JP2016214174A - Container assembly set - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a container assembly set that can seal/store a plurality of fluids by separate containers, in which connection of each container is easy, sealing is released when connected, and a flow channel where the plurality of fluids are arranged can be formed.SOLUTION: A container assembly set according to the present invention includes: a first container having an insertion part which forms a first flow channel for allowing a first fluid to flow, a cylindrical part formed around the insertion part, and a first film which seals the cylindrical part to seal the first fluid; and a second container having a receiving part which forms a second flow channel for allowing a second fluid to flow and receives the insertion part, and a second film which seals the receiving part to seal the second fluid, in which the first film and the second film are broken when the first container and the second container are connected, and the first flow channel and the second flow channel communicate with each other.SELECTED DRAWING: Figure 15

Description

  The present invention relates to a container assembly 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 PCR 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. Further, in general outpatients at medical institutions, the time that can be spent on examinations is limited to a short time because the examination time is limited. For this reason, for example, the inspection of influenza is carried out in a short time by a simple inspection such as immunochromatography at the expense of the accuracy of the inspection. Further, in quarantine at an airport or the like, since it is necessary to obtain a result quickly, it is desired to increase the speed of PCR.

  In recent years, as a device used for PCR, etc., nucleic acid is purified by alternately laminating an aqueous liquid layer and a water-insoluble gel layer in a capillary and passing magnetic particles to which the nucleic acid is attached. A device has been proposed (see Patent Document 1).

International Publication No. 2012/086243

  However, when the device disclosed in Patent Document 1 is stored for a long time, the components of the aqueous liquid layer may diffuse through the gel layer, and one aqueous liquid layer may be contaminated with the components of the other aqueous liquid layer. there were. Therefore, when storing a device for a long period of time, it is desirable to store at least the aqueous liquid layer separately in separate containers.

  When dividing each liquid into a plurality of containers, it is necessary to seal each container, and each container is connected by a simple operation as much as possible, and a predetermined aqueous phase and oil phase are alternately arranged. It is required to form a cut capillary.

  One of the objects according to some aspects of the present invention is that a plurality of fluids can be sealed and stored in separate containers, and the containers can be easily combined, and the seals are released when they are combined. Another object of the present invention is to provide a container assembly set capable of forming a flow path in which a plurality of fluids are arranged.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following aspects or application examples.

[Application Example 1] One aspect of a container assembly set according to the present invention is as follows.
The insertion part which forms the 1st flow path which distribute | circulates a 1st fluid, the cylindrical part formed around the said insertion part, and the 1st film which seals the said cylindrical part and seals the said 1st fluid A first container having,
A second container having a receiving portion that forms a second flow path through which the second fluid flows and receives the insertion portion; and a second film that seals the receiving portion and seals the second fluid;
Including
When the first container and the second container are connected, the first film and the second film are broken and the first flow path and the second flow path are communicated.

  According to the container assembly set of this application example, the first fluid and the second fluid can be sealed in the first container and the second container, respectively, and can be stored separately. In addition, the first container and the second container can be easily connected, and the first fluid and the second fluid can be easily arranged in the flow path that is communicated by the connection. Furthermore, even if the film that seals the fluid is broken by the connection of the container, it is difficult to block or narrow the communication channel.

[Application Example 2] In Application Example 1,
The first film and the second film may include an aluminum layer.

  According to such a container assembly set, the fluid sealed in each container is less likely to diffuse to the outside, and other substances are less likely to enter the container from the outside, and long-term storage is easier.

[Application Example 3] In Application Example 1 or Application Example 2,
At least one of the first film and the second film may have a multilayer structure in which two or more layers are laminated.

  According to such a container assembly set, the fluid sealed in each container is less likely to diffuse to the outside, and other substances are less likely to enter the container from the outside, and long-term storage is easier.

[Application Example 4] In any one of Application Examples 1 to 3,
The first film may have a three-layer structure of a polypropylene layer, an aluminum layer, and a polyethylene terephthalate layer in order from the side in contact with the first fluid.

  According to such a container assembly set, when the material of the first container is polypropylene, the adhesive strength between the first container and the first film is high and adhesion (fusion) is easy. Further, the strength and stiffness of the first film can be increased by the polyethylene terephthalate layer.

[Application Example 5] In any one of Application Examples 1 to 4,
The second film may have a three-layer structure of a polypropylene layer, an aluminum layer, and a polyethylene terephthalate layer in order from the side in contact with the second fluid.

  According to such a container assembly set, when the material of the second container is polypropylene, the adhesive strength between the second container and the second film is high and adhesion (fusion) is easy. The polyethylene terephthalate layer can increase the strength and stiffness of the second film.

[Application Example 6] In any one of Application Examples 1 to 5,
The thickness of the first film may be not less than 30 μm and not more than 100 μm.

According to such a container assembly set, the first film is not easily broken by an unintended force, and the user can break the first film without requiring a strong force. The first container and the second container Can be assembled (joined) easily.

[Application Example 7] In any one of Application Examples 1 to 6,
The thickness of the second film may be not less than 30 μm and not more than 100 μm.

  According to such a container assembly set, the second film is not easily broken by an unintended force, and the user can break the second film without requiring a strong force. The first container and the second container Can be assembled (joined) easily.

[Application Example 8] In any one of Application Examples 1 to 7,
When the first container and the second container are connected, the insertion part may be inserted into the receiving part, and the insertion part may have a shape that narrows toward the receiving part.

  According to such a container assembly set, the film can be more easily broken by the insertion portion, and the broken film can be further prevented from remaining in the flow path communicating by connection.

[Application Example 9] In any one of Application Examples 1 to 8,
When the first container and the second container are connected, the force required between the insertion part being inserted into the receiving part and the connection between the first flow path and the second flow path May be 0.1N or more and 10N or less.

  According to such a container assembly set, the film is not easily broken by an unintended force, and the user can assemble (couple) the first container and the second container without requiring a strong force.

[Application Example 10] In any one of Application Examples 1 to 9,
At least one of the first film and the second film may have a multilayer structure in which two or more layers are laminated, and at least one of the multilayer structures may have a notch.

  According to such a container assembly set, since the film is easily torn from the portion where the cutout exists, the position at which the film starts to tear can be controlled. As a result, the torn film can be further prevented from remaining in the flow path communicating by connection.

[Application Example 11] In any one of Application Examples 1 to 10,
The opposite side of the first channel to the insertion portion and the opposite side of the second channel to the receiving portion may be sealed.

  According to such a container assembly set, when the first container and the second container are connected, the internal pressure of each container increases until the film is broken. Therefore, the elongation until the film is torn is kept small. Thereby, the flow path is not easily blocked by the torn film.

Application Example 12 In Application Example 11,
When the first container and the second container are connected, the first flow path may be filled with the first fluid, and the second flow path may be filled with the second fluid. .

According to such a container assembly set, since each container does not contain bubbles or the like and is filled with a low-compressible fluid, when the first container and the second container are connected, until the film is torn, The internal pressure of each container can be further increased. Therefore, the elongation until the film is torn is kept small. Thereby, the flow path is not easily blocked by the torn film.

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. It is sectional drawing of the container assembly set 7 which concerns on embodiment. It is sectional drawing of the 2nd temporary assembly 612 which concerns on embodiment. It is sectional drawing of the 3rd washing container 230 and the elution container 300 which concern on embodiment. It is sectional drawing which shows the state which the 3rd washing container 230 and the elution container 300 which concern on embodiment joined. It is a schematic diagram which shows the state immediately before a film is torn when the 1st container and 2nd container which concern on embodiment are joined. It is a schematic diagram explaining the film 516 which concerns on 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 container assembly set according to the present invention is applicable to a biological material purification cartridge (container assembly), can be used as a cartridge for performing PCR, and can purify the biological material. Here, the biological substance is a substance related to the living body, such as nucleic acid (DNA, RNA), biological polymer such as polypeptide, protein, polysaccharide, protein, enzyme, peptide, nucleotide, amino acid, vitamin, etc. Including low molecular organic compounds derived from living organisms and inorganic compounds. In the following embodiments, description will be made using a nucleic acid as an example of a biological substance.

  The container assembly set according to the present invention may include a substance-binding solid phase carrier on which a biological substance is adsorbed. Here, the substance-binding solid phase carrier is a substance capable of holding a biological substance by adsorption, that is, by reversible physical bonding. The shape of the substance-binding solid phase carrier is preferably fine particles, but is not limited thereto, and may be fine fibers or nets, and is not particularly limited. The substance-binding solid phase carrier preferably has magnetism in order to move it in a desired direction while adsorbing the biological substance. In the following embodiment, a description will be given using a magnetic bead 30 (see FIGS. 7 and 8 described later) that adsorbs a nucleic acid as a substance-binding solid phase carrier.

The container assembly set according to the present embodiment seals the insertion portion that forms the first flow path through which the first fluid flows, the cylindrical portion that is formed around the insertion portion, and the cylindrical portion. A first container having a first film for sealing the first fluid; a second flow path through which a second fluid flows; and a receiving portion for receiving the insertion portion; and the receiving portion for sealing the receiving portion A second container having a second film for sealing the second fluid, and the first film and the second film are broken when the first container and the second container are connected to each other. One channel and the second channel communicate with each other.

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 container 400 includes a reaction receiving portion 404 formed at the open end, a bottom portion 402 formed at the other closed end, and a reservoir 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 406 having a predetermined space is provided around the reaction receiving unit 404. The reservoir 406 has a volume capable of receiving the liquid overflowing from the reaction container 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,
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 ionic agents and anionic surfactants such as sodium N-uraroyl sarcosine (SDS) can be mentioned. It is preferable to use so that it may become a range. 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 concentration of these chaotropic substances used varies depending on each substance. For example, when guanidine thiocyanate is used, the concentration ranges from 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 it is adsorbed to a solid rather than being 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 Device A PCR device 50 that performs nucleic acid elution processing and PCR using the container assembly 1 will be described with reference to FIGS. 9 and 10. 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. Next, the container assembly set according to this embodiment will be described with reference to the drawings. FIG. 11 is a cross-sectional view schematically showing the container assembly set 7 according to the embodiment. FIG. 11 shows the same cross section as FIG. The container assembly 1 can be obtained by assembling the container assembly set 7.

  In the container assembly set 7, as shown in FIG. 11, the flow path 2 of the adsorption container 100, the flow path 2 of the cleaning containers 210, 220, and 230, and the flow path 2 of the elution container 300 are not in communication.

  In the container assembly set 7, a film 502 is attached to one end (upper end) side of the adsorption container 100 (one end side of the plunger part 130), and the other end (lower end) side (the other end of the syringe part 120) of the adsorption container 100. A film 504 is attached to the side. The adsorption container 100 stores the adsorbed liquid 10 and the first oil 20 in a sealed manner.

  A film 506 is attached to one end side of the first cleaning container 210, and a film 508 is attached to the other end side of the first cleaning container 210. The first cleaning container 210 seals and stores the first cleaning liquid 12 and the oils 20 and 22.

  A film 510 is attached to one end side of the second cleaning container 220, and a film 512 is attached to the other end side of the second cleaning container 210. The second cleaning container 220 seals and stores the second cleaning liquid 14, the oils 22 and 24, and the magnetic beads 30.

  A film 514 is attached to one end side of the third cleaning container 230, and a film 516 is attached to the other end side of the third cleaning container 230. The third cleaning container 230 contains the third cleaning liquid 16 and the oils 24 and 26 in a sealed manner.

  A film 518 is attached to one end side of the elution container 300, and a film 520 is attached to the other end side of the elution container 300. The elution container 300 contains the eluate 32 and the fourth oil 26 in a sealed manner.

  A film 522 is attached to one end side of the reaction vessel 400. The reaction container 400 contains the reagent 34 and the fourth oil 26 in a sealed manner.

  The films 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522 will be described later. For convenience, in FIG. 11, the thicknesses of the films 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522 are shown relatively large relative to the sizes of the other members. Show.

  In the illustrated example, the adsorption container 100 and the cleaning containers 210 and 220 form a first temporary assembly 610, and the third cleaning container 230 and the elution container 300 form a second temporary assembly 620. .

  The container assembly set 7 includes a first container having a first flow path for containing a first fluid and a second container having a second flow path for containing a second fluid. Hereinafter, the first container is described as the third cleaning container 230, and the second container is described as the elution container 300.

5-1. First Container and Second Container FIG. 12 is a cross-sectional view schematically showing the third cleaning container 230 (first container) and the elution container 300 (second container), and forms a second temporary assembly 612. It shows the state. FIG. 13 is an enlarged view of the periphery of the third insertion portion 232 of the third cleaning container 230 and the elution receiving portion 304 of the elution container 300, and shows a state where the second temporary assembly 612 is released. FIG. 14 shows an enlarged view of a state in which the third cleaning container 230 and the elution container 300 are joined. 12 to 14 show the same cross section as FIG.

  As described in “2-2. Cleaning Container” above, the third cleaning container 230 is provided around the third insertion part 232 inserted into the elution receiving part 304 and the opening 233 of the third insertion part 232. And an annular third cover portion (first annular wall portion) 236.

  As shown in FIG. 12, in the second temporary assembly 612, the third insertion part 232 of the third cleaning container 230 is not inserted into the elution receiving part 304 of the elution container 300. That is, the flow path 2 of the third cleaning container 230 and the flow path 2 of the elution container 300 are not in communication. In the second temporary assembly 612, the inner wall 236 a of the third cover part 236 is in contact with the flange 308 of the elution container 300. Due to the friction between the third cover part 236 and the flange 308, the third cleaning container 230 moves in the vertical direction (the insertion direction A of the third insertion part 232 and the direction opposite to the direction A) with respect to the elution container 300. It is temporarily fixed to the elution container 300 in a difficult state. Similarly, in the first temporary assembly 610, the adsorption container 100 is temporarily fixed to the first cleaning container 210, and the first cleaning container 210 is temporarily fixed to the second cleaning container 220.

  As shown in FIG. 13, the third cover portion 236 of the third cleaning container 230 has an upper end (end opposite to the insertion direction A) connected to the outer wall of the third insertion portion 232 and a lower end (insertion direction A side). Extends beyond the third insertion portion 232. The inner wall 236a of the third cover portion 236 has an annular stepped portion 236b that increases in diameter downward (in the insertion direction A). The step portion 236b is positioned slightly below the lower end of the third insertion portion 232 (than the opening 233 of the third insertion portion 232).

  A film 514 is attached to the upper end side of the third cleaning container 230 as shown in FIG. As shown in FIGS. 12 and 13, a film 516 (first film) is attached to the inner wall 236 a of the third cover portion 236. Specifically, the film 516 is attached to the step portion 236b.

  The third cleaning container 230 is a film 514, 516, the first fluid (the third cleaning liquid 16 for cleaning the magnetic beads 30 on which the nucleic acid has been adsorbed), and the third fluid (oils 24, 26) that is immiscible with the first fluid. Is sealed and stored.

  The elution container 300 has the elution receiving part 304 into which the third insertion part 232 is inserted as described in “2-3. Elution container” above. An opening 305 is provided on the upper end side of the elution receiving portion 304, and the opening 305 is sealed with a film 518. In the example shown in FIG. 13, the elution receiving portion 304 has an annular second annular wall portion 320 provided around the film 518. That is, the film 518 is provided inside the second annular wall portion 320.

As shown in FIG. 12, the elution container 300 has an annular elution cover portion 306 provided around the opening of the elution insertion portion 302. The elution cover portion 306 has an upper end connected to the outer wall of the elution insertion portion 302 and a lower end extending beyond the elution insertion portion 302. The inner wall 306a of the elution cover portion 306 has an annular step portion 306b whose diameter increases downward. The step portion 306 b is located slightly below the lower end of the elution insertion portion 302. Elution insertion part 302
A film 520 is attached.

  The elution container 300 is a film 518, 520 that seals and stores the second fluid (eluent 32 that elutes nucleic acids from the magnetic beads 30) and the fourth fluid (fourth oil 26) that is immiscible with the second fluid. ing.

5-2. Joining of the first container and the second container When joining the third cleaning container 230 (first container) and the elution container 300 (second container), as shown in FIG. The third cleaning container 230 is moved in the insertion direction A. By the movement of the third cleaning container 230, the film 516 (first film) and the film 518 (second film) are broken by the third insertion portion 232 and / or the elution receiving portion 304, and the third insertion portion 232 is subsequently eluted. It is inserted into the receiving unit 304. The flow path 2 (first flow path) of the third cleaning container 230 and the flow path 2 (second flow path) of the elution container 300 communicate with each other. In this way, as shown in FIG. 14, the third cleaning container 230 and the elution container 300 can be joined. Therefore, the first container and the second container can break the two films and connect the flow paths by one operation (operation) of connecting the first container and the second container.

  The force applied when joining the 3rd washing container 230 (1st container) and the elution container 300 (2nd container) is not specifically limited. However, from the viewpoint that the film is not easily broken by an unintended force and the user can assemble (join) the first container and the second container without requiring a strong force, the first container and the second container. As the force required between the insertion portion being inserted into the receiving portion and the communication between the first flow path and the second flow path is 0.01N or more and 20N or less, preferably 0.05N or more 15N or less, more preferably 0.1N or more and 10N or less, and further preferably 1N or more and 8N or less.

  The force when the first container and the second container are connected can be measured, for example, using a force gauge. For example, two containers are arranged coaxially on the trajectory of a device that slides left and right (or up and down), and a digital force gauge (for example, SIMPO; FGP-50) is brought into contact with one container. The load at that time can be measured while sliding in the connecting direction, and the maximum value can be stored.

  Here, the first container is described as the third cleaning container 230, and the second container is described as the elution container 300. However, the first container and the second container are the other containers constituting the container assembly sets 7, 8. It may be.

  For example, the first container may be the adsorption container 100 and the second container may be the first cleaning container 210. In this case, the first fluid and the third fluid sealed and accommodated in the adsorption container 100 are the adsorption liquid 10 and the oil 20 for adsorbing the nucleic acid on the magnetic beads 30, respectively. The second fluid and the fourth fluid sealed and accommodated in the first cleaning container 210 are the first cleaning liquid 12 and the oils 20 and 22 for cleaning the magnetic beads 30 to which the nucleic acid has been adsorbed, respectively. The film 504 (first film) and the film 506 (second film) are broken by the first receiving part 214 and / or the suction insertion part 122, and the suction container 100 and the first cleaning container 210 are joined. Is done.

For example, the first container may be the first cleaning container 210 and the second container may be the second cleaning container 220. In this case, the first fluid and the third fluid sealed and accommodated in the first cleaning container 210 are the first cleaning liquid 12 and the oils 20 and 22 for cleaning the magnetic beads 30 to which the nucleic acid is adsorbed, respectively. The second fluid and the fourth fluid sealed and accommodated in the second washing container 220 are the second washing liquid 14 and the oils 22 and 24 for washing the magnetic beads 30 to which the nucleic acid has been adsorbed, respectively. Then, the film 508 is formed by the second receiving unit 224 and / or the first insertion unit 212.
The (first film) and the film 510 (second film) are broken, and the first cleaning container 210 and the second cleaning container 220 are joined.

  For example, the first container may be the second cleaning container 220, and the second container may be the third cleaning container 230. In this case, the first fluid and the third fluid sealed and accommodated in the second cleaning container 220 are the second cleaning liquid 14 and the oils 22 and 24 for cleaning the magnetic beads 30 to which the nucleic acid has been adsorbed, respectively. The second fluid and the fourth fluid sealed and accommodated in the third washing container 230 are the second washing liquid 16 and the oils 24 and 26 for washing the magnetic beads 30 to which the nucleic acid has been adsorbed, respectively. Then, the film 512 (first film) and the film 514 (second film) are broken by the third receiving part 234 and / or the second insertion part 222, and the second cleaning container 220 and the third cleaning container 230 are joined. Is done.

  Further, for example, the first container may be the elution container 300 and the second container may be the reaction container 400. In this case, the first fluid and the third fluid sealed and accommodated in the elution container 300 are the eluent 32 and the oil 26 that elute the nucleic acid from the magnetic beads 30, respectively. The second fluid to be sealed and stored in the reaction vessel 400 is the oil 26 that is immiscible with the reagent 34 for performing the nucleic acid amplification reaction. Then, the film 520 (first film) and the film 522 (second film) are broken by the reaction receiving unit 404 and / or the elution insertion unit 302, and the elution container 300 and the reaction container 400 are joined.

  As above-mentioned, although the 1st film and the 2nd film are affixed on the 1st container and the 2nd container, respectively, it is pierced by an insertion part and / or a receiving part, or when an affixing part peels off The respective flow paths of the first container and the second container are communicated and joined. Then, as shown in FIG. 14, after the first container and the second container are joined, the first film and the second film block the flow path 2 in the gap α between the first container and the second container. It is stored without.

  When the first container and the second container are joined, the flow path 2 is not blocked and is not sandwiched by the seal part S1 between the third insertion part 232 and the elution receiving part 304. As one of the conditions required for the film, the maximum length from the breaking point to the pasting portion when it is torn is the length that does not reach the seal portion S1 (not reach the flow path 2). It is. Moreover, in this condition, it is more preferable to consider the elongation when the film is torn.

  In the container assembly set 7 of the present embodiment, when the first container and the second container are connected, the insertion part of the first container is inserted into the receiving part of the second container. In the present embodiment, as illustrated, the third insertion portion 232 (insertion portion) has a shape that becomes narrower toward the elution reception portion 304 (reception portion). Thereby, the first film and the second film are more easily broken by the insertion portion. Thereby, the elongation when the film is torn is suppressed to be small, and it is difficult to be pinched by the seal between the insertion portion and the receiving portion.

  Furthermore, in the container assembly set 7 of this embodiment, no matter how the first container and the second container are selected, the two films are broken by one operation and both are joined to form a flow path. Can do. Further, as described above, both ends of each container are sealed with a film. Therefore, no matter what order the containers are joined together, when joining, the opposite side of the flow path communicating with the insertion section and the receiving section to be joined is sealed with a film.

  FIG. 15 is a schematic diagram showing a state immediately before the film is broken when the first container and the second container are joined. FIG. 15A shows a state where the internal pressure in each container does not increase (a state where the end opposite to the joining end is open, for example), and FIG. 15B shows that the internal pressure in each container increases. The state (state in which the edge part on the opposite side of a joining end is sealed) is shown.

  As shown in FIG. 15 (a), when the film is pierced by the insertion portion and the receiving portion, the two films are broken in a stretched state if the internal pressure in each container is not increased. However, in this case, the film is designed to be located at a position out of the flow path regardless of the position where the film starts to tear.

  On the other hand, as shown in FIG. 15 (b), when the internal pressure in each container is increased, the expansion of the film is suppressed due to the influence of the internal pressure (the volume inside the container is less likely to decrease than the film). The As a result, the elongation of the two films breaks in a smaller state. Thereby, since the maximum length from the breaking point at the time of tearing to the affixed portion can be shortened, the torn film is more difficult to reach the seal portion S1 (see FIG. 14), and the first is more reliably performed. The torn film can be stored in the gap α between the container and the second container without blocking the flow path.

  Here, a fluid such as oil has a low compressibility, whereas a gas is a fluid having a compressibility higher than that of oil (more easily compressed). Therefore, when the end opposite to the joining end of the first container and the second container is sealed, if it is sealed so that no gas exists in the flow path of each container, the insertion part and the receiving part are received. Since the internal pressure rises sharply when the film is pierced by the portion, the elongation of the ruptured film can be easily reduced. That is, when the first container and the second container are connected, if the first flow path is filled with the first fluid and the second flow path is filled with the second fluid, It is possible to securely store the gap α between the first container and the second container without blocking the flow path.

5-4. Films 504, 506, 508, 510, 512, 514, 516, 518, 520, 522 are all films that are torn when the container assembly set 7 is put into use. These films are not particularly limited as long as a predetermined fluid is sealed in a container and can be broken by a predetermined operation. These films may have either a single layer structure or a multilayer structure. A multilayer structure is more preferable because various functions can be assigned to each layer. Examples of such functions include gas barrier properties, adhesiveness, weldability, strength (rigidity), antibacterial properties, and visibility. The material of the film is also not particularly limited, and examples thereof include polymers such as polyester, polyolefin, polyvinyl alcohol, polyethylene glycol, polyvinyl acetate, polycarbonate, acrylic resin, and copolymers thereof, aluminum, titanium, silicon, and the like. Metals and colorants such as oxides, pigments, and dyes thereof can be appropriately included in accordance with a predetermined function.

  FIG. 16 is an external perspective view and a schematic cross-sectional view of an example of a film. The films 504, 506, 508, 510, 512, 514, 516, 518, 520, and 522 may have the same configuration or different configurations. Here, the film 516 will be described as an example. By describing the film 516, description of substantially similar films 504, 506, 508, 510, 512, 514, 518, 520, 522 is omitted.

  The film 516 includes an aluminum layer 516a (Al layer 516a), a polyethylene terephthalate layer 516b (PET layer 516b) disposed on one surface side of the Al layer 516a, and a polypropylene disposed on the other surface side of the Al layer 516a. A multilayer structure including the layer 516c.

The Al layer 516a is, for example, an aluminum vapor deposition film. Since the Al layer 516a has gas barrier properties, when the film 516 includes the Al layer 516a, the first fluid sealed in the third cleaning container 230 (the third cleaning liquid 16 that cleans the magnetic beads 30 on which the nucleic acid has been adsorbed). ) Can be made difficult to diffuse outside, and other substances can be made difficult to enter the third cleaning container 230 from the outside. Therefore, in a state where the first fluid (the third cleaning liquid 16 for cleaning the magnetic beads 30 to which the nucleic acid has been adsorbed) and the third fluid (oils 24 and 26) that are immiscible with the first fluid are sealed, the third cleaning container is sealed. It becomes easy to store 230 for a long time. The thickness of the Al layer 516a is not particularly limited, and is, for example, 10 μm or more and 50 μm or less, preferably 15 μm or more and 45 μm or less, and more preferably 20 μm or more and 40 μm or less. When the thickness of the Al layer 516a is within this range, better gas barrier properties can be imparted.

  The film 516 has a PET layer 516b and a PP layer 516c. Examples of the function of the PET layer 516b include giving the film 516 mechanical strength and stiffness and protecting the Al layer 516a. The Al layer 516a may be formed by depositing aluminum on the PET layer 516b. Examples of the function of the PP layer 516c include providing the film 516 with mechanical strength and protecting the Al layer 516a. In addition, when the material of the container to which the film 516 is attached is PP, it is easy to fuse the film to the container. In this case, if the other layer is made of a material having a higher melting point than PP, the film and the container can be easily fused. In addition, as an aspect which affixes a film to a container, it is not limited to a melt | fusion, You may use an appropriate adhesive agent etc. In the illustrated example, the container is sealed so that the PP layer 516c side is in contact with the container. In the example of the film 516, a three-layer structure is used, but the number of layers is not limited, and a two-layer structure or a multilayer structure of four or more layers may be used as long as it has a predetermined function.

  The thickness of the PET layer 516b is not particularly limited, and is, for example, 5 μm to 40 μm, preferably 10 μm to 30 μm. When the thickness of the PET layer 516b is within this range, sufficient strength can be imparted to the film 516. Further, the thickness of the PP layer 516c is not particularly limited, and is, for example, 5 μm or more and 40 μm or less, preferably 10 μm or more and 40 μm or less.

  Further, the total thickness of the film 516 is such that the film is not easily broken by an unintended force, and the user can assemble (join) the first container and the second container without requiring a strong force. Therefore, the thickness is 15 μm or more and 200 μm or less, preferably 20 μm or more and 150 μm or less, more preferably 30 μm or more and 100 μm or less.

On the other hand, the film 516 may have a function of determining a position at which tearing starts (controlling a break starting position). In the example of FIG. 16, a notch 516d is formed in the PP layer 516c. The notch 516d can be a thin portion of the PP layer 516c and does not necessarily pass through the PP layer 516c. In any case, the strength of the PP layer 516c is small at the notch 516d, and the strength of the entire film 516 is weakened at this position. Therefore, by forming the notch 516d, the break start position in the film 516 can be controlled. Although not shown, when the film 516 has a layer structure of four or more layers, if the cutout 516d is formed in at least one layer, the position of the cutout 516d is set to the break start point. Can do. That is, when the film 516 has a multilayer structure, at least one layer of the multilayer structure may have a notch (or a partially thin area).
The planar position where the notch 516d is formed is not particularly limited, and can be set to an appropriate position according to the shape of the first container and the second container. In addition, the shape of the notch 516d is a linear groove in a plan view in the illustrated example. However, if the strength of the film 516 can be reduced and the film can be easily broken from the vicinity of the notch 516d. Any shape may be used. In the example shown in the drawing, the notch 516d is a groove having a depth enough to expose the Al layer 516a, but it is not necessary to penetrate the thickness direction of the PP layer 516c, and the PP layer 516c is thinned. Only the effect as the notch 516d can be obtained.

  In the illustrated example, the notch 516d is formed in the PP layer 516c of the three-layer structure, but it may be formed in any layer or a plurality of layers. In the illustrated film 516, since the Al layer 516a functions as a gas barrier layer, the cutout 516d is more preferably formed in the PP layer 516c and / or the PET layer, but not in the Al layer 516a. Further, when the film 516 is handled with vacuum tweezers or the like when the film 516 is attached to the container, it is more preferable that the notch 516d is not formed on the surface that adsorbs the vacuum tweezers.

  Further, in the illustrated example, the notch 516d has a shape in which two linear grooves as viewed in a plane intersect with each other near the center of the film 516. If it does in this way, while the break start point of the film 516 can be set to the part which the groove | channel crossed, the film 515 can be broken along the groove | channel on a line. Therefore, in addition to the control of the break start point, the break shape (break direction) can also be controlled. In the illustrated example, when the film 516 is torn, it is broken into four fan-shaped shapes, and the length of the torn film 516 along the longitudinal direction of the container is a pasted region (stepped portion) from the center of the film 516. 236b) (see FIG. 15). In the example shown in the figure, two linear grooves are formed as the notch 516d. However, when the notch 516d is a linear groove, the number of grooves may be one or three or more. Good.

  The break starting point can be set at an appropriate position in plan view, but by setting it near the center of the film 516, the broken film 516 is prevented from becoming too long along the longitudinal direction of the container. For example, the length of the container can be reduced. Here, the vicinity of the center of the film 516 refers to a position within 50, preferably within 40, more preferably within 30 from the center of gravity when the distance from the center of gravity to the end of the film 516 is 100 in plan view. Refers to that.

  The above-described embodiments and modifications are merely examples, and the present invention is not limited to these. For example, it is possible to appropriately combine each embodiment and each modification.

  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, 3 ... Magnet, 5 ... Nucleic acid purification device, 7, 8 ... Container assembly set, 10 ... Adsorption liquid, 11 ... Air, 12 ... 1st washing | cleaning liquid, 14 ... 2nd washing | cleaning liquid , 16 ... the third cleaning solution, 20 ... the first oil, 22 ... the second oil, 24 ... the third oil, 26 ... the fourth oil, 30 ... the magnetic beads, 32 ... the eluate, 34 ... the reagent, 36 ... the droplet, DESCRIPTION OF SYMBOLS 50 ... PCR apparatus, 55 ... Fluorescence measuring device, 60 ... Rotation mechanism, 61a ... 1st inside surface, 61b ... 2nd inside surface, 65 ... Heater, 66 ... Motor for rotation, 70 ... Magnet moving mechanism, 80 ... Pressing mechanism , 90 ... Controller, 100 ... Suction container, 110 ... Cap, 112 ... Ventilation part, 120 ... Syringe part, 122 ... Suction insertion part, 126 ... Suction cover part, 130 ... Plunger part, 131 ... Cylindrical part, 132 ... Rod-shaped part, 134 ... tip part, 00 ... Cleaning container, 210 ... First cleaning container, 212 ... First insertion part, 214 ... First receiving part, 216 ... First cover part, 220 ... Second cleaning container, 222 ... Second insertion part, 224 ... First 2 receiving part, 226 ... 2nd cover part, 230 ... 3rd washing container, 232
3rd insertion part, 233 ... opening, 234 ... 3rd receiving part, 236 ... 3rd cover part, 236a ... inner wall, 236b ... step part, 300 ... elution container, 302 ... elution insertion part, 304 ... elution receiving part, 305 ... Opening, 306 ... Elution cover part, 306a ... Inner wall, 306b ... Step part, 310 ... End face, 320 ... Second annular wall part, 400 ... Reaction vessel, 402 ... Bottom part, 404 ... Reaction receiving part, 406 ... Reservoir part , 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522 ... film, 516a ... Al layer, 516b ... PET layer, 516c ... PP layer, 516d ... notch, 610 ... first provisional Assembly, 612 ... second temporary assembly, S1 ... seal part, α ... gap

Claims (12)

The insertion part which forms the 1st flow path which distribute | circulates a 1st fluid, the cylindrical part formed around the said insertion part, and the 1st film which seals the said cylindrical part and seals the said 1st fluid A first container having,
A second container having a receiving portion that forms a second flow path through which the second fluid flows and receives the insertion portion; and a second film that seals the receiving portion and seals the second fluid;
Including
A container assembly set, wherein when the first container and the second container are connected, the first film and the second film are broken and the first flow path and the second flow path are communicated. In claim 1,
The container assembly set, wherein the first film and the second film include an aluminum layer. In claim 1 or claim 2,
At least one of the first film and the second film is a container assembly set having a multilayer structure in which two or more layers are laminated. In any one of Claims 1 to 3,
The container assembly set, wherein the first film has a three-layer structure of a polypropylene layer, an aluminum layer, and a polyethylene terephthalate layer in order from the side in contact with the first fluid. In any one of Claims 1 thru | or 4,
The container assembly set, wherein the second film has a three-layer structure of a polypropylene layer, an aluminum layer, and a polyethylene terephthalate layer in order from the side in contact with the second fluid. In any one of Claims 1 thru | or 5,
The container assembly set, wherein the first film has a thickness of 30 μm or more and 100 μm or less. In any one of Claims 1 thru | or 6,
The container assembly set, wherein the second film has a thickness of 30 μm or more and 100 μm or less. In any one of Claims 1 thru | or 7,
When the first container and the second container are connected, the insertion portion is inserted into the receiving portion, and the insertion portion has a shape that narrows toward the receiving portion. In any one of Claims 1 thru | or 8,
When the first container and the second container are connected, the force required between the insertion part being inserted into the receiving part and the connection between the first flow path and the second flow path Is a container assembly set that is 0.1N or more and 10N or less. In any one of Claims 1 thru | or 9,
At least one of the first film and the second film has a multilayer structure in which two or more layers are laminated, and at least one of the multilayer structures has a notch. In any one of Claims 1 thru | or 10,
The container assembly set in which the side opposite to the insertion portion of the first channel and the side opposite to the receiving portion of the second channel are sealed. In claim 11,
When the first container and the second container are connected, the first assembly is filled with the first fluid, and the second passage is filled with the second fluid. Solid set.