Device and method for producing device

Abstract

This device includes an internal space capable of storing a sample liquid containing a substance to be detected and coupled to magnetic particles, and is used for purifying the substance by moving the substance in the internal space using the magnetic particles. The device includes an inlet for introducing a sample liquid to a central part of the internal space. In the internal space, a cleaning liquid for removing impurities contained in the sample liquid and an eluent for eluting the substance are stored separately from each other by a hydrophobic liquid in an outer peripheral part surrounding the central part. The internal space is filled with a liquid containing the cleaning liquid, the eluent, and the hydrophobic liquid.

Description

Device and method for manufacturing the same

[0001] The present disclosure relates to devices and methods for manufacturing devices.

[0002] In the testing of specimens collected from living organisms, in order to detect the target substance with high sensitivity, a purification procedure to remove the influence of contaminants contained in the specimen is required. A known purification method uses magnetic particles whose surfaces are modified with a binding substance that binds to the target substance, and traps the target substance on the magnetic particles via the binding substance to remove the contaminants.

[0003] Japanese Patent Application Laid-Open No. 2013-130548 proposes a chip device that enables purification operations using magnetic particles.

[0004] The chip device disclosed in JP 2013-130548 A includes a chip having an internal space defined by a groove formed in a substrate, in which a gel phase and an aqueous liquid phase are alternately accommodated in the longitudinal direction of the chip, and a magnetic field application means for applying a magnetic field to the chip and moving it. In this chip device, by moving the magnetic field along the longitudinal direction of the chip groove, it is possible to move the gel phase and the aqueous liquid phase, in which magnetic particles are alternately arranged.

[0005] The chip described in JP 2013-130548 A can separate and hold different types of aqueous liquid phases with a gel phase, and by moving the magnetic particles in the longitudinal direction, the magnetic particles can be brought into contact with different types of aqueous liquid phases in sequence. On the other hand, in the device described in JP 2013-130548 A, the magnetic particles are configured to move only along the longitudinal direction of the groove, and the degree of freedom of movement of the magnetic particles is limited.

[0006] The present disclosure has been made in consideration of the above circumstances, and provides a device for purifying a substance to be detected using magnetic particles that bind to the substance, in which the magnetic particles have a high degree of freedom of movement, and a method for manufacturing the device.

[0007] The device of the present disclosure has an internal space capable of containing a sample liquid containing a target substance to be detected that binds to magnetic particles, and is a device in which the target substance is purified by moving the target substance within the internal space using the magnetic particles, and has an inlet in the center of the internal space for introducing the sample liquid, and in the internal space, a washing liquid for removing impurities contained in the sample liquid and an elution liquid for eluting the target substance to be detected are contained in the outer periphery surrounding the central part, separated from each other by a hydrophobic liquid, and the internal space is filled with a liquid containing the washing liquid, the elution liquid, and the hydrophobic liquid.

[0008] A magnetic particle solution containing magnetic particles may be accommodated in the center of the internal space.

[0009] The thickness of the central portion of the internal space is preferably greater than the thickness of the outer periphery.

[0010] In the inner space, a reagent for the substance to be detected is preferably accommodated in the outer peripheral portion, separated from the washing liquid and the elution liquid by a hydrophobic liquid.

[0011] The eluate may contain a reagent for the substance to be detected.

[0012] The inner space may be provided with a processing region on the outer periphery for mixing the reagent and the substance to be detected.

[0013] The inner space may be provided with a detection region at its outer periphery, which contains the reagent and the substance to be detected that have been mixed in the processing region, and in which the substance to be detected is optically detected from the outside.

[0014] It is preferable that the thickness of the processing region of the internal space is thinner than the thickness of other regions.

[0015] It is preferable that the internal space has a narrowed portion between the processing region and the other region.

[0016] The device of the present disclosure may further include a reagent storage section connected to the internal space via a valve and containing a reagent for the substance to be detected, and a processing chamber located farther from the center than the storage section, into which a liquid containing the reagent and the substance to be detected is delivered and into which the substance to be detected is optically detected from the outside.

[0017] The internal space is preferably formed by a main body member having a recess on one surface thereof and a thin film bonded to the main body member in a manner that covers the recess on one surface thereof.

[0018] A pillar may be provided in the internal space to prevent the thin film from coming into contact with the bottom surface of the recess.

[0019] It is preferable that at least one of the region for storing the cleaning solution, the region for storing the eluate, and the central portion of the internal space is subjected to a hydrophilic treatment.

[0020] The hydrophobic liquid preferably has a lower specific gravity than the washing liquid and the elution liquid.

[0021] The hydrophobic liquid may be silicone oil.

[0022] The thickness of the internal space is preferably 1 mm or less.

[0023] The device of the present disclosure may be provided with a detachable lid for the insertion port, and the lid preferably includes a plug to be inserted into the insertion port, the plug having an air hole for discharging air from the internal space to the outside when the plug is inserted into the insertion port, and a sealing member for blocking the air hole.

[0024] The method for manufacturing a device disclosed herein is a method for manufacturing a device in which a cleaning solution and an elution solution are placed at positions separated from each other on the outer periphery of the internal space using centrifugal force generated by rotating the device around different centers of rotation.

[0025] According to the present disclosure, it is possible to provide a device having a function of performing purification using magnetic particles, in which the magnetic particles have a high degree of freedom of movement, and a method for manufacturing the device.

[0026] 15. A perspective view schematically showing a device of an embodiment. A cross-sectional view taken along line II-II of FIG. 1. An exploded perspective view of a device of an embodiment. A perspective view of a main body member viewed from one side. A plan view showing the internal space of a device of an embodiment. A schematic configuration diagram of an inspection apparatus. A diagram showing the trajectories of magnetic particles in the internal space of a device. A perspective view of a main body member of a modified example. A perspective view of a main body member of a modified example. A plan view of a device of a modified example. A diagram showing a liquid disposing process in the internal space of a device. A diagram showing a liquid disposing process in the internal space of a device. An explanatory diagram of a method of sealing the internal space. An explanatory diagram of a method of sealing the internal space. A plan view of the internal space of a device of a modified example. A diagram showing the trajectories of magnetic particles in the internal space of the device shown in FIG. 15 (part 1). A diagram showing the trajectories of magnetic particles in the internal space of the device shown in FIG. 15 (part 2). A diagram showing a liquid disposing process in the internal space of the device shown in FIG. 15 (part 1). A diagram showing a liquid disposing process in the internal space of the device shown in FIG. 15 (part 2).

[0027] Hereinafter, embodiments of the present disclosure will be described in detail with reference to the drawings. Note that the scale of each component in the drawings has been appropriately changed from the actual scale. In each drawing, the same components are assigned the same reference numerals.

[0028] "Device" Fig. 1 is a perspective view schematically showing a device 10 of one embodiment, and Fig. 2 is a cross-sectional view taken along line II-II in Fig. 1. Fig. 3 is an exploded perspective view of the device 10.

[0029] 1 includes an internal space 12 capable of accommodating a sample liquid containing a target substance to be detected that binds to magnetic particles. In the device 10, the target substance is purified by moving the target substance within the internal space 12 using the magnetic particles.

[0030] The particle size of the magnetic particles is, for example, 0.1 μm to 500 μm. The surfaces of the magnetic particles are provided with modifiers such as silica or functional groups that specifically bind to the target substance.

[0031] First, a brief description of the purification method for the analyte is provided. Magnetic particles bearing modifiers that specifically bind to the analyte are prepared, and a magnetic particle solution containing these magnetic particles is mixed with a sample solution. If the analyte is contained in the sample solution, the analyte binds to the modifiers on the magnetic particles. The magnetic particles that have captured the analyte are trapped with a magnet and transferred from the mixture of the sample solution and magnetic particle solution into a washing solution for washing. This allows impurities adhering to the magnetic particles to be removed by the washing solution. The magnetic particles that have captured the analyte are then transferred into an elution solution. The elution solution has the function of breaking down the bond between the analyte and the modifier, and the analyte is released from the magnetic particles in the elution solution. After the analyte is released from the magnetic particles, a purified extract of the analyte is obtained by transferring only the magnetic particles from the elution solution.

[0032] In the device 10, a purification process for the object to be detected is carried out, and an extract of the object to be detected can be obtained.

[0033] As shown in FIGS. 1 to 3, device 10 comprises a body member 20, a membrane 30, and a lid 40.

[0034] Fig. 4 is a perspective view of the main body member 20 viewed from one surface 20a (the surface that corresponds to the back surface in Fig. 3). As shown in Fig. 4, the main body member 20 has a recess 21 on one surface 20a, and the thin film 30 is bonded to this surface 20a of the main body member 20 in a manner that covers the recess 21. The recess 21 of the main body member 20 and the thin film 30 that covers the recess 21 form an internal space 12. The internal space 12 is a thin-plate-like space whose surface perpendicular to the thickness direction is sufficiently wide relative to its thickness.

[0035] The material of the main body member 20 can be any known resin molded plastic material without any particular restrictions, but polycarbonate, polypropylene, cycloolefin or silicone resin is preferred from the viewpoint of heat resistance and transparency.

[0036] As with the main body member 20, the thin film 30 can be made of any known plastic material without restriction, but is preferably made of the same material as the main body member 20 in terms of adhesion to the main body member 20. The main body member 20 is rigid, and the thin film 30 is flexible. In the following description, for convenience of explanation, the thin film 30 side is referred to as the bottom surface 10a of the device 10, and the side with the lid 40 is referred to as the top surface 10b of the device 10.

[0037] As shown in Fig. 1, the device 10 has, as an example, a circular outer shape in plan view. In this case, as shown in Fig. 3, the body member 20 and the thin film 30 also have a circular outer shape in plan view.

[0038] 4, the recess 21 provided on one surface 20a of the main body member 20 includes a circular portion 21a, a first region 21b and a second region 21c spaced apart from each other on the outer edge of the circular portion 21a, and a through-hole 22 at the center of the circular portion 21a.

[0039] 2, a cylindrical boss 23 is provided on the other surface 20b of the main body member 20, opposite the surface 20a where the recess 21 is provided, with the edge of the through-hole 22 protruding from the surface. The sample liquid is introduced into the internal space 12 from the boss 23 side through the through-hole 22. That is, the opening of the boss 23, which is one end of the through-hole 22, is an inlet 24 for introducing the sample liquid (see FIG. 3). The inlet 24 is located in the center 13a of the internal space 12 in a plan view (see FIG. 5), and is configured so that the sample liquid is introduced into the center 13a of the internal space 12.

[0040] The lid 40 is provided detachably to the inlet 24. Before use of the device 10, the lid 40 is attached to the inlet 24. A user removes the lid 40 from the inlet 24 and inserts a sample liquid into the internal space 12 through the inlet 24. After the sample liquid has been inserted, the user attaches the lid 40 to the inlet 24. The device 10 is then loaded into a testing apparatus 100 (see FIG. 6 ), which will be described later, and a purification process is performed in the testing apparatus 100. With the lid 40 attached to the inlet 24, the internal space 12 is formed by the main body member 20, the thin film 30, and the lid 40.

[0041] 5 is a plan view showing the internal space 12 of the device 10. As described above, the internal space 12 is defined by the recess 21 formed on one surface 20a of the main body member 20, the thin film 30, and the lid 40, and the shape of the internal space 12 in a plan view substantially coincides with the shape of the recess 21 in a plan view. The internal space 12 includes a circular portion 13 defined by the circular portion 21a of the recess 21 and the thin film 30, a cleaning liquid storage region 14 defined by the first region 21b of the recess 21 and the thin film 30, and an eluate storage region 15 defined by the second region 21c of the recess 21 and the thin film. The internal space 12 is geometrically composed of three parts: a circular portion 13, a cleaning liquid storage area 14, and an eluate storage area 15. However, functionally, the circular portion 13 is separated into a central portion 13a (the portion indicated by dotted hatching in FIG. 5 ), which is an area that includes the center of the circular portion 13 and is smaller than the circular portion 21a, and an annular portion 13b (the portion indicated by gray in FIG. 5 ) that surrounds the central portion 13a. The annular portion 13b, the cleaning liquid storage area 14, and the eluate storage area 15 correspond to the "periphery surrounding the central portion" in the technology of the present disclosure.

[0042] In this example, the recess depth of the second region 21c is shallower than that of the circular region 21a and the first region 21b. Note that the internal space 12 is formed by the recess 21 and the thin film 30 covering the recess 21, and therefore the recess depth corresponds to the thickness of the internal space 12. That is, in the internal space 12, the thickness of the eluate storage region 15 is thinner than that of the circular region 13 and the cleaning liquid storage region 14.

[0043] The thickness of the internal space 12 is preferably 1 mm or less throughout, more preferably less than 1 mm, and even more preferably 0.8 mm or less.

[0044] Four pillars 25 are provided in the circular portion 21a of the recess 21 of the main body member 20 at positions surrounding the through-hole 22. The thickness of the pillars 25 is equal to the depth of the recess 21. The pillars 25 are provided to prevent the thin film 30 from being depressed into the recess due to its flexibility and coming into contact with the bottom surface of the recess 21.

[0045] The lid 40 includes a plug 42 inserted into the insertion port 24 and a sealing member 46. The plug 42 has an air hole 43 that discharges air from the internal space 12 to the outside when inserted into the insertion port 24. The air hole 43 is located in the center of a cylindrical protrusion 44 of the plug 42 that is inserted into the insertion port 24. A groove is formed along the circumference of the cylindrical protrusion 44 of the plug 42, and an O-ring 45 is provided in the groove. In this example, the sealing member 46 has a protrusion 46a (see FIG. 2) that blocks the air hole. A circular groove centered on the protrusion 46a is formed on the surface of the sealing member 46 that contacts the plug 42, and an O-ring 47 is provided in the groove. Both the O-rings 45 and 47 are provided to seal the internal space 12. Note that the sealing member 46 does not necessarily have to have the protrusion 46a and may be formed of a flat plate, a thin film, or the like. Furthermore, the sealing member 46 may be configured to be detachable from the plug 42 as desired. Furthermore, the sealing member 46 and the plug 42 may be integrated via a hinge or the like.

[0046] 5 , in the device 10, a magnetic particle solution 32 containing magnetic particles is contained in the central portion 13a, a hydrophobic liquid 38 is contained in the annular portion 13b surrounding the central portion 13a, a cleaning solution 34 is contained in the cleaning solution containing region 14, and an elution solution 36 is contained in the elution solution containing region 15. In FIG. 5 , the magnetic particle solution 32 is indicated by dotted hatching, the cleaning solution 34 by diagonal hatching downward to the right, the elution solution 36 by diagonal hatching downward to the left, and the hydrophobic liquid 38 by gray. In this way, the internal space 12 is filled with the magnetic particle solution 32, the cleaning solution 34, the elution solution 36, and the hydrophobic liquid 38, and the magnetic particle solution 32, the cleaning solution 34, and the elution solution 36 are contained while being separated from one another by the hydrophobic liquid 38.

[0047] The magnetic particle solution 32, the cleaning solution 34, and the elution solution 36 are mixtures containing water as a primary component and are liquids that remain separate from the hydrophobic liquid 38. Water or mixtures containing water as a primary component are hereinafter referred to as aqueous liquids. The aqueous liquids are contained within the internal space 12, separated by the hydrophobic liquid 38. Since the thin internal space 12 is filled with these liquids, the movement of each liquid is suppressed and each liquid is maintained in its respective position. It is preferable that at least one of the central portion 13a containing the magnetic particle solution 32, the cleaning solution containing region 14 containing the cleaning solution 34, and the elution solution containing region 15 containing the elution solution 36 be subjected to a hydrophilic treatment. It is more preferable that all of the central portion 13a, the cleaning solution containing region 14, and the elution solution containing region 15 be subjected to a hydrophilic treatment. The hydrophilic treatment may be applied to portions of the recess 21 formed in the main body member 20 corresponding to each region, or to portions of the thin film 30 corresponding to each region, or to both. If the areas that contain the water-based liquid are subjected to a hydrophilic treatment, the water-based liquid can be more easily contained in the respective positions, and migration to other areas can be restricted.

[0048] The hydrophobic liquid 38 preferably has a specific gravity smaller than that of the cleaning liquid 34 and the elution liquid 36. Silicon oil is suitable as the hydrophobic liquid 38.

[0049] The eluate 36 may contain a reagent for the target substance. When the eluate 36 contains a reagent, the eluate storage area 15 also functions as a processing area for mixing the target substance with the reagent. In this case, the eluate storage area 15 may be used as a detection area for optically detecting the target substance from the outside.

[0050] A method for testing an object to be detected, including a purification process for the object to be detected, using the device 10 will be described. The device 10 is loaded into a testing apparatus 100, the schematic configuration of which is shown in Figure 6, and testing of the object to be detected is carried out. The testing apparatus 100 is, as an example, a nucleic acid testing apparatus, and the object to be detected is a specific nucleic acid. The device 10 contains, as reagents in the eluate 36, primers and fluorescent probes for nucleic acid amplification.

[0051] The inspection device 100 shown in FIG. 6 includes a rotation mechanism 110, a magnet moving mechanism 120, a heating unit 130, and a light detection unit 140.

[0052] The rotation mechanism 110 includes a rotation drive unit 112 such as a motor, a mounting unit 113 to which the device 10 is mounted, and a rotation coupler 114 connected to the shaft of the rotation drive unit 112 and rotatably supporting the mounting unit 113. The mounting unit 113 is provided with a chuck mechanism that chucks the device 10. The chuck mechanism is, for example, a claw that fits into a recess (not shown) provided in the device 10. By driving the rotation drive unit 112, the mounting unit 113 is rotated about a rotation axis A, and thereby the device 10 is rotated.

[0053] The magnet moving mechanism 120 is disposed opposite the rotation mechanism 110 across the device 10. The magnet moving mechanism 120 includes a magnet 122 and a moving unit 124 that moves the magnet 122 in a radial direction from a central position P0 that coincides with the rotation axis A. The magnet 122 may be a permanent magnet or an electromagnet, but a permanent magnet is preferred due to its simple configuration. The moving unit 124 is, for example, a linear actuator that moves the magnet 122 linearly in the radial direction. In this example, the magnet 122 can be moved from the central position P0 to an outer peripheral position P2. Note that the magnet 122 may also be configured to move to P3, which is beyond the outer peripheral position of the device 10.

[0054] The heating unit 130 is provided so as to be movable between a position in contact with the bottom surface of the eluate storage area 15 of the device 10 attached to the testing apparatus 100 and a position spaced apart from it. The heating unit 130 is disposed at a position spaced apart from the device 10 while the device 10 is being rotated by the rotation mechanism 110, and is disposed at a position in contact with the bottom surface of the eluate storage area 15 when the device 10 is stationary. The heating unit 130 heats the liquid contained in the eluate storage area 15. Here, the eluate storage area 15 contains a purified target substance to be detected and an eluate containing a reagent.

[0055] The heating unit 130 is equipped with a Peltier element or the like and is capable of controlling the temperature, and performs a temperature cycle for nucleic acid amplification.

[0056] The light detection unit 140 optically detects whether or not a detection target substance is contained. The light detection unit 140 is disposed opposite the heating unit 130 across the device 10. Light detection by the light detection unit 140 is also performed when the rotation of the device 10 by the rotation mechanism 110 is stopped. The light detection unit 140 irradiates the eluate storage area 15 with excitation light L1 of a specific wavelength and detects fluorescence L2 emitted from the fluorescent probe upon excitation by the excitation light L1. The excitation light L1 is selected according to the excitation wavelength of the fluorescent probe.

[0057] The light detection unit 140 includes a light source such as an LED or laser, a wavelength selection filter that transmits only light emitted from the light source with a wavelength corresponding to the excitation wavelength of the probe, and a condenser lens as a mechanism for irradiating excitation light L1. The light detection unit 140 also includes a photodetector such as a photodiode or a photomultiplier tube as a mechanism for detecting fluorescence. The light detection unit 140 may use an image sensor such as a CMOS (Complementary Metal Oxide Semiconductor) or a CCD (Charge Coupled Device) as the photodetector to acquire a two-dimensional image of the fluorescence emitted from the fluorescent probe.

[0058] The inspection device 100 includes a control unit (not shown) that controls the overall operation of the device. The control unit is implemented by a computer including a processor configured with a central processing unit (CPU), non-volatile memory (NVM), random access memory (RAM), etc.

[0059] The user removes the lid 40 of the device 10 from the input port, inputs the sample liquid through the input port, and then attaches the lid 40 to the input port. For example, the sample liquid may be obtained by dissolving a sample collected from the nasal cavity, pharynx, oral cavity, or affected area of ​​a subject using a swab or the like into a solution, or a body fluid such as a nasal cavity, pharynx, or oral cavity wash, saliva, urine, or blood.

[0060] The sample liquid is introduced into the central portion 13a of the internal space 12 through an introduction port. A magnetic particle solution 32 is contained in the central portion 13a, and the sample liquid is mixed with the magnetic particle solution 32 in the central portion 13a. The substance to be detected in the sample liquid binds to the modifiers that modify the magnetic particles, and is thereby captured by the magnetic particles.

[0061] The user mounts the device 10 on the mounting section 113 of the inspection device 100. The following processing is performed within the inspection device 100.

[0062] Initially, the magnet 122 is positioned in a position facing the central portion 13a of the internal space 12 when the device 10 is attached. When the device 10 is attached to the attachment portion 113, the magnetic particles are attracted to and trapped by the magnet 122. When the magnet 122 is moved, the magnetic particles move along with the movement of the magnet 122.

[0063] The rotation mechanism 110 and the magnet moving mechanism 120 are controlled to follow the trajectory shown by the dashed arrow in Fig. 7. By combining the rotation of the device 10 by the rotation mechanism 110 and the movement of the magnet 122 in the radial direction B by the magnet moving mechanism 120, the magnetic particles can be moved along the dashed arrow shown in Fig. 7.

[0064] For example, while rotating the device 10, the magnet 122 is moved from the central position P0 to the outer peripheral position P2, and then the magnet 122 is moved back and forth multiple times between a position P1 between the outer peripheral position P2 and the central position P0 and the outer peripheral position P2, thereby moving the magnetic particles back and forth multiple times between the cleaning liquid 34 and the hydrophobic liquid 38 in contact with the cleaning liquid 34, thereby washing the magnetic particles.

[0065] Thereafter, when the magnet 122 is stopped at position P1, the magnet 122 moves in an arc relative to the device 10 as the device 10 rotates. Furthermore, when the eluate storage area 15 approaches the magnet 122, the magnet 122 is moved from position P1 to outer peripheral position P2, thereby moving the magnetic particles within the device 10 along the trajectory shown in Fig. 7. Here, the rotation of the device 10 is stopped.

[0066] When the magnetic particles are moved into the elution solution 36, the nucleic acids bound to the magnetic particles are released and eluted into the elution solution 36. The magnetic particles are moved to the central portion 13a of the device 10 by moving the magnet 122 from the outer peripheral position P2 to the central position P0. This removes the magnetic particles from the elution solution 36, leaving the purified nucleic acids as the detection target released from the magnetic particles in the elution solution 36. In this state, the device 10 is rotated so that the elution solution storage area 15 of the device 10 coincides with the processing position where the heating unit and the light detection unit are arranged.

[0067] In this example, the eluate 36 contains a nucleic acid amplification reagent and a fluorescent probe, and nucleic acid amplification processing and fluorescence detection are carried out in the eluate storage area 15 .

[0068] The nucleic acid amplification process amplifies a specific nucleic acid, which is a detection target substance. The heating unit 130 is moved to a position where it contacts the bottom surface of the eluate storage area 15, and the heating unit 130 performs a temperature cycle on the liquid in the eluate storage area 15, thereby amplifying the nucleic acid. While the nucleic acid amplification process is not limited to this, examples include the polymerase chain reaction (PCR) method and the reverse transcription PCR (RT-PCR) method. When the PCR method is used, the following steps are repeated: a thermal denaturation step in which double-stranded DNA (deoxyribonucleic acid) is dissociated into single-stranded DNA at high temperature; an annealing step in which the temperature is then lowered to allow a primer to bind to the single-stranded DNA; and an extension step in which new double-stranded DNA is synthesized using a polymerase with the single-stranded DNA as a template. An example of the temperature cycle for the thermal denaturation step, annealing step, and extension step is one cycle of 94°C for 1 minute, 50-60°C for 1 minute, and 72°C for 1-5 minutes, repeated 20-50 times. Alternatively, the thermal denaturation step and annealing step may be performed at a single temperature. An example of such a temperature cycle is one cycle of 94°C for 1 minute and 60°C for 1 minute, repeated 20-50 times. There are no particular limitations on the temperature and time of the temperature cycle in the amplification step, and these may be selected as desired depending on the performance of the polymerase and primers.

[0069] Fluorescence detection is performed by the light detection unit 140. The light detection unit 140 irradiates the eluate storage area 15 with excitation light L1 of a specific wavelength and detects fluorescence L2 emitted from the fluorescent probe upon excitation by the excitation light L1. For example, when the nucleic acid amplification process is a PCR method, fluorescence detection is performed for each temperature cycle described above to monitor the amplification status in real time.

[0070] If a specific nucleic acid is present in the sample solution, the nucleic acid is amplified in the amplification step, and fluorescence L2 is detected when excitation light L1 is irradiated onto the fluorescent probe labeled with this specific nucleic acid. On the other hand, if the specific nucleic acid sequence is not present in the sample solution, fluorescence L2 is not detected even when excitation light L1 is irradiated. This makes it possible to determine whether or not the specific nucleic acid is present.

[0071] As described above, the device 10 of this embodiment includes an internal space 12 and an inlet 24 in the central portion 13a of the internal space 12. The washing solution 34 and the elution solution 36 are contained in the peripheral portion 16 surrounding the central portion 13a, separated from each other by the hydrophobic liquid 38. This configuration allows the magnetic particles to be freely moved from the central portion 13a toward the washing solution 34 or the elution solution 36. The transport path of the magnetic particles within the internal space 12 can be appropriately changed depending on the test item or specimen type. As the magnetic particles pass through the interface between the washing solution 34 and the hydrophobic liquid 38 and the hydrophobic liquid 38, moisture containing impurities present around the nucleic acid is removed due to contact resistance with the hydrophobic liquid. Therefore, passing the magnetic particles through the interface multiple times can enhance the cleaning effect. Furthermore, because the internal space 12 is filled with liquid, fluctuations in the positions of the washing solution 34, the elution solution 36, and the hydrophobic liquid 38 are suppressed.

[0072] In the device 10 of the above embodiment, the magnetic particle solution 32 is contained in the central portion 13a of the internal space 12. However, the central portion 13a may contain an aqueous liquid such as a buffer solution that does not contain magnetic particles. In this case, the magnetic particle solution 32 and the sample liquid are mixed outside the device 10, and the mixed liquid is then introduced into the internal space. The aqueous liquid contained in the central portion 13a may be any liquid that does not have the effect of breaking up the bond between the magnetic particles and the target substance to be detected, but it is more preferable that the aqueous liquid has the effect of stabilizing the bond between the magnetic particles and the target substance to be detected. If the magnetic particle solution 32 is contained in the device 10 in advance, the user does not need to mix the magnetic particle solution 32 and the sample liquid outside the device, thereby reducing the burden on the user.

[0073] As described above, in the device 10, if the thickness of the internal space 12 is 1 mm or less, it is easy to maintain the aqueous liquid and the hydrophobic liquid in their respective positions. The thickness of the internal space 12 is preferably less than 1 mm, and may be 0.8 mm or less, 0.7 mm or less, or even 0.4 mm or less. The magnetic particles used should be smaller than the thickness of the internal space 12.

[0074] Furthermore, in the device 10 of this embodiment, the thickness of the processing region (here, the eluate storage region) is formed thinner than the other regions in the internal space 12. This configuration enables high-speed temperature control in processes such as nucleic acid amplification.

[0075] The device 10 of this embodiment is provided with pillars 25 in the internal space 12 that prevent the thin film 30 from contacting the bottom surface of the recess 21 of the main body member 20. This configuration prevents the thin film 30 from recessing into the recess and contacting the bottom surface of the recess 21, thereby blocking the transport path for the magnetic particles. Depending on the shape of the internal space 12 and the tension of the thin film 30 when joined to the main body member 20, there may be cases where the thin film 30 does not come into contact with the bottom surface of the recess, and in such cases the pillars 25 may not be required.

[0076] On the other hand, because the thickness of the internal space 12 is very thin, preferably about 1 mm or less, the flexible thin film 30 easily dents toward the recessed portion. Therefore, the embodiment is not limited to the four columnar pillars 25 shown in FIG. 4 , and a configuration including more pillars 25A, such as a modified main body member 20A shown in FIG. 8 , may be used. In the example shown in FIG. 8 , the pillars 25A are polygonal, and nine pillars 25A are arranged in a ring shape surrounding the through-hole 22. Furthermore, one pillar 25B is also arranged near the boundary between the first region 21b and the circular portion 21a. This means that the pillar 25B is arranged near the boundary between the cleaning liquid storage region 14 and the circular portion 13 in the internal space 12.

[0077] 8, the recess 21 may have a narrowed portion 21d between the second region 21c and the circular portion 21a. This means that the internal space 12 may have a narrowed portion between the eluate storage region 15 and the circular portion 13. When the eluate storage region 15 is used as a processing region for mixing a reagent and a detection target, the temperature control effect in processing such as nucleic acid amplification can be improved by providing a narrowed portion between the processing region and other regions.

[0078] 9, the depth of the central portion 21a1 of the circular portion 21a of the recess 21 may be deeper than the depth of the outer peripheral portion 21a2. This means that the thickness of the central portion 13a in the internal space 12 may be thicker than the thickness of the annular portion 13b. By providing a step at the boundary between the central portion 13a containing the magnetic particle solution 32 and the annular portion 13b containing the hydrophobic liquid 38 and narrowing the annular portion 13b containing the hydrophobic liquid 38, the liquid is less likely to flow from the narrow space toward the wider space due to capillary force, and the hydrophobic liquid 38 contained in the annular portion 13b can be prevented from returning to the central portion 13a.

[0079] In the device 10 of the above embodiment, the eluate storage area 15 doubles as a processing area for mixing the reagent and the target substance to be detected, and a detection area for containing the reagent and the target substance mixed in the processing area and optically detecting the target substance from the outside, and an example has been described in which the eluate storage area 15 contains the reagent along with the eluate 36. However, a processing area and a detection area may be provided separately from the eluate storage area 15 on the outer periphery of the internal space 12. Furthermore, the outer periphery of the internal space 12 may further include a reagent storage area for containing a reagent solution containing the reagent. By using the eluate storage area 15 as both a processing area and a detection area, a device with a simple configuration can be realized.

[0080] Alternatively, the device 10 may not include a processing region or a detection region and may only purify and extract the target substance. The eluate containing the purified target substance may be removed from the device 10 and mixed with a reagent and subjected to a detection process in a separate testing device.

[0081] 10 , the device 11 may further include a reagent storage section 52 connected to the internal space 12 via a valve 50, and a processing chamber 54 disposed farther from the central section 13a than the reagent storage section 52. The reagent storage section 52 stores a reagent 39 for the test object. In the processing chamber 54, a process of mixing the reagent and the detection object is performed, and the detection chamber 54 is subjected to a detection process of optically detecting the detection object from outside.

[0082] The reagent storage section 52 is disposed on the outer periphery of the eluate storage area 15. As shown in Figure 10, the eluate storage area 15, the reagent storage section 52, and the treatment chamber 54 are disposed linearly in a direction radially away from the center of the circular section 13. In this example, the valve 50 is a rotary valve that contains the reagent storage section 52. The valve 50 in this example can be switched between a first state in which the reagent storage section 52 is connected only to the eluate storage area 15, a second state in which the reagent storage section 52 is connected only to the treatment chamber 54, and a third state in which the reagent storage section 52 is not connected to either the eluate storage area 15 or the treatment chamber 54.

[0083] During the purification process, the second or third state is set, and the connection between the eluate storage area 15 and the reagent storage section 52 is kept closed.

[0084] The mixing of the purified reagent and the target substance using the device 11 and the detection process are carried out in the following manner.

[0085] With the extraction liquid of the test object contained in the eluate containing region 15 as a result of the purification process, the valve 50 is switched to the first state to connect the eluate containing region 15 to the reagent containing section 52. Here, the device 11 is rotated by the rotation mechanism 110 to generate centrifugal force, thereby sending the extraction liquid in the eluate containing region 15 to the reagent containing section 52. After the extraction liquid has been contained in the reagent containing section 52, the valve 50 is switched to the second state to connect the reagent containing section 52 to the processing chamber 54. The device 11 is again rotated by the rotation mechanism 110 to generate centrifugal force, thereby sending the mixture of the extraction liquid and the reagent 39 in the reagent containing section 52 to the processing chamber 54. After the mixture has been contained in the processing chamber 54, the valve 50 is switched to the first state or the third state to close the processing chamber 54.

[0086] The processing chamber 54 contains a mixed solution, and in a closed state, a process such as a heating process is carried out to cause a reaction between the test object and the reagent, and a process for detecting the detection object substance by the light detection unit is carried out. If the test object is a nucleic acid, the same processes as the nucleic acid amplification process and fluorescence detection process in the above-mentioned testing method are carried out.

[0087] As described above, the device 11 includes the reagent storage section 52 and the treatment chamber 54 independent of the internal space 12, and the connection state between the reagent storage section 52 and the internal space 12 can be switched by the valve 50. Therefore, the reagent storage section 52 can be closed off from the internal space 12 by the valve 50, allowing the reagent to be stored separately from the liquid in the internal space 12, making it possible to use a freeze-dried reagent that has high storage stability. Furthermore, the device 11 can close the treatment chamber 54 during heat treatment, thereby improving the temperature control effect when controlling the temperature of the treatment chamber 54.

[0088] Next, a method for manufacturing the device 10 will be described. In particular, a liquid disposing process step for accommodating the cleaning liquid 34, the elution liquid 36, the hydrophobic liquid 38, and the magnetic particle solution 32 in the internal space 12 of the device 10, and a liquid filling process step for filling the internal space 12 with liquid will be described. Figures 11A and 11B are diagrams showing the liquid disposing process step.

[0089] In step S1, the eluate 36 is placed in the eluate storage area 15. First, the eluate 36 is placed into the central portion 13a of the internal space 12 through the inlet 24. The device 10 is centrifuged around a point C1, which is located on an extension of a line connecting the eluate storage area 15 and the center C0 of the circular portion 13 and sandwiches the center C0 between the eluate storage area 15 and the central portion 13a. The eluate 36 placed in the central portion 13a moves in the direction indicated by the solid arrow due to centrifugal force and is sent from point C1, the center of rotation, to the eluate storage area 15, which is the outermost periphery. As a result, the eluate 36 is placed in the eluate storage area 15.

[0090] Next, in step S2, the cleaning liquid 34 is placed in the cleaning liquid storage area 14. The cleaning liquid 34 is placed into the central portion 13a of the internal space 12 from the inlet 24 of the device 10, with the eluate 36 stored in the eluate storage area 15. The device 10 is centrifuged around a point C2, which is located on an extension of the line connecting the cleaning liquid storage area 14 and the center C0 and sandwiches the center C0 between the device 10 and the cleaning liquid storage area 14. The cleaning liquid 34 placed in the central portion 13a moves in the direction indicated by the solid arrow due to centrifugal force, and is sent from point C2, the center of rotation, to the cleaning liquid storage area 14, which is the outermost side. As a result, the cleaning liquid 34 is placed in the cleaning liquid storage area 14.

[0091] Next, in step S3, the hydrophobic liquid 38 is placed in the annular portion 13b. The hydrophobic liquid 38 is placed in the central portion 13a of the internal space 12 through the inlet 24 of the device 10, with the elution liquid 36 contained in the elution liquid storage region 15 and the cleaning liquid 34 contained in the cleaning liquid storage region 14. The device 10 is centrifuged around the center C0 as the rotation center. In this example, a liquid having a lower specific gravity than the elution liquid 36 and the cleaning liquid 34 is used as the hydrophobic liquid 38. The hydrophobic liquid 38 is moved radially outward by centrifugal force, but because its specific gravity is lower than the elution liquid 36 and the cleaning liquid 34, it is placed in the annular portion 13b, which is more inward than the elution liquid 36 and the cleaning liquid 34.

[0092] Next, in step S4, the magnetic particle solution 32 is placed in the central portion 13a. The magnetic particle solution 32 is introduced into the central portion 13a of the internal space 12 through the inlet 24 of the device 10, which has the elution solution 36 contained in the elution solution storage region 15, the cleaning solution 34 contained in the cleaning solution storage region 14, and the hydrophobic liquid 38 contained in the annular portion 13b. If the inner wall surface of the central portion 13a has been subjected to a hydrophilic treatment, the magnetic particle solution 32 will quickly spread within the central portion 13a.

[0093] By the above steps S1 to S4, the elution liquid 36 can be placed in the elution liquid storage area 15 of the internal space 12 of the device 10, the cleaning liquid 34 in the cleaning liquid storage area 14, the hydrophobic liquid 38 in the annular portion 13b, and the magnetic particle solution 32 in the central portion 13a.

[0094] In this way, the cleaning liquid 34, the elution liquid 36, and the hydrophobic liquid 38 can be placed at mutually separated positions on the outer periphery of the internal space 12 by centrifugal force generated by rotating the device 10 about different points C1, C2, and C0 as the rotation center. Furthermore, because centrifugal force is used, the liquids can be placed at the respective positions in the internal space 12 without containing air bubbles.

[0095] As a method for changing the position of the center of rotation, the attachment position of the device 10 relative to the rotation axis of the centrifuge may be changed so that points C1, C2, and C0 become the centers of rotation for the centrifuge, or the attachment position on the device 10 side may be fixed, and a first centrifuge in which the center of rotation of the attached device 10 is point C1, a second centrifuge in which the center of rotation is point C2, and a third centrifuge in which the center of rotation is point C0 may be used in sequence.

[0096] As described above, the elution liquid 36 is placed in the elution liquid storage area 15 of the internal space 12 of the device 10, the cleaning liquid 34 is placed in the cleaning liquid storage area 14, the hydrophobic liquid 38 is placed in the annular portion 13b, and the magnetic particle solution 32 is placed in the central portion 13a, and then the lid 40 is attached to the inlet 24, thereby sealing the internal space 12 while it is filled with liquid.

[0097] A method for sealing the internal space 12 by attaching the lid 40 will be described with reference to FIG.

[0098] 12 is a cross-sectional view of the vicinity of the central portion 13a in a state after step S4 in Fig. 11B, in which the elution liquid 36 is placed in the elution liquid storage region 15 of the internal space 12, the cleaning liquid 34 is placed in the cleaning liquid storage region 14, the hydrophobic liquid 38 is placed in the annular portion 13b, and the magnetic particle solution 32 is placed in the central portion 13a. At this point, the inlet 24, which is the end of the through-hole 22 in the central portion 13a, is in an open state.

[0099] 12 , the convex portion 44 of the plug 42 of the lid 40 is inserted into the inlet 24. At this time, air between the convex portion 44 and the magnetic particle solution 32 is discharged to the outside through the air hole 43, as shown by the arrow. As shown in step S12, the plug 42 is pushed into the inlet 24 until the magnetic particle solution 32 enters the plug 42.

[0100] Thereafter, as shown in step S13, the protrusion 46a of the sealing member 46 is inserted into the air hole 43, thereby sealing the internal space 12 with the lid 40.

[0101] By attaching the lid to the insertion port 24 in this way, the internal space 12 becomes substantially filled with liquid. Although a small gap 49 remains in part of the air hole 43, this does not interfere with the test.

[0102] 13 , the magnetic particle solution 32 may be poured into the internal space 12, and when the plug 42 is inserted into the inlet 24, the magnetic particle solution 32 may be allowed to overflow from the air hole 43, and in the overflowing state, the protrusion 46a of the sealing member 46 may be inserted into the air hole 43, as shown in step S23. This procedure allows the internal space 12 to be filled with liquid without leaving any gaps in the air hole 43. After the sealing member 46 is attached, any magnetic particle solution 32 that has overflowed onto the side surface of the lid 40 is wiped off.

[0103] 14 shows another embodiment of a method for sealing the internal space 12 by attaching the lid 40. As shown in step S31 of FIG. 14 , the device 10 before the lid 40 is attached is placed in a reduced pressure atmosphere. Subsequently, as shown in step S32, the lid 40 with the plug 42 and sealing member 46 set therein is inserted into the inlet 24 in a reduced pressure atmosphere. Under the reduced pressure atmosphere, the lid 40 is not pushed into the inlet 24, but is instead opened to the atmosphere in the next step, step S33. The gap between the magnetic particle solution 32 and the tip of the plug 42 is reduced in pressure, and the lid 40 is pushed toward the through-hole 22 by atmospheric pressure. As a result, as shown in step S34, the internal space 12 can be substantially filled with liquid, leaving only a very small gap 51 in a portion of the air hole 43.

[0104] The smaller the voids in the internal space 12, i.e., the fewer air bubbles, the more effectively the movement of the liquids placed in each region can be suppressed. If air bubbles are present in the internal space 12, the bubbles may move within the internal space 12 due to vibration or impact, which may cause the liquids contained in the internal space 12 to shift from their respective positions. However, if the internal space 12 is filled with liquid, fluctuations in the placement of the liquids within the internal space 12 can be suppressed. Note that even if a void 49 remains in the air hole 43 of the plug 42 as shown in step S13 of FIG. 12 and step S34 of FIG. 14 , as long as the liquid is filled to the point where the tip surface of the convex portion 44 of the plug 42 is in contact with the magnetic particle solution 32, this is considered to be a state in which "the internal space is filled with liquids including a cleaning liquid, an elution liquid, and a hydrophobic liquid" in the technology of the present disclosure.

[0105] "Modified Device" A modified device 210 will now be described with reference to FIG. 15. FIG. 15 is a plan view of an internal space 212 of the modified device 210. The shape of the internal space 212 of the device 210 is different from the shape of the internal space 12 of the device 10 described with reference to FIGS. 1 to 5. The difference between the internal space 212 and the internal space 12 means that the shape of the recess 21 provided in the main body member 20 is different. The configuration of the device 210 is the same as that of the device 10, except for the shape of the recess 21.

[0106] In the device 210, the internal space 212 is generally circular in plan view, and is a generally disk-shaped space. A partition wall 228 is provided within the internal space 212 to separate the cleaning solution storage area 214, the elution solution storage area 215, and the reagent storage area 216. The partition wall 228 is formed to extend from the outer periphery toward the center. A through-hole 22 is provided in the central portion 213 of the internal space 212, and one end of the through-hole 22 serves as an inlet 24, similar to the device 10.

[0107] The washing liquid containing area 214, the eluate containing area 215, and the reagent containing area 216 are arranged at positions spaced apart from one another along the outer periphery of the internal space 212. The washing liquid containing area 214 is an area along a sectorial arc with a central angle of approximately 90°. The eluate containing area 215 and the reagent containing area 216 are each arranged at a position facing a partition wall 228 that partitions the washing liquid containing area 214.

[0108] The washing liquid storage area 214 stores a washing liquid 34, the elution liquid storage area 215 stores an elution liquid 36, and the reagent storage area 216 stores a reagent solution 39. Furthermore, the central portion 213 of the internal space 212 stores a magnetic particle solution 32, and the magnetic particle solution 32, washing liquid 34, elution liquid 36, and reagent solution 39 are separated by a hydrophobic liquid 38. Here, in addition to the magnetic particle solution 32, washing liquid 34, and elution liquid 36, the reagent solution 39 is also an aqueous liquid. The internal space 212 is filled with the magnetic particle solution 32, washing liquid 34, elution liquid 36, reagent solution 39, and hydrophobic liquid 38.

[0109] The modified device 210 also has an inlet 24 in the central portion 213 of the internal space 212, and the cleaning liquid 34 and the elution liquid 36 are contained in the internal space 212 in an outer peripheral portion surrounding the central portion 213, separated from each other by a hydrophobic liquid 38. Therefore, similar to the device 10, the magnetic particles can be freely moved from the central portion 13a toward the cleaning liquid 34 or the elution liquid 36, and similar effects can be obtained. Furthermore, in this device 210, the reagent solution 39 is contained separated from the cleaning liquid 34 and the elution liquid 36 by the hydrophobic liquid 38, so the magnetic particles can also be moved toward the reagent solution 39. Storing the reagent solution 39 separated from the elution liquid improves the stability of the reagent solution 39.

[0110] 16 and 17, a brief description will be given of an inspection method using the device 210. The device 210 is also loaded into the inspection apparatus 100 shown in FIG. 6, and an inspection of the object to be detected is carried out.

[0111] The device 210 containing the sample liquid is loaded into the testing apparatus 100 so that the center of the circular internal space 212 of the device 210 coincides with the rotation axis A of the rotation mechanism 110 .

[0112] By combining the rotation of the device 210 by the rotation mechanism 110 and the movement of the magnet by the magnet moving mechanism 120, the magnetic particles can be moved along the trajectory shown by the dashed line in FIG.

[0113] In this example, the magnetic particles bound to nucleic acid are also moved along the trajectory shown by the dashed line in Figure 16. The magnetic particles are moved back and forth across the interface between the washing solution 34 and the hydrophobic liquid 38 multiple times, and then moved to the elution solution storage area 215. The nucleic acid is released from the magnetic particles in the elution solution 36 and eluted into the elution solution 36. The magnetic particles from which the nucleic acid has been released are then moved to the reagent storage area 216. In this example, the magnetic particles are modified not only with modifiers that bind to nucleic acid, but also with modifiers that bind to reagents. Alternatively, the magnetic particles may include magnetic particles modified with modifiers that bind to nucleic acid and magnetic particles modified with modifiers that bind to reagents (here, primers and fluorescent probes).

[0114] 17 , the magnetic particles bound to the reagent are moved from the reagent storage area 216 to the eluate storage area 215. The reagent bound to the magnetic particles is released in the eluate 36 and eluted into the eluate 36. The magnetic particles from which the reagent has been released are evacuated to the center section 213, and in the eluate storage area 215, nucleic acid amplification processing and fluorescence detection processing are carried out.

[0115] In the above example, the magnetic particles are moved from the washing solution storage area to the elution solution storage area to elute the nucleic acid, and then the magnetic particles are moved to the reagent storage area. However, it is also possible to move the magnetic particles with nucleic acid bound to them from the washing solution storage area to the reagent storage area, bind primers and fluorescent probes to the magnetic particles in the reagent storage area, and then move the magnetic particles with nucleic acid bound to them and the magnetic particles with primers and fluorescent probes bound to them to the elution solution storage area 215.

[0116] A method for manufacturing the device 210 will now be described. In particular, a liquid placement process for accommodating a cleaning liquid 34, an elution liquid 36, a hydrophobic liquid 38, a reagent solution 39, and a magnetic particle solution 32 in the internal space 12 of the device 210 will be described with reference to FIGS. 18 and 19 .

[0117] In the device 210, in the first step S51, the hydrophobic liquid 38 is filled into the internal space 212. First, the hydrophobic liquid 38 is poured into the central portion 213 of the internal space 212 through the inlet 24. The device 210 is centrifuged around the center C10 of the internal space 212 as the center of rotation, thereby filling the hydrophobic liquid 38 throughout the entire internal space 212.

[0118] Next, in step S52, the reagent solution 39 is placed in the reagent containing region 216. The reagent solution 39 is poured into the central portion 213 of the device 210, which is filled with the hydrophobic liquid 38. The device 210 is centrifuged around a point C11, which is located on the periphery of the internal space 212 and is a point on an extension of a line connecting the reagent containing region 216 and the center C10, sandwiching the center C10 between the reagent containing region 216 and the central portion 213. The reagent solution 39 poured into the central portion 213 is transported from the center C11 to the reagent containing region 216, which is the outermost reagent containing region 216, due to centrifugal force generated in the direction indicated by the dashed arrows. As a result, the reagent solution 39 is placed in the reagent containing region 216.

[0119] Next, in step S53, the eluate 36 is placed in the eluate storage area 215. First, the eluate 36 is placed into the center 213 of the internal space 212 through the inlet 24. The device 210 is centrifuged around a point C12, which is located on an extension of a line connecting the eluate storage area 215 and the center C10, and which is located on the circumference of the internal space 212 and sandwiches the center C10 between the eluate storage area 215 and the device 210. The eluate 36 placed in the center 213 is transported from the center C12 to the eluate storage area 215, which is the outermost periphery, due to centrifugal force generated in the direction indicated by the dashed arrow. As a result, the eluate 36 is placed in the eluate storage area 215. Note that during this centrifugation, centrifugal force is also applied to the reagent stored in the reagent storage area 216, but the partition 228 that partitions the reagent storage area 216 prevents the reagent solution 39 from splashing out of the reagent storage area 216.

[0120] Next, in step S54, the cleaning liquid 34 is placed in the cleaning liquid storage area 214. The cleaning liquid 34 is placed into the central portion 213 of the internal space 212 through the inlet 24. The device 210 is centrifuged around the intersection C13 of the extension of the line connecting the cleaning liquid storage area 214 and the center C10 with the straight line connecting the eluate storage area 215 and the reagent storage area 216 as the center of rotation. The cleaning liquid 34 placed in the central portion 213 is sent from point C13, the center of rotation, to the cleaning liquid storage area 214, which is the outermost side, by centrifugal force generated in the direction indicated by the dashed arrow. As a result, the cleaning liquid 34 is placed in the cleaning liquid storage area 214.

[0121] Next, in step S55, the positions of the reagent solution 39, the elution liquid 36, and the cleaning liquid 34 are adjusted, and then the magnetic particle solution 32 is placed in the central portion 213. Specifically, first, the device 210, in which the reagent solution 39 is accommodated in the reagent accommodation region 216, the elution liquid 36 is accommodated in the elution liquid accommodation region 215, the cleaning liquid 34 is accommodated in the cleaning liquid accommodation region 214, and the hydrophobic liquid 38 is accommodated in another region, is centrifuged around the center C10 of the internal space 12 as the rotation center, thereby adjusting the positions of the hydrophobic liquid 38, the reagent solution 39, the elution liquid 36, and the cleaning liquid 34. Thereafter, the magnetic particle solution 32 is introduced into the central portion 213 of the internal space 212 through the inlet 24 of the device 210.

[0122] By performing the above steps S51 to S55, the internal space 212 of the device 210 can be filled with a hydrophobic liquid 38, and then the reagent solution 39 can be placed in the reagent storage area 216, the elution liquid 36 in the elution liquid storage area 215, the cleaning liquid 34 in the cleaning liquid storage area 214, and the magnetic particle solution 32 in the central portion 213.

[0123] In this way, by applying centrifugal forces generated by rotating the device 10 about different points C11, C12, and C13 as the rotation centers to the reagent solution 39, the elution liquid 36, and the cleaning liquid 34, it is possible to arrange the reagent solution 39, the elution liquid 36, and the cleaning liquid 34 at positions separated from each other by the hydrophobic liquid 38 on the outer periphery of the internal space 212. Furthermore, because centrifugal force is used, the liquids can be arranged at their respective positions in the internal space 12 without containing air bubbles.

[0124] The following additional notes are further disclosed regarding the above embodiment.

[0125] <Supplementary Note 1> A device comprising an internal space capable of accommodating a sample liquid containing a target substance that binds to magnetic particles, wherein the target substance is purified by moving the target substance within the internal space using the magnetic particles, the device comprising: an inlet for introducing the sample liquid at a center of the internal space; a washing liquid for removing impurities contained in the sample liquid and an elution liquid for eluting the target substance are contained in an outer peripheral portion of the internal space surrounding the center, the washing liquid being separated from each other by a hydrophobic liquid; and the internal space being filled with a liquid containing the washing liquid, the elution liquid, and the hydrophobic liquid. <Supplementary Note 2> The device according to Supplementary Note 1, wherein a magnetic particle solution containing magnetic particles is contained in the internal space at a center portion. <Supplementary Note 3> The device according to Supplementary Note 1 or Supplementary Note 2, wherein the thickness of the internal space at the center portion is greater than the thickness of the outer peripheral portion. <Supplementary Note 4> The device according to any one of Supplements 1 to 3, wherein a reagent for the target substance is contained in the internal space at an outer peripheral portion, the reagent being separated from the washing liquid and the elution liquid by a hydrophobic liquid. <Appendix 5> The device according to any one of Appendices 1 to 3, wherein the eluate contains a reagent for the substance to be detected. <Appendix 6> The device according to Appendices 4 or 5, wherein the internal space has, at its outer periphery, a processing region for mixing the reagent and the substance to be detected. <Appendix 7> The device according to Appendices 6, wherein the internal space has, at its outer periphery, a detection region for accommodating the reagent and the substance to be detected mixed in the processing region and for optically detecting the substance to be detected from the outside. <Appendix 8> The device according to Appendices 6 or 7, wherein the thickness of the processing region in the internal space is thinner than the thickness of other regions. <Appendix 9> The device according to any one of Appendices 6 to 8, wherein the internal space has a narrowed portion between the processing region and other regions. <Supplementary Note 10> The device according to any one of Supplementary Notes 1 to 9, comprising: a reagent storage section connected to the internal space via a valve and storing a reagent for the substance to be detected; and a processing chamber arranged farther from the center than the storage section, to which a liquid containing the reagent and the substance to be detected is delivered and to which the substance to be detected is optically detected from the outside. <Supplementary Note 11> The device according to any one of Supplementary Notes 1 to 10, wherein the internal space is formed by a main body member having a recess on one surface and a thin film joined to one surface in a manner covering the recess.<Appendix 12> The device according to Appendix 11, wherein the internal space is provided with pillars that prevent contact with the bottom surface of the recess of the thin film. <Appendix 13> The device according to any one of Appendixes 1 to 12, wherein the internal space is subjected to a hydrophilic treatment in at least one of the region for storing a cleaning liquid, the region for storing an elution liquid, and the central portion. <Appendix 14> The device according to any one of Appendixes 1 to 13, wherein the hydrophobic liquid has a specific gravity lower than that of the cleaning liquid and the elution liquid. <Appendix 15> The device according to Appendix 14, wherein the hydrophobic liquid is silicone oil. <Appendix 16> The device according to any one of Appendixes 1 to 15, wherein the thickness of the internal space is 1 mm or less. <Appendix 17> The device according to any one of Appendixes 1 to 16, further comprising a detachable lid for the insertion port, wherein the lid is a plug to be inserted into the insertion port, the plug having an air hole for discharging air in the internal space to the outside when the plug is inserted into the insertion port, and a sealing member for closing the air hole. <Supplementary Note 18> A method for manufacturing a device according to any one of Supplementary Note 1 to Supplementary Note 17, wherein the cleaning solution and the elution solution are disposed at positions separated from each other on the outer periphery of the internal space by using centrifugal forces generated by rotating the device about different centers of rotation.

[0126] The disclosure of Japanese Patent Application No. 2023-145446, filed on September 7, 2023, is incorporated herein by reference in its entirety. All documents, patent applications, and technical standards mentioned herein are incorporated herein by reference to the same extent as if each individual document, patent application, and technical standard was specifically and individually indicated to be incorporated by reference.

Claims

1. A device having an internal space capable of containing a specimen liquid containing a detection target that binds to magnetic particles, and in which the detection target is purified by moving the detection target within the internal space using the magnetic particles, the device having an inlet for introducing the specimen liquid in the center of the internal space, and in the internal space, a washing liquid for removing impurities contained in the specimen liquid and an elution liquid for eluting the detection target are contained in the outer periphery surrounding the center, separated from each other by a hydrophobic liquid, and the internal space is filled with a liquid containing the washing liquid, the elution liquid, and the hydrophobic liquid.

2. The device of claim 1, wherein a magnetic particle solution containing the magnetic particles is contained in the central portion of the internal space.

3. The device according to claim 1 or 2, wherein the thickness of the central portion of the internal space is greater than the thickness of the outer periphery.

4. The device according to claim 1, wherein a reagent for the substance to be detected is contained in the outer periphery of the internal space and separated from the washing liquid and the elution liquid by the hydrophobic liquid.

5. The device according to claim 1, wherein the eluate contains a reagent for the substance to be detected.

6. The device according to claim 4 or 5, wherein the internal space is provided at the outer periphery with a processing region for mixing the reagent and the substance to be detected.

7. The device according to claim 6, wherein the internal space is provided at the outer periphery with a detection area that contains the reagent and the substance to be detected that are mixed in the processing area, and in which the substance to be detected is optically detected from the outside.

8. The device according to claim 6, wherein the internal space has a thickness in the processing region that is thinner than the thickness in other regions.

9. The device of claim 6, further comprising a constriction in said interior space between said treatment region and another region.

10. The device described in claim 1, comprising: a reagent storage section connected to the internal space via a valve and containing a reagent for the detection target; and a processing chamber located farther from the central section than the reagent storage section, into which a liquid containing the reagent and the detection target is delivered and into which the detection target is optically detected from the outside.

11. The device according to claim 1, wherein the internal space is formed by a body member having a recess on one surface and a thin film bonded to the one surface in a manner covering the recess.

12. The device according to claim 11, wherein the internal space is provided with pillars that prevent the thin film from contacting the bottom surface of the recess.

13. The device according to claim 1, wherein at least one of the region for storing the cleaning liquid, the region for storing the elution liquid, and the central portion of the internal space is subjected to a hydrophilic treatment.

14. The device of claim 1, wherein the hydrophobic liquid has a lower specific gravity than the cleaning liquid and the elution liquid.

15. The device of claim 14, wherein the hydrophobic liquid is silicone oil.

16. The device of claim 1, wherein the thickness of the internal space is 1 mm or less.

17. The device according to claim 1, further comprising a detachable cover for said insertion port, said lid including: a plug to be inserted into said insertion port, said plug having an air hole for discharging air within said internal space to the outside when said plug is inserted into said insertion port; and a sealing member for closing said air hole.

18. A method for manufacturing a device as described in claim 1, comprising: disposing the cleaning solution and the elution solution at positions separated from each other on the outer periphery of the internal space by using centrifugal forces generated by rotating the device about different centers of rotation.

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