Test container and nucleic acid test method

Abstract

A test container that is attached to a centrifuge, and in which a sample solution is sent in sequence from a first chamber to a second chamber by the centrifugal force caused by the rotation of the centrifuge, said test container comprising: a solution storage part that stores, in advance of the first chamber, the sample solution input from an input port; a valve that has a solution accommodation part for temporarily accommodating the sample solution sent from the first chamber toward the second chamber, said valve being capable of switching among a first state in which the first chamber is in communication with the solution accommodation part and the second chamber is not in communication with the solution accommodation part, a second state in which the second chamber is in communication with the solution accommodation part and the first chamber is not in communication with the solution accommodation part, and a third state in which both the first chamber and the second chamber are not in communication with the solution accommodation part; and a sealing part that prevents backflow of the sample solution after the sample solution has been sent from the solution storage part to the first chamber.

Description

Test container and nucleic acid testing method

[0001] The present invention relates to a test container and a nucleic acid testing method.

[0002] Test containers, such as test cartridges or analytical chips, are known for performing various analyses on specimens extracted from biological samples. For example, the test containers include multiple chambers (liquid storage sections) for storing liquids and flow paths connecting the chambers. In the test containers, specimen liquid is transported from chamber to chamber using external forces such as electromagnetic force, centrifugal force, or pressure.

[0003] JP 2019-515308 A discloses a microfluidic chip in which a sample (analyte liquid) is introduced through a sample inlet (inlet) provided at one end of a sample chamber and transferred from the sample chamber to a mixing chamber by centrifugal force. The microfluidic chip of JP 2019-515308 A includes a circuit configured to provide an air connection between the sample chamber and the mixing chamber, i.e., a vent flow path, enabling smooth liquid transfer.

[0004] In the microfluidic chip of JP 2019-515308 A, after fluid is transferred from the upstream sample chamber to the downstream mixing chamber, the flow path between the two chambers remains open. In the case of an analysis that requires a heating temperature of 90°C or higher in the downstream chamber, the sample liquid may vaporize and flow back into the upstream chamber, resulting in a test error due to an insufficient amount of sample liquid in the downstream chamber. JP 2019-515308 A discloses that a heating process is performed in the downstream chamber to which the sample liquid is transferred for pretreatment, etc. However, because the heating temperature is low at around 65°C, the problem of vaporized sample liquid flowing back into the upstream chamber is not taken into consideration.

[0005] The present disclosure has been made in consideration of the above circumstances, and aims to provide a test container and a nucleic acid testing method that can suppress backflow of sample liquid from a downstream chamber to an upstream chamber in a test container that uses centrifugal force to transport liquid.

[0006] The test container of the present disclosure has an inlet through which a sample liquid is introduced, a first chamber that contains the sample liquid delivered from the inlet, and a second chamber that contains the sample liquid delivered from the first chamber, and is attached to a centrifuge, so that the sample liquid is delivered from the first chamber to the second chamber in sequence by centrifugal force generated by the rotation of the centrifuge; the test container comprises: a removable lid that covers the inlet; a liquid reservoir that stores the sample liquid introduced from the inlet just before the first chamber; and a valve that is disposed between the first chamber and the second chamber and has a liquid storage section that temporarily stores the sample liquid delivered from the first chamber to the second chamber, the valve being switchable between a first state in which the first chamber communicates with the liquid storage section but the second chamber does not communicate with the liquid storage section; a second state in which the second chamber communicates with the liquid storage section but the first chamber does not communicate with the liquid storage section; and a third state in which both the first chamber and the second chamber do not communicate with the liquid storage section. and a sealing portion that prevents backflow of the sample liquid after the sample liquid has been transferred from the liquid reservoir portion to the first chamber.

[0007] The sealing portion is preferably made of a hydrophobic material having a specific gravity lower than that of the sample liquid.

[0008] The hydrophobic material is preferably a low polarity solvent or a thermoplastic material.

[0009] The hydrophobic material is preferably provided in the reservoir or the first chamber.

[0010] The centrifuge may further include a hydrophobic substance storage section for storing a hydrophobic substance, the hydrophobic substance storage section being located closer to the rotation axis of the centrifuge than the liquid reservoir section.

[0011] When the volume of the liquid reservoir is V1, the volume of the first chamber is V2, the volume of the liquid storage portion is V3, and the volume of the second chamber is V4, it is preferable that the relationship be V1 ≥ V2 ≥ V3 ≥ V4.

[0012] A reagent to be reacted with the sample liquid may be contained in the first chamber, the liquid containing portion, or the second chamber.

[0013] The reagents can be reagents that include amplification reagents for amplifying specific nucleic acid sequences and probes for nucleic acid sequencing.

[0014] The probe may be a fluorescent probe.

[0015] a valve in the first state or the third state in an initial state; a sample liquid is introduced into the inlet of the test container while the valve is in the initial state, and the sample liquid is contained in the liquid reservoir; the inlet is sealed with the lid; the sample liquid is sent from the liquid reservoir to the first chamber by centrifugal force; the first chamber is heated to heat the sample liquid while the sealing part prevents backflow of the sample liquid from the first chamber to the liquid reservoir; the valve is switched to the first state; the sample liquid is sent from the first chamber to the liquid reservoir of the valve by centrifugal force, and the sample liquid and the reagent are mixed in the liquid reservoir; the valve is switched to the second state; the mixed liquid of the sample liquid and the reagent is sent from the liquid reservoir to the second chamber by centrifugal force; the valve is switched to the third state or the first state; the mixed liquid in the second chamber is temperature-regulated to amplify a specific nucleic acid sequence in the mixed liquid; The mixture is irradiated with excitation light, and the fluorescence emitted from the fluorescent probe is detected to determine the presence or concentration of a specific nucleic acid sequence.

[0016] According to the test container and nucleic acid testing method of the present disclosure, it is possible to prevent the backflow of the specimen liquid from the downstream chamber to the upstream chamber.

[0017] 1A and 1B are perspective views showing a schematic configuration of a cuvette 10 according to an embodiment. FIG. 2A is a cross-sectional view of the cuvette 10, and FIG. 2B is a plan view of the flow path structure. FIG. 3A is a cross-sectional view of the cuvette 10, and FIG. 3B is a plan view of the flow path structure, showing a state switched to a first state by a valve 20. FIG. 4A is a cross-sectional view of the cuvette 10, and FIG. 4B is a plan view of the flow path structure, showing a state switched to a second state by a valve 20. FIG. 1B is a cross-sectional view of the cuvette 10, and FIG. 1C is a plan view of the flow path structure. FIG. 1C is a diagram showing an embodiment of the cuvette 10. FIG. 1D is a diagram showing before and after the transfer of specimen liquid from a reservoir to a first chamber in a flow path structure 11. FIG. 1E is a diagram showing before and after the transfer of specimen liquid from a reservoir to a first chamber in a flow path structure 11A of a modified example. FIG. 1F is a diagram showing a cuvette provided with a reagent. FIG. 1G is a plan view of a cuvette 50 of a modified example. FIG. 1G is a plan view of a cuvette 52 of a modified example. FIG. 1H is a diagram showing before and after the transfer of specimen liquid from a reservoir to a first chamber in a flow path structure 11B of the cuvette 52. FIG. 1 is a schematic plan view of a test container 53 of a modified example. FIG. 2 is a diagram showing the state before and after sample liquid is sent from the liquid reservoir to the first chamber in a test container 54 of a modified example. FIG. 3 is a diagram showing the general configuration of a test device 100 of an embodiment. FIG. 4 is a diagram showing the general configuration of a test device 101 of a modified example. FIG. 5 is a diagram showing the general configuration of a test device 102 of a modified example. FIG. 6 is a diagram showing the steps of a nucleic acid test (part 1). FIG. 7 is a diagram showing the steps of a β-glucan test (part 1). FIG. 8 is a diagram showing the steps of a β-glucan test (part 2).

[0018] 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.

[0019] "Cuir" Fig. 1 is a perspective view that schematically shows a cuvette 10 of one embodiment. Fig. 2A is a schematic cross-sectional view of the cuvette 10, and Fig. 2B is a schematic plan view that simply shows a flow path structure 11 of the cuvette 10.

[0020] The test vessel 10 is a microfluidic device having a channel structure 11 therein including a plurality of chambers and channels connecting the chambers. The test vessel 10 is attached to a centrifuge, and the liquid inside is transferred from chamber to chamber by centrifugal force generated by the rotation of the centrifuge. The test vessel 10 in this example is loaded into a testing device 100 (see FIG. 14 ) equipped with a centrifuge, and is used for biochemical analysis testing.

[0021] The cuvette 10 is composed of a main body member 10A having recesses and holes that form part of a flow path structure 11 including a flow path and a chamber, and a bottom member 10B that forms the bottom surface of the flow path (see Figure 2).

[0022] The main body member 10A can be made of 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.

[0023] The bottom member 10B is formed, for example, from a thin plate or film. Any known resin-molded plastic material can be used as the bottom member 10B without any particular restrictions, but from the viewpoint of adhesion with the main body member 10A, it is preferable that the bottom member 10B be made of the same material as the main body member 10A. As will be described in detail later, from the viewpoint of increasing adhesion with the heating means and enabling efficient heating when heating the first chamber 15 and the second chamber 16, the bottom member 10B is preferably made of a film. The portions of the main body member 10A that form the first chamber 15 and the second chamber 16 on the upper surface side may also be made of a film.

[0024] The cuvette 10 comprises an inlet 12, a lid 12A, a liquid reservoir 13, a first chamber 15, a second chamber 16, a valve 20 arranged between the first chamber 15 and the second chamber 16, flow paths 18a to 18c connecting these, and a sealing part 19. The multiple chambers comprised by the cuvette 10 are the liquid reservoir 13, the first chamber 15, the second chamber 16, and a liquid storage part 24. Hereinafter, when it is not necessary to distinguish between the liquid reservoir 13, the first chamber 15, the second chamber 16, and the liquid storage part 24, they may be collectively referred to as chambers.

[0025] The inlet 12 is an opening for introducing the sample liquid 40. The lid 12A covers the inlet 12 and is detachable from the opening of the inlet 12. In this example, the lid 12A is formed so as to be threadably engaged with the end of the tubular portion 14 that forms the inlet 12. There are no particular limitations on the method of attachment or detachment; for example, the lid 12A may be attached to or detached from the tubular portion 14 using a snap-type cap structure or an adhesive. The lid 12A opens the inlet 12 when introducing the sample liquid, but closes the inlet 12 at other times to prevent contamination from the outside and to prevent evaporation of the sample liquid 40 from the inside. The sample liquid 40 is, for example, a liquid in which nucleic acids have been extracted from a sample collected from the nasal cavity, pharynx, oral cavity, or affected area of ​​a subject.

[0026] In the following description, the surface on which the inlet 12 is provided will be referred to as the upper surface of the cuvette 10, and the side facing the bottom member 10B will be referred to as the lower surface of the cuvette 10. Here, the upper surface of the main body member 10A is the same as the upper surface of the cuvette 10, the lower surface of the main body member 10A is the surface that contacts the upper surface of the bottom member 10B, and the lower surface of the bottom member 10B is the same as the lower surface of the cuvette 10.

[0027] The liquid reservoir 13 is provided at a position opposite the inlet 12 and stores the sample liquid 40 dropped from the inlet 12. There are no particular restrictions on the shape of the liquid reservoir 13, and it can be arbitrarily selected from a columnar, conical, frustum, or other shapes.

[0028] In the cuvette 10, the inlet 12 is formed by an opening of a cylindrical portion 14 that penetrates the main body member 10A in the thickness direction and protrudes from the upper surface of the main body member 10A, and the liquid reservoir 13 is formed by the inner part of the cylindrical portion 14 in the main body member 10A and the bottom member 10B. The liquid reservoir 13 is a space that stores the sample liquid 40 introduced from the inlet 12 just before the first chamber 15.

[0029] First chamber 15 is a chamber capable of containing a liquid, and is a chamber that contains sample liquid 40 that is delivered from inlet 12 through liquid reservoir 13. First chamber 15 is formed by a recess provided on the lower surface of main body member 10A and bottom member 10B.

[0030] The first chamber 15 is, for example, a chamber in which pre-processing for inspection is carried out, and in an inspection device 100 (see FIG. 14) described later, a heating process is carried out by a first heating unit 120.

[0031] The second chamber 16 is a chamber capable of containing a liquid, and is a chamber that contains the specimen liquid 40 transferred from the first chamber 15. In this example, the specimen liquid 40 is transferred from the first chamber 15 to the second chamber 16 via a liquid storage section 24 of the valve 20, which will be described later. The second chamber 16 is formed by a recess provided on the lower surface of the main body member 10A and a bottom member 10B.

[0032] The second chamber 16 is, for example, a chamber in which reaction and detection processes for testing are carried out, and in the testing device 100 (see FIG. 14) described later, a heating process is carried out by a second heating section 122.

[0033] As shown in FIG. 2A , the valve 20 has a liquid storage portion 24 that temporarily stores the sample liquid 40 transferred from the first chamber 15 to the second chamber 16. The valve 20 switches the connection state between the first chamber 15 and the second chamber 16 and the liquid storage portion 24 between a first state, a second state, and a third state. In the first state, the first chamber 15 and the liquid storage portion 24 are connected, but the second chamber 16 and the liquid storage portion 24 are not connected. In the second state, the second chamber 16 and the liquid storage portion 24 are connected, but the first chamber 15 and the liquid storage portion 24 are not connected. In the third state, neither the first chamber 15 nor the second chamber 16 are connected, but the liquid storage portion 24 is not connected.

[0034] Fig. 2A shows a state in which neither the first chamber 15 nor the second chamber 16 communicates with the liquid storage portion 24. In the schematic diagram showing the flow path structure in Fig. 2B, an x ​​shown above flow path 18b schematically indicates a closed state in which the first chamber 15 and the liquid storage portion 24 do not communicate with each other. Similarly, an x ​​shown above flow path 18c schematically indicates a closed state in which the liquid storage portion 24 and the second chamber 16 do not communicate with each other. In the following drawings, an x ​​shown above a flow path also indicates that the chambers connected via the flow path do not communicate with each other.

[0035] The valve 20 in this cuvette 10 has a rotating member 25 disposed in a tubular portion 22 consisting of a cylindrical recess provided on the upper surface of the main body member 10A and a portion surrounding the recess and protruding from the upper surface of the main body member 10A. The rotating member 25 is a cylindrical member whose outer diameter is slightly smaller than the inner diameter of the tubular portion 22, and is rotatably installed within the tubular portion 22. A liquid containing portion 24 is provided inside the rotating member 25, and one hole 24a communicating with the liquid containing portion 24 is provided on the bottom surface. The bottom surface of the tubular portion 22 has a hole 22a communicating with the flow path 18b and a hole 22b communicating with the flow path 18c. O-rings 26 are provided in the holes 22a and 22b. A knob 25a used to rotate the rotating member 25 is provided on the top surface of the rotating member 25.

[0036] The O-ring 26 aligns the hole 24a on the bottom surface of the rotating member 25 with either the hole 22a or the hole 22b on the bottom surface of the cylindrical portion 22, thereby preventing leakage from the connection between the hole 24a and the hole 22a or the hole 22b when the liquid storage portion 24 is connected to the flow path 18b or the flow path 18c. As a sealing means for preventing leakage, in addition to the O-ring 26, known sealing materials such as packings and gaskets can be used without particular limitation. Examples of materials for the sealing means include rubber materials such as nitrile rubber, urethane rubber, silicone rubber, and fluororubber, as well as known elastomers such as olefin and urethane. Furthermore, the sealing portion may be molded integrally with the rotating member 25 or the cylindrical portion 22 of the main body member 10A.

[0037] By rotating the rotating member 25, it is possible to achieve a state in which the hole 24a on the bottom surface of the rotating member 25 is aligned with the hole 22a on the bottom surface of the tubular portion 22, a state in which the hole 24a is aligned with the hole 22b, or a state in which the hole 24a is not aligned with either the hole 22a or the hole 22b.

[0038] Each of the flow paths 18a to 18c is formed by a recess provided in the lower surface of the main body member 10A and the bottom member 10B. The flow path 18a connects the liquid reservoir 13 and the first chamber 15. The flow path 18b connects the first chamber 15 and the liquid storage portion 24 of the valve 20. The flow path 18b communicates with the hole 22a in the bottom surface of the cylindrical portion 22 of the valve 20. The flow path 18c connects the second chamber 16 and the liquid storage portion 24 of the valve 20. The flow path 18c communicates with the hole 22b in the bottom surface of the cylindrical portion 22. The cross-sectional size of the flow paths 18a to 18c is, for example, about 0.5 mm to 1 mm in width and about 0.5 mm in depth.

[0039] 3 and 4 are diagrams for explaining the first state and the second state switched by the valve 20. Fig. 3 is an explanatory diagram of the cuvette 10 in the first state, Fig. 3A is a schematic cross-sectional view of the cuvette 10, and Fig. 3B is a schematic plan view of the flow path structure 11. Fig. 4 is an explanatory diagram of the cuvette 10 in the second state, Fig. 4A is a schematic cross-sectional view of the cuvette 10, and Fig. 4B is a schematic plan view of the flow path structure 11.

[0040] 3, the hole 24a in the bottom surface of the rotating member 25 is aligned with the hole 22a in the bottom surface of the cylindrical portion 22. At this time, the first chamber 15 and the liquid storage portion 24 are in communication with each other, and the second chamber 16 and the liquid storage portion 24 are not in communication with each other, which is a first state.

[0041] 4, the hole 24a in the bottom surface of the rotating member 25 is aligned with the hole 22b in the bottom surface of the cylindrical portion 22. At this time, the second chamber 16 and the liquid storage portion 24 are in communication with each other, and the first chamber 15 and the liquid storage portion 24 are not in communication with each other, creating a second state.

[0042] As shown in Figure 2, the state in which the hole 24a on the bottom surface of the rotating member 25 does not coincide with either the hole 22a or the hole 22b on the bottom surface of the cylindrical portion 22 is the third state in which neither the first chamber 15 nor the second chamber 16 is connected to the liquid storage portion 24.

[0043] In this way, valve 20 is a rotary valve that can be switched between a first state, a second state, and a third state by rotating rotary member 25 within cylindrical portion 22 of main body member 10A. Note that valve 20 is not limited to a rotary valve having the above structure as long as it can realize the first to third states.

[0044] As shown in FIG. 2 , the sealing portion 19 is provided in a part of the flow path structure 11 to prevent backflow of the sample liquid 40 after the sample liquid 40 has been sent from the liquid reservoir 13 to the first chamber 15 .

[0045] In the example shown in FIG. 2, the sealing portion 19 is made of a hydrophobic material having a specific gravity lighter than that of the sample liquid 40 (hereinafter referred to as the hydrophobic material 19).

[0046] The hydrophobic substance 19 is a low-polarity solvent or a thermoplastic substance. The hydrophobic substance 19 may be a substance that is liquid at room temperature, or a substance that is solid at room temperature but becomes liquid when heated. Examples of low-polarity solvents include silicone oil, hydrocarbon solvents, fluorine-based solvents, and carbon tetrachloride. Here, low polarity means that the substance has a polarity that does not mix with the sample liquid 40. Examples of thermoplastic substances include various waxes and adhesives.

[0047] In the cuvette 10 having this configuration, the hydrophobic substance 19 is provided in the liquid reservoir 13 or the first chamber 15. In the example shown in FIG.

[0048] As an example, the cuvette 10 is set in a loading section 111A of a disk-shaped support 111 as shown in Fig. 5, and rotated around an axis A in a testing device 100 (see Fig. 13) equipped with a centrifuge 110. The cuvette 10 is set in the centrifuge 110 so that the liquid reservoir 13, the first chamber 15, the liquid storage section 24, and the second chamber 16 are arranged in this order from the inner diameter side closest to the axis A, which is the center of rotation.

[0049] The sample liquid 40 introduced from the inlet 12 is transferred from the liquid reservoir 13 to the first chamber 15, from the first chamber 15 to the liquid storage section 24, and from the liquid storage section 24 to the second chamber 16, each of which is achieved by centrifugal force generated by the rotation of the centrifuge 110.

[0050] Here, the method of sending the specimen liquid 40 from the reservoir 13 to the first chamber 15 and the function of the hydrophobic substance 19 will be described with reference to Fig. 6. Fig. 6 shows the state of the flow path structure 11 before and after sending the specimen liquid 40 from the reservoir 13 to the first chamber 15.

[0051] The introduction of the sample liquid 40 is performed in a state where the first chamber 15 is not in communication with the liquid storage portion 24 of the valve 20 by the valve 20. In the example of Fig. 6, the third state is set in which neither the first chamber 15 nor the second chamber 16 is in communication with the liquid storage portion 24, but the second state may also be used.

[0052] As shown in the left diagram of Figure 6 , immediately after the specimen liquid 40 is introduced through the inlet 12, the specimen liquid 40 is stored in the reservoir 13. At this point, the hydrophobic substance 19 is contained in the first chamber 15. When the sample is rotated in this state by a centrifuge, the specimen liquid 40, which has a relatively high specific gravity, moves toward the outer diameter away from the center, and the hydrophobic substance 19, which has a relatively low specific gravity, moves toward the inner diameter, as shown in the right diagram of Figure 6 . As a result, the specimen liquid 40 is sent from the reservoir 13 to the first chamber 15.

[0053] As described above, the cuvette 10 of this embodiment is provided with a sealing part 19 (hydrophobic material 19 in this example) that prevents backflow of the specimen liquid 40 after the specimen liquid 40 is transferred from the reservoir 13 to the first chamber 15. With this configuration, when the first chamber 15 is heated for pre-treatment of the specimen liquid 40 while the specimen liquid 40 is contained in the first chamber 15, for example, when the first chamber 15 is heated to a high temperature of 90° C. or higher, it is possible to prevent the specimen liquid 40 from flowing back into the reservoir 13.

[0054] When the hydrophobic substance 19 is provided as the sealing portion as in this embodiment, after the sample liquid 40 is delivered to the first chamber 15, the sample liquid 40 and the hydrophobic substance 19 are switched, and the hydrophobic substance 19 is contained in the liquid reservoir 13. Because the hydrophobic substance 19 is contained in the liquid reservoir 13, it is possible to prevent the vaporized sample liquid 40 from flowing back into the liquid reservoir 13, compared to when the hydrophobic substance 19 is not provided and only air is present in the liquid reservoir 13. The backflow of the sample liquid 40 from the first chamber 15 to the liquid reservoir 13 can be prevented without providing a mechanical structure, and the sealing portion can be realized very simply and at low cost.

[0055] It is preferable that the hydrophobic substance 19 be provided in a volume sufficient to fill the reservoir 13. If the hydrophobic substance 19 fills the reservoir 13 after the sample liquid 40 is delivered to the first chamber 15, backflow of the sample liquid 40 from the first chamber 15 to the reservoir 13 can be more effectively suppressed.

[0056] When the hydrophobic substance 19 is a low-polarity solvent, it does not mix with the sample liquid 40, and therefore the sample liquid 40 can be prevented from flowing back into the liquid reservoir 13.

[0057] When the hydrophobic substance 19 is a thermoplastic substance, the hydrophobic substance 19 is liquefied, and then the specimen liquid 40 is sent to the liquid reservoir 13, where it is cooled and solidified. This effectively prevents the specimen liquid 40 from flowing back into the liquid reservoir 13.

[0058] In the above embodiment, the hydrophobic substance 19 is provided in the liquid reservoir 13 or the first chamber 15 before the sample liquid 40 is introduced. However, the portion that provides the hydrophobic substance 19 is not limited to these. As shown in FIG. 7 , a modified flow path structure 11A may include a hydrophobic substance storage section 17 that is provided closer to the rotation axis of the centrifuge than the liquid reservoir 13, and the hydrophobic substance 19 may be stored in the hydrophobic substance storage section 17.

[0059] In the test container equipped with the modified flow path structure 11A, when the specimen liquid 40 is introduced into the inlet 12 and contained in the reservoir 13, the hydrophobic substance 19 is contained in the hydrophobic substance containing portion 17. By rotating the container in this state using a centrifuge, the specimen liquid 40 is sent from the reservoir 13 to the first chamber 15, and the hydrophobic substance 19 is sent to the reservoir 13 side, as shown in the right diagram of Fig. 7 .

[0060] In this way, even when a hydrophobic substance 19 is provided in the hydrophobic substance storage section 17, which is located closer to the rotation axis of the centrifuge than the liquid reservoir section 13, the hydrophobic substance 19 is stored in the liquid reservoir section 13 after the sample liquid 40 is delivered, so that backflow of the sample liquid 40 into the liquid reservoir section 13 can be suppressed.

[0061] In flow path structure 11 (or flow path structure 11A), when the volume of liquid reservoir 13 is V1, the volume of first chamber 15 is V2, the volume of liquid storage portion 24 is V3, and the volume of second chamber 16 is V4, it is preferable that the relationship be V1 ≥ V2 ≥ V3 ≥ V4. In other words, it is preferable that the volumes of adjacent chambers are equal, or that among multiple chambers arranged from the inner diameter side to the outer diameter side of the centrifugal center, the chambers arranged closer to the outer diameter have smaller volumes.

[0062] As an example, the volume V1 of the liquid reservoir 13 is greater than 100 μL to 200 μL, the volume V2 of the first chamber 15 is 100 μL, the volume V3 of the liquid storage section 24 is 50 μL, and the volume V4 of the second chamber 16 is 30 μL.

[0063] If the volume V1 of the liquid reservoir 13 is larger than that of the first chamber 15, then the liquid reservoir 13 can store more sample liquid 40 than the volume V2 of the first chamber 15. If the relationship between the volumes of the multiple chambers arranged from the central inner diameter side to the outer diameter side of the centrifuge satisfies V1 ≥ V2 ≥ V3 ≥ V4, then when the sample liquid 40 is delivered from the chamber arranged on the central inner diameter side to the chamber arranged on the outer diameter side of the centrifuge, the chamber on the outer diameter side (i.e., downstream) can be filled with sample liquid 40. This prevents a shortage of sample liquid 40 in the second chamber 16, which is ultimately used for detection.

[0064] As shown in Fig. 8, the test container 10 may include a reagent 32 in the flow path structure 11 to react with the sample liquid 40. For example, when the test container 10 is used for nucleic acid detection, the reagent 32 is a reagent containing an amplification reagent for amplifying a specific nucleic acid sequence and a fluorescent probe for determining the nucleic acid sequence. The reagent 32 may be provided in any of the liquid reservoir 13, the first chamber 15, the liquid storage section 24, and the second chamber 16. The reagent 32 is preferably provided in the first chamber 15 or the liquid storage section 24, and particularly preferably in the liquid storage section 24.

[0065] A stirring rod for promoting mixing of the sample liquid 40 and the reagent 32 may be contained in the chamber containing the reagent 32 or in a chamber downstream of the chamber.

[0066] The cuvette 10 of the above embodiment is provided with one flow path structure 11 from the inlet 12 to the second chamber 16. As a modified cuvette 50, as shown in Fig. 9, one cuvette 50 may be provided with a plurality of flow path structures 11. The cuvette 50 shown in Fig. 9 includes a disk-shaped main body member and a bottom member, and includes four flow path structures 11 arranged radially around the rotation axis A. Of course, the number of flow path structures 11 provided in one cuvette 50 is not limited to four.

[0067] Furthermore, as in a modified cuvette 52 shown in FIG. 10 , the flow path structure 11B may include a vent flow path 35. In this example, the vent flow path 35 is a flow path connecting the first chamber 15 and the liquid reservoir 13. The vent flow path 35 is a flow path through which gas passes. The inclusion of the vent flow path 35 facilitates the transfer of the sample liquid 40 from the liquid reservoir 13 to the first chamber 15, enabling efficient liquid transfer. On the other hand, when the vent flow path 35 is included, if the first chamber 15 is heated to a high temperature for pretreatment after the sample liquid 40 has been transferred to the first chamber 15, the vaporized sample liquid 40 may return to the liquid reservoir 13 through the vent flow path 35. However, in this embodiment, a hydrophobic substance is contained in the liquid reservoir 13, thereby preventing backflow of the sample liquid 40 from the vent flow path 35.

[0068] 11 shows the states before and after liquid transfer in the flow path structure 11B of the cuvette 52. As shown in Fig. 11, after the sample liquid 40 is transferred to the first chamber 15, the hydrophobic substance 19 fills the liquid reservoir 13, and some of it remains in the first chamber 15. If the hydrophobic substance 19 is configured to block the inlet of the vent flow path 35 of the first chamber 15, backflow of the sample liquid 40 into the vent flow path 35 can be more effectively prevented.

[0069] However, considering that the vent flow path 35 may not be completely blocked, it is more preferable not to provide a vent flow path like the cuvette 10 .

[0070] Alternatively, multiple second chambers 16a-16c may be provided, as in the modified test container 53 shown in FIG. 12 . Providing multiple second chambers 16a-16c allows for the same test to be performed on a single specimen in multiple second chambers 16a-16c, thereby improving test accuracy. Providing multiple second chambers 16a-16c also allows for parallel testing of multiple test substances on a single specimen. For example, the first second chamber 16a may be used to test for influenza, the second second chamber 16b for COVID-19, and the third second chamber 16c for respiratory syncytial virus (RS) virus. While the example shown in FIG. 12 features three second chambers 16a-16c, there is no limit to the number of second chambers. However, when multiple second chambers 16a-16c are provided, it is desirable for the total volume of all second chambers 16a-16c to be smaller than the volume of the liquid storage portion 24 of the valve 20.

[0071] In the above embodiment, the sealing portion is provided with a hydrophobic material 19, but the sealing portion may be any portion that functions to prevent backflow of the specimen liquid 40 after the specimen liquid 40 is transferred from the liquid reservoir 13 to the first chamber 15, and may be provided with a structure such as a check valve between the liquid reservoir 13 and the first chamber 15.

[0072] Fig. 13 is a schematic cross-sectional view of a modified cuvette 54. Fig. 13 also shows the states of the flow path structure 11 before and after the sample liquid 40 is sent from the liquid reservoir 13 to the first chamber 15.

[0073] In the cuvette 54, a flow path 29a connecting the liquid reservoir 13 and the first chamber 15 is formed in a tapered shape with a diameter gradually increasing from the liquid reservoir 13 side toward the first chamber 15 side. A sphere 29b is provided within the flow path 29a. The diameter of the sphere 29b is larger than the diameter of the flow path 29a at the side closest to the liquid reservoir 13. A stopper 29c is provided at a portion of the flow path 29a on the first chamber 15 side to limit the movement of the sphere 29b toward the first chamber 15. In this example, the flow path 29a, the sphere 29b, and the stopper 29c form a sealing part 29 that prevents backflow of the sample liquid 40 after the sample liquid 40 has been sent from the liquid reservoir 13 to the first chamber 15.

[0074] Before the liquid is delivered to the test container 54, the sphere 29b is movably disposed within the flow path 29a. In the upper diagram of FIG. 13 , the sample liquid 40 is introduced through the inlet 12 and stored in the liquid reservoir 13. As in the previously described embodiment, the test container 54 is rotated by a centrifuge with the space between the first chamber 15 and the liquid storage section 24 closed. As a result, as shown in the lower diagram of FIG. 13 , the sample liquid 40 is delivered to the first chamber 15 by centrifugal force. As the first chamber 15 is filled with the sample liquid 40, the sphere 29b is gradually pushed toward the liquid reservoir 13 by the sample liquid 40 and moves to a portion having a diameter equivalent to that of the sphere 29b. As a result, the sphere 29b acts as a check valve that blocks the flow path 29a.

[0075] According to this configuration, when first chamber 15 is heated for pre-processing of specimen liquid 40 while it is contained therein, for example, when first chamber 15 is heated to a high temperature of 90° C. or higher, specimen liquid 40 vaporizes and the pressure inside first chamber 15 increases, so that sphere 29 b is pressed further toward reservoir 13, sealing flow path 29 a. Therefore, it is possible to prevent specimen liquid 40 from flowing back into reservoir 13.

[0076] In addition, the sealing portion may be configured such that the bottom member 10B that forms the bottom surface of the flow path 18c connecting the liquid reservoir 13 and the first chamber 15 is made of a film, and the flow path 18c can be blocked by pressing the bottom member 10B from the outside.

[0077] The flow path structures 11, 11A, 11B in each of the cuvettes 10, 50, 52, 54 described above may further include a deformable chamber communicating with the second chamber 16. By deforming the deformable chamber by, for example, pressing it from the outside, it is possible to pressurize the deformable chamber and the second chamber 16 communicating with it. By pressurizing the inside of the second chamber 16, it is possible to improve the heating and cooling properties of the liquid contained in the second chamber 16. Furthermore, by pressurizing the inside of the second chamber 16, it is possible to suppress evaporation of the liquid inside when heated.

[0078] "Testing Device" A testing device 100 in which the test container 10 is loaded and which performs testing will be described with reference to Fig. 14. Fig. 14 is a diagram showing a schematic configuration of the testing device 100. The testing device 100 is, for example, a nucleic acid testing device.

[0079] The inspection device 100 includes a centrifuge 110 , a first heating unit 120 , a second heating unit 122 , a valve control unit 124 , and a detection unit 126 .

[0080] The centrifuge 110 includes a base 112 equipped with a rotation mechanism such as a motor, a disk-shaped support 111 constituting a turntable having a loading section 111A (see FIG. 5) for loading the cuvette 10, and a rotary coupler 114 connected to the axis of the rotation mechanism and rotatably supporting the disk-shaped support 111. By driving the rotation mechanism, the disk-shaped support 111 is rotated around the axis A. The centrifugal force accompanying this rotation realizes liquid transfer in the flow path structure within the cuvette 10. In the example shown in FIG. 13, the disk-shaped support 111 has a presser 111B for pressing the cuvette 10 set in the loading section 111A.

[0081] The first heating unit 120 is provided so as to be movable between a position in contact with the bottom surface of the first chamber 15 of the cuvette 10 set on the disk-shaped support 111 and a position spaced apart from it. The first heating unit 120 is disposed at a position spaced apart from the bottom surface of the first chamber 15 while the disk-shaped support 111 is rotating, and is disposed at a position in contact with the bottom surface of the first chamber 15 when the disk-shaped support 111 is stationary. The first heating unit 120 heats the liquid contained in the first chamber 15. Here, the liquid contained in the first chamber 15 is a sample liquid 40. The first heating unit 120 heats the sample liquid 40 to a high temperature, for example, 90° C. or higher, for pretreatment.

[0082] The second heating unit 122 is provided so as to be movable between a position in contact with the bottom surface of the second chamber 16 of the cuvette 10 set on the disk-shaped support 111 and a position spaced apart from it. The second heating unit 122 is disposed at a position spaced apart from the bottom surface of the second chamber 16 while the disk-shaped support 111 is rotating, and is disposed at a position in contact with the bottom surface of the second chamber 16 when the disk-shaped support 111 is stationary. The second heating unit 122 heats the liquid contained in the second chamber 16. Here, the liquid contained in the second chamber 16 is, for example, a mixture of a sample liquid 40 and a reagent 32. The second heating unit 122 heats the mixture of the sample liquid 40 and the reagent 32 to promote nucleic acid amplification.

[0083] The second heating unit 122 is equipped with a Peltier element or the like, and is capable of temperature control, and performs temperature cycling in the nucleic acid amplification process. On the other hand, the first heating unit 120 does not require temperature cycling like the second heating unit 122, and is composed of, for example, a heater. The heating mechanisms used in the first heating unit 120 and the second heating unit 122 can be known heating mechanisms and are not particularly limited.

[0084] Valve control unit 124 includes a mechanism for controlling valve 20 to switch the communication state between first chamber 15, second chamber 16, and liquid storage unit 24 among a first state, a second state, and a third state. Valve control unit 124 includes, for example, a gripping unit that grips knob 25 a of rotating member 25, a rotation mechanism that rotates the gripping unit while knob 25 a is gripped by the gripping unit, and a processor that drives the rotation mechanism as necessary during the inspection processing steps.

[0085] The detection unit 126 detects whether or not the sample liquid 40 in the second chamber 16 contains a detection target. As shown in FIG. 17 , the detection unit 126 includes a light source 126a, a wavelength-selecting filter 126b, and a photodetector 126c. The detection unit 126 is disposed above the second chamber 16 of the cuvette 10. Detection by the detection unit 126 is also performed while the rotation of the centrifuge 110 is stopped. The light source 126a irradiates the second chamber 16 with excitation light L1 of a specific wavelength via the wavelength-selecting filter 126b. The photodetector 126c 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. If necessary, the detection unit 126 may also include a filter for adjusting the intensity or light amount, a lens for converging the excitation light L1, or an optical system for focusing the fluorescence L2 derived from the detection probe onto the photodetector 126c.

[0086] The light source 126a may be an LED or a laser. The wavelength selection filter 126b is a filter that transmits only light emitted from the light source 126a with a wavelength corresponding to the excitation wavelength of the probe. The photodetector 126c may be, for example, a photodiode or a photomultiplier tube.

[0087] The cuvette 10 can be used in an inspection device such as the inspection device 100 shown in Fig. 14, the inspection device 101 shown in Fig. 15, or the inspection device 102 shown in Fig. 16. In Figs. 15 and 16, the same components as those in the inspection device 100 shown in Fig. 14 are designated by the same reference numerals, and detailed description thereof will be omitted.

[0088] The inspection device 101 and the inspection device 102 are respectively equipped with centrifuges 110A and 110B having a structure different from that of the centrifuge 110 of the inspection device 100.

[0089] 15, the inspection device 101 is configured such that a centrifuge 110A rotates the disk-shaped support 111 with the surface of the support 111 tilted from the horizontal plane. The rotation axis A of the centrifuge 110A is tilted from the direction perpendicular to the horizontal plane.

[0090] 16, a centrifuge 110B includes an umbrella-shaped support 131 instead of the disk-shaped support 111. The umbrella-shaped support 131 includes a horizontal portion supported by a rotary coupler 114 and an inclined portion surrounding the horizontal portion, the outer periphery of which is positioned vertically downward relative to the horizontal portion, and the cuvette 10 is set on the inclined portion.

[0091] The cuvette 10 can be applied to any of the various types of centrifuges 110A, 110B, and 110C shown in FIGS.

[0092] In the above, the testing devices 100, 101, and 102 are described as nucleic acid testing devices used for nucleic acid testing involving nucleic acid amplification processing. In the testing devices 100, 101, and 102, the first heating unit 120, the second heating unit 122, and the detection unit 126 can be configured to be suitable for various tests, making them devices for various tests other than nucleic acid testing.

[0093] "Nucleic Acid Testing Method" A nucleic acid testing method according to one embodiment using the test container 10 of the above embodiment will be described with reference to Figures 17 and 18. Here, a test container 10 (see Figure 8) is used in which the first chamber 15 is provided with a hydrophobic material 19 as a sealing part and the liquid storage part 24 is provided with a reagent 32 for nucleic acid testing.

[0094] First, as a preliminary step, as shown in Figure 17, a specimen 40a collected from a living body using a collection tool 60 such as a swab is immersed in an extraction solution 62 containing a surfactant and ProK (proteinase K) to extract nucleic acids and remove impurities. Hereinafter, the liquid containing the specimen 40a mixed with the extraction solution 62 and having undergone nucleic acid extraction in the extraction solution 62 will be referred to as specimen liquid 40.

[0095] The specimen 40a is collected, for example, from the nasal cavity, pharynx, or oral cavity of the subject, or from an affected area, using a collection tool. Alternatively, the specimen may be a body fluid such as a nasal cavity, pharynx, or oral cavity wash, saliva, urine, or blood. Any known nucleic acid extraction method can be used without particular limitation. Examples include methods using surfactants or chaotropic substances, and methods that apply physical shearing, such as ultrasonic waves or a bead mill.

[0096] In the initial state of the test container 10 before the specimen liquid 40 is poured into it, the valve 20 is in the second state or the third state in which at least the first chamber 15 and the liquid storage portion 24 are not in communication. In this embodiment, the third state is used in which neither the first chamber 15 nor the second chamber 16 is in communication with the liquid storage portion 24. In the test, first, with the valve 20 in the initial state as described above, a drip cap 66 equipped with a filter 64 for removing coarse contaminants is attached to a container 63 containing specimen liquid 40 containing specimen 40a whose nucleic acid has been extracted in an extract solution, and the specimen liquid 40 filtered through the filter 64 is poured into the inlet 12 of the test container 10 (step ST1). The specimen liquid 40 is stored in the liquid reservoir 13.

[0097] After the sample liquid 40 has been poured into the test container 10, the inlet 12 is closed with the lid 12A to seal the internal space of the test container 10 (step ST2). The test container 10 is loaded into the test device 100 with the sample liquid 40 poured into it and the inlet 12 closed with the lid 12A. The following process is performed within the test device 100.

[0098] The test container 10 is rotated by the centrifuge 110 of the testing device 100, and the specimen liquid 40 is sent from the liquid reservoir 13 to the first chamber 15 (step ST3). As the specimen liquid 40 is sent to the first chamber 15, the hydrophobic substance 19 contained in the first chamber 15 is moved to the liquid reservoir 13.

[0099] With the hydrophobic material 19 preventing backflow of the specimen liquid 40 from the first chamber 15 to the liquid reservoir 13, the first chamber 15 is heated by the first heating unit 120 to perform pretreatment by heating the specimen liquid 40 (step ST4). This heating treatment deactivates ProK. The heating temperature is preferably 80°C or higher, more preferably 85°C or higher, and even more preferably 90°C or higher. The upper limit temperature is preferably 100°C or lower, more preferably 95°C or lower, from the viewpoint of evaporation of the specimen liquid.

[0100] After the pre-processing, the valve control unit 124 (see FIG. 14 ) switches the valve 20 to the first state (step ST5), i.e., the first chamber 15 and the liquid storage unit 24 are in communication with each other, and the liquid storage unit 24 and the second chamber 16 are not in communication with each other.

[0101] Next, the test container 10 is rotated by the centrifuge 110 of the testing device 100, and the specimen liquid 40 is sent from the first chamber 15 to the liquid storage section 24 of the valve 20 (step ST6). The liquid storage section 24 stores a reagent 32, and the sent specimen liquid 40 and the reagent 32 are mixed in the liquid storage section 24.

[0102] After the sample liquid 40 and the reagent 32 are mixed, the valve 20 is switched to the second state by the valve control unit 124 (see FIG. 14 ) (step ST7), i.e., the second chamber 16 and the liquid storage unit 24 are in communication with each other, but the liquid storage unit 24 and the first chamber 15 are not in communication with each other.

[0103] Then, the cuvette 10 is rotated by the centrifuge 110 of the testing device 100, and the mixture 42 of the sample liquid 40 and the reagent 32 is sent from the liquid storage portion 24 of the valve 20 to the second chamber 16 (step ST8).

[0104] After the mixed liquid 42 has been sent to the second chamber 16, the valve control unit 124 (see FIG. 14 ) switches the valve 20 to the third state (step ST9). Here, at least the second chamber 16 is not in communication with the liquid storage unit 24. In this example, neither the first chamber 15 nor the second chamber 16 is in communication with the liquid storage unit 24, but the first state may also be used as long as the second chamber 16 is not in communication with the liquid storage unit 24.

[0105] Here, in a state where the second chamber 16 and the liquid storage portion 24 are not in communication with each other, the nucleic acid amplification process and the fluorescence detection are carried out on the mixed liquid 42 (step ST10).

[0106] The nucleic acid amplification process involves heating the mixed solution 42 in the second chamber 16 using the second heating unit 122 to amplify a specific nucleic acid sequence. The nucleic acid amplification process is not limited to this process, but may be, for example, RT-PCR or PCR. When PCR is used, the process involves repeatedly dissociating double-stranded DNA into single-stranded DNA at high temperature (thermal denaturation process), then lowering the temperature to allow primers to bind to the single-stranded DNA (annealing process), and synthesizing new double-stranded DNA using polymerase as a template (extension process). An example of a temperature cycle for the thermal denaturation process, annealing process, and extension process is one in which one cycle consists of 94°C for 1 minute, 50-60°C for 1 minute, and 72°C for 1-5 minutes, repeated 20-50 times. The thermal denaturation process and annealing process may also be performed at a single temperature. An example of such a temperature cycle is one in which one cycle consists of 1 minute at 94° C. and 1 minute at 60° C., and this cycle is repeated 20 to 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 appropriate depending on the performance of the polymerase and primers.

[0107] Fluorescence detection is performed by a detection unit 126 disposed above the second chamber 16. Excitation light L1 of a specific wavelength is irradiated from a light source 126a through a wavelength-selective filter 126b into the second chamber 16. Fluorescence L2 emitted from the fluorescent probe upon excitation by the excitation light L1 is detected by a photodetector 126c. For example, if 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.

[0108] If a specific nucleic acid sequence is present in the sample solution 40, the nucleic acid sequence 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 sequence. On the other hand, if the specific nucleic acid sequence is not present in the sample solution 40, 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 sequence is present.

[0109] According to the testing method using the cuvette 10, the specimen liquid 40 sent from the liquid reservoir 13 to the first chamber 15 is pre-treated in the first chamber 15 by high-temperature heating, but the cuvette 10 is provided with a sealing part (hydrophobic material 19 in this example) that prevents the specimen liquid 40 from flowing back into the liquid reservoir 13, thereby suppressing backflow of the specimen liquid 40. This makes it possible to suppress a decrease in the amount of specimen liquid 40 that is ultimately processed in the second chamber 16, thereby improving testing accuracy.

[0110] In the testing method using the testing device 100, the presence or absence of nucleic acid is determined by a fluorescence method using a fluorescent probe, but the method for detecting the presence or absence of nucleic acid is not limited to the fluorescence method, and other detection methods such as nucleic acid chromatography, light scattering, sequencing, and electrochemical methods may also be used. These can be achieved by appropriately modifying the detection unit, and the test container may be provided with a reagent containing a probe appropriate for each detection method, instead of the fluorescent probe.

[0111] In the above, nucleic acid testing has been cited as an example of testing using the test container 10, but the test container 10 can also be applied to other genetic testing or microorganism testing in addition to nucleic acid testing.

[0112] As an example of a microorganism test, an example in which the test container 10 is used for a β-glucan test will be described with reference to Figures 19 and 20. Here, a test container 10 (see Figure 8) is used in which the first chamber 15 is provided with a hydrophobic material 19 as a sealing portion, and the liquid storage portion 24 is provided with an LAL (Limulus Amebocyte Lysate) reagent as the reagent 32. In this example, a test device with a configuration similar to that of the test device 100 can be used.

[0113] The specimen and the pretreatment liquid are mixed to prepare the specimen liquid 40. A drip cap 66 equipped with a filter 64 is attached to a container 63 containing the specimen liquid 40, and the specimen liquid 40 filtered through the filter 64 is poured into the inlet of the test container 10 (step ST11). The specimen liquid 40 is stored in the liquid reservoir 13.

[0114] After the sample liquid 40 has been poured into the test container 10, the inlet 12 is closed with the lid 12A to seal the internal space of the test container 10 (step ST12). The test container 10 is loaded into the testing device 100 with the sample liquid 40 poured into it and the inlet 12 closed with the lid 12A. The following process is performed within the testing device 100.

[0115] The test container 10 is rotated by the centrifuge 110 of the testing device 100, and the specimen liquid 40 is sent from the liquid reservoir 13 to the first chamber 15 (step ST13). As the specimen liquid 40 is sent to the first chamber 15, the hydrophobic substance 19 contained in the first chamber 15 is moved to the liquid reservoir 13.

[0116] With the hydrophobic substance 19 preventing the backflow of the specimen liquid 40 from the first chamber 15 to the liquid reservoir 13, the first heating unit 120 heats the first chamber 15 to raise the temperature of the specimen liquid 40 to 70°C for 10 minutes, and then cools it to 3°C (step ST14). The pretreatment performed here is a process of heating the specimen liquid 40 to stop the endotoxin reaction cascade.

[0117] After the pre-processing, the valve control unit 124 (see FIG. 14 ) switches the valve 20 to the first state (step ST15), i.e., the first chamber 15 and the liquid storage unit 24 are in communication with each other, and the liquid storage unit 24 and the second chamber 16 are not in communication with each other.

[0118] Next, the cuvette 10 is rotated by the centrifuge 110 of the testing device 100, and the specimen liquid 40 is sent from the first chamber 15 to the liquid storage section 24 of the valve 20 (step ST16). A reagent 32 is stored in the liquid storage section 24, and the sent specimen liquid 40 and the reagent 32 are mixed in the liquid storage section 24. The reagent 32 is a freeze-dried reagent, and is dissolved in the specimen liquid 40 and mixed.

[0119] After the sample liquid 40 and the reagent 32 are mixed, the valve 20 is switched to the second state by the valve control unit 124 (see FIG. 14 ) (step ST17), i.e., the second chamber 16 and the liquid storage unit 24 are in communication with each other, but the liquid storage unit 24 and the first chamber 15 are not in communication with each other.

[0120] Then, the cuvette 10 is rotated by the centrifuge 110 of the testing device 100, and the mixture 42 of the sample liquid 40 and the reagent 32 is sent from the liquid storage portion 24 of the valve 20 to the second chamber 16 (step ST18).

[0121] After the mixed liquid 42 has been sent to the second chamber 16, the valve control unit 124 (see FIG. 14 ) switches the valve 20 to the third state (step ST19). Here, at least the second chamber 16 is not in communication with the liquid storage unit 24. In this example, the first chamber 15 and the second chamber 16 are not in communication with the liquid storage unit 24, but the first state may also be used as long as the second chamber 16 is not in communication with the liquid storage unit 24.

[0122] Here, with the second chamber 16 and the liquid storage section 24 not in communication with each other, the LAL reaction in the mixed liquid 42 is detected by colorimetry (step ST20). The colorimetric detection is performed by a detection section 126 disposed above the second chamber 16. Irradiation light L of a specific wavelength is irradiated into the second chamber 16 from a light source 126a via a wavelength selection filter 126b. The LAL reaction is detected by detecting the absorbance (or transmittance) of the irradiation light L using a photodetector 126c.

[0123] In this way, in the β-glucan test, the test container 10 can be used in the same way as in the nucleic acid test, and the same effects can be obtained.

[0124] The disclosure of Japanese Patent Application No. 2022-057525, filed on March 30, 2022, 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 test container having an inlet for introducing a specimen liquid, a first chamber for accommodating the specimen liquid delivered from the inlet, and a second chamber for accommodating the specimen liquid delivered from the first chamber, the test container being attached to a centrifuge, and the specimen liquid being delivered from the first chamber to the second chamber in sequence by centrifugal force generated by rotation of the centrifuge, A removable lid portion for covering the insertion port; a liquid reservoir that stores the sample liquid introduced through the introduction port before the first chamber; a valve disposed between the first chamber and the second chamber, the valve having a liquid storage portion for temporarily storing the sample liquid transferred from the first chamber to the second chamber, the valve being switchable between a first state in which the first chamber communicates with the liquid storage portion and the second chamber does not communicate with the liquid storage portion, a second state in which the second chamber communicates with the liquid storage portion and the first chamber does not communicate with the liquid storage portion, and a third state in which neither the first chamber nor the second chamber communicates with the liquid storage portion; a sealing portion that prevents backflow of the specimen liquid after the specimen liquid is delivered from the liquid reservoir portion to the first chamber.

2. The cuvette according to claim 1 , wherein the sealing portion is made of a hydrophobic material having a specific gravity lower than that of the sample liquid.

3. 3. The inspection vessel according to claim 2, wherein the hydrophobic material is a low polarity solvent or a thermoplastic material.

4. The cuvette according to claim 2 or 3, wherein the hydrophobic material is provided in the liquid reservoir or the first chamber.

5. 4. The cuvette according to claim 2, further comprising a hydrophobic material containing section for containing the hydrophobic material, the hydrophobic material containing section being provided closer to a rotation shaft of the centrifuge than the liquid reservoir section.

6. The test container according to any one of claims 1 to 3, wherein when the volume of the liquid reservoir is V1, the volume of the first chamber is V2, the volume of the liquid storage portion is V3, and the volume of the second chamber is V4, the relationship is V1 ≧ V2 ≧ V3 ≧ V4.

7. The cuvette according to claim 1 , wherein a reagent to be reacted with the sample liquid is contained in the first chamber, the liquid containing portion, or the second chamber.

8. 8. The test container according to claim 7, wherein the reagent comprises an amplification reagent for amplifying a specific nucleic acid sequence and a probe for determining the nucleic acid sequence.

9. The test vessel according to claim 8 , wherein the probe is a fluorescent probe.

10. 10. A nucleic acid testing method using the test container according to claim 9, wherein the reagent is contained in the liquid containing section, the valve is initially in the second state or the third state, and while the valve is in the initial state, the sample liquid is poured into the inlet of the test container to accommodate the sample liquid in the liquid reservoir; The insertion port is sealed by the lid portion, The sample liquid is transferred from the liquid reservoir to the first chamber by centrifugal force; heating the first chamber to heat the specimen liquid while preventing backflow of the specimen liquid from the first chamber to the liquid reservoir by the sealing portion; Switching the valve to the first state; The specimen liquid is sent from the first chamber to the liquid storage portion of the valve by centrifugal force, and the specimen liquid and the reagent are mixed in the liquid storage portion; Switching the valve to the second state; A mixture of the sample liquid and the reagent is transferred from the liquid storage portion to the second chamber by centrifugal force; Switching the valve to the third state or the first state; In the second chamber, the temperature of the mixture is adjusted to amplify the specific nucleic acid sequence in the mixture; Irradiating the mixture with excitation light and detecting the fluorescence emitted from the fluorescent probe; A nucleic acid testing method for determining the presence or absence or concentration of the specific nucleic acid sequence.