly 1. Is preferred because it facilitates this. The material of the container assembly 1 can be, for example, polypropylene resin.

  The adsorption container 100 includes a cylindrical syringe part 120 that accommodates an adsorbing liquid (not shown), a plunger part 130 that is a movable pusher inserted into the syringe part 120, and one of the plunger parts 130. And a cap 110 that is fixed to the end of the head. The adsorption container 100 moves the cap 110 with respect to the syringe part 120 to slide the plunger part 130 on the inner surface of the syringe part 120, and pushes an unillustrated adsorbing liquid stored in the syringe part 120 to the cleaning container 200. be able to. The adsorbing liquid will be described later.

  The cleaning container 200 is obtained by joining and assembling the first to third cleaning containers 210, 220, and 230. The first to third cleaning containers 210, 220, and 230 each have one or more cleaning liquid layers partitioned by an oil layer (not shown). Then, by joining the first to third cleaning containers 210, 220, and 230, the cleaning container 200 has a plurality of cleaning liquid layers partitioned by a plurality of oil layers (not shown) inside. In the cleaning container 200 of the present embodiment, the example using the three cleaning containers including the first to third cleaning containers 210, 220, and 230 has been described. However, the present invention is not limited thereto, and the number may be appropriately increased or decreased according to the number of cleaning liquid layers. be able to. The cleaning liquid will be described later.

  The elution container 300 is joined to the third cleaning container 230 of the cleaning container 200 and accommodates the eluate therein so that the shape of the plug can be maintained. Here, the “plug” means a liquid when a specific liquid occupies one section in the flow path. More specifically, the plug of a specific liquid refers to a columnar shape in which only the specific liquid occupies the inside in the longitudinal direction of the flow path, and a certain space inside the flow path is formed by the liquid plug. Shows a state where is marked. The expression “substantially” here means that a small amount (for example, a thin film) of another substance (liquid or the like) may be present around the plug, that is, on the inner wall of the flow path. The eluate will be described later.

  The nucleic acid purification device 5 includes an adsorption container 100, a cleaning container 200, and an elution container 300.

  The reaction container 400 is a container that is joined to the elution container 300 and receives the liquid pushed out from the elution container 300, and is a container that stores droplets of the eluate containing the specimen during the thermal cycle process. Moreover, the reaction container 400 stores a reagent (not shown). The reagent will be described later.

2. Detailed Structure of Container Assembly Next, the detailed structure of the container assembly 1 will be described with reference to FIGS. FIG. 5 is a cross-sectional view taken along the line AA in FIG. 3 of the container assembly 1 according to the embodiment. FIG. 6 is a cross-sectional view taken along the line CC of FIG. 3 of the container assembly 1 according to the embodiment. Actually, the container assembly 1 is assembled in a state in which the contents such as the cleaning liquid are filled. However, in FIGS. 5 and 6, the contents are described to explain the structure of the container assembly 1. Is omitted.

2-1. Adsorption container In the adsorption container 100, the plunger part 130 is inserted from one opening end of the syringe part 120, and the cap 110 is inserted in the opening end of the plunger part 130. The cap 110 has a ventilation portion 112 at the center thereof, and the change in the internal pressure of the plunger portion 130 can be suppressed by the ventilation portion 112 when the plunger portion 130 is operated.

The plunger portion 130 is a substantially cylindrical pusher that slides on the inner peripheral surface of the syringe portion 120. The plunger portion 130 has an opening end portion into which the cap 110 is inserted and a bottom portion facing the opening end portion of the syringe portion 120. It has the rod-shaped part 132 extended in a longitudinal direction, and the front-end | tip part 134 of the front-end | tip of the rod-shaped part 132. The rod-shaped portion 132 protrudes from the center of the bottom portion of the plunger portion 130, and a through-hole is formed around the rod-shaped portion 132 so that the plunger portion 130 and the syringe portion 120 communicate with each other.

  The syringe part 120 constitutes a part of the flow path 2 of the container assembly 1, and includes a large diameter part that accommodates the plunger part 130, a small diameter part having an inner diameter smaller than the large diameter part, and a large diameter part to a small diameter part. A reduced diameter portion that reduces the inner diameter of the lip, a suction insertion portion 122 at the tip of the small diameter portion, and a cylindrical suction cover portion 126 that covers the periphery of the suction insertion portion 122. The large diameter portion, the small diameter portion, and the suction insertion portion 122 that are part of the flow path 2 of the container assembly 1 are substantially cylindrical.

  The tip part 134 of the plunger part 130 seals the small diameter part of the syringe part 120 and partitions the large diameter part, the reduced diameter part, and the small diameter part at the time of provision to the operator to form two compartments.

  The suction insertion part 122 of the syringe part 120 is inserted and fitted into the first receiving part 214 which is one open end part of the first cleaning container 210 in the cleaning container 200, so that the syringe part 120 and the first cleaning container are fitted. 210 is joined. The outer peripheral surface of the suction insertion portion 122 and the inner peripheral surface of the first receiving portion 214 are in close contact with each other to prevent the liquid as the contents from leaking to the outside.

2-2. Cleaning Container The cleaning container 200 is an assembly that forms part of the flow path 2 of the container assembly 1 and includes the first to third cleaning containers 210, 220, and 230. Since the basic structures of the first to third cleaning containers 210, 220, and 230 are the same, the structure of the first cleaning container 210 will be described, and the description of the second and third cleaning containers 220 and 230 will be omitted. To do.

  The first cleaning container 210 has a substantially cylindrical shape extending in the longitudinal direction of the container assembly 1, and includes a first insertion portion 212 formed at one opening end and a first opening formed at the other opening end. It has a receiving part 214 and a cylindrical first cover part 216 that covers the periphery of the first insertion part 212.

  The outer diameter of the first insertion part 212 is substantially the same as the inner diameter of the second receiving part 224. Further, the inner diameter of the first receiving portion 214 is substantially the same as the outer diameter of the suction insertion portion 122.

  By inserting and fitting the first insertion portion 212 of the first cleaning container 210 into the second receiving portion 224 of the second cleaning container 220, the outer periphery of the first insertion portion 212 is aligned with the inner periphery of the second receiving portion 224. The first cleaning container 210 and the second cleaning container 220 are joined together while closely sealing. Similarly, the first to third cleaning containers 210, 220, and 230 are connected to form the cleaning container 200. Here, “sealing” means sealing so that at least liquid or gas contained in a container or the like does not leak to the outside, including sealing that liquid or gas enters from the outside to the inside. Good.

2-3. The elution container The elution container 300 has a substantially cylindrical shape extending in the longitudinal direction of the container assembly 1 and constitutes a part of the flow path 2 of the container assembly 1. The elution container 300 has an elution insertion portion 302 formed at one opening end portion and an elution receiving portion 304 formed at the other opening end portion.

  The inner diameter of the elution receiving part 304 is substantially the same as the outer diameter of the third insertion part 232 of the third cleaning container 230. By inserting and fitting the third insertion portion 232 into the elution receiving portion 304, the outer periphery of the third insertion portion 232 is in close contact with the inner periphery of the elution receiving portion 304 and is sealed with the third cleaning container 230. The container 300 is joined.

2-4. Reaction container The reaction container 400 has a substantially cylindrical shape extending in the longitudinal direction of the container assembly 1 and constitutes a part of the flow path 2 of the container assembly 1. The reaction vessel 400 has a reaction receiving portion 404 formed at the open end, a bottom portion 402 formed at the other closed end, and a reservoir portion 406 that covers the reaction receiving portion 404.

  The inner diameter of the reaction receiving unit 404 is substantially the same as the outer diameter of the elution insertion unit 302 of the elution container 300. The elution container 300 and the reaction container 400 are joined by inserting and fitting the elution insertion part 302 into the reaction receiving part 404.

  A reservoir unit 406 having a predetermined space is provided around the reaction receiving unit 404. The reservoir unit 406 has a volume capable of receiving the liquid overflowing from the reaction vessel 400 due to the movement of the plunger unit 130.

3. Contents of Container Assembly and Operation of Container Assembly Next, the contents of the container assembly 1 will be described with reference to FIG. 7A, and the operation of the container assembly 1 will be described with reference to FIGS. explain. Drawing 7 is a mimetic diagram explaining operation of container assembly 1 concerning an embodiment. Drawing 8 is a mimetic diagram explaining operation of container assembly 1 concerning an embodiment. 7 and 8, each container is represented by a flow path 2 and the outer shape and the joining structure are omitted in order to explain the state of the contents.

3-1. Contents FIG. 7A shows the state of the contents in the flow path 2 in the state of FIG. The contents in the flow path 2 are, in order from the cap 110 toward the reaction vessel 400, the adsorbed liquid 10, the first oil 20, the first cleaning liquid 12, the second oil 22, the second cleaning liquid 14, the third oil 24, Magnetic beads 30, third oil 24, third cleaning liquid 16, fourth oil 26, eluent 32, fourth oil 26, and reagent 34.

  In the flow channel 2, portions having a large cross-sectional area (a thick portion of the flow channel 2) and small portions (a thin portion of the flow channel 2) are alternately arranged on a surface orthogonal to the longitudinal direction of the container assembly 1. Part or all of the first to fourth oils 20, 22, 24, 26 and the eluate 32 are accommodated in a narrow portion of the flow path 2. The cross-sectional area of the narrow part of the flow path 2 is maintained stably when the interface of adjacent liquids (which may be fluids; the same applies hereinafter) is disposed in the narrow part of the flow path 2. Has a possible area. Therefore, the liquid disposed in the narrow portion of the flow path 2 can stably maintain the positional relationship between the liquid and the other liquid disposed above and below the liquid. Even if the interface between the liquid disposed in the narrow portion of the flow path 2 and the other liquid disposed in the thick portion of the flow path 2 is formed in the thick portion of the flow path 2, Even if the interface is disturbed, the interface is stably formed at a predetermined position by placing the interface in a stationary state.

  The narrow portion of the flow path 2 is formed inside the adsorption insertion portion 122, the first insertion portion 212, the second insertion portion 222, the third insertion portion 232, and the elution insertion portion 302. In the elution container 300, the elution insertion portion 302 is formed. Extends upward beyond. In addition, the liquid accommodated in the thin part of the flow path 2 is stably maintained even before the container is assembled.

3-1-1. Oil The first to fourth oils 20, 22, 24, and 26 are all made of oil, and exist as plugs between liquids before and after each oil in the state of FIG. Since the first to fourth oils 20, 22, 24, and 26 exist as plugs, the liquids adjacent to each other before and after each oil are selected as liquids that are phase-separated from each other, that is, liquids that are not miscible. The oils constituting the first to fourth oils 20, 22, 24, 26 may be different types of oils. Examples of the oil that can be used for these include one selected from silicone oil such as dimethyl silicone oil, paraffin oil, mineral oil, and mixtures thereof.

3-1-2. Adsorbing liquid The adsorbing liquid 10 refers to a liquid that serves as a field for adsorbing nucleic acids to the magnetic beads 30 and is, for example, an aqueous solution containing a chaotropic substance. As the adsorbing liquid 10, for example, 5M guanidine thiocyanate, 2% Triton X-100, 50 mM Tris-HCl (pH 7.2) can be used. The adsorbing liquid 10 is not particularly limited as long as it contains a chaotropic substance, but the adsorbing liquid 10 may contain a surfactant for the purpose of disrupting the cell membrane or denaturing proteins contained in the cells. The surfactant is not particularly limited as long as it is generally used for nucleic acid extraction from cells or the like. Specifically, a Triton surfactant such as Triton-X or a tween surfactant such as Tween 20 is used. Nonionic surfactants such as agents, and anionic surfactants such as sodium N-uraroyl sarcosine (SDS) can be mentioned, with nonionic surfactants in particular in the range of 0.1 to 2% It is preferable to use so that it becomes. Furthermore, it is preferable to contain a reducing agent such as 2-mercaptoethanol or dithiothreitol. The lysis solution may be a buffer solution, but is preferably neutral at pH 6-8. Taking these into account, specifically, 3M-7M guanidine salt, 0% -5% nonionic surfactant, 0 mM-0.2 mM EDTA, 0M-0.2M reducing agent, etc. It is preferable to contain.

  Here, the chaotropic substance has a function of generating chaotropic ions (a monovalent anion having a large ionic radius) in an aqueous solution and increasing the water solubility of the hydrophobic molecule. If it contributes to adsorption | suction, it will not specifically limit. Specific examples include guanidine hydrochloride, sodium iodide, sodium perchlorate, and the like. Among these, guanidine thiocyanate or guanidine hydrochloride having a strong protein-modifying action is preferable. The specified concentration of these chaotropic substances varies depending on each substance. For example, when guanidine thiocyanate is used, it is within a range of 3M to 5.5M, and when guanidine hydrochloride is used, it is used at 5M or more. Is preferred.

  Since the chaotropic substance is present in the aqueous solution, the nucleic acid in the aqueous solution is thermodynamically advantageous when adsorbed on a solid rather than surrounded by water molecules. Will be adsorbed.

3-1-3. Cleaning Liquid The first to third cleaning liquids 12, 14, and 16 are for cleaning the magnetic beads 30 to which the nucleic acid is bound.

  The first cleaning liquid 12 is a liquid that is phase-separated from both the first oil 20 and the second oil 22. The first washing liquid 12 is preferably water or a low salt concentration aqueous solution, and in the case of a low salt concentration aqueous solution, it is preferably a buffer solution. The salt concentration of the low salt concentration aqueous solution is preferably 100 mM or less, more preferably 50 mM or less, and most preferably 10 mM or less. Moreover, the 1st washing | cleaning liquid 12 may contain surfactant as mentioned above, and pH is not specifically limited. Although the salt for using the 1st washing | cleaning liquid 12 as a buffer solution is not specifically limited, Salts, such as a tris, hepes, pipes, and phosphoric acid, are preferable. Furthermore, the first washing solution 12 preferably contains alcohol in an amount that does not inhibit the adsorption of the nucleic acid to the carrier, the reverse transcription reaction, the PCR reaction, or the like. In this case, the alcohol concentration is not particularly limited.

  The first cleaning liquid 12 may contain a chaotropic substance. For example, when guanidine hydrochloride is contained in the first washing liquid 12, the magnetic beads 30 and the like can be washed while maintaining or enhancing the adsorption of the nucleic acid adsorbed to the magnetic beads 30 and the like.

  The second cleaning liquid 14 is a liquid that is phase-separated from both the second oil 22 and the third oil 24. The second cleaning liquid 14 may basically have the same or different composition as the first cleaning liquid 12, but is preferably a solution that does not substantially contain a chaotropic substance. This is to eliminate the introduction of chaotropic substances into the later solution. As the 2nd washing | cleaning liquid 14, you may consist of 5 mM Tris hydrochloric acid buffer, for example. As described above, the second cleaning liquid 14 preferably contains alcohol.

  The third cleaning liquid 16 is a liquid that is phase-separated from both the third oil 24 and the fourth oil 26. The third cleaning liquid 16 may basically have the same or different composition as the second cleaning liquid 14, but does not contain alcohol. The third cleaning liquid 16 may include citric acid to prevent alcohol from being brought into the reaction vessel 400.

3-1-4. Magnetic Beads The magnetic beads 30 are beads that adsorb nucleic acids and preferably have relatively strong magnetism so that they can be moved by the magnet 3 outside the container assembly 1. The magnetic beads 30 may be, for example, silica beads or silica-coated beads. The magnetic beads 30 may be preferably silica-coated beads.

3-1-5. The eluate 32 is a liquid that is phase-separated from the fourth oil 26, and exists as a plug sandwiched between the fourth oils 26 and 26 in the flow path 2 in the elution container 300. The eluate 32 is a liquid for eluting the nucleic acid adsorbed on the magnetic beads 30 from the magnetic beads 30 into the eluate 32. Further, the eluate 32 becomes droplets in the fourth oil 26 by heating. For the eluent 32, for example, pure water can be used. Here, the “droplet” is a liquid surrounded by a free surface.

3-1-6. Reagent Reagent 34 contains components necessary for the reaction. When the reaction in the reaction vessel 400 is PCR, the reagent 34 is an enzyme such as a DNA polymerase for amplifying the target nucleic acid (DNA) eluted in the droplet 36 (see FIG. 8) of the eluate, and At least one of a primer (nucleic acid) and a fluorescent probe for detecting an amplification product can be included, and here, all of the primer, enzyme, and fluorescent probe are included. The reagent 34 is incompatible with the fourth oil 26 and dissolves and reacts when it comes into contact with the droplet 36 of the eluate 32 containing nucleic acid, and is the lowest part in the gravity direction of the flow path 2 in the reaction container 400. Exists in a solid state in the region. For example, the reagent 34 may be freeze-dried (freeze-dried).

3-2. Operation of Container Assembly An example of the operation of the container assembly 1 will be described with reference to FIGS.

The operation of the container assembly 1 is as follows:
(A) a step of assembling the container assembly 1 by joining the adsorption container 100, the cleaning container 200, the elution container 300, and the reaction container 400;
(B) introducing a sample containing nucleic acid into the adsorption container 100 in which the adsorbing liquid 10 is stored;
(C) a step of moving the magnetic beads 30 from the second cleaning container 220 to the adsorption container 100;
(D) swinging the adsorption container 100 to adsorb the nucleic acid to the magnetic beads 30;
(E) From the adsorption container 100, the first oil 20, the first cleaning liquid 12, the second oil 22, the second cleaning liquid 14, the third oil 24, the third cleaning liquid 16, and the fourth oil 26 are passed in this order, and the elution container Moving the magnetic beads 30 adsorbed with nucleic acids to 300;
(F) a step of eluting the nucleic acid from the magnetic beads 30 with respect to the eluate 32 in the elution container 300;
(G) contacting the droplet containing the nucleic acid with the reagent 34 in the reaction vessel 400;
including.

  Hereinafter, each process is demonstrated in order.

(A) Step of assembling container assembly 1 As shown in FIG. 7A, the step of assembling is a flow path that joins from the adsorption vessel 100 to the reaction vessel 400 and continues from the adsorption vessel 100 to the reaction vessel 400. Assemble container assembly 1 to form 2. In FIG. 7A, the cap 110 is attached to the adsorption container 100, but the cap 110 is attached to the plunger portion 130 after the step (B).

  More specifically, the elution insertion section 302 of the elution container 300 is inserted into the reaction receiving section 404 of the reaction container 400, and the third insertion section 232 of the third cleaning container 230 is inserted into the elution receiving section 304 of the elution container 300. The second insertion part 222 of the second cleaning container 220 is inserted into the third receiving part 234 of the third cleaning container 230, and the first insertion part of the first cleaning container 210 is inserted into the second receiving part 224 of the second cleaning container 220. 212 is inserted, and the suction insertion part 122 of the suction container 100 is inserted into the first receiving part 214 of the first cleaning container 210.

(B) Step of introducing the sample In the step of introducing, for example, a cotton swab to which the sample is attached is inserted into the adsorbing liquid 10 from the opening where the cap 110 of the adsorption container 100 is attached, and this is immersed in the adsorbing liquid 10. Do. More specifically, a cotton swab is inserted from an opening at one end of the plunger portion 130 in a state of being inserted into the syringe portion 120 of the adsorption container 100. Next, the cotton swab is taken out from the adsorption container 100 and the cap 110 is attached. This is the state shown in FIG. Further, the specimen may be introduced into the adsorption container 100 by a pipette or the like. In addition, if the specimen is in a paste form or a solid form, the specimen may be attached to or put into the inner wall of the plunger unit 130 with a scissors or tweezers, for example. As shown in FIG. 7A, the adsorbing liquid 10 is partially filled in the syringe part 120 and the plunger part 130, but a space is left on the opening side where the cap 110 is attached. .

  The sample contains the target nucleic acid. Hereinafter, this may be simply referred to as a target nucleic acid. The target nucleic acid is, for example, DNA or RNA (DNA: Deoxyribonucleic Acid and / or RNA: Ribonucleic Acid). The target nucleic acid is extracted from the specimen, eluted in an eluate 32 described later, and then used as a PCR template, for example. Examples of the specimen include blood, nasal mucus, oral mucosa, and other various biological samples.

(C) Step of moving the magnetic beads The step of moving the magnetic beads 30 is a magnet that is sandwiched between the third oils 24 and 24 of the second cleaning container 220 and exists in a plug shape as shown in FIG. The bead 30 is performed by moving the magnet 3 toward the adsorption container 100 in a state where the magnetic force of the magnet 3 arranged outside the container is applied.

  The cap 110 and the plunger part 130 are moved in the direction of extracting from the syringe part 120 in accordance with the movement of the magnetic beads 30 or earlier, and the sample in the adsorbed liquid 10 is moved from the plunger part 130 to the syringe part 120. Move in. Due to the movement of the plunger part 130, the flow path 2 that has been blocked by the tip part 134 communicates with the adsorbing liquid 10.

  The magnetic beads 30 ascend in the flow path 2 as the magnet 3 moves, and reach the adsorbing liquid 10 with the specimen as shown in FIG.

(D) Step of adsorbing nucleic acid to magnetic beads The step of adsorbing nucleic acid is performed by swinging the adsorption container 100. This step can be efficiently performed because the opening of the adsorption container 100 is sealed by the cap 110 so that the adsorbed liquid 10 does not leak out. By this step, the target nucleic acid is adsorbed on the surface of the magnetic bead 30 by the action of the chaotropic agent. In this step, nucleic acids and proteins other than the target nucleic acid may be adsorbed on the surface of the magnetic beads 30.

  As a method of swinging the adsorption container 100, a known device such as a vortex shaker may be used, or shaking may be performed by an operator's hand. Further, the magnetism of the magnetic beads 30 may be used to swing the adsorption container 100 while applying a magnetic field from the outside.

(E) Step of moving magnetic beads adsorbed with nucleic acid The step of moving magnetic beads 30 adsorbed with nucleic acid moves while applying the magnetic force of the magnet 3 from the outside of the adsorption vessel 100, the washing vessel 200, and the elution vessel 300. As a result, the magnetic beads 30 are moved in the adsorbing liquid 10, the first to fourth oils 20, 22, 24, 26 and the first to third cleaning liquids 12, 14, 16.

  For example, a permanent magnet or an electromagnet can be used as the magnet 3. Moreover, the magnet 3 may be moved by an operator's hand or may be performed using a mechanical device or the like. Since the magnetic beads 30 have a property of being attracted by a magnetic force, the relative arrangement of the magnets 3 is changed to the adsorption vessel 100, the cleaning vessel 200, and the elution vessel 300 by using this property. The inside of the flow path 2 is moved. The speed at which the magnetic beads 30 pass through each cleaning liquid is not particularly limited, and the magnetic beads 30 may be moved so as to reciprocate along the longitudinal direction of the flow path 2 in the same cleaning liquid. In addition, when moving particles other than the magnetic bead 30 in a tube, this can be performed using gravity or a potential difference, for example.

(F) Step of Eluting Nucleic Acid In the step of eluting nucleic acid, the nucleic acid is eluted from the magnetic beads 30 in the elution liquid droplets 36 in the elution container 300. The eluate 32 in FIG. 7 was present as a plug in the narrow portion of the flow path of the elution container 300. However, the content liquid can be obtained by heating the reaction container 400 while moving the magnetic beads 30 as described above. It expands and moves upward in the elution container 300 as a droplet 36 as shown in FIG. Then, as shown in FIG. 8A, when the magnetic beads 30 reach the eluate droplets 36 in the elution container 300, the target nucleic acid adsorbed on the magnetic beads 30 is dissolved into the eluate by the action of the eluate. Elution into the liquid droplet 36.

(G) The process of making it contact with the reagent 34 The process of making it contact with the reagent 34 makes the droplet 36 containing a nucleic acid contact the reagent 34 in the lowest part in the reaction container 400. FIG. Specifically, as shown in FIG. 8B, the magnetic beads 30 to which the magnetic force of the magnet 3 is applied by pressing the cap 110 and pressing down the first oil 20 by the tip portion 134 of the plunger portion 130. Is maintained at a predetermined position, the droplet 36 of the eluate from which the target nucleic acid is eluted moves to the reaction vessel 400 and contacts the reagent 34 at the bottom of the reaction vessel 400. The reagent 34 in contact with the droplet 36 melts and mixes with the target nucleic acid in the eluate, and for example, PCR using thermal cycling can be performed.

4). PCR apparatus PCR which performs nucleic acid elution processing and PCR using container assembly 1 using FIG.9 and FIG.10.
The device 50 will be described. FIG. 9 is a schematic configuration diagram of the PCR device 50. FIG. 10 is a block diagram of the PCR device 50.

  The PCR device 50 includes a rotation mechanism 60, a magnet moving mechanism 70, a pressing mechanism 80, a fluorescence measuring instrument 55, and a controller 90.

4-1. Rotation mechanism The rotation mechanism 60 includes a rotation motor 66 and a heater 65, and rotates the container assembly 1 and the heater 65 by driving the rotation motor 66. When the rotating mechanism 60 rotates the container assembly 1 and the heater 65 to turn upside down, the droplet containing the target nucleic acid moves in the flow path of the reaction container 400, and thermal cycle processing is performed.

  The heater 65 includes a plurality of heaters (not shown), and can include elution, high temperature, and low temperature heaters, for example. The elution heater heats the plug-like eluate of the container assembly 1 and promotes elution of the target nucleic acid from the magnetic beads to the eluate. The high temperature heater heats the liquid upstream of the flow path of the reaction vessel 400 to a temperature higher than that of the low temperature heater. The low temperature heater heats the bottom 402 of the flow path of the reaction vessel. A temperature gradient can be formed in the liquid in the flow path of the reaction vessel 400 by the high temperature heater and the low temperature heater. The heater 65 is provided with a temperature control device, and the liquid in the container assembly 1 can be set to a temperature suitable for processing in accordance with a command from the controller 90.

  The heater 65 has an opening through which the outer wall of the bottom 402 of the reaction vessel 400 is exposed. The fluorescence measuring device 55 measures the luminance of the droplet of the eluate from the opening.

4-2. Magnet moving mechanism The magnet moving mechanism 70 is a mechanism for moving the magnet 3. The magnet moving mechanism 70 draws the magnetic beads in the container assembly 1 toward the magnet 3 and moves the magnetic beads in the container assembly 1 by moving the magnet 3. The magnet moving mechanism 70 includes a pair of magnets 3, an elevating mechanism, and a swing mechanism.

  The swing mechanism is a mechanism that swings the pair of magnets 3 in the left-right direction in FIG. 9 (may be the front-rear direction in FIG. 9). The pair of magnets 3 is disposed so as to sandwich the container assembly 1 mounted on the PCR device 50 from the left and right directions (see FIGS. 7 and 8), and is perpendicular to the flow path of the container assembly 1 (here, The distance between the magnetic beads and the magnet 3 can be made closer in the left-right direction in FIG. Therefore, when the pair of magnets 3 are swung in the left-right direction as indicated by arrows, the magnetic beads in the container assembly 1 move in the left-right direction in accordance with the movement. The elevating mechanism can move the magnet 3 in the vertical direction and move the magnetic beads in the vertical direction in FIG. 9 in accordance with the movement of the magnet 3.

4-3. Pressing mechanism The pressing mechanism 80 is a mechanism that presses the plunger portion of the container assembly 1. When the plunger portion is pressed by the pressing mechanism 80, the droplet in the elution container 300 is pushed into the reaction container 400, PCR can be performed in the reaction vessel 400.

  In FIG. 9, the pressing mechanism 80 is shown arranged above the upright container assembly 1, but the direction in which the pressing mechanism 80 pushes the plunger portion is not the vertical direction in FIG. 9, for example, the vertical direction It may be inclined 45 degrees with respect to. By doing in this way, it becomes easy to arrange | position the press mechanism 80 in the position which does not interfere with the magnet moving mechanism 70. FIG.

4-4. Fluorescence measuring instrument The fluorescence measuring instrument 55 is a measuring instrument that measures the brightness of the droplets in the reaction vessel 400. The fluorescence measuring device 55 is disposed at a position facing the bottom portion 402 of the reaction vessel 400. Note that the fluorescence measuring device 55 is preferably capable of detecting luminance in a plurality of wavelength regions so as to be compatible with multiplex PCR.

4-5. Controller The controller 90 is a control unit that controls the PCR apparatus 50. The controller 90 includes a processor such as a CPU and a storage device such as a ROM and a RAM. Various programs and data are stored in the storage device. Further, the storage device provides an area for developing a program. Various processes are realized by the processor executing the program stored in the storage device.

  For example, the controller 90 controls the rotation motor 66 to rotate the container assembly 1 to a predetermined rotation position. The rotation mechanism 60 is provided with a rotation position sensor (not shown), and the controller 90 drives and stops the rotation motor 66 according to the detection result of the rotation position sensor.

  Further, the controller 90 controls the heater 65 to turn on / off the heater to generate heat, thereby heating the liquid in the container assembly 1 to a predetermined temperature.

  Further, the controller 90 controls the magnet moving mechanism 70 to move the magnet 3 in the vertical direction, and swings the magnet 3 in the horizontal direction in FIG. 9 according to the detection result of a position sensor (not shown).

  In addition, the controller 90 controls the fluorescence measuring device 55 to measure the luminance of the droplet in the reaction container 400. This measurement result is stored in a storage device (not shown) of the controller 90.

  The container assembly 1 is attached to the PCR device 50, and the above steps 3-2 (C) to (G) can be performed, and further PCR can be performed.

5. Nucleic Acid Amplification Reaction Cartridge Set A nucleic acid amplification reaction cartridge set 500 according to this embodiment will be described with reference to the drawings. FIG. 11 is a perspective view of the elution container 300. FIG. 12 is a perspective view of the reaction vessel 400. FIG. 13 is a longitudinal sectional view schematically showing the nucleic acid amplification reaction cartridge set 500 according to the present embodiment in the BB section of FIGS. 11 and 12. The nucleic acid amplification reaction cartridge set 500 is a part of the container assembly 1 described with reference to FIGS.

  The nucleic acid amplification reaction cartridge set 500 (FIG. 13) described here becomes a nucleic acid amplification reaction cartridge 502 (FIG. 14) by assembling a plurality of containers (100, 200, 300, 400), and further, the adsorption container 100 and the cleaning container. By joining 200 (FIG. 1 and the like) to the nucleic acid amplification reaction cartridge 502, the above-described container assembly 1 can be obtained. In addition, the joining process order of each container can be changed suitably. The container assembly 1 can be mounted on a nucleic acid amplification reaction device (PCR device 50: FIGS. 9 and 10).

As shown in FIG. 13, the nucleic acid amplification reaction cartridge set 500 includes an elution container 300 that is a first container having a first channel 2a in which a first fluid is sealed and a first container joined to the first container. And a reaction vessel 400 which is a second vessel having a second flow channel 2b in which a second fluid is sealed and accommodated, which communicates with the flow channel 2a. As described above, since the first fluid and the second fluid are sealed and accommodated in separate containers, the movement of the fluid between the containers can be suppressed until the containers are joined to each other. Here, although the combination using the elution container 300 as the first container and the reaction container 400 as the second container will be described, the combination is not limited to the elution container 300 and the reaction container 400. For example, you may combine with the reaction container 400 by using what the elution container 300 and the washing | cleaning container 200 (FIG. 1 etc.) mentioned above and / or the adsorption | suction container 100 were joined as a 1st container.

  Hereinafter, the elution container 300 and the reaction container 400 will be described in detail as the nucleic acid amplification reaction cartridge set 500, and then the nucleic acid amplification reaction cartridge 502 will be described.

5-1. Elution Container The first container shown in FIGS. 11 and 13 is an elution container 300 that stores the eluate 32 as a first fluid, and is the container described in “2-3. Elution Container” above. The elution container 300 includes a substantially cylindrical body 308 that forms a part of the first flow path 2a and extends in the axial direction of the first flow path 2a, and one end of a second flow path 2b of the reaction container 400 described later. And an elution insertion portion 302 which is an insertion portion to be inserted into the.

  The trunk portion 308 has the first flow path 2a formed therein, and has a plurality of flange portions outside. The body 308 has an elution insertion portion 302 at one end, and an elution receiving portion 304 at the other end. The end portion of the body portion 308 on the elution receiving portion 304 side has an annular step portion 306 on the inside, and a film 322 is attached to the step portion 306. In FIG. 11, the film 322 is omitted to explain the stepped portion 306 and the first flow path 2a.

  Among the plurality of flange portions, the flange portion 310 closest to the elution insertion portion 302 is a cylindrical portion that is a cylindrical portion formed around the first flow path 2a (or the elution insertion portion 302) on one surface. 312 and a projection 318 that protrudes in the direction intersecting the insertion direction S for inserting the elution insertion portion 302 into the reaction receiving portion 404 on the other surface. The flange portion 310 is a sealing portion that comes into contact with a cylindrical portion 407 of a reservoir portion 406 to be described later and seals the opening of the cylindrical portion 407 when the elution container 300 and the reaction container 400 are joined. The outer periphery of the flange portion 310 is formed in the same shape as the inner periphery of the reservoir portion 406 or a slightly similar shape. The flange portion 310 has a disk shape when the cylindrical portion 407 of the reservoir portion 406 is cylindrical.

  The cylindrical portion 312 opens at the end opposite to the portion connected to the flange portion 310, that is, the end on the insertion direction S side. A contact portion whose movement is restricted by contact with a movement restriction portion 408 of the reaction container 400 described later is formed at the open end of the cylindrical portion 312. An annular step 307 is provided inside the opening end of the cylindrical portion 312, and a film 320 is attached to the step 307 (FIG. 13). Accordingly, the first flow path 2 a is sealed by the films 320 and 322 attached to both ends thereof, and contains the eluate 32 and the fourth oil 26. The eluent 32 is described in “3-1-5. Eluent”, and the fourth oil 26 is described in “3-1-1. Oil”, and thus the overlapping description is omitted.

  When the elution container 300 and the reaction container 400 are joined, the contact part 314 contacts the movement restriction part 408 in a direction parallel to the insertion direction S or is opposed to the movement restriction part 408 with a predetermined interval.

  The abutting portion 314 is a cutting hole that forms a communication hole that connects the inside and the outside of the cylindrical portion 312 in a state where the elution container 300 and the reaction vessel 400 are joined and the abutting portion 314 is in contact with the movement restricting portion 408. It has a notch 316.

The notches 316 are a plurality of grooves formed at the opening end of the cylindrical portion 312 and extend from the inside of the cylindrical portion 312 to the outside. Therefore, the opening end portion of the cylindrical portion 312 formed between the adjacent notch portions 316 becomes the contact portion 314. Although the number of the contact part 314 may be one, multiple may be arrange | positioned around the 2nd flow path 2b mentioned later. A plurality of contact portions 314 arranged in contact with the movement restricting portion 408 in a direction parallel to the insertion direction S or are opposed to each other with a predetermined interval.

  The protrusion 318 is formed to protrude outward from the outer periphery of the flange portion 310. The protrusions 318 are formed at two locations facing each other across the first flow path 2a. The number of protrusions 318 is not limited to two.

  The elution insertion portion 302 is a portion that is inserted into a reaction receiving portion 404 of the reaction vessel 400 described later, and has a shape that is easy to break through the film 320 and the film 414 of the reaction vessel 400 with a tapered tip.

5-2. Reaction Container The second container shown in FIGS. 12 and 13 is a reaction container 400 in which the fourth oil 26 as the second fluid is sealed and accommodated, and the container described in “2-4. Reaction container” above. . The second flow path 2 b includes a reagent 34 that reacts with the nucleic acid eluted in the eluate 32. The reaction vessel 400 has a cylindrical body 403 that forms a second flow path 2b inside and forms a second flow path 2b, and a cylindrical shape that opens around the reaction receiving part 404 and is formed around the body 403. Reservoir portion 406. The reaction container 400 includes a reaction receiving unit 404 that receives the elution insertion unit 302 in the second flow path 2b, and a movement restriction unit 408 that restricts the elution insertion unit 302 from moving beyond a predetermined position in the insertion direction S. Have.

  When the elution container 300 is joined to the reaction container 400, the movement restricting unit 408 prevents the elution insertion part 302 from being erroneously inserted into the second flow path 2b more than a predetermined position (over-insertion). By preventing over-insertion of the elution insertion portion 302, it is possible to prevent the pressure in the flow paths 2a and 2b from becoming too high or causing the thin elution vessel 300 or the reaction vessel 400 to be deformed. The shape of the movement restricting portion is not limited as long as the elution insertion portion 302 can be prevented from being inserted too much into the second flow path 2b from a predetermined position, and is formed as a part of the reservoir portion as will be described later. In addition, when it is provided as a flange around the body portion 403, it can be realized as a rod-shaped member disposed from the body portion 403 toward the periphery.

  The trunk portion 403 has a cylindrical shape extending in the axial direction of the second flow path 2 b, one end portion is closed by the bottom portion 402, and the other end portion opens as the reaction receiving portion 404. In the body 403, the axial direction of the second flow path 2b is the longitudinal direction. The film 414 is attached to the opening end of the body 403 to seal the second flow path 2b, and the fourth oil 26 and the reagent 34 are sealed and stored in the second flow path 2b.

  The reservoir portion 406 includes a cylindrical tube portion 407, and a connection portion that connects the tube portion 407 and the body portion 403 and functions as the movement restricting portion 408. The cylinder part 407 is cylindrical. The movement restricting portion 408 extends from the other end of the tube portion 407 to the body portion 403. The reservoir unit 406 is a container formed around the second flow path 2b (or the reaction receiving unit 404). The reservoir unit 406 has a capacity for receiving fluid flowing out from the elution container 300 and the reaction container 400 between the reservoir unit 406 and the body 403 when the elution container 300 and the reaction container 400 are joined. Therefore, the fourth oil 26 that has flowed out from each container when the elution container 300 and the reaction container 400 are joined can be received by the reservoir 406.

  The movement restriction unit 408 contacts the elution container 300 and restricts the movement of the elution insertion unit 302 in the insertion direction S. When a part of the elution container 300 comes into contact with the movement restriction unit 408, the movement of the elution insertion unit 302 can be reliably restricted. Here, the movement restriction part 408 restricts the movement of the elution insertion part 302 by contacting the contact part 314 of the elution container 300.

The movement restricting portion 408 is an annular portion that is formed around the second flow path 2 b and connects the inner periphery to the outer surface of the trunk portion 403 and connects the outer periphery to the lower end of the cylindrical portion 407. The movement restriction unit 408 is circular when viewed from the insertion direction S in which the elution insertion unit 302 is inserted into the reaction receiving unit 404. Further, the outer peripheral shape of the movement restricting portion 408 may have a shape according to the shape of the tube portion 407 as long as the tube portion 407 is a cylinder other than a cylinder. Since the movement restricting portion 408 is annular or circular, it is ensured that the elution insertion portion 302 is inserted into the second flow path 2b beyond the predetermined position at any position around the second flow path 2b. Can be arranged.

  The tubular portion 407 has a through hole 412 that penetrates in the direction intersecting the insertion direction S. The through hole 412 is provided in the retaining portion 410 that protrudes in the direction opposite to the insertion direction S from the opening end of the cylindrical portion 407. The retaining portions 410 are provided at two opposing locations. Further, the through-hole 412 may be formed on the side surface of the cylindrical portion 407 as long as the through-hole 412 is on the opening end side with respect to the portion with which the flange portion 310 is in contact.

5-3. Nucleic Acid Amplification Reaction Cartridge A nucleic acid amplification reaction cartridge 502 and its assembly procedure will be described with reference to FIGS. FIG. 14 is a BB cross-sectional view of the nucleic acid amplification reaction cartridge 502 according to the embodiment. In FIG. 14, a part of the elution container 300 is omitted.

  The nucleic acid amplification reaction cartridge 502 is obtained by assembling the nucleic acid amplification reaction cartridge set 500 described with reference to FIGS. The specific assembly procedure is as follows.

  First, the elution insertion part 302 at the tip of the first flow path 2 a of the elution container 300 is inserted into the reaction receiving part 404 of the reaction container 400. At that time, the film 320 and the film 414 shown in FIG. 13 are broken by the tips of the elution insertion portion 302 and the reaction receiving portion 404, and the first flow path 2a and the second flow path 2b communicate with each other.

  When the film 320 is broken, the fourth oil 26 leaks into the reservoir 406 from the first flow path 2a. Further, when the elution insertion portion 302 is pushed into the second flow path 2b, the fourth oil 26 overflows into the reservoir section 406 from the second flow path 2b. The reservoir unit 406 has a capacity to receive the fluid (fourth oil 26) flowing out from the elution container 300 and the reaction container 400 with the body 403 at least when the elution container 300 and the reaction container 400 are joined. The fluid flowing out from each container can be received between the reservoir 406 and the body 403.

  When the flange portion 310 of the elution container 300 enters the reservoir portion 406, the flange portion 310 comes into contact with the tube portion 407 and seals the opening of the tube portion 407. Specifically, the outer periphery of the flange portion 310 slides while being in close contact with the inner peripheral surface of the cylindrical portion 407. Accordingly, the inside of the cylindrical portion 407 is sealed by the flange portion 310, and the fourth oil 26 is securely held in the reservoir portion 406, and is prevented from leaking from the inside of the reservoir portion 406 to the outside.

  The contact portion 314 of the elution container 300 is arranged at a plurality of locations around the second flow path 2b when the elution container 300 and the reaction container 400 are joined, and is limited in movement in a direction parallel to the insertion direction S. The part 408 is disposed in contact with or facing the part 408. It is not essential that the contact part 314 is in contact with the movement restriction part 408. The contact portion 314 only needs to come into contact with the movement restricting portion 408 when the elution insertion portion 302 at the predetermined position in the second flow path 2b is about to be inserted excessively. Thus, by providing the contact part 314 and the movement restricting part 408, it is possible to reliably prevent the elution insertion part 302 from being inserted beyond the predetermined position with respect to the second flow path 2b.

  The contact portion 314 is formed at the opening end of the cylindrical portion 312 formed around the first flow path 2a, and the movement limiting portion 408 is an annular flat surface formed around the second flow path 2b. The movement can be limited in a predetermined state without the first flow path 2a being inclined with respect to the two flow paths 2b.

  When the elution container 300 and the reaction container 400 are joined at a predetermined position, the protrusion 318 of the elution container 300 is fitted into the through hole 412 of the reaction container 400, and the elution insertion part 302 is removed from the reaction receiving part 404 (insertion direction). The movement in the opposite direction of S) is restricted. Thereby, the joining state of the elution container 300 and the reaction container 400 can be reliably maintained.

  The notch 316 forms a communication hole that communicates the inside and the outside of the cylindrical portion 312 even when the contact portion 314 is in contact with the movement restricting portion 408. Therefore, even when the contact portion 314 and the movement restricting portion 408 are in contact with each other, the fourth oil 26 flowing out from each container can pass through the communication hole, and the fourth oil 26 located inside the cylinder portion 312 is used. However, when the elution container 300 and the reaction container 400 are joined, there is no resistance, and the user can easily assemble the containers. In addition, the plurality of communication holes formed by the notches 316 allow the droplets of the eluate 32 to move to the reagent 34 side in the steps (F) and (G) of “3-2. Operation of container assembly”. When extruding, the fourth oil 26 pushed out from the elution container 300 and the reaction container 400 is allowed to flow into the reservoir section 406.

  The present invention is not limited to the above-described embodiments, and various modifications can be made. For example, the present invention includes configurations that are substantially the same as the configurations described in the embodiments (for example, configurations that have the same functions, methods, and results, or configurations that have the same objects and effects). In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that exhibits the same operational effects as the configuration described in the embodiment or a configuration that can achieve the same object. In addition, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

DESCRIPTION OF SYMBOLS 1 ... Container assembly, 2 ... Flow path, 2a ... 1st flow path, 2b ... 2nd flow path, 3 ... Magnet, 5 ... Nucleic acid purification device, 10 ... Adsorption liquid, 12 ... 1st washing | cleaning liquid, 14 ... 2nd Cleaning fluid, 16 ... third cleaning fluid, 20 ... first oil, 22 ... second oil, 24 ... third oil, 26 ... fourth oil, 30 ... magnetic beads, 32 ... eluent, 34 ... reagent, 36 ... droplet , 50 ... PCR device, 55 ... fluorescence measuring device 60 ... rotation mechanism, 65 ... heater, 66 ... rotation motor, 70 ... magnet movement mechanism, 80 ... pressing mechanism, 90 ... controller, 100 ... adsorption container, 110 ... cap, DESCRIPTION OF SYMBOLS 112 ... Ventilation part, 120 ... Syringe part, 122 ... Suction insertion part, 126 ... Suction cover part, 130 ... Plunger part, 132 ... Bar-shaped part, 134 ... Tip part, 200 ... Cleaning container, 210 ... First cleaning container, 212 ... 1st insertion part, 21 ... 1st receiving part, 216 ... 1st cover part, 220 ... 2nd washing container, 222 ... 2nd insertion part, 224 ... 2nd receiving part, 226 ... 2nd cover part, 230 ... 3rd washing container, 232 ... 3rd insertion part, 234 ... 3rd receiving part, 236 ... 3rd cover part, 300 ... elution container, 302 ... elution insertion part, 304 ... elution receiving part, 306 ... step part, 307 ... step part, 308 ... trunk | drum , 310 ... Flange part, 312 ... Cylinder part, 314 ... Contact part, 316 ... Notch part, 318 ... Projection part, 320, 322 ... Film, 400 ... Reaction vessel, 402 ... Bottom part, 403 ... Body part, 404 ... Reaction accepting unit, 406 ... reservoir unit, 407 ... cylinder unit, 408 ... movement restricting unit, 410 ... prevention unit, 412 ... through hole, 414 ... film, 500 ... nucleic acid amplification reaction cartridge set, 502 ... nucleic acid amplification reaction cartridge, ... the insertion direction

Claims (9)

A nucleic acid amplification reaction cartridge set comprising a first container having a first flow path in which a first fluid is sealed and housed, and a second container having a second flow path in which a second fluid is sealed and housed. ,
The first container has an insertion portion that is inserted into one end of the second flow path,
The second container includes a receiving part that receives the insertion part in the second flow path, and a movement restriction part that restricts the insertion part from moving beyond a predetermined position in the insertion direction. Cartridge set. In claim 1,
The movement restriction unit is a nucleic acid amplification reaction cartridge set that contacts the first container and restricts movement of the insertion unit in the insertion direction. In claim 1 or 2,
The first container is an elution container that stores an eluate as the first fluid,
The nucleic acid amplification reaction cartridge set, wherein the second container is a reaction container including a reagent that reacts with nucleic acid eluted in the eluate in the second flow path. In any one of Claims 1 thru | or 3,
The movement restriction part is formed around the second flow path,
The first container has an abutment portion that is restricted from moving in the insertion direction of the insertion portion by contacting the movement restriction portion,
A plurality of the contact parts are arranged around the second flow path, and contact the movement restriction part in a direction parallel to the insertion direction when the first container and the second container are joined. Or a nucleic acid amplification reaction cartridge set that is opposed to each other with a predetermined interval. In claim 4,
The movement restriction part is a nucleic acid amplification reaction cartridge set that is circular when viewed from the insertion direction. In any one of Claims 1 thru | or 3,
The movement restriction part is formed around the second flow path,
The first container has an abutment portion that is restricted from moving in the insertion direction of the insertion portion by contacting the movement restriction portion,
The contact portion is formed at an opening end of a cylindrical portion formed around the first flow path,
The contact portion is a communication hole that connects the inside and the outside of the cylindrical portion in a state where the first container and the second container are joined and the contact portion is in contact with the movement restricting portion. A cartridge set for nucleic acid amplification reaction, having a notch that forms a surface. In any one of Claims 1 thru | or 6,
The second container has a cylindrical body part that forms the second flow path, and a reservoir part that is opened on the receiving part side and formed around the body part,
The reservoir portion includes a cylindrical tube portion, a connection portion that connects the tube portion and the body portion, and functions as the movement restriction portion,
A nucleic acid amplification reaction cartridge having a capacity for receiving fluid flowing out of the first container and the second container between the reservoir section and the body section when the first container and the second container are joined. set. In claim 7,
The nucleic acid amplification reaction cartridge set, wherein the first container has a sealing part that contacts the cylindrical part and seals the opening of the cylindrical part when the first container and the second container are joined. In claim 7 or 8,
The cylindrical portion has a through hole penetrating in a direction intersecting the insertion direction,
The first container has a protrusion protruding in a direction intersecting the insertion direction,
When the first container and the second container are joined, it is possible to restrict the protrusion from fitting into the through-hole and moving the insertion part in a direction of coming out of the receiving part. Nucleic acid amplification reaction cartridge set.