Dilution device and inspection device

Abstract

The dilution device includes a microchannel chip at least in a part thereof and can dilute a specimen after reacting the specimen with a reagent or the like.SOLUTION: The diluting device 10 includes a diluting liquid outflow part 30 for flowing out at least the diluting liquid 17 when the fluid 16 is introduced, and a specimen diluting part 40 for introducing the specimen 18 and diluting the specimen 18 with the diluting liquid 17 flowing out from the diluting liquid outflow part 30. The diluent outflow part 30 includes a flow path groove 12 formed on at least one surface of the substrate 11, a covering material that covers the flow path groove 12, a fluid receiving part 31 that communicates with the flow path groove 12 and into which the fluid 16 is introduced, a diluent receiving part 33 that communicates with the flow path groove 12 and into which the diluent 17 is introduced, a fluid control mechanism that controls the outflow of the diluent 17 from the diluent receiving part 33, and an outflow port 15 that is located downstream of the flow path groove 12 and through which at least the diluent 17 flows out. When the liquid mixture of the diluting solution 17 and the sample 18 reaches a predetermined liquid level height H1, the sample diluting unit 40 dilutes the sample 18 to a predetermined magnification.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a dilution device and an inspection device. [Background technology]

[0002] Flow channel devices are known that perform biochemical measurements and chemical synthesis by flowing fluids through channels formed on a substrate. In particular, microanalysis devices and microreaction devices fabricated using microfabrication technology are preferred for their miniaturization, portability, small sample volumes, small reagent volumes, small waste volumes, and speed.

[0003] Conventionally, techniques for performing operations required for sample detection on such microfluidic devices have been proposed. For example, Japanese Patent Laid-Open Publication No. 2006-266974 (Patent Document 1) proposes a test chip for liquid samples, which has, on the upper surface of a substrate, a sample reservoir for storing a liquid sample, a diluent injector into which a diluent is injected, a mixing pot where the liquid sample and the diluent are joined, a flow path extending from the sample reservoir and the diluent injector to the mixing pot, and a microflow path through which a diluted sample flows from the mixing pot, and the area of ​​the substrate including the mixing pot, the flow path, and the microflow path is covered from the upper side with a membrane, and the mixing pot houses a rotor having a magnetic body therein that rotates in response to changes in a magnetic field, and a dilution device on which the test chip is mounted for diluting a sample.

[0004] Meanwhile, in recent years, test kits using nucleic acid amplification have become commercially available as a rapid diagnostic method for viruses and bacteria such as the novel coronavirus. Nucleic acid amplification is performed using the following steps: (1) A nucleic acid amplification reagent that acts to amplify part of the viral gene is prepared, and a sample (saliva, nasopharyngeal swab, etc.) is added to the nucleic acid amplification reagent. (2) The nucleic acid amplification reagent amplifies the viral gene (nucleic acid) in the sample, yielding an amplification reaction solution. (3) The viral gene (gene amplification product), which has been amplified tens of thousands of times by reaction with the nucleic acid amplification reagent, is detected from the amplification reaction solution. The PCR method mentioned above is also a type of nucleic acid amplification method. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2006-266974 Summary of the Invention [Problem to be solved by the invention]

[0006] In test kits using the nucleic acid amplification method described above, a high sample concentration is desirable at the amplification reaction stage (2) above, and the amplification reaction solution (3) above is desirably diluted to optimal properties (viscosity, concentration, etc.) for detecting gene amplification products. On the other hand, the test chip of Patent Document 1 injects a reagent containing a fluorescent labeling substance into a diluted sample on the test chip, and analyzes the sample's components through fluorescence detection. Unlike the nucleic acid amplification method, in which the collected sample is reacted with a reaction reagent to obtain an amplification reaction solution, which is then diluted. Furthermore, the test chip of Patent Document 1 requires an external stirring mechanism to stir the tiny rotor in the mixing pod, which can easily make the device complex.

[0007] Therefore, there is a demand for a dilution device that includes at least a part of a microchannel chip and that can dilute a specimen after reacting it with a reaction reagent, etc., and a testing device that employs the dilution device. [Means for solving the problem]

[0008] The dilution device according to the present invention is a dilution device comprising: a diluent outlet section that discharges at least a diluent when a fluid is introduced therein; and a specimen dilution section that receives an introduced specimen and dilutes the specimen with the diluent discharged from the diluent outlet section, wherein the diluent outlet section comprises a flow channel formed on at least one surface of a substrate; a coating material that covers the flow channel; a fluid receiving section that communicates with the flow channel and into which the fluid is introduced; a diluent receiving section that communicates with the flow channel and into which the diluent is introduced; a fluid control mechanism that controls the discharge of the diluent from the diluent receiving section; and an outlet located downstream of the flow channel and through which at least the diluent discharges; wherein the dilution device is configured such that when the fluid is introduced into the fluid receiving section and the diluent is introduced into the diluent receiving section, at least the diluent discharges from the outlet; and the specimen dilution section is configured such that when a mixture of the diluent and the specimen reaches a predetermined liquid level, the specimen is diluted to a predetermined ratio.

[0009] a fluid receiving section communicating with the flow channel and into which the fluid is introduced; a diluent receiving section communicating with the flow channel and into which the diluent is introduced; a fluid control mechanism for controlling the outflow of the diluent from the diluent receiving section; and an outlet located downstream of the flow channel and through which at least the diluent flows; wherein the testing device according to the present invention is configured so that when the fluid is introduced into the fluid receiving section and the diluent is introduced into the diluent receiving section, at least the diluent flows out from the outlet; and the specimen dilution section is configured so that when a mixture of the diluent and the specimen reaches a predetermined liquid level or above, the mixture reaches the detection section.

[0010] According to these configurations, the microchannel chip is provided with at least a diluent outlet, and the diluent flowing out from the diluent outlet dilutes the specimen in the specimen dilution section, allowing a grace period before the specimen and the diluent mix together, and allowing the specimen to react with the reaction reagent in advance in the specimen dilution section. Furthermore, because the specimen liquid containing the specimen does not pass through the fluid control mechanism, the properties of the specimen that can be used are not limited by the fluid control mechanism.

[0011] According to the testing device of the present invention, a sample liquid diluted to a predetermined dilution rate or more can be introduced into the detection section, making it easier to detect the sample in the detection section.

[0012] Preferred embodiments of the present invention will be described below, but the scope of the present invention is not limited to the preferred embodiments described below.

[0013] In the dilution device of the present invention, it is preferable that the flow path groove has a regulated section in which the time required for the fluid to pass through is regulated, and that when the fluid and the diluent are introduced, at least the diluent flows out after a predetermined time.

[0014] According to this configuration, the flow channel has a regulated section in which the time required for the fluid to pass through is regulated, so that the diluent can flow out after a predetermined time.

[0015] In the dilution device according to the present invention, the restriction section preferably includes a section in which the flow channel has a width of 0.8 to 2.0 mm and a depth of 70 to 100 μm.

[0016] According to this configuration, by having a section within the above range, it is possible to reduce the variation in the time required for the fluid to pass through the regulated section.

[0017] In the dilution device according to the present invention, it is preferable that the substrate is erected so that the outflow direction downstream of the flow channel faces downward.

[0018] This configuration makes it easier to quickly discharge the diluent from the outlet.

[0019] In the dilution device according to the present invention, it is preferable that the diluent receiving portion is formed in a pocket shape on at least one surface of the base plate.

[0020] According to this configuration, the diluent receiving portion is formed in a pocket shape, which makes it easy to increase the volume of the diluent and to hold the diluent easily when the substrate is placed upright.

[0021] In the dilution device of the present invention, the dilution liquid outflow section comprises a first flow path formed by the flow path groove and the covering material, and a second flow path fluidly connected to the dilution liquid receiving section and fluidly connected to at least one of the side surface and bottom surface of the first flow path, and the fluid control mechanism is configured so that the surface of the first flow path has a first property which is either hydrophobic or hydrophilic, the surface of the covering material of the first flow path facing the flow path groove has a second property which is the other of hydrophobic and hydrophilic, and the second flow path has the second property, and has a guide section that guides the dilution liquid introduced into the dilution liquid receiving section to a connection portion between the first flow path and the second flow path, and when the dilution liquid is introduced into the dilution liquid receiving section, the dilution liquid guided by the guide section forms droplets at the connection portion.

[0022] According to this configuration, an autonomously diluting device can be realized.

[0023] In the dilution device of the present invention, the diluent outlet portion is preferably configured such that the fluid is the diluent, and the diluent reservoir serves as both the fluid receiving portion and the diluent receiving portion, and the diluent reservoir is fluidically connected to each of the first flow path and the second flow path, and when the diluent is introduced from the diluent reservoir into the first flow path, the diluent that flows out from the diluent reservoir through the second flow path flows out from the outlet.

[0024] According to this configuration, the fluid is a diluent, and a single reservoir can serve as both the fluid receiving portion and the diluent receiving portion, making it easy to make a simple and compact device.

[0025] According to the testing device of the present invention, it is preferable that the specimen dilution section includes at least a part of the substrate, the detection section is housed in a through-hole formed in the substrate or a recess formed on at least one surface of the substrate, and at least one surface of the substrate further includes a detection flow channel connecting the specimen dilution section and the detection section, and the mixed liquid in the specimen dilution section flows out to the detection section via the detection flow channel.

[0026] This configuration makes it easy to mount the specimen dilution unit and the detection unit on one substrate.

[0027] Further features and advantages of the present invention will become more apparent from the following description of exemplary, non-limiting embodiments, which proceeds with reference to the drawings. [Brief explanation of the drawings]

[0028] [Figure 1] FIG. 1 is a front view showing an inspection device according to a first embodiment of the present invention; [Figure 2] Right side view of the inspection device in Figure 1 [Figure 3] FIG. 1 is a schematic diagram illustrating an inspection device according to a first embodiment of the present invention. [Figure 4] FIG. 1 is a schematic diagram illustrating an inspection device according to a first embodiment of the present invention. [Figure 5] FIG. 1 is a schematic diagram illustrating an inspection device according to a first embodiment of the present invention. [Figure 6] FIG. 2 is a front view showing the circuit board of the inspection device of FIG. [Figure 7] FIG. 2 is a perspective view showing a pocket-shaped member of the inspection device of FIG. 1; [Figure 8] Cross section of line BB in Figure 6 [Figure 9] Cross section of line CC in Figure 6 [Figure 10] Enlarged view of the main part of the cross section of line DD in Figure 6 [Figure 11] FIG. 1 is a diagram illustrating a fluid control mechanism of an inspection device according to a first embodiment of the present invention. [Figure 12] Enlarged view of the main part of the cross section of line EE in Figure 11 [Figure 13] FIG. 1 is a diagram illustrating a fluid control mechanism of an inspection device according to a first embodiment of the present invention. [Figure 14] Enlarged view of the main part of the FF cross section in Figure 13 [Figure 15] FIG. 1 is a diagram illustrating a fluid control mechanism of an inspection device according to a first embodiment of the present invention. [Figure 16] FIG. 1 is a perspective view of an inspection device according to a modified example of the first embodiment of the present invention. [Figure 17] Enlarged view of the main part of the G-G cross section of the inspection device in Figure 16 [Figure 18] FIG. 10 is a perspective view showing a dilution fluid outlet and a specimen dilution unit of a testing device according to a second embodiment of the present invention. [Figure 19] Enlarged view of the main part of the HH cross section of the inspection device in Figure 18 [Figure 20] FIG. 20 is a diagram illustrating the state in which the diluent has flowed in in FIG. 19. [Figure 21] FIG. 10 is a diagram illustrating a method of using an inspection device according to a third embodiment of the present invention. [Figure 22] FIG. 10 is a schematic diagram illustrating an inspection apparatus according to a third embodiment of the present invention. [Figure 23] FIG. 10 is a schematic diagram illustrating an inspection apparatus according to a third embodiment of the present invention. [Figure 24] FIG. 10 is a schematic diagram illustrating an inspection apparatus according to a third embodiment of the present invention. [Figure 25] FIG. 10 is a schematic diagram illustrating an inspection apparatus according to a third embodiment of the present invention. [Figure 26] FIG. 10 is a schematic diagram illustrating an inspection apparatus according to a third embodiment of the present invention. [Figure 27] 10A and 10B are diagrams showing other examples of connection parts in the fluid control mechanism of the present invention; [Figure 28] Enlarged view of the main part of cross section II in Figure 27 [Figure 29] 10A and 10B are diagrams showing other examples of connection parts in the fluid control mechanism of the present invention; [Figure 30] Enlarged view of the main part of the JJ cross section in Figure 29 DETAILED DESCRIPTION OF THE INVENTION

[0029] An inspection device 1 according to a first embodiment of the present invention will be described with reference to Figures 1 to 15. When terms indicating directions are used in the following description of this specification, unless otherwise specified, the vertical direction is the up-down direction on the plane of each drawing, and the horizontal direction is the left-right direction on the plane of each drawing that is perpendicular to the plane of each drawing. Furthermore, the up-down direction, depth direction, and height direction are also the up-down direction on the plane of each drawing.

[0030] [Overview of Inspection Device 1] The testing device 1 according to the present invention is used as a test kit for testing for infectious diseases caused by, for example, viruses, bacteria, etc., and includes a specimen preparation unit 10 (an example of a dilution device according to the present invention) and a detection unit 20. The specimen preparation unit 10 reacts a specimen such as saliva or a nasopharyngeal swab with a nucleic acid amplification reagent (an example of a reaction reagent), and has the function of diluting the specimen liquid 18B after the reaction before introducing it into the detection unit 20. In this specification, the terms "specimen" and "specimen liquid" are used regardless of whether they are before or after the reaction or the degree of progress of the reaction.

[0031] [Configuration of the sample preparation section] As shown in Figures 1 to 5, the specimen preparation section 10 is composed of a diluent outlet section 30 that discharges diluent 17 into a specimen dilution tank 40 (an example of a specimen dilution section), and the specimen dilution tank 40 that reacts with the introduced specimen and dilutes the specimen after the reaction.

[0032] (Configuration of diluent outflow section) The diluent outlet section 30 according to this embodiment is formed on a microchannel chip substrate (substrate 11). As shown in cross-sectional views such as Fig. 1 and Fig. 8, the diluent outlet section 30 according to this embodiment includes a first flow channel 12 formed on at least one surface of the substrate 11, a first coating material 13 covering the first flow channel 12, a fluid inlet 31 (an example of a fluid receiving section) that communicates with the first flow channel 12 and into which a fluid 16 is introduced, and a diluent reservoir 35 (an example of a diluent receiving section) that communicates with the first flow channel 12 and into which a diluent 17 is introduced. The fluid inlet 31 is located upstream of the first flow channel 12, and the diluent reservoir 35 is located downstream of the fluid inlet 31 and downstream of the first flow channel 12.

[0033] In this embodiment, the substrate 11 is erected approximately vertically so that the outflow direction downstream of the first flow channel 12 faces vertically downward (the direction of the arrow in Figure 1) and the longitudinal direction of the substrate 11 is positioned in the up-down direction.

[0034] As shown in the side view of Fig. 2, the diluent reservoir 35 is formed in the shape of a pocket on one surface of the substrate 11. More specifically, a fixing portion 36A of a pocket-shaped member 36 having the shape shown in Fig. 7 is fixed to an attachment portion 11A on one surface of the substrate 11 shown in Fig. 6, thereby forming a diluent storage portion for the diluent 17 in the diluent reservoir 35. The diluent reservoir 35 is in fluid communication with the second flow channel 34 via a diluent inlet 33 formed as a through-hole in the substrate 11.

[0035] As shown in FIG. 11 , in a front view (partially enlarged view) of the upright substrate 11, the second flow channel 34, which connects the diluent inlet 33 and the first flow channel 12, intersects with the first flow channel 12 at a right angle. The first flow channel 100, formed by the first flow channel 12 and the first covering material 13, and the second flow channel 200, formed by the second flow channel 34 and the second covering material 32, have a connection portion X. In this embodiment, as shown in FIG. 1 , the first flow channel 12 constituting the first flow channel 100 has five parallel straight sections and four arc-shaped curved sections connecting the five straight sections. The fluid 16 introduced into the fluid inlet 31 makes four U-turns on the substrate 11 before reaching the connection portion X. That is, the second flow channel 200 intersects the fifth straight section of the first flow channel 100 at a substantially right angle. An outlet 15, through which at least the diluent 17 flows, is provided downstream of the first flow channel 12.

[0036] As will be described in detail later, the diluent outlet unit 30 according to this embodiment includes a fluid control mechanism that controls the outflow of the diluent 17 from the diluent reservoir 35. As shown in FIG. 13 , the diluent 17 introduced into the diluent reservoir 35 fills the second flow channel 34 and is configured to remain at the connection portion X. When the fluid 16 introduced into the fluid inlet 31 reaches the connection portion X, the second flow channel 200 is opened, and as shown in FIG. 15 , the diluent 17 flows out of the diluent reservoir 35 and reaches the outlet 15. Because the diluent outlet unit 30 includes the fluid control mechanism, when the fluid 16 is introduced into the fluid inlet 31 and the diluent 17 is introduced into the diluent reservoir 35, at least the diluent 17 autonomously flows out from the outlet 15.

[0037] Furthermore, as shown in FIG. 1, the diluent outflow section 30 according to this embodiment has a regulated section 14 in which the time required for the fluid 16 to pass through the first flow channel 12 is regulated. That is, the fluid 16 passes through the regulated section 14 of the first flow channel 12 for a time corresponding to a predetermined time. Specifically, the time required for the fluid 16 to pass through is adjusted by the length, width, depth, etc. of the first flow channel 12. As shown in FIG. 10, the first flow channel 12 is composed of a first flow channel 12A and a first flow channel 12B, which have different depths, with point P as the boundary. The regulated section 14 will be described in detail below.

[0038] In this way, the diluent outflow section 30 of this embodiment has a time control function that controls the outflow time of the diluent 17 and a fluid control function that controls the outflow of the diluent 17 and autonomously causes the diluent 17 to flow out, and is therefore configured to autonomously cause the diluent 17 to flow out from the outlet 15 a predetermined time after the fluid 16 is introduced.

[0039] (Overview of the sample dilution tank) When specimen liquid 18A, which is a reaction liquid obtained by mixing a nucleic acid amplification reagent and a specimen, is introduced into specimen dilution tank 40 according to this embodiment as shown in Fig. 3, a nucleic acid amplification reaction of the virus in the specimen proceeds, and specimen liquid 18B containing a gene amplification product is obtained as shown in Fig. 4. In this way, specimen dilution tank 40 functions as a specimen reaction tank.

[0040] In this embodiment, as described above, restriction section 14 is set so that the time it takes for diluent 17 to flow from diluent outlet 30 into specimen dilution tank 40 is a predetermined time. Therefore, as shown in Fig. 3, fluid 16 is introduced into fluid inlet 31 at the timing when specimen liquid 18A, which serves as a reaction liquid, is introduced into specimen dilution tank 40 to start the reaction between the specimen and the nucleic acid amplification reagent. As a result, fluid 16 reaches connection part X as shown in Fig. 4, and after a predetermined time, diluent 17 autonomously flows into specimen dilution tank 40. When specimen liquid 18B containing the gene amplification product in specimen dilution tank 40 becomes mixed liquid 19 diluted to a predetermined dilution ratio, mixed liquid 19 is introduced into detection unit 20 as shown in Fig. 5.

[0041] That is, as diluent 17 flows out of outlet 15 of diluent outlet 30 after a predetermined time and is gradually introduced into specimen dilution tank 40, the liquid level rises, and diluent 17 and specimen liquid 18B after the nucleic acid amplification reaction are mixed at a predetermined ratio to form mixture 19. In this embodiment, the amount of specimen liquid 18A introduced into specimen dilution tank 40 is set to a predetermined amount, and detection unit 20 is installed so that its lower end is positioned at liquid level H1 at which specimen liquid 18B after the nucleic acid amplification reaction has a predetermined dilution ratio (preferably a dilution ratio between 10 and 50, e.g., 20). Therefore, when mixture 19 reaches the predetermined liquid level H1, specimen liquid 18B is diluted to the predetermined dilution ratio. In testing device 1, when mixture 19 of diluent 17 and specimen liquid 18B reaches or exceeds the predetermined liquid level H1, mixture 19 reaches the lower end of detection unit 20, and mixture 19 is quickly introduced into detection unit 20.

[0042] [Detection unit configuration] The detection unit 20 detects the sample introduced from the sample dilution tank 40. In this embodiment, a long, narrow piece of chromatography paper with a conjugate pad (e.g., a nucleic acid chromatography strip) is used as the detection unit 20, and the vicinity of the upper end of the detection unit 20 is fixed to the substrate 11 so that the lower end of the detection unit 20 is located at the liquid level height H1. Such a detection unit 20 absorbs the mixed liquid 19 containing the sample, making it easy for the detection unit 20 to quickly detect the sample (target substance). The detection unit 20 is not limited to the above, and a detection unit of a commercially available test kit for immunochromatography or the like can be suitably used.

[0043] With the above configuration, the testing device 1 is configured such that by introducing the fluid 16 at approximately the same time as the start of the nucleic acid amplification reaction in the specimen dilution tank 40, the specimen liquid 18B that has been reacted for a predetermined time is diluted to properties (viscosity, concentration, etc.) suitable for detection, and the target substance (gene amplification product) in the specimen liquid 18B is autonomously detected. The test result is confirmed, for example, by whether or not both detection line C and detection line T appear in the detection unit 20, as shown in Figure 5. The appearance of both detection line C and detection line T in the detection unit 20 indicates a positive result, and the appearance of only detection line C indicates a negative result.

[0044] The configurations of the fluid control mechanism, the restriction section 14, the diluent outlet section 30, and the specimen dilution tank 40 will be described in detail below.

[0045] (Fluid control mechanism) First, the fluid control mechanism will be described. As shown in FIGS. 11 to 15, the fluid control mechanism includes a first flow path 100, a second flow path 200, and an outlet 15 fluidly connected to a connection portion X between the first flow path 100 and the second flow path 200. A pocket-shaped member 36 constituting a diluent reservoir 35 is located in front of (on the front side of the page) the diluent inlet 33 in FIGS. 11, 13, and 15, and above the diluent inlet 33 in FIGS. 12 and 14, and stores the diluent 17 therein; however, this is not shown for ease of explanation. As shown in FIGS. 11 and 12, the first flow path 100 and the second flow path 200 are fluidly connected to each other at the connection portion X, where a portion of the first flow path groove 12 and a portion of the second flow path groove 34 intersect.

[0046] The fluid control mechanism in the diluent outlet 30 has the function of releasing the diluent 17, which has been introduced into the diluent reservoir 35 and remains in the diluent reservoir 35 and the second flow path 200, by the action of the fluid 16 that has flowed through the first flow path 100, and causing it to flow out to the outlet 15. In other words, the fluid control mechanism in the diluent outlet 30 has the function of a passive valve that opens the second flow path 200 in response to the action of the fluid 16.

[0047] In this embodiment, a passive valve is configured by utilizing surface tension due to the hydrophobic and water-repellent properties of the flow path surface. Specifically, in this embodiment, the fluid control mechanism has a first flow path 100 having a first property, either hydrophobic or hydrophilic, a surface of the first coating material 13 of the first flow path 100 facing the first flow path groove 12 having a second property, either hydrophobic or hydrophilic, and a second flow path 200 having the second property. The fluid control mechanism also has a guide portion that guides the diluent 17 introduced into the diluent reservoir 35 to a connection portion X between the first flow path 100 and the second flow path 200. When the diluent 17 is introduced into the diluent reservoir 35 and the diluent inlet 33, the diluent 17 guided by the guide portion forms droplets 17A at the connection portion X.

[0048] In this embodiment, a substrate 11 made of a hydrophobic resin is used, and in the arrangement shown in Figure 12, the surfaces (side surface 12D and top surface 12E) of the first flow channel 12 in the first flow channel 100 are hydrophobic (first property). The resin component constituting the substrate 11 is not particularly limited, but in this embodiment, it is selected from one or more hydrophobic resins selected from the group consisting of polycarbonate, cycloolefin copolymer, cycloolefin polymer, polymethylpentene, polystyrene, polymethyl (meth)acrylate, and polyethylene terephthalate. The resin constituting the substrate 11 is preferably one that has either heat resistance or transparency, or both.

[0049] The surface of the first covering material 13 covering the first flow channel 12 is hydrophilic (second property). The first covering material 13 may have at least the surface facing the first flow channel 12 as hydrophilic. In this embodiment, a hydrophilic resin film, such as a hydrophilic acrylic resin, is used. The resin constituting the first covering material 13 is preferably one that has either heat resistance or transparency, or both. A hydrophilic portion may be provided by applying a hydrophilic coating to a covering material made of a hydrophobic resin, or the surface may be made hydrophilic by subjecting the covering material made of a hydrophobic resin to a hydrophilic treatment such as plasma treatment. In this embodiment, the first covering material 13 covers the entire back surface (lower surface in FIG. 12 ) of the erected substrate 11, including the first flow channel 12.

[0050] The thickness of first covering material 13 is not particularly limited, but can be, for example, 0.05 mm to 2 mm. When the thickness is 0.05 mm or more, wrinkles are less likely to occur during bonding, and first flow channel 12 can be easily sealed. Furthermore, when the thickness is 2 mm or less, good conformability to the irregularities of substrate 11 can be easily obtained.

[0051] The bonding between the first covering material 13 and the substrate 11 may be performed by providing an adhesive layer on the first covering material 13 side to serve as a bonding layer with the substrate 11, by bonding the substrate 11 and the first covering material 13 together with an adhesive or the like, or by bonding the substrate 11 and the first covering material 13 together by thermocompression bonding.

[0052] The second flow channel 34 according to this embodiment is formed so as to open on the surface of the substrate 11 opposite to the first flow channel 12 (the upper surface in FIG. 12 ). The second flow channel 34 is covered with a second coating material 32 to form a second flow channel 200. The first flow channel 12 is covered with a coating material 13 (referred to as a first coating material) as described above to form a first flow channel 100.

[0053] The second covering material 32 is disposed at least in a portion covering the second flow channel 34 on the surface of the substrate 11 opposite to the first covering material 13. In this embodiment, the second covering material 32 is disposed so as to overlap the connection portion X in a front view of the substrate 11. The second covering material 32 may extend so as to protrude from the second flow channel 34 toward the diluent inlet 33.

[0054] In this embodiment, the second flow channel 34 is a linear groove that is shallower and shorter than the first flow channel 12B. Specifically, the length of the second flow channel 34 is set so that the length of the first flow channel 100 from the outlet of the fluid inlet 31 to the connection portion X (referred to as flow channel length M1, not shown) is longer than the length of the second flow channel 200 from the outlet of the diluent inlet 33 to the connection portion X (referred to as flow channel length M2) (FIG. 11). In this embodiment, the flow channel length M1 is set sufficiently long, for example, to be 10 times or more longer than the flow channel length M2.

[0055] The width and depth of the second flow channel 34 are not particularly limited, but in this embodiment, the width is set to be in the range of 0.8 to 2.0 mm, and the depth is set to be in the range of 0.1 to 0.5 mm, for example.

[0056] The second flow channel 34 is formed on the same substrate 11 as the first flow channel 12, and therefore the surfaces (side surface 34D and bottom surface 34E) of the second flow channel 34 are hydrophobic (first property). In this embodiment, the second covering material 32 uses the same hydrophilic resin film as the first covering material 13, and the surface 32A of the second covering material 32 is hydrophilic (second property).

[0057] The diluent 17 introduced into the diluent reservoir 35 is an aqueous solution such as water or a buffer solution, and the fluid 16 introduced into the fluid inlet 31 is an aqueous solution such as water or a buffer solution. Since both the fluid 16 and the diluent 17 are hydrophilic liquids, the fluid 16 is repelled by the surface of the first flow channel 12 in the first flow channel 100 and exhibits the property of wetting and spreading on the surface of the first covering material 13. Similarly, the diluent 17 is repelled by the surface of the first flow channel 12 and the surface of the second flow channel 34 in the second flow channel 200 and exhibits the property of wetting and spreading on the surfaces of the first covering material 13 and the second covering material 32.

[0058] Therefore, when a fluid 16 is introduced into the first flow path 100 from the fluid inlet 31, the fluid 16 flows along the hydrophilic surface of the first covering material 13 and into the first flow path 100. Similarly, when a diluent 17 is introduced into the second flow path 200 from the diluent reservoir 35, the diluent 17 flows along the hydrophilic surface 32A of the second covering material 32 and into the second flow path 200.

[0059] The first covering material 13 and the second covering material 32 may be different in material, thickness, etc., as long as their surfaces are hydrophilic. Furthermore, the method for bonding the substrate 11 and the second covering material 32 may be different from the method for bonding the substrate 11 and the first covering material 13. In this embodiment, since the area of ​​the second covering material 32 is smaller than the area of ​​the first covering material 13, the first covering material 13 may be bonded by thermocompression bonding, and the second covering material 32 may be bonded by an adhesive layer or bonding layer.

[0060] Although this is an example of one embodiment and is not limiting, the thickness of the substrate 11 in this embodiment is approximately 0.75 mm. The depth D1 of the first flow channel 12A is 0.1 mm, the depth D2 of the first flow channel 12B is approximately 0.4 mm, and the depth D3 of the second flow channel 34 is approximately 0.35 mm. Therefore, the groove bottoms (12E and 34E) of the first flow channel 100 and the second flow channel 200 are on approximately the same plane, and the groove bottoms of the first flow channel 12B and the second flow channel 34 overlap, providing fluid communication between them. With this configuration, a through-hole is formed in the substrate 11 at the connection portion X, and the first coating material 13 and the second coating material 32 are located on either side of the through-hole.

[0061] Fluid inlet 31 is a through-hole formed in substrate 11, and although there are no limitations on its shape or size as long as it can introduce fluid 16 into first flow channel 100, in this embodiment it is cylindrical with a diameter of 2 to 5 mm (for example, 4 mm). One side of the through-hole of fluid inlet 31 is open, and the other side is covered with first covering material 13 extending from first flow channel 100.

[0062] Like fluid inlet 31, diluent inlet 33 is a through-hole formed in substrate 11, and although there are no limitations on its shape or size as long as it can introduce diluent 17 into second flow path 200, in this embodiment it is cylindrical with a diameter of 2 to 5 mm (for example, 4 mm). One side of the through-hole of diluent inlet 33 opens to pocket-shaped member 36 (diluent reservoir side), and the other side is covered with first covering material 13.

[0063] Outlet 15 indicates the region downstream from connection portion X and includes the downstream end portion of first flow channel 100 (having first flow channel groove 12B with depth D2) and notch 15A that is fluidly connected to the downstream end portion of first flow channel 100. Notch 15A opens on one side of substrate 11 and penetrates through substrate 11 in the thickness direction. In this embodiment, a specimen dilution tank 40 is attached to the lower end portion of notch 15A. The sidewall of notch 15A is hydrophobic, similar to substrate 11. The open side of notch 15A is covered with a hydrophilic first coating material 13 that extends from first flow channel 100.

[0064] The outer shape and size of substrate 11 can be set appropriately taking into consideration ease of handling, etc. For example, if it is quadrilateral (square or rectangle), it is preferable that each side is 10 mm or more and 200 mm or less, and more preferably 10 mm or more and 100 mm or less. The outer shape of substrate 11 is not particularly limited, and may be other polygonal, circular, elliptical, or the like. The thickness of substrate 11 is also not particularly limited, and can be, for example, 5 to 20 mm.

[0065] The resin substrate 11 can be produced by a resin molding process technique, such as injection molding, transfer molding, or extrusion molding.

[0066] In this embodiment, the first property is hydrophobic and the second property is hydrophilic. However, the first property is a property that prevents the fluid used from wetting and spreading (i.e., a repelling property), and the second property is a property that the fluid used spreads and wetting, and the degree and physical value of the property are not particularly limited. In this embodiment, the surface of substrate 11, which is hydrophobic (first property), has a contact angle with water of, for example, 60° or more and 100° or less. The lower limit of the contact angle of substrate 11 with water is preferably 70° or more, and more preferably 80° or more. The upper limit of the contact angle of substrate 11 with water is preferably 95° or less, and more preferably 90° or less. Furthermore, the surfaces of first coating material 13 and second coating material 32, which are hydrophilic (second property), have a contact angle with water of, for example, 0° or more and 40° or less. The upper limit of the contact angle of first coating material 13 and second coating material 32 is preferably 20° or less, and more preferably 10° or less. The contact angle with water in this embodiment is a value measured at 25°C using a commercially available contact angle meter.

[0067] The specimen dilution tank 40 is not particularly limited as long as it has a capacity large enough to accommodate the diluent 17 and the specimen liquid 18 (18A, 18B). In this embodiment, the specimen dilution tank 40 is made of a resin microtube, which is fixed to the lower end of the notch 15A of the substrate 11. Specifically, two notches extending vertically are made at opposing positions on the upper opening of the microtube, and the substrate 11 is inserted and fixed into the notches so as to receive the lower end of the notch 15A of the substrate 11, allowing the diluent 17 flowing out of the outlet 15 to be stored.

[0068] It is preferable that specimen dilution vessel 40 is configured to be kept at a constant temperature (for example, a predetermined temperature between 25 and 65°C) while functioning as a specimen reaction vessel for nucleic acid amplification reaction in particular, so that the reaction can be carried out at a constant temperature. Specimen dilution vessel 40 can be kept warm or heated by placing the entire testing device 1 in a thermostatic bath or room, or by placing a heating device such as a heater only around specimen dilution vessel 40.

[0069] (Restricted section) As described above, the flow channel 12 associated with the diluent outlet 30 has a regulated section 14 in which the time required for the fluid 16 to pass through is regulated. In the regulated section 14, the fluid 16 passes through the flow channel 12 of the regulated section 14 for a predetermined time, for example, within a range of 5 to 30 minutes, such as 5, 10, or 20 minutes. In this embodiment, the time required for the regulated section 14 is appropriately set according to the reaction time required for the specimen in the specimen liquid 18A to react with the reaction reagent, and is desirably set within a tolerance of ±10%, preferably ±5%. With this configuration, the diluent 17 can be discharged from the diluent outlet 30 after the predetermined time.

[0070] The restriction section 14 only needs to restrict the time required for the fluid 16 to pass through, and the restriction section may be set afterwards by measuring the time required for the flow channel 12 that has already been created, or the time required for the fluid 16 to pass through may be set by adjusting the length, width, depth, etc. of the first flow channel 12 to predetermined dimensions.

[0071] In the restriction section 14 according to this embodiment, the first flow path 100 includes a first section L1 (not shown). In this embodiment, the first section L1 starts from a starting point S and extends to a point P midway along the fifth straight section of the first flow path 100. The first section L1 is a section in which the width W1 of the first flow path groove 12A is 0.8 to 2.0 mm and the depth D1 is 0.07 to 0.10 mm (70 to 100 μm). Setting the width W1 and depth D1 of the first flow path groove 12A within this range can suppress variations in the flow velocity of the fluid flowing through the flow path.

[0072] The width W1 of the first flow channel 12A is preferably 0.8 to 1.5 mm, and more preferably 0.9 to 1.2 mm, and the depth D1 of the first flow channel 12A is preferably 0.09 to 0.10 mm, and more preferably 0.095 to 0.100 mm.

[0073] In this embodiment, as shown in FIG. 6, a second section L2, which is deeper than the first flow channel 12A in the first section L1, is provided adjacent to the first section L1 (not shown). In this embodiment, the depth D2 of the first flow channel 12B in the second section L2 is set to 2 to 10 times the depth W1 of the first flow channel 12A in the first section L1 (particularly, 4 times in this embodiment). The width W2 of the first flow channel 12B in the second section is set to the same as the width W1 of the first flow channel 12A in the first section. By providing the second section L2, the flow rate can be reduced, allowing for longer measurement times without increasing the dimensions of the device. Therefore, the predetermined time can be set to a long time, such as 10 to 30 minutes.

[0074] Point P, which is the transition point from first flow channel 12A in first section L1 to first flow channel 12B in second section L2, has an inclined surface 12C as shown in FIG. 10 . In this embodiment, the angle θ between top surface 12E of first flow channel 12B and inclined surface 12C in FIG. 10 is 124°. At point P, first flow channel 12A in first section L1 suddenly expands from depth D1 to depth D2 of first flow channel 12B in second section L2. This is a so-called sudden expansion pipe. When fluid 16 enters first flow channel 12B, the flow of fluid 16 cannot immediately follow the flow channel shape, and a vortex is formed at the sudden expansion portion, drawing in surrounding fluid 16. This vortex is a flow that remains in place, resulting in pressure loss. However, in this embodiment, this pressure loss is utilized to reduce the fluid flow velocity. The angle θ formed between the top surface 12E of the first flow channel 12B and the inclined surface 12C is not particularly limited, but is preferably 110° to 135°.

[0075] The ratio of the lengths of the first section L1 and the second section L2 (length of L1: length of L2) is not particularly limited, but is preferably 9:1 to 5:5. With such a ratio, the effect of suppressing time variations in the first section L1 can be easily obtained.

[0076] In this embodiment, the cross-sectional shape of the first flow channel 12 (12A, 12B) is quadrangular (particularly rectangular), but the shape is not limited thereto. The cross-sectional shape of the first flow channel 12 (12A, 12B) may be trapezoidal or semicircular. In particular, if the cross-sectional shape of the first flow channel 12 (12A, 12B) is a trapezoid in which the side on the opening side (the side of the first covering material 13) is larger than the side on the groove bottom of the first flow channel 12 (12A, 12B), the first flow channel 12 can be easily manufactured using a mold. In the case of such a shape, the widths W1 and W2 and the depths D1 and D2 of the first flow channel 12 (12A, 12B) refer to the maximum width and maximum depth of the first flow channel 12 (12A, 12B).

[0077] In this embodiment, the first flow path 100 is configured to have a greater conduit resistance than the second flow path 200. The conduit resistance of a flow path is determined by various conditions, and generally, the first flow path 100 will have a greater conduit resistance than the second flow path 200 when the first flow path 100 is longer, has a greater conduit friction coefficient, is narrower, or has a higher flow rate than the second flow path 200. In this embodiment, as described above, the flow path length M1 from the fluid inlet 31 to the connection portion X is sufficiently longer than the flow path length M2 from the diluent inlet 33 to the connection portion X, and the conduit resistance of the first flow path 100 is greater than the conduit resistance of the second flow path 200.

[0078] [Inspection method using inspection device 1] Next, an inspection method using the inspection device 1 will be described in relation to the functions and actions of each part of the inspection device 1.

[0079] Sample liquid 18A, fluid 16, and diluent 17 are prepared in advance. Sample liquid 18A is, for example, a sample such as saliva or a nasopharyngeal swab collected from a human or animal mixed with a reaction reagent such as a nucleic acid amplification reagent (pre-reaction liquid). Fluid 16 is a fluid control fluid for causing diluent 17 remaining in second flow path 200 and diluent inlet 33 to flow out to outlet 15. Fluid 16 regulates the predetermined time based on the time required for the fluid to flow through regulation section 14 of first flow path 100. For this reason, fluid 16 is a liquid with small viscosity variation, such as water or a buffer solution, and it has been confirmed in advance that the time required for fluid 16 introduced into fluid inlet 31 to reach connection portion X is within a tolerance of ±5% of the predetermined time. Diluent 17 is a fluid for diluting sample liquid 18B after the nucleic acid amplification reaction, and is a liquid such as water or a buffer solution. The sample liquid 18 (18A, 18B), the fluid 16 and the diluent 17 are all hydrophilic.

[0080] The order in which the specimen liquid 18A, fluid 16, and diluent 17 are introduced into the testing device 1 is not limited. For example, the diluent 17 may be introduced into the diluent reservoir 35 first, the specimen liquid 18A (specimen and reaction reagent) may then be introduced into the specimen dilution tank 40, and the fluid 16 may then be introduced into the fluid inlet 31. The specimen liquid 18A and the fluid 16 are preferably introduced simultaneously, but may be introduced in a slightly different order, and the order of introduction is not particularly limited. The diluent 17 may be introduced as shown in FIG. 3 by the time the fluid 16 reaches the connection portion X. As shown in FIGS. 13 and 14 , the surface 32A of the second coating material 32 in the second flow path 200 serves as a guide, causing a portion of the diluent 17 to flow out of the diluent inlet 33, fill the second flow path 200, and be further guided to the connection portion X. The diluent 17 then forms a droplet 17A at the connection portion X. In this embodiment, droplet 17A spreads over surface 32A of second covering material 32 above connection portion X in FIG. 14 and is held on surface 32A of second covering material 32 in a drooping state due to surface tension and gravity. At this time, droplet 17A does not contact first covering material 13. Even if droplet 17A contacts side surface 12D and top surface 12E of first flow channel 12 or side surface 34D and bottom surface 34E of second flow channel 34, droplet 17A does not spread because these surfaces are all hydrophobic, and instead becomes a rounded droplet due to surface tension. This configuration can prevent diluent 17 from flowing out of outlet 15 before droplet 17A contacts fluid 16.

[0081] 4, when fluid 16 is introduced into fluid inlet 31, it starts from starting point S and flows through first flow path 100 (12A, 12B) over a predetermined time (e.g., a preset time such as 20 minutes) until it reaches connecting point X. First flow path 100 according to this embodiment can regulate the predetermined time by the time required for the fluid to pass through first flow path 100, and the reaction between the specimen and the reaction reagent in specimen liquid 18A progresses during this predetermined time.

[0082] Next, the fluid 16 that has passed through the first flow path 100 over a predetermined time period reaches the connection portion X and comes into contact with the droplets 17A. Then, as shown in FIGS. 5 and 15 , at least the diluent 17 flows out from the outlet 15. In this manner, the fluid 16 that has passed through the first flow path 100 acts to cause the diluent 17 that has remained in the second flow path 200 and the diluent inlet 33 to flow out to the outlet 15. The fluid 16 that has passed through the long first flow path 100 (12A, 12B) loses pressure (propulsion force) due to friction with the wall surfaces within the flow path and the like before reaching the connection portion X. Therefore, at the connection portion X, the outflow of the diluent 17 from the second flow path 200 takes precedence over the outflow of the fluid 16 from the first flow path 100. The diluent outlet 30 can also be called a "surface tension valve" that utilizes surface tension, since at the connection point X where two flow paths are connected, the fluid that has passed through one flow path breaks the balance of the surface tension of the fluid in the other flow path, thereby opening the other flow path.

[0083] According to the diluent outlet 30 of this embodiment, the diluent 17 in the second flow path 200 can be discharged to the outlet 15 by the action of the fluid 16 introduced into the first flow path 100 formed in the same substrate 11, without requiring any external operation to open the second flow path 200. Furthermore, in the diluent outlet 30 of this embodiment, as shown in FIG. 11 , the second covering material 32 at least partially overlaps the connection portion X in the front view of the substrate 11, so that the surface 32A of the second covering material 32 can guide the sample liquid 18 to the connection portion X. That is, in the diluent outlet 30, the surface 32A of the second covering material 32 in the second flow path 200 serves as a guide that guides the diluent 17 to the connection portion X.

[0084] In the diluent outlet section 30 according to this embodiment, the flow path length M1 of the first flow path 100 from the fluid inlet 31 to the connection portion X is greater than the flow path length M2 of the second flow path 200 from the diluent inlet 33 to the connection portion X, and the first flow path 100 has a greater conduit resistance than the second flow path 200. Therefore, when the fluid 16 passing through the first flow path 100 comes into contact with the droplet 17A of the diluent 17 formed at the connection portion X, the diluent 17 naturally flows out of the outlet 15 and into the specimen dilution tank 40. One side of the through-hole in the substrate 11 corresponding to the notch 15A of the outlet 15 is covered with a first coating material 13 having a hydrophilic surface extending from the first flow path 100. The substrate 11 is erected so that the outflow direction downstream of the first flow channel 12 faces downward. Therefore, the hydrophilic diluent 17 travels through the first coating material 13 and is rapidly introduced into the specimen dilution tank 40. Through the above steps, the diluent 17 can be introduced into the specimen dilution tank 40 after a predetermined time.

[0085] In specimen dilution tank 40, the reaction between the specimen in specimen liquid 18A and the reaction reagent (nucleic acid amplification reagent) progresses until diluent 17 is introduced after a predetermined time, resulting in specimen liquid 18B containing a gene amplification product after the nucleic acid amplification reaction, to which diluent 17 is introduced. When diluent 17 flows in and mixed liquid 19 of diluent 17 and specimen liquid 18B reaches a liquid level H1 as shown in Figure 5, mixed liquid 19 comes into contact with the lower end of detection unit 20, and mixed liquid 19 is quickly absorbed by the chromatography paper of detection unit 20.

[0086] The test result in the test device 1 is confirmed by whether or not both detection line C and detection line T appear in the detection unit 20, as shown in an example in Figure 5. If both detection line C and detection line T appear in the detection unit 20, it is positive, and if only detection line C appears, it is negative.

[0087] When the fluid 16 reaches the connection part X, the diluent outflow part 30 of the testing device 1 causes the diluent 17 to naturally flow out to the outlet 15. In this way, the diluent outflow part 30 opens the second flow path 200 in response to the action of the fluid 16 and discharges the diluent 17, even without external control to open the second flow path 200.

[0088] In the diluent outlet 30 according to this embodiment, the fluid 16 introduced into the fluid inlet 31 passes through the restricted section 14 of the first flow path 100 over a predetermined time, causing the diluent 17 to flow out after the predetermined time. Therefore, according to the testing device 1 according to this embodiment, the diluent 17 is introduced into the sample liquid 18B that has been reacted for a predetermined time, and the sample can be detected after diluting it to a predetermined dilution ratio. Consequently, the diluent outlet 30 functions as a timer that measures the reaction time of the sample liquid 18A. That is, according to the testing device 1, the sample liquid 18A containing the sample before reaction introduced into the sample dilution tank 40 becomes the sample liquid 18B after the nucleic acid amplification reaction after a predetermined time. The sample liquid 18B is then autonomously diluted with the diluent 17, and the mixture 19 at the predetermined dilution ratio is autonomously introduced into the detection unit 20. Therefore, while the dilution mechanism is capable of diluting the sample liquid 18B after the nucleic acid amplification reaction to a concentration suitable for detection in the detection unit 20, variation in test results due to variation in reaction time can be suppressed. Therefore, the testing kit can be configured to enable simple and rapid detection while improving testing accuracy.

[0089] 6 and 11, etc., in the front view, the second flow path 200 is provided so as to protrude into the first flow path 100. That is, in the front view, the second flow path 200 is in a state of protruding into the first flow path 100. Therefore, it is possible to easily provide the connection portion X while suppressing the outflow of the diluent 17 from the second flow path 200 to the first flow path 100.

[0090] According to the testing device 1 of this embodiment, the post-reaction specimen liquid 18B diluted to a predetermined dilution ratio or greater can be introduced into the detection unit 20, facilitating detection of the target substance (gene amplification product) in the detection unit 20. The testing device 1 includes at least a diluent outlet 30 on the microchip. The diluent 17 flowing out from the diluent outlet 30 dilutes the specimen liquid 18B in the specimen dilution tank 40. This allows a grace period before the specimen liquid 18A and the diluent 17 mix, allowing the specimen to react with the reaction reagent in advance in the specimen dilution tank 40. Furthermore, according to the testing device 1 of this configuration, the outflow of the diluent 17, not the specimen liquid 18, is controlled by the fluid control mechanism. Since the specimen liquid 18 (18A, 18B) does not pass through the fluid control mechanism, the properties of the specimen that can be used are not limited by the fluid control mechanism. For example, a specimen containing a surfactant can be diluted. In other words, the inclusion of a surfactant in the specimen liquid 18 makes it possible to extract and amplify genes from bacteria or viruses with thick cell walls.

[0091] In the testing device 1 according to this embodiment, the substrate 11 is set upright, so that the diluent 17 is quickly discharged from the outlet 15 and easily introduced into the specimen dilution tank 40. This reduces the likelihood of errors in measuring the required time. In addition, the specimen liquid 18 and the diluent 17 introduced into the specimen dilution tank 40 are easily mixed.

[0092] In the testing device 1 according to this embodiment, the diluent reservoir 43 is formed in a pocket shape, so that it is easy to hold a large volume of diluent while the substrate 11 is in an upright position. In addition, by forming the diluent reservoir 43 with a specified cross-sectional area, etc., it is easy to measure the diluent 17.

[0093] Next, an inspection device 1B, which is a modified example of the first embodiment of the present invention, will be described with reference to FIGS.

[0094] As shown in Fig. 16, the testing device 1B differs significantly from the testing device 1 in that the substrate 11 is not placed upright but is placed substantially horizontally. In the testing device 1B, the second flow path 200 and the diluent inlet 33 are provided closer to the notch 15A of the outlet 15 than in the testing device 1 of the first embodiment, thereby shortening the length of the outlet 15. Furthermore, in the testing device 1B, as shown in the cross-sectional view of Fig. 17, the first coating material 13 placed on the underside of the outlet 15 of the substrate 11 is peeled off, and the peeled first coating material 13 is placed so as to hang down into the specimen dilution tank 40 placed below the substrate 11.

[0095] In the testing device 1B, the substrate 11 is not provided upright, and therefore the pocket-shaped diluent reservoir 35 as in the testing device 1 is not provided. In addition, the detection unit 20 is fixed to the edge of the specimen dilution tank 40 at a position where the lower end thereof is at the liquid level height H2.

[0096] 16 shows the state immediately after fluid 16 has been introduced into fluid inlet 31, diluent 17 has been introduced into diluent inlet 33, and a pre-measured amount of specimen liquid 18A has been introduced into specimen dilution tank 40. Even with this configuration, diluent 17 can be discharged from outlet 15 along first covering material 13, and can be introduced into specimen dilution tank 40. Testing device 1B introduces diluent 17 into specimen dilution tank 40 more slowly than testing device 1, but otherwise achieves substantially the same effects as testing device 1. Note that in testing device 1B, instead of diluent reservoir 35 of testing device 1, the capacity of diluent inlet 33 may be increased, or a separate diluent reservoir fluidly connected to diluent inlet 33 may be provided.

[0097] Next, an inspection device 2 according to a second embodiment of the present invention will be described with reference to FIGS.

[0098] The testing device 2 according to the second embodiment differs from the testing device 1B according to the modified example of the first embodiment in the configuration of the sample preparation unit 10. Therefore, only the sample preparation unit 10 will be described. In the testing device 2, the sample dilution tank 60 is fixed to the downstream end of the first flow channel 12 and to the underside of the substrate 11. That is, the diluent 17 flows directly into the sample dilution tank 60 from the first flow channel 12B. Figures 18 and 19 show the state immediately after fluid 16 has been introduced into the fluid inlet 31, diluent 17 has been introduced into the diluent inlet 33, and a pre-measured sample liquid 18A has been introduced into the sample dilution tank 60.

[0099] The specimen dilution tank 60 has a diluent outlet 39 formed as a through-hole in the substrate 11 at its upper portion. A cylindrical container 61 having a bottom 61A and the same inner diameter as the diluent outlet 39 is connected below the diluent outlet 39. With this configuration, the inner surface of the diluent outlet 39 is continuous with the inner wall 61B of the cylindrical container 61, forming the inner wall of the specimen dilution tank 60. The specimen dilution tank 60 has a bottom 61A that is continuous with the lower end of the inner wall 61B of the cylindrical container 61 and is horizontal with the substrate 11. The inner wall 61B of the specimen dilution tank 60 has an outlet 61C and is fluidly connected to the first flow channel 12B on the substrate 11. The outlet 61C is located at a height H3 at which a predetermined volume ratio is achieved between the internal volume of the diluent inlet 33 (which functions as a diluent reservoir in the testing device 2) and the volume of the specimen liquid 18A introduced into the specimen dilution tank 60.

[0100] In the testing device 2, the bottom surface (bottom 61A) of the specimen dilution tank 60 is located below the substrate 11. Therefore, in the specimen dilution tank 60, the diluent 17 stored in the diluent inlet 33 autonomously flows into the specimen liquid 18B after the nucleic acid amplification reaction. This simplifies the structure. Furthermore, in the specimen dilution tank 60, the diluent 17 and the specimen liquid 18B are mixed due to the difference in height to form the mixed liquid 19 as shown in FIG. 20, so that the diluent 17 and the specimen liquid 18B are easily mixed. The mixed liquid 19, which has reached a predetermined height H3 and is discharged from the outlet 61C, is introduced into the detection unit 20 via the pipe 62 connected to the outlet 61C. The detection unit and other components may be configured similarly to those of the first embodiment, and the detection unit may be disposed horizontally (not shown).

[0101] Next, an inspection device 3 according to a third embodiment of the present invention will be described with reference to FIGS.

[0102] [Configuration of inspection device 3] The testing device 3 according to the third embodiment is a modification of the testing device 1, in which the diluent outflow unit 30, specimen dilution tank 88, and detection unit 20 are integrated into one cassette. As shown in Fig. 21, the testing device 3 is used by being set upright in a warming device 70 (S01). The warming device 70 has a window 70a through which the test results can be visually observed.

[0103] 22, the testing device 3 includes a main body board 81 on which the diluent outflow unit 30, the specimen dilution tank 88, and the detection unit 20 are mounted, a front cover 82 that covers the front of the main body board 81, and a diluent introduction button 83 for starting the introduction of the fluid and diluent. The front cover 82 is provided with a window 82a for viewing the test results at a position overlapping with the window 70a of the warming device 70.

[0104] The inspection device 3 according to this embodiment differs significantly from the inspection device 1 according to the first embodiment in the following points.

[0105] In the testing device 3, the diluent 17, which is both a fluid and a diluent, is supplied from a single diluent reservoir 84. That is, in the testing device 3, the fluid 16 and the diluent 17 are the same liquid (water or buffer solution), the fluid 16 is the diluent 17, and the main substrate 81 is provided with the diluent reservoir 84, which serves as both the fluid inlet and the diluent inlet in the testing device 1.

[0106] In the testing device 3, the diluent reservoir 84 is configured to be fluidly connected to the first flow path 100 and the second flow path 200 when the diluent introduction button 83 is pressed, and the diluent 17 flows out into the first flow path 100 and the second flow path 200. For example, when the diluent introduction button 83 shown in Figure 21 is pressed (S03), the bottom surface 84A of the diluent reservoir 84 is opened, as shown in Figures 23 and 24, and the diluent 17 flows out.

[0107] The testing device 3 includes a first communication path 85 and a second flow path 200 branching from a diluent reservoir 84. The first communication path 85 connects the diluent reservoir 84 and the first flow path 100. When the diluent 17 is introduced from the diluent reservoir 84 through the first communication path 85 into the first flow path 100, the diluent 17 passes through the second flow path 200 and forms a droplet 17A at the connection portion X while passing through the restriction section 14 of the first flow path 100. Similar to the testing device 1, the testing device 3 is configured such that the diluent 17 flowing out from the diluent reservoir 84 through the second flow path 200 flows out from the outlet 87 using a fluid control mechanism. At least a portion of the cross section of the outlet 87 in the outflow direction of the diluent 17 is hydrophilic. The restriction section 14 and the fluid control mechanism have the same configurations as those of the testing device 1 according to the first embodiment.

[0108] In the testing device 3, the specimen dilution tank 88 is configured to include at least a part of the main body substrate 81. In this embodiment, as shown in S02 in Fig. 21 , when a container 90 containing specimen liquid 18A, which is a mixture of a specimen and a reaction reagent, is set in a specimen container receptacle 91 configured to include the main body substrate 81, the bottom of the container 90 is opened and the specimen liquid 18A flows into the specimen dilution tank 88 (Fig. 23). This configuration is one example, and specimen liquid 18A may be introduced directly into the specimen dilution tank 88, or a freeze-dried reaction reagent may be sealed in the specimen dilution tank 88 in advance, and the specimen or a nucleic acid extract extracted from the specimen may be introduced directly into the specimen dilution tank 88.

[0109] In the testing device 3, the detection unit 20 is housed in a through-hole 81B formed in the main body substrate 81. At least one surface of the main body substrate 81 is provided with a detection flow channel 89 that connects the specimen dilution tank 88 and the detection unit 20. The mixed solution 19 in the specimen dilution tank 88 flows out to the detection unit 20 via the detection flow channel 89.

[0110] According to the testing device 3, since the diluent 17 is used as the fluid, one reservoir can serve as both the fluid receiving section and the diluent receiving section, making it easy to make the device simple and compact.

[0111] [Inspection method using inspection device 3] Next, a series of inspection methods using the inspection device 3 will be described.

[0112] First, as shown in S01 of FIG. 21, the testing device 3 is inserted into a recess in the heating device 70. As shown in FIG. 22, the diluent 17 is pre-filled in the diluent reservoir 84 in the testing device 3. The interior of the heating device 70 is set to a predetermined temperature of, for example, 60 to 68°C in the case of the LAMP method. Next, as shown in S02 of FIG. 21, a container 90 containing a specimen liquid 18A (a mixture of a specimen and a reaction reagent) is set in the testing device 3. Then, as shown in FIG. 23, the bottom of the container 90 breaks, and the specimen liquid 18A is introduced into the specimen dilution tank 88. Next, as shown in S03 of FIG. 21, the diluent introduction button 83 is pressed, and the diluent 17 is introduced from the diluent reservoir 84 into the first communication path 85 and the second flow path 200. The diluent 17 introduced into the second flow path 200 forms a droplet 17A at the connection portion X. Diluent 17 introduced into first communicating passage 85 passes through restricted section 14 over a predetermined time period, as shown in Fig. 24. Restricted section 14 is set so that this predetermined time period corresponds to the reaction time of the nucleic acid amplification reaction of specimen liquid 18A in specimen dilution tank 88. While diluent 17 passes through restricted section 14, the nucleic acid amplification reaction of specimen liquid 18A progresses, and specimen liquid 18A becomes specimen liquid 18B containing a nucleic acid amplification product after the reaction.

[0113] Next, when the diluent 17 that has passed through the regulated section 14 in the first flow path 100 comes into contact with the droplet 17A at the connection part X, the diluent 17 flows out of the diluent reservoir 84, and as shown in Figure 25, the diluent 17 flows into the specimen dilution tank 88 through the outlet 87.

[0114] As diluent 17 is introduced into specimen dilution tank 88 containing specimen liquid 18B, diluent 17 and specimen liquid 18 form mixed liquid 19, and the liquid level rises. As shown in FIG. 26, when mixed liquid 19 reaches a predetermined liquid level H4, mixed liquid 19 containing the gene amplification product after the reaction reaches detection unit 20 via detection flow channel 89. The gene amplification product is then detected in detection unit 20, and a test result, positive or negative, is displayed (FIG. 26). As shown in S04 in FIG. 21, the test result can be visually confirmed through viewing window 82a of front cover 82 of testing device 3 and window 70a of warming device 70.

[0115] [Other embodiments] In the above embodiment, an example has been described in which the dilution device according to the present invention is applied as a specimen preparation unit of the testing devices 1 to 3. However, the present invention is not limited to such a configuration, and may be used for other purposes requiring a dilution device.

[0116] In the above embodiment, an example has been described in which the first flow channel and the first flow channel groove 12 (12A, 12B) constituting the first flow channel have the restriction section 14. However, the present invention is not limited to such a configuration, and the restriction section 14 may not be included. Furthermore, the restriction section 14 is not limited to the form of the flow channel groove described in the above embodiment. For example, an example has been described in which the restriction section 14 includes a first section L1 and a second section L2 in this order from the starting point S. However, the present invention is not limited to such a configuration, and the second section L2 may be located before the first section L1, or the second section L2 may be sandwiched between the first sections L1, or the first section L1 may be sandwiched between the second sections L2. Furthermore, an example has been described in which the first section L1 is longer than the second section L2, but the present invention is not limited to such a configuration, and the second section may be longer than the first section L1. Furthermore, the present invention is not limited to such a configuration, and the width of the second section L2 may be larger than the width of the first section L1. For example, the width of the second section L2 may be two to four times the width of the first section L1.

[0117] In the above embodiments, the test devices 1 to 3 are described as being used as test kits for testing infectious diseases caused by viruses, bacteria, etc. However, the test devices are not limited to such a configuration and may be used for tests other than those described above.

[0118] In the above embodiment, an example has been described in which a surface tension valve is used as a configuration for realizing the fluid control mechanism of the inspection devices 1 to 3. However, the present invention is not limited to such a configuration, and other fluid control devices may also be used.

[0119] In the above-described embodiments, in the fluid control mechanisms of the testing devices 1 to 3, as shown in FIGS. 11 and 12, the first flow path 100 and the second flow path 200 overlap at the bottoms of their respective flow path grooves, and the second flow path 200 protrudes into the first flow path 100 in a front view, forming a connection portion X. However, the connection portion X is not limited to such a configuration, as long as the first flow path 100 and the second flow path 200 are fluidically connected. For example, as shown in FIGS. 27 and 28, the second flow path 200 may be provided so as to abut against the side wall of the first flow path 100, and the first flow path 100 and the second flow path 200 may be fluidically connected at the side wall portions of the respective flow path grooves, thereby forming the connection portion X. In this case, the depth of the second flow path groove is D4, which is deeper than D3 in the above-described embodiments. Alternatively, as shown in FIGS. 29 and 30, the connection portion X may be formed by providing the second flow path 200 so as to cross the first flow path 100 in a front view. In this case, it is not necessary to precisely control the processing dimensions of the connection portion X between the first flow channel 12 and the second flow channel 34 in a microchannel device having a fine flow channel, and the diluent outflow portion 30 is easy to manufacture.

[0120] In the third embodiment described above, an example has been described in which the detection unit 20 is accommodated in a through-hole 51B formed in the main body substrate 51. However, the present invention is not limited to such a configuration, and the detection unit 20 may be accommodated in a recess formed on at least one surface of the main body substrate 51. Furthermore, in the third embodiment described above, as shown in FIGS. 22 to 26 , an example has been described in which the diluent 17 flows from the diluent reservoir 84 through one flow path, and the first communicating path 85 and the second flow path 200 partially overlap. However, the first communicating path 85 and the second flow path 200 may not overlap, and the diluent 17 may flow from the diluent reservoir 84 through two flow paths, the first communicating path 85 and the second flow path 200, respectively.

[0121] Regarding other configurations, it should be understood that the embodiments disclosed in this specification are illustrative in all respects and that the scope of the present invention is not limited thereby. Those skilled in the art will easily understand that appropriate modifications are possible without departing from the spirit of the present invention. Therefore, other embodiments modified without departing from the spirit of the present invention are naturally included in the scope of the present invention. [Industrial Applicability]

[0122] The present invention can be used, for example, as a dilution device on a microchannel chip and a test kit using the same. [Explanation of symbols]

[0123] 1, 1B, 2, 3: Inspection equipment 10: Sample preparation section (dilution device) 11, 81: Circuit board (main board) 12: Flow channel (first flow channel) 13: Coating material (first coating material) 14: Restricted section 15: Outlet 16 :Fluid 17: Diluted solution 17A:Droplet 18, 18A, 18B: Sample solution 20: Detection unit 30: Diluted liquid outflow section 31: Fluid inlet (fluid receiving portion) 32: Second coating material 33: Diluent inlet 34: Second flow channel 35: Diluent reservoir (diluent receiving part) 36: Pocket-shaped member 40, 60: Sample dilution tank (sample dilution section) 70:Heating device 83: Diluent introduction button 84: Diluent reservoir 87: Outlet 88: Sample dilution tank (sample dilution section) 89: Detection flow channel groove 100: First flow path 200: Second flow path H1~H4: Liquid level X: Connection part

Claims

1. A dilution device comprising: a diluent outlet portion that discharges at least a diluent when a fluid is introduced; and a specimen dilution portion that receives a specimen and dilutes the specimen with the diluent that has discharged from the diluent outlet portion, The diluent outlet is a flow channel formed on at least one surface of the substrate; a covering material that covers the flow channel; a fluid receiving portion that communicates with the flow channel and into which the fluid is introduced; a diluent receiving portion communicating with the flow channel and into which the diluent is introduced; a fluid control mechanism for controlling the outflow of the diluent from the diluent receiving portion; an outlet located downstream of the flow channel and through which at least the dilution liquid flows out, When the fluid is introduced into the fluid receiving portion and the diluent is introduced into the diluent receiving portion, at least the diluent flows out from the outlet, The sample dilution unit is configured to dilute the sample to a predetermined ratio when the mixture of the diluent and the sample reaches a predetermined liquid level.

2. The dilution device according to claim 1, wherein the flow channel has a regulated section in which the time required for the fluid to pass through is regulated, and when the fluid and the diluent are introduced, at least the diluent flows out after a predetermined time.

3. The dilution device according to claim 2, wherein the regulated section includes a section in which the flow channel has a width of 0.8 to 2.0 mm and a depth of 70 to 100 μm.

4. The dilution device according to claim 1 , wherein the substrate is erected so that the outflow direction downstream of the flow channel faces downward.

5. The dilution device according to claim 4 , wherein the dilution liquid receiving portion is formed in the shape of a pocket on at least one surface of the base plate.

6. The diluent outlet is a first flow path formed by the flow path groove and the covering material; a second flow path in fluid communication with the diluent receiving portion and in fluid communication with at least one of a side surface and a bottom surface of the first flow path; The fluid control mechanism includes: the surface of the first flow path has a first property that is either hydrophobic or hydrophilic; a surface of the coating material of the first flow path on the side of the flow path groove has a second property which is the other of hydrophobicity and hydrophilicity; the second flow path has the second property and includes a guide portion that guides the diluent introduced into the diluent receiving portion to a connection portion between the first flow path and the second flow path; The dilution device according to any one of claims 1 to 5, wherein when the diluent is introduced into the diluent receiving section, the diluent guided by the guide section forms droplets at the connecting portion.

7. the diluent outlet portion includes a diluent reservoir serving as both the fluid receiving portion and the diluent receiving portion, the diluent reservoir being in fluid communication with each of the first flow path and the second flow path; The dilution device according to claim 6, wherein when the diluent is introduced into the first flow path from the diluent reservoir, the diluent that flows out from the diluent reservoir through the second flow path flows out from the outlet.

8. A testing device comprising: a diluent outlet portion that discharges at least a diluent when a fluid is introduced; a specimen dilution portion that receives a specimen and dilutes the specimen with the diluent that has discharged from the diluent outlet portion; and a detection portion that can detect the specimen that has discharged from the specimen dilution portion, The diluent outlet is a flow channel formed on at least one surface of the substrate; a covering material that covers the flow channel; a fluid receiving portion that communicates with the flow channel and into which the fluid is introduced; a diluent receiving portion communicating with the flow channel and into which the diluent is introduced; a fluid control mechanism for controlling the outflow of the diluent from the diluent receiving portion; an outlet located downstream of the flow channel and through which at least the dilution liquid flows out, When the fluid is introduced into the fluid receiving portion and the diluent is introduced into the diluent receiving portion, at least the diluent flows out from the outlet, The specimen dilution unit is configured so that the mixed liquid of the dilution liquid and the specimen reaches the detection unit when the mixed liquid reaches a predetermined liquid level or higher.

9. the specimen dilution unit is configured to include at least a part of the substrate; the detection unit is accommodated in a through hole formed in the substrate or a recess formed in at least one surface of the substrate, a detection channel on at least one surface of the substrate, the detection channel communicating the specimen dilution unit and the detection unit; The testing device according to claim 8 , wherein the mixed solution in the specimen dilution section flows out to the detection section via the detection flow channel.

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