Detector, detection device, and detection method

Abstract

The detector (1, 1A, 1B, 1C, 1D) has a sensor housing portion (3), a sample loading portion (2), a moving flow path (7), and a change substance supply portion (11, 11A). The sensor housing portion houses a sensor for measuring ions. The sample loading portion is for loading a sample. The moving flow path connects the sample loading portion and the sensor housing portion, and supplies the sample to the sensor housing portion. The change substance supply portion supplies an ion concentration change substance (C) that changes the ion concentration of the sample to the sensor housing portion.

Description

Cross-reference of related applications

[0001] This international application claims priority based on Japanese Patent Application No. 2024-033954, filed with the Japan Patent Office on March 6, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This disclosure relates to detectors, detection devices, and detection methods. Background Technology

[0003] Patent Document 1 discloses a technique for highly sensitively detecting changes in the ion concentration of a sample in a solution by using a low-concentration (i.e., low-buffering-capacity) buffer solution or by using a solution without buffering material.

[0004] Existing technical documents Patent documents Patent Document 1: International Publication No. 2014 / 031783 Summary of the Invention

[0005] However, the inventors’ detailed research revealed the following problem in the technology of Patent Document 1: when the change in ion concentration from the sample is small, it is sometimes impossible to detect the change in ion concentration sufficiently.

[0006] According to one aspect of this disclosure, a technique is preferably provided to improve the detection sensitivity of changes in ion concentration in a sample.

[0007] One embodiment of this disclosure is a detector comprising a sensor housing, a sample introduction unit, a flow path, and a variable substance supply unit. The sensor housing houses a sensor for measuring ions. The sample introduction unit introduces a sample. The flow path connects the sample introduction unit and the sensor housing, supplying the sample to the sensor housing. The variable substance supply unit supplies an ion concentration-changing substance that alters the ion concentration of the sample to the sensor housing.

[0008] This structure can improve the detection sensitivity of changes in ion concentration in the sample.

[0009] One aspect of this disclosure is a detection apparatus comprising a detector and a control unit. The detector comprises a sensor housing, a sample introduction unit, a flow path, and a variable substance supply unit. The sensor housing houses a sensor for measuring ions. The sample introduction unit introduces a sample. The flow path connects the sample introduction unit and the sensor housing, supplying the sample to the sensor housing. The variable substance supply unit supplies an ion concentration changing substance, which changes the ion concentration of the sample, to the sensor housing. The control unit comprises a sample supply unit, an ion concentration changing substance addition unit, and a measuring unit. The sample supply unit supplies the introduced sample to the sensor housing. The ion concentration changing substance addition unit supplies the ion concentration changing substance housed in the variable substance housing to the sensor housing. The measuring unit measures the ion concentration in the sample.

[0010] This structure can improve the detection sensitivity of changes in ion concentration in the sample.

[0011] One aspect of this disclosure is a detection apparatus comprising a detector and a control unit. The detector comprises a sensor housing, a sample input unit, a flow path, a variable substance supply unit, and a mixing unit. The sensor housing houses a sensor for measuring ions. The sample input unit supplies a sample. The flow path connects the sample input unit and the sensor housing, supplying the sample to the sensor housing. The variable substance supply unit supplies an ion concentration changing substance that changes the ion concentration of the sample to the sensor housing. The mixing unit mixes beads, a linker-enzyme fusion compound, and a substrate with the sample before the sample from the sample input unit reaches the sensor housing. The control unit comprises a bead supply unit, a linker-enzyme fusion compound supply unit, a substrate supply unit, a reaction unit, an ion concentration changing substance addition unit, and a measurement unit. The bead supply unit supplies beads to the input sample, generating a first conjugate formed by the sample and the beads. The linker-enzyme fusion compound supply unit supplies the linker-enzyme fusion compound to the first conjugate, generating a second conjugate formed by the first conjugate and the linker-enzyme fusion compound. The substrate supply unit supplies the substrate to the second conjugate. The reaction unit supplies the second conjugate and substrate to the sensor housing, causing the enzyme in the second conjugate to produce a chemical reaction that consumes the substrate. The ion concentration changing substance addition unit supplies the ion concentration changing substance contained in the changing substance housing to the sensor housing. The measurement unit measures the ion concentration in the sample.

[0012] This structure can improve the detection sensitivity of changes in ion concentration in the sample.

[0013] One aspect of this disclosure is a detection method using a detector. The detector includes a sensor housing, a sample introduction unit, a flow path, and a variable substance supply unit. The sensor housing houses a sensor for measuring ions. The sample introduction unit provides for sample introduction. The flow path connects the sample introduction unit and the sensor housing, supplying the sample to the sensor housing. The variable substance supply unit supplies an ion concentration changing substance, which changes the ion concentration of the sample, to the sensor housing. The detection method includes the following steps: introducing a sample into the sample introduction unit; supplying the introduced sample to the sensor housing; supplying the sensor housing with an ion concentration changing substance housed in the variable substance housing; and measuring the ion concentration in the sample.

[0014] By implementing such a method, the detection sensitivity of changes in ion concentration in the sample can be improved.

[0015] One aspect of this disclosure is a detection method using a detector. The detector includes a sensor housing, a sample introduction unit, a flow path, a variable substance supply unit, and a mixing unit. The sensor housing houses a sensor for measuring ions. The sample introduction unit provides the sample. The flow path connects the sample introduction unit and the sensor housing, supplying the sample to the sensor housing. The variable substance supply unit supplies an ion concentration-changing substance that changes the ion concentration of the sample to the sensor housing. The mixing unit mixes beads, a linker-enzyme fusion compound, and a substrate with the sample before the sample from the sample introduction unit reaches the sensor housing. The detection method comprises the following steps: inserting a sample into a sample input section; supplying beads to the inserted sample to prepare a first conjugate formed by combining the sample and the beads; supplying a linker-enzyme fusion compound to the first conjugate to prepare a second conjugate formed by combining the first conjugate and the linker-enzyme fusion compound; supplying a substrate to the second conjugate; supplying the second conjugate and the substrate to a sensor receiving section, causing the enzyme contained in the second conjugate to undergo a chemical reaction that consumes the substrate; supplying a substance containing a change in ion concentration contained in a substance receiving section to the sensor receiving section; and measuring the ion concentration in the sample.

[0016] By implementing such a method, the detection sensitivity of changes in ion concentration in the sample can be improved. Attached Figure Description

[0017] Figure 1A This is a schematic diagram of the second composite disclosed herein. Figure 1B This is another schematic diagram of the second assembly disclosed herein.

[0018] Figure 2 These represent experimental results obtained by using acid as a substance with varying ion concentration to confirm the effect of the substance with varying ion concentration on the enzyme-substrate reaction disclosed herein.

[0019] Figure 3These represent experimental results obtained by using air (i.e., CO2) as a substance with varying ion concentration to confirm the effect of the substance with varying ion concentration on the enzyme-substrate reaction of this disclosure.

[0020] Figure 4 This represents experimental results obtained by confirming the change in sensor output value caused by the addition of an ion concentration-changing substance corresponding to the enzyme-substrate reaction of this disclosure when using acid as the ion concentration-changing substance.

[0021] Figure 5 This is a block diagram showing the structure of the detector in the first embodiment.

[0022] Figure 6 This is a schematic diagram showing the structure of the detection device in the first embodiment.

[0023] Figure 7 This is a flowchart of the processing of the detection device in the first embodiment.

[0024] Figure 8 This is a block diagram illustrating the structure of the detector in the second embodiment.

[0025] Figure 9 This is a block diagram showing the structure of the detector in the third embodiment.

[0026] Figure 10 This is a block diagram showing the structure of the detector in the fourth embodiment.

[0027] Figure 11 This is a block diagram showing the structure of the detector in the fifth embodiment.

[0028] Figure 12 This is a schematic diagram showing the modified material supply unit in other embodiments.

[0029] Figure 13A , Figure 13B , Figure 13C It is a schematic diagram showing the changes in the mixing section. Detailed Implementation

[0030] Hereinafter, embodiments of the present disclosure will be described with reference to the accompanying drawings.

[0031] [1. Detection Method] A summary of the detection methods used in this disclosure is provided.

[0032] The detection method disclosed herein can be used to detect changes in the ion concentration of a sample. Furthermore, the detection method disclosed herein can be used to detect objects contained within the sample. The sample can be a liquid or a solid. In the case of a solid sample, the detection method disclosed herein can employ a mixture of the solid and any liquid. As an example, the object may contain a virus (e.g., SARS-CoV-2).

[0033] The main principle of the detection method disclosed herein is to detect changes in ion concentration caused by an enzyme-substrate reaction. Therefore, to perform the detection method of this disclosure, at least an enzyme and a substrate are required. That is, it can be configured to add an enzyme and a substrate to a sample and detect changes in ion concentration resulting from the chemical reaction in which the enzyme consumes the substrate. However, for a more suitable implementation of the detection method of this disclosure, the detection method shown below can be used.

[0034] First, the beads are bound to the sample to prepare a first conjugate. The beads can be substances that specifically bind to the target in the sample. For example, the beads can bind to antibodies that specifically recognize the surface of a virus (e.g., the spike protein in SARS-CoV-2). Furthermore, the antibody can specifically bind to viral proteins in the sample. Additionally, various types of beads can be used as long as they bind to the target in the sample. As an example, the beads can be magnetic beads.

[0035] Next, the adapter-enzyme fusion body, formed by the fusion of the adapter and the enzyme, is combined with the first conjugate to prepare the second conjugate. Figure 1A , Figure 1B A schematic diagram showing the second composite. Figure 1A In this process, for the viral surface protein VP in the sample, the antibody IB, which specifically binds to VP, binds to the bead MB, which acts as a magnetic bead. That is, Figure 1A In this mixture, bead MB, antibody IB, and virus constitute the first conjugate A. Furthermore, the linker LK, fused with enzyme EZ, binds to protein VP on the surface of the virus, which belongs to the first conjugate A, thereby preparing the second conjugate B. Alternatively, the linker-enzyme fusion can also exist as a solution. Additionally, the linker LK can be a substance that specifically binds to a target in the sample. Examples of linker LK include DNA aptamers, RNA aptamers, peptide aptamers, antibodies, antibody fragments, modified antibodies, peptides, and nucleic acids. Examples of enzyme EZ include alkaline phosphatase (hereinafter referred to as ALP).

[0036] In addition, such as Figure 1B As shown, Figure 1AThe antibody IB in the second conjugate B shown can be replaced with the linker LK. That is, in this disclosure, the bead MB can bind to the linker LK, which can bind to the protein VP on the viral surface. Furthermore, the second conjugate B can be prepared by further binding the linker LK fused with enzyme EZ to the protein VP on the viral surface.

[0037] Next, after removing excess solution except for the second conjugate B, substrate solution is added to the second conjugate B. Then, an enzyme present in the second conjugate B undergoes a chemical reaction that consumes the substrate. Through this chemical reaction, the ion concentration in the reaction solution changes. Examples of substrates include p-nitrophenyl phosphate.

[0038] The sensor is an ion sensor used to measure ions. The sensor can be a semiconductor sensor that measures the ion concentration (e.g., hydrogen ion concentration) in a sample. Specifically, as described above, the sensor can be configured to detect the hydrogen ion concentration, which changes due to a chemical reaction involving the consumption of the substrate, produced by an enzyme present in the second conjugate B containing the sample, as the ion concentration in the sample. Furthermore, it is preferable to perform sensor-based measurements after minimizing external factors that may affect the ion concentration of the sample. For example, sensor-based measurements can be performed in a nitrogen environment, which is inert to both the sample and the substrate.

[0039] Furthermore, a washing solution can be used in the stages of preparing the first conjugate A and the second conjugate B, and in the stage of adding the substrate solution to the second conjugate B. The washing solution is used to remove components that are not used as constituent elements of the first conjugate A and the second conjugate B. Examples of washing solutions include phosphate buffer.

[0040] Furthermore, a buffer solution can be used in the above-described detection method. A buffer solution is a solution used to stabilize the ion concentration. Various buffer solutions (e.g., acetate buffer, phosphate buffer, citrate buffer, borate buffer, and Tris buffer) can be used as long as they can stabilize the ion concentration. Additionally, a buffer solution with any buffer range suitable for the detection method of this disclosure can be used. The buffer solution can be mixed with the sample to form a mixture. Alternatively, the buffer solution can be mixed with the reaction solution obtained by adding a substrate solution to the second conjugate B. By using the buffer solution, various reactions can be carried out under more stable conditions.

[0041] For the reaction solution obtained by adding substrate solution to the second conjugate B, an ion concentration changing substance C is further added. Ion concentration changing substance C is a substance that changes the ion concentration of the sample by mixing with the sample. Specifically, it is a substance that acts in a direction that further increases the change in hydrogen ion concentration generated in the reaction solution by the chemical reaction of the enzyme and substrate described above. Thus, the hydrogen ion concentration in the reaction solution changes significantly further. Examples of ion concentration changing substances C include acids, bases, and air (i.e., CO2). Alternatively, for example, when a buffer solution is used in the above detection method, the addition of the ion concentration changing substance C can also be achieved by mixing the ion concentration changing substance C into the mixture of buffer solution and sample, thereby changing the ion concentration of the sample contained in the mixture to outside the buffer range. Alternatively, the addition of the ion concentration changing substance C can also be achieved by mixing the ion concentration changing substance C into the reaction solution obtained by adding substrate solution to the second conjugate B mixed with buffer solution, thereby changing the ion concentration of the sample contained in the reaction solution to outside the buffer range.

[0042] As described above, the detection method disclosed herein can also be used to determine whether a sample contains an object (e.g., a virus).

[0043] Specifically, the presence of an object contained in the sample can be detected by comparing the measured ion concentration in the reaction solution obtained by adding a substrate solution to the second conjugate B, and / or the measured ion concentration in the reaction solution after adding a substance C that changes the ion concentration, with reference data. The reference data are the measured ion concentrations of a reference sample without the object under the same conditions.

[0044] Regarding the detection method described above, the following experiment was conducted to verify the effect of changes in ion concentration of substance C.

[0045] [Experimental Example 1] An enzyme was added to the substrate to initiate the chemical reaction. p-Nitrophenyl phosphate was used as the substrate, and ALP was used as the enzyme. Enzyme concentrations were set at 0 pM, 0.01 pM, 0.1 pM, 1 pM, and 10 pM. For each case, the change in hydrogen ion concentration produced by the chemical reaction between the enzyme and the substrate was measured using a sensor. Figure 2 As shown, hydrogen ions produced by a low concentration of enzyme alone could not be detected as a change in hydrogen ion concentration. However, by adding acid to the reaction solution of the enzyme and substrate, the hydrogen ion concentration shifts to a steeper region of the pH titration curve, allowing even minute changes in hydrogen ion concentration to be amplified and detected. Furthermore, whenever the sensor signal stabilizes, further addition of acid to the reaction solution can further amplify and detect changes in hydrogen ion concentration.

[0046] [Experimental Example 2] An enzyme was added to the substrate to initiate the chemical reaction. p-Nitrophenyl phosphate was used as the substrate, and ALP was used as the enzyme. Enzyme concentrations were set to 0 pM, 0.1 pM, and 10 pM. For each case, the change in hydrogen ion concentration produced by the chemical reaction between the enzyme and the substrate was measured using a sensor. Figure 3 As shown, hydrogen ions produced solely by a low concentration of the enzyme could not be detected as a change in hydrogen ion concentration. However, by exposing the reaction solution of the enzyme and substrate to air, CO2 from the air dissolved in, reducing the ion concentration. Consequently, the hydrogen ion concentration shifted to a steeper region of the pH titration curve, amplifying and detecting even minute changes in hydrogen ion concentration.

[0047] [Experiment Example 3] An enzyme was added to the substrate to initiate the chemical reaction. p-Nitrophenyl phosphate was used as the substrate, and ALP was used as the enzyme. Enzyme concentrations were set at 0 pM, 0.01 pM, 0.1 pM, 1 pM, and 10 pM. For each case, acid was added to the reaction mixture of enzyme and substrate, and changes in hydrogen ion concentration were detected using a sensor. Acid was further added to the reaction mixture whenever the sensor signal stabilized.

[0048] like Figure 4 As shown, the sensor output value increases with the amount of acid added, resulting in an increase in the detected hydrogen ion concentration. Using samples with known enzyme concentrations as standard samples, standard titration curves representing the relationship between enzyme concentration and sensor output value were created by adding multiple amounts of acid to these samples.

[0049] Such a titration curve can be used when detecting objects contained in a sample according to the detection method of this disclosure. Specifically, the enzyme concentration in the sample can be quantitatively evaluated by fitting the sensor output value obtained when a specified amount of acid is added to the sample to the standard titration curve. Thus, the enzyme concentration in the sample, i.e., the amount of object in the sample bound to the enzyme, can be quantitatively and accurately evaluated.

[0050] Furthermore, the sensor output values ​​obtained when multiple amounts of acid are added to the standard sample can be stored on any recording medium, or they can be used as learning data for machine learning to generate a learned model. By using such sensor output values, for example, when performing the detection method of this disclosure later, even with a smaller amount of acid added to the standard sample (e.g., one sample), a standard titration curve can be generated. Additionally, sensor calibration can be performed based on the difference between such sensor output values ​​and newly obtained sensor output values ​​when performing the detection method of this disclosure later.

[0051] [2. First Implementation Method] The detector 1 and the detection device 100 that implement the above measurement method will be described.

[0052] [2-1. Structure of Detector 1] use Figure 5 The structure of detector 1 will be described.

[0053] like Figure 5 As shown, detector 1 includes substrate 10. Detector 1 is also referred to as a housing. Substrate 10 is a plate-shaped component with a rectangular plate surface. Furthermore, when detector 1 is mounted on detection device 100, the upper plate surface is referred to as the surface of substrate 10, and the lower plate surface is referred to as the back surface of substrate 10. The material of substrate 10 is, for example, acrylic resin, polycarbonate resin, or polystyrene resin.

[0054] The detector 1 has a sample loading section 2 and a sensor receiving section 3.

[0055] The sample insertion section 2 has a hole on the surface of the substrate 10 for inserting a sample.

[0056] The sensor housing 3 is formed within the substrate 10 and serves as a space for holding the object to be detected by the sensor. A semiconductor sensor 31 for measuring ion concentration is disposed in the sensor housing 3.

[0057] The detector 1 has a pressure inlet 4, a pressure outlet 5, a first flow path 6, a second flow path 7, a third flow path 8, and a waste liquid flow path 9.

[0058] The first flow path 6 is a passageway formed in the substrate 10 to allow fluid to flow, which connects the pressure inlet 4 and the sample inlet 2.

[0059] The second flow path 7 is a passageway formed within the substrate 10 to allow fluid to flow, connecting the sample input section 2 and the sensor receiving section 3. The second flow path 7 serves to supply the sample input into the sample input section 2 to the sensor receiving section 3.

[0060] The third flow path 8 is a passageway formed in the substrate 10 to allow fluid to flow, in a manner that connects the sensor housing 3 to the pressure outlet 5.

[0061] Waste liquid flow path 9 is a passage for discharging fluids (hereinafter referred to as waste liquid) that are not needed in the second flow path 7, etc., to the outside of the detector 1.

[0062] The pressure inlet 4 has an opening at its first end 10A along the length of the plate. The pressure inlet 4 is a portion that receives fluid pressurized by a pump 104 or similar device located outside the detector 1 (described later) and directs it toward the first flow path 6. The fluid flowing in the first flow path 6 may be, for example, a washing liquid or an inert gas.

[0063] Like the pressure inlet 4, the pressure outlet 5 has an opening at its first end 10A along the length of the plate. The pressure outlet 5 is a portion for discharging fluid that has reached it via the third flow path 8 to the outside of the substrate 10. Negative pressure can be applied to the pressure outlet 5 by means of a pump 104 or the like, which is located outside the detector 1 and will be described later.

[0064] The first flow path 6 has a bead housing section 61. The magnetic beads are housed in the magnetic bead housing section 61.

[0065] The second flow path 7 includes a first mixing section 71, a holding section 72, a second mixing section 73, a separating section 74, and an adjusting section 75.

[0066] The first mixing section 71 is a passageway formed in a bent shape. The first mixing section 71 utilizes the turbulence generated when the fluid passes through the section that is bent at a right angle to fully mix the magnetic beads and the sample supplied from the sample input section 2.

[0067] The holding section 72 is a space that temporarily holds the substance mixed by the first mixing section 71. The holding section 72 is connected to the waste liquid flow path 9 via the first discharge valve 92. The holding section 72 is connected to the second mixing section 73 via the first control valve 76.

[0068] The second mixing section 73, like the first mixing section 71, is formed as a zigzag passage. The second mixing section 73 thoroughly mixes the substance supplied from the holding section 72.

[0069] The separation section 74 is a space that temporarily holds the material mixed by the second mixing section 73. The separation section 74 is connected to the waste liquid flow path 9 via the second discharge valve 93. The separation section 74 is connected to the adjustment section 75 via the second control valve 77.

[0070] The adjustment section 75 is a straight passage leading to the sensor housing section 3. The adjustment section 75 can be set to obtain the length of time required for the mixture supplied from the separation section 74 to react fully (i.e., enzyme-substrate reaction).

[0071] Waste liquid flow path 9 has a waste liquid receiving section 91. The waste liquid receiving section 91 is a space formed within the substrate 10 for temporarily holding waste liquid. In the waste liquid flow path 9, waste liquid discharged from the holding section 72 via the first discharge valve 92 and waste liquid discharged from the separating section 74 via the second discharge valve 93 are received in the waste liquid receiving section 91. The waste liquid receiving section 91 is connected to the third flow path 8 via the third discharge valve 94.

[0072] The sensor housing 3 is connected to the third flow path 8 via the fourth discharge valve 95.

[0073] The third flow path 8 is a passage leading to the pressure discharge port 5 formed in the substrate 10. The third flow path 8 is configured to discharge waste liquid discharged from the waste liquid receiving section 91 via the third discharge valve 94 and waste liquid discharged from the sensor receiving section 3 via the fourth discharge valve 95 from the pressure discharge port 5.

[0074] The first discharge valve 92, the second discharge valve 93, the third discharge valve 94, the fourth discharge valve 95, the first control valve 76, and the second control valve 77 are all solenoid valves that are opened and closed by electrical signals.

[0075] The detector 1 includes a first operating unit 11, a second operating unit 12, a third operating unit 13, a fourth operating unit 14, and a fifth operating unit 15. Each of the first operating unit 11, second operating unit 12, third operating unit 13, fourth operating unit 14, and fifth operating unit 15 has a space formed within the substrate 10 and a connection flow path 17. The first operating unit 11, second operating unit 12, third operating unit 13, fourth operating unit 14, and fifth operating unit 15 are configured such that when pressure is applied from the surface side of the substrate 10, the contents are supplied to the connection destination of the connection flow path 17.

[0076] The first operating unit 11 contains the substance C with varying ion concentration and is connected to the sensor housing unit 3 via the connecting flow path 17.

[0077] The second operating section 12 houses the connector-enzyme fusion body and is connected to the holding section 72 via the connecting flow path 17.

[0078] The third operating section 13 contains the substrate liquid and is connected to the separation section 74 via the connecting flow path 17.

[0079] The fourth operating section 14 contains the washing liquid and is connected to the holding section 72 via the connecting flow path 17.

[0080] The fifth operating section 15 contains the washing liquid and is connected to the separation section 74 via the connecting flow path 17.

[0081] Furthermore, in this disclosure, when using a buffer solution, the buffer solution can be contained in at least one of the sample insertion unit 2, the sensor receiving unit 3, the first operating unit 11, the second operating unit 12, the third operating unit 13, the fourth operating unit 14, and the fifth operating unit 15. For example, the buffer solution can be contained in at least one of the sample insertion unit 2 and the sensor receiving unit 3.

[0082] Furthermore, while the detector 1 disclosed herein can be discarded after a single use, it can also be reused. In the detector 1, the sample loading section 2, the sensor housing section 3, each operating section 11-15, and each flow path 6-9 are washed, and the required substance or liquid is refilled in a state that eliminates the influence of the previous use, thereby enabling reuse.

[0083] [2-2. Structure of the detection device 100] use Figure 6 The structure of the detection device 100 will be described.

[0084] The detection device 100 has a slit-shaped insertion part 101 for inserting the detector 1. The detection device 100 has a pressure port 102 and a discharge port 103 on the inner side of the insertion part 101. The pressure port 102 is connected to the pressure input part of the detector 1, and the discharge port 103 is connected to the pressure discharge part of the detector 1.

[0085] The detection device 100 includes a pump 104, a heating unit 105, a control unit 106, a magnetic force generating unit 107, and a pressing operation unit 108.

[0086] Pump 104 pressurizes the fluid introduced into pressure inlet 4 and outputs it from pressure inlet 102. Additionally, pump 104 can generate a negative pressure applied to pressure outlet 5 to draw fluid from outlet 103. Pump 104 operates according to instructions from control unit 106.

[0087] The heating unit 105 heats one or more parts of the detection device 100. The heating unit 105 operates according to instructions from the control unit 106. For example, the control unit 106 controls the heating unit 105 to maintain the temperature of one or more parts of the detection device 100 within an appropriate temperature range (e.g., around 25°C).

[0088] The magnetic force generating unit 107 is positioned opposite the part of the detector 1 mounted on the insertion unit 101 where magnetic force needs to be generated. Figure 6 For ease of observation, only one magnetic force generating unit 107 is shown in the accompanying drawings. However, in reality, there are two locations where magnetic force needs to be generated: the holding part 72 and the separating part 74, and the magnetic force generating unit 107 is arranged at each of them. The magnetic force generating unit 107 is an electromagnet that generates magnetic force by being energized according to the instruction from the control unit 106. Hereinafter, the magnetic force generating unit 107 arranged opposite to the holding part 72 will also be referred to as the first electromagnet M1, and the magnetic force generating unit 107 arranged opposite to the separating part 74 will also be referred to as the second electromagnet M2. Furthermore, the first electromagnet M1 and the second electromagnet M2 are collectively referred to as the magnetic force generating unit 107.

[0089] For example, when the first electromagnet M1, which is positioned opposite the holding section 72, is activated, the magnetic bead is attracted by the first electromagnet M1. Therefore, even if the first discharge valve 92 or the first control valve 76 is open, the magnetic bead remains in the holding section 72, and only the fluid supplied to the holding section 72 along with the magnetic bead is discharged through the first discharge valve 92. If the first electromagnet M1 is stopped, the magnetic bead is released from the magnetic force of the first electromagnet M1 and can be discharged from the first control valve 76. Furthermore, the situation is the same when the second electromagnet M2, which is positioned opposite the separating section 74, is activated.

[0090] The pressing operation section 108 is positioned opposite the surface of the substrate 10 where the pressing operation is required in the detector 1 mounted on the insertion section 101. Figure 6 In order to facilitate observation of the accompanying drawings, similar to the magnetic force generating unit 107, only one pressing operation unit 108 is shown. However, in reality, as the parts requiring pressing operation, there are a first operation unit 11, a second operation unit 12, a third operation unit 13, a fourth operation unit 14, and a fifth operation unit 15. The pressing operation unit 108 has a mechanism that allows each operation unit 11-15 to be pressed individually using a pressing member according to the instructions from the control unit 106.

[0091] The control unit 106 is centered around a microcomputer including a CPU 110, ROM, and RAM. Various functions of the microcomputer are implemented by the CPU 110 executing programs stored on a non-transitory physical recording medium. In this example, the ROM is equivalent to a non-transitory physical recording medium storing the program. Furthermore, by executing the program, methods corresponding to the program are executed. Moreover, some or all of the functions executed by the CPU 110 can be configured in hardware using one or more ICs. Additionally, the number of microcomputers constituting the control unit 106 can be one or more.

[0092] The control unit 106 performs at least the detection process for substances that have become the target of detection from the sample detection. During the detection process, control signals for operating the detector 1 are generated sequentially to control the operation of the pump 104, heating unit 105, magnetic force generating unit 107, pressing operation unit 108, first discharge valve 92, second discharge valve 93, third discharge valve 94, first control valve 76, and second control valve 77. This enables the semiconductor sensor 31 to achieve highly sensitive detection of changes in ion concentration.

[0093] [2-3. Processing] Next, use Figure 7The flowchart illustrates the processing performed by the detection device 100. Furthermore, the following processing can be performed by the CPU 110 of the detection device 100. In the initial state, the first discharge valve 92, the second discharge valve 93, the third discharge valve 94, the first control valve 76, and the second control valve 77 are all closed, and the pump 104, the magnetic force generating unit 107, and the pressing operation unit 108 are all stopped. Moreover, the term "initial state" in this disclosure refers, for example, to the state before the detection device 100 is used for ion concentration measurement.

[0094] In S10, a sample is placed into the sample placement unit 2. The placement of the sample can be performed by the operator.

[0095] In S20, CPU 110 performs a first mixing process. This first mixing process involves mixing the magnetic beads with the sample to generate a first conjugate A and holding it in the holding section 72. In this first mixing process, CPU 110 activates the first electromagnet M1, opens the first discharge valve 92, and drives the pump 104. The magnetic beads housed in the bead receiving section 61 pass through the first flow path 6 and, together with the sample introduced into the sample input section 2 in S10, pass through the first mixing section 71. Thus, the magnetic beads and the sample are mixed to generate the first conjugate A. The first conjugate A moves towards the holding section 72. Because the magnetic beads are attracted to the first electromagnet M1, the first conjugate A is held within the holding section 72. Components that do not constitute the first conjugate A move through the waste liquid flow path 9 to the waste liquid receiving section 91. After a predetermined time has elapsed since the pump 104 was started, CPU 110 closes the first discharge valve 92 and stops the pump 104.

[0096] In S30, CPU 110 performs a first washing process. This first washing process washes the holding section 72. During this process, CPU 110 operates the pressing operation unit 108, pressing the fourth operation unit 14. As a result, the washing liquid contained in the fourth operation unit 14 is supplied to the holding section 72 through the connecting flow path 17. The supplied washing liquid washes the holding section 72. Afterward, CPU 110 opens the first discharge valve 92 and drives the pump 104. As a result, the components that do not constitute the first compound A move towards the waste liquid receiving section 91 through the waste liquid flow path 9. After a predetermined time has elapsed since the pump 104 was started, CPU 110 closes the first discharge valve 92, stopping the first electromagnet M1 and the pump 104.

[0097] In S40, CPU110 performs a second mixing process. This second mixing process involves mixing the first conjugate A with the adapter-enzyme fusion compound to generate a second conjugate B, which is then held in the separation section 74. In this second mixing process, CPU110 activates the second electromagnet M2, opens the first control valve 76 and the second discharge valve 93, and activates the pressing component to press the second operating section 12. As a result, the adapter-enzyme fusion solution contained in the second operating section 12 is supplied to the holding section 72 through the connecting flow path 17. Then, CPU110 drives the pump 104. The first conjugate A and the adapter-enzyme fusion solution present in the holding section 72 move through the second mixing section 73 to the separation section 74. Here, the first conjugate A and the adapter-enzyme fusion compound are mixed to generate the second conjugate B. Because the magnetic beads are adsorbed onto the second electromagnet M2, the second conjugate B is held within the separation section 74. Components that do not constitute the second conjugate B move through the waste liquid flow path 9 to the waste liquid receiving section 91. After a predetermined time has elapsed since the pump 104 was started, the CPU 110 closes the first control valve 76 and the second discharge valve 93, causing the pump 104 to stop.

[0098] In S50, CPU 110 performs a second washing process. This second washing process washes the separation section 74. During this process, CPU 110 operates the pressing operation unit 108, pressing the fifth operation unit 15. As a result, the washing liquid contained in the fifth operation unit 15 is supplied to the separation section 74 through the connecting flow path 17. The supplied washing liquid washes the separation section 74. Afterward, CPU 110 opens the second discharge valve 93 and drives the pump 104. As a result, the components that do not constitute the second compound B move through the waste liquid flow path 9 to the waste liquid collection section 91. After a predetermined time has elapsed since the pump 104 was started, CPU 110 closes the second discharge valve 93, stops the second electromagnet M2, and stops the pump 104.

[0099] In S60, CPU 110 performs a third mixing process. This third mixing process involves mixing the second conjugate B with the substrate solution to induce a chemical reaction between the enzyme and the substrate. In this third mixing process, CPU 110 opens the second control valve 77, activating the pressing operation unit 108 and pressing the third operation unit 13. As a result, the substrate solution contained in the third operation unit 13 is supplied to the separation unit 74 via the connecting flow path 17. Then, CPU 110 drives pump 104. As a result, the second conjugate B and the substrate solution present in the separation unit 74 move towards the sensor housing unit 3 via the adjustment unit 75. During this movement, the second conjugate B and the substrate solution are mixed, thereby activating a chemical reaction that consumes the substrate using the enzyme present in the second conjugate B. Through this chemical reaction, the hydrogen ion concentration in the mixture of the second conjugate B and the substrate solution changes. After a predetermined time has elapsed since the pump 104 was started, CPU 110 closes the second control valve 77, stopping pump 104.

[0100] In S70, CPU 110 performs a first ion concentration measurement process. CPU 110 measures the ion concentration using semiconductor sensor 31. Alternatively, the first ion concentration measurement process can be omitted if necessary.

[0101] In S80, CPU 110 performs ion concentration change processing. After a predetermined time has elapsed since the start of the first ion concentration measurement process, CPU 110 activates the pressing operation unit 108 to press the first operation unit 11. As a result, the acid, which is the ion concentration change substance C, contained in the first operation unit 11, is supplied to the sensor housing unit 3 through the connection flow path 17.

[0102] In S90, CPU 110 performs a second ion concentration measurement process. CPU 110 measures the ion concentration using semiconductor sensor 31. After the second ion concentration measurement process, CPU 110 opens the fourth discharge valve 95 and drives pump 104. As a result, the liquid in sensor housing 3 is discharged through third flow path 8.

[0103] Furthermore, by comparing the results obtained from the first ion concentration measurement process in S70 and / or the second ion concentration measurement process in S90 with reference data, the presence of an object contained in the sample can be detected. To obtain reference data, two identical structures can be set up within the same detector 1 for use in sample determination and for a reference sample not containing an object. The aforementioned processing can be performed in each set. Alternatively, to obtain reference data, two detectors 1 can be used, one for sample determination and the other for a reference sample, with the aforementioned processing performed independently in each.

[0104] Furthermore, when using a buffer solution in this disclosure, the buffer solution can be mixed with the sample at any point in time during the process from when the sample is inserted into the sample insertion section 2 until it moves to the sensor receiving section 3 (S10~S60). Alternatively, the buffer solution can be mixed with the mixture of the second conjugate B and the substrate solution at any point in time after the sample has moved to the sensor receiving section 3 (S70~S90).

[0105] [2-4. Effects] According to the first embodiment described above, the following effects are achieved.

[0106] (2a) According to the above embodiment, in detector 1, ion concentration is measured in a sample after the addition of ion concentration-changing substance C. Ion concentration-changing substance C is a substance that causes a change in the ion concentration of the sample by mixing with it. Therefore, the change in ion concentration in the sample is greater. As a result, the detection sensitivity of the change in ion concentration in the sample can be improved.

[0107] (2b) Furthermore, according to the above embodiment, an acid is used as the substance C that changes the ion concentration. Therefore, the ion concentration of the sample can be changed in a simple way. As a result, the detection sensitivity of the change in ion concentration in the sample can be appropriately improved.

[0108] (2c) Furthermore, according to the above embodiment, a first conjugate A is prepared by binding a magnetic bead to a sample, and a second conjugate B is prepared by binding a linker-enzyme fusion compound to the first conjugate A. A substrate solution is added to the second conjugate B, and a chemical reaction consuming the substrate occurs using the enzyme present in the second conjugate B, causing a change in the hydrogen ion concentration in the reaction solution. After adding an ion concentration-changing substance C, this change in hydrogen ion concentration is measured as the ion concentration in the sample. Therefore, the ion concentration of a specific object bound to the linker in the sample is measured. This appropriately improves the detection sensitivity of changes in ion concentration. Furthermore, by comparing the measured ion concentration with reference data, it is possible to determine whether the sample contains the object.

[0109] (2d) Furthermore, according to the above embodiment, the detector 1 includes a sample insertion section 2, a sensor receiving section 3, a first operation section 11, a second operation section 12, a third operation section 13, a fourth operation section 14, and a fifth operation section 15, as well as the aforementioned flow paths, within the substrate 10. Therefore, if the detector 1 of this disclosure is used, it is not necessary to change the container for each step, and ion concentration measurement can be performed easily and conveniently. In addition, the detector 1 can be miniaturized.

[0110] (2e) Furthermore, according to the above embodiments, in this disclosure, when using a buffer solution, the ion concentration of the sample can be changed to outside the buffer range by mixing the substance C, which changes the ion concentration, with the sample. Therefore, outside the buffer range, the change in ion concentration in the sample becomes larger. As a result, the detection sensitivity of the change in ion concentration can be appropriately improved.

[0111] [3. Second Implementation] [3-1. Differences from the first embodiment] The basic structure of the second embodiment is the same as that of the first embodiment; therefore, the differences will be described below. Furthermore, the same reference numerals as in the first embodiment denote the same structures, as described previously.

[0112] The second embodiment differs in the structure of the first flow path 6, the structure of the waste liquid flow path 9, and the installation positions of the second operation unit 12, the third operation unit 13, the fourth operation unit 14, and the fifth operation unit 15.

[0113] like Figure 8 As shown, in detector 1, the first flow path 6 omits the bead receiving section 61 and instead has a sixth operation section 16.

[0114] The sixth operating unit 16 contains a magnetic bead. Like the other operating units 11-15, the sixth operating unit 16 supplies its contents to the connection destination of the connecting flow path 17 by pressing. The sixth operating unit 16 is connected to the first flow path 6 via the connecting flow path 17.

[0115] The second operation unit 12, the third operation unit 13, the fourth operation unit 14 and the fifth operation unit 15 are all connected to the first flow path 6 via the connecting flow path 17.

[0116] In the second flow path 7, the holding part 72 omits the connection to the first discharge valve 92 and the waste liquid flow path 9, and also omits the first control valve 76.

[0117] In the waste liquid flow path 9, the third discharge valve 94 is omitted.

[0118] In the detection device 100, the pressing operation unit 108 is provided with a mechanism for pressing each operation unit 11 to 16 at a position opposite to the first operation unit 11, the second operation unit 12, the third operation unit 13, the fourth operation unit 14, the fifth operation unit 15 and the sixth operation unit 16.

[0119] The magnetic force generating unit 107 includes a second electromagnet M2 disposed opposite to the separation unit 74. The first electromagnet M1 disposed opposite to the holding unit 72 is omitted. However, in this second embodiment, the magnetic force generating unit 107 may include the first electromagnet M1 in addition to the second electromagnet M2, disposed opposite to the holding unit 72.

[0120] That is, in the detection device 100, the pressing operation unit 108 is arranged according to the position of the first to sixth operation units 11 to 16 in the detector 1A, and the magnetic force generating unit 107 is arranged at the position where magnetic force needs to be generated. The same applies in the following embodiments.

[0121] [3-2. Processing] Next, based on Figure 7 The process described in the first embodiment will be used to explain the processing performed by the detection device 100 of the second embodiment. Furthermore, processes common to the first embodiment will be omitted from the description.

[0122] In S20, CPU 110 activates the pressing operation unit 108, pressing the sixth operation unit 16. As a result, the magnetic bead housed in the sixth operation unit 16 is supplied to the first flow path 6 via the connecting flow path 17. Then, CPU 110 drives the pump 104. The magnetic bead passes through the first flow path 6 and, together with the sample introduced into the sample introduction unit 2 in S10, passes through the first mixing unit 71. Thus, the magnetic bead and the sample are mixed to form a first composite A. The first composite A moves towards the holding unit 72. After a predetermined time has elapsed since the pump 104 was started, CPU 110 stops the pump 104.

[0123] In step S30, CPU 110 activates the pressing operation unit 108, pressing the fourth operation unit 14. As a result, the washing liquid contained in the fourth operation unit 14 is supplied to the first flow path 6 via the connecting flow path 17. Then, CPU 110 drives pump 104. The washing liquid passes through the first flow path 6 and the first mixing unit 71, and is supplied to the holding unit 72. The supplied washing liquid washes the holding unit 72. After a predetermined time has elapsed since pump 104 was started, CPU 110 stops pump 104.

[0124] In S40, CPU 110 activates the second electromagnet M2, opening the second discharge valve 93 and activating the pressing operation unit 108, pressing the second operation unit 12. As a result, the connector-enzyme fusion solution contained in the second operation unit 12 is supplied to the first flow path 6 via the connecting flow path 17. Then, CPU 110 drives pump 104. The connector-enzyme fusion solution is supplied to the holding unit 72 via the first flow path 6 and the first mixing unit 71. Furthermore, the first conjugate A and the connector-enzyme fusion solution present in the holding unit 72 move to the separation unit 74 via the second mixing unit 73. Other processes in S40 are the same as in the first embodiment.

[0125] In S50, CPU 110 activates the pressing operation unit 108, pressing the fifth operation unit 15. As a result, the washing liquid contained in the fifth operation unit 15 is supplied to the first flow path 6 via the connecting flow path 17. Then, CPU 110 opens the second discharge valve 93, driving the pump 104. The washing liquid is supplied to the separation unit 74 via the first flow path 6, the first mixing unit 71, the holding unit 72, and the second mixing unit 73. The supplied washing liquid washes the separation unit 74. Furthermore, other processes in S50 are the same as in the first embodiment.

[0126] In S60, CPU 110 activates the pressing operation unit 108, pressing the third operation unit 13. As a result, the connector-enzyme fusion solution housed in the second operation unit 12 is supplied to the first flow path 6 via the connecting flow path 17. Then, CPU 110 drives the pump 104. The substrate solution is supplied to the separation unit 74 via the connecting flow path 17, through the first flow path 6, the first mixing unit 71, the holding unit 72, and the second mixing unit 73. Furthermore, the second conjugate B and the substrate solution present in the separation unit 74 move towards the sensor housing unit 3 via the adjustment unit 75. Other processes in S60 are the same as in the first embodiment.

[0127] [3-3. Effect] In addition to achieving the effects of the first embodiment described above, the second embodiment also achieves the following effects.

[0128] (3a) The second embodiment differs in the structure of the first flow path 6, the structure of the waste liquid flow path 9, and the installation positions of the second operation unit 12, the third operation unit 13, the fourth operation unit 14, and the fifth operation unit 15. Thus, even with differences in the structure and installation position of the detector 1A, it achieves the same effect as the first embodiment described above.

[0129] (3b) In addition, the structure of detector 1A can be simplified compared to detector 1, depending on the configuration of the structure.

[0130] [4. Third Implementation Method] [4-1. Differences from the first embodiment] The basic structure of the third embodiment is the same as that of the first embodiment; therefore, the differences will be described below. Furthermore, the same reference numerals as in the first embodiment denote the same structures, as described previously.

[0131] The structure of the second flow path 7 and the waste liquid flow path 9 of the detector 1B in the third embodiment are different from those in the first embodiment.

[0132] Furthermore, in detector 1B, the first operation unit 11, the second operation unit 12, and the third operation unit 13 are omitted, and the connection destinations of the fourth operation unit 14 and the fifth operation unit 15 are different from those in the first embodiment.

[0133] like Figure 9 As shown, the specimen input section 2 is connected to the waste liquid flow path 9 via the fifth discharge valve 96.

[0134] The second flow path 7 includes a first connecting part 711, a connector-enzyme containing part 712, a second connecting part 713, a substrate liquid containing part 714, and an adjustment part 75.

[0135] The first connecting part 711 is a flow path that connects the sample input part 2 and the connector-enzyme receiving part 712, and a third control valve 78 is provided at the upstream end connected to the sample input part 2.

[0136] The connector-enzyme containment section 712 is a space that contains the connector-enzyme fusion body in its initial state. The connector-enzyme containment section 712 is connected to the waste liquid flow path 9 via the sixth discharge valve 97.

[0137] The second connection part 713 is a flow path that connects the connector-enzyme receiving part 712 and the substrate liquid receiving part 714, and a fourth control valve 79 is provided on the upstream end connected to the connector-enzyme receiving part 712.

[0138] In the initial state of detector 1B, the substance C with changing ion concentration is contained in sensor housing 3.

[0139] The substrate liquid containment section 714 is a space that contains the substrate liquid in its initial state.

[0140] The adjustment section 75 is a flow path that connects the substrate liquid receiving section 714 and the sensor receiving section 3. The length of the flow path is set in the same way as that of the adjustment section 75 described in the first embodiment.

[0141] In the detection device 100, the magnetic force generating unit 107 is provided with a third electromagnet M3 at a position opposite to the sample input unit 2, and a fourth electromagnet M4 at a position opposite to the adapter-enzyme receiving unit 712.

[0142] [4-2. Processing] Next, based on Figure 7 The process described in the first embodiment will be used to explain the processing performed by the detection device 100 of the third embodiment. Furthermore, processes common to the first embodiment will be omitted from the description.

[0143] In S20, CPU 110 activates the third electromagnet M3, opens the fifth discharge valve 96, and drives pump 104. Magnetic beads housed in the bead receiving section 61 move through the first flow path 6 to the sample input section 2. The magnetic beads mix with the sample input to the sample input section 2, forming a first conjugate A. Because the magnetic beads are attracted to the third electromagnet M3, the first conjugate A is maintained within the sample input section 2. Components not constituting the first conjugate A move through the waste liquid flow path 9 to the waste liquid receiving section 91. After a predetermined time has elapsed since pump 104 was started, CPU 110 closes the fifth discharge valve 96, stopping pump 104.

[0144] In step S30, CPU 110 activates the pressing operation unit 108, pressing the fourth operation unit 14. As a result, the washing liquid contained in the fourth operation unit 14 is supplied to the sample placement unit 2 through the connecting flow path 17. The supplied washing liquid washes the sample placement unit 2. Afterwards, CPU 110 opens the fifth discharge valve 96, driving the pump 104. After a predetermined time has elapsed since the pump 104 was started, CPU 110 closes the fifth discharge valve 96, stopping the third electromagnet M3 and stopping the pump 104.

[0145] In step S40, CPU 110 activates the fourth electromagnet M4, opening the third control valve 78 and the sixth discharge valve 97, thus driving pump 104. The first conjugate A, present in the sample input section 2, moves via the first connection section 711 to the adapter-enzyme containing section 712. The second conjugate B mixes with the adapter-enzyme fusion body, forming a second conjugate B. Because the magnetic bead is attracted to the fourth electromagnet M4, the second conjugate B is held within the adapter-enzyme containing section 712. After a predetermined time has elapsed since pump 104 was started, CPU 110 closes the third control valve 78 and the sixth discharge valve 97, stopping pump 104.

[0146] In S50, CPU 110 activates the pressing operation unit 108, pressing the fifth operation unit 15. As a result, the washing solution contained in the fifth operation unit 15 is supplied to the connector-enzyme containing unit 712 through the connecting flow path 17. The supplied washing solution washes the connector-enzyme containing unit 712. Afterwards, CPU 110 opens the sixth discharge valve 97, driving pump 104. After a predetermined time has elapsed since pump 104 was started, CPU 110 closes the sixth discharge valve 97, stopping the fourth electromagnet M4 and stopping pump 104.

[0147] In step S60, CPU 110 opens the fourth control valve 79, activating pump 104. As a result, the second conjugate B, present in the adapter-enzyme housing 712, moves via the second connection 713 to the substrate solution housing 714, and together with the substrate solution, moves via the adjustment section 75 to the sensor housing 3. Thereafter, they mix with the acid, which is a substance C representing a change in ion concentration, housed in the sensor housing 3. After a predetermined time has elapsed since pump 104 was activated, CPU 110 closes the fourth control valve 79, stopping pump 104.

[0148] In the third embodiment, the processes S70 and S80 are omitted.

[0149] [4-3. Effect] According to the third embodiment detailed above, in addition to achieving the effects of the first embodiment described above, it also achieves the following effects.

[0150] (4a) In the detector 1B of the third embodiment, the first operation unit 11, the second operation unit 12, and the third operation unit 13 are omitted, and the operation of pressing these operation units 11 to 13 by the CPU 110 is also omitted. As a result, the construction and processing of the detector 1B can be simplified compared to the detector 1 in this disclosure.

[0151] [5. Fourth Implementation Method] [5-1. Differences from the third embodiment] The basic structure of the fourth embodiment is the same as that of the third embodiment; therefore, the differences will be explained below. Furthermore, the same reference numerals as in the third embodiment denote the same structures, as described previously.

[0152] like Figure 10 As shown, in the detector 1C of the fourth embodiment, the bead receiving section 61 is omitted from the first flow path 6. In the detector 1, the magnetic bead is received in the sample input section 2 in the initial state.

[0153] [5-2. Processing] Next, regarding the processing performed by the detection device 100 of the fourth embodiment, the differences from the first and third embodiments will be explained below.

[0154] In the detector 1 of the fourth embodiment, the magnetic bead receiving section 61 is omitted from the first flow path 6, and the magnetic bead is received in the sample input section in the initial state. Therefore, in the detector 1 of the fourth embodiment, in S10, when the sample is inserted into the sample input section 2, the magnetic bead is mixed with the magnetic bead received in the sample input section 2 to generate a first composite A formed by the combination of the two.

[0155] Other processing is the same as in the third embodiment described above.

[0156] [5-3. Effect] According to the fourth embodiment described above, in addition to achieving the effects of the first embodiment and the third embodiment, it also achieves the following effects.

[0157] (5a) In the detector 1C of the fourth embodiment, the bead housing 61 is omitted. Therefore, the detector 1C can be provided with a simpler structure compared to the detector 1.

[0158] [6. Fifth Implementation Method] [6-1. Differences from the first embodiment] The basic structure of the fifth embodiment is the same as that of the first embodiment; therefore, the differences will be described below. Furthermore, the same reference numerals as in the first embodiment denote the same structures, as described previously.

[0159] like Figure 11 As shown, in the detector 1D of the fifth embodiment, the third operation unit 13 is omitted. In the initial state of the detector 1, the substrate liquid is contained in the sensor housing unit 3.

[0160] [6-2. Processing] Next, based on Figure 7 The process described in the first embodiment will be used to explain the processing performed by the detection device 100 in the second embodiment. Furthermore, processes common to the first embodiment will be omitted from the description.

[0161] In step S60, CPU 110 opens the second control valve 77, activating pump 104. As a result, the second conjugate B, present in the separation section 74, moves towards the sensor housing 3 via the adjustment section 75. In the sensor housing 3, the second conjugate B and the substrate liquid are mixed, thereby utilizing the enzyme present in the second conjugate B to undergo a chemical reaction that consumes the substrate. After a predetermined time has elapsed since pump 104 was activated, CPU 110 closes the second control valve 77, stopping pump 104.

[0162] Other processing is the same as in the third embodiment described above.

[0163] [6-3. Effect] According to the fifth embodiment detailed above, in addition to achieving the effects of the first embodiment described above, it also achieves the following effects.

[0164] (6a) In the detector 1D of the fifth embodiment, the third operation unit 13 is omitted. Therefore, compared with the detector 1, the detector 1D can be provided with a simpler structure.

[0165] [7. Other Implementation Methods] The embodiments of this disclosure have been described above, but this disclosure is not limited to the above embodiments and can be implemented in various ways.

[0166] (7a) The detector 1 may also be a detector 1 having a sample insertion unit 2, a sensor receiving unit 3 and a first operation unit 11. That is, the second operation unit 12, the third operation unit 13, the fourth operation unit 14 and the fifth operation unit 15 may also be omitted.

[0167] As an example, firstly, the sample can be inserted into the sample insertion unit 2. Next, the CPU 110 can drive the pump 104. Thus, the sample inserted into the sample insertion unit 2 can move to the sensor receiving unit 3 via the second flow path 7. Furthermore, the CPU 110 can arbitrarily measure the ion concentration of the sample using the semiconductor sensor 31. Afterwards, the CPU 110 can activate the pressing operation unit 108 to press the first operation unit 11, thereby supplying the acid, which is the substance C representing the change in ion concentration, contained in the first operation unit 11, to the sensor receiving unit 3 via the connecting flow path 17. Then, the CPU 110 can measure the ion concentration of the sample using the semiconductor sensor 31. Therefore, even with a minimal structure, the detection sensitivity for changes in ion concentration in the sample can be improved in the detector 1.

[0168] (7b) In the above embodiment, an acid is used as the ion concentration changing substance C, but for example, a base or air (i.e., CO2) can also be used as the ion concentration changing substance C. When air is used as the ion concentration changing substance C, the detector 1 can have an ion concentration changing substance supply passage 11A. Specifically, as... Figure 12 As shown, the ion concentration change substance supply passage 11A can be a passage extending from the sensor housing 3 to the surface of the substrate 10. Alternatively, an opening and closing mechanism 11B (e.g., a valve 11B) for the ion concentration change substance supply passage 11A can be provided on the surface of the substrate 10 to which the ion concentration change substance supply passage 11A is connected. Furthermore, it can be configured such that, as an ion concentration change process, by opening the opening and closing mechanism 11B from the surface side of the substrate 10, air is drawn into the ion concentration change substance supply passage 11A from outside the detector 1, and this air is supplied to the sensor housing 3.

[0169] (7c) In the above embodiment, a pressurized fluid is used as a method for moving the sample, etc., within the detector 1. The method for moving the sample, etc., is not limited to this; particularly when the sample is mixed with a magnetic bead, a magnetic force attracting the magnetic bead can be generated in a manner that moves along the flow path, thereby causing the sample, etc., to move together with the magnetic bead. In addition to magnetic force, the sample, etc., can also be moved by combining it with a negative pressure applied to the pressure outlet 5.

[0170] (7d) In the above embodiments, several forms have been shown for accommodating the magnetic beads, the adapter-enzyme fusion compound, the substrate solution, and the substance C with varying ion concentration in the initial state. These forms include spaces, sample insertion units, and sensor receiving units, located in each of the operation units 11-16, or in the spaces within the first flow path 6 and the second flow path 7. However, these structures are not limited to the forms described above, and various forms can be adopted.

[0171] (7e) In the above embodiment, the first mixing section 71 and the second mixing section 73, which serve as the second flow path 7, use a bent flow path, but are not limited to this. For example, they can be as follows: Figure 13A , Figure 13B , Figure 13C As shown, turbulence is generated in the flow path by erecting pillars or setting tiny ribs, thereby mixing the solution. Alternatively, the solution can be mixed using a vibrator mechanism that applies vibration to detector 1, or a vortex mechanism that generates eddies in the flow path. Alternatively, a rotor with a built-in magnet can be mixed with the solution, or the solution can be mixed using a stirrer mechanism that uses magnetic force to rotate the rotor. Furthermore, in detector 1, ultrasonic waves can also be generated to mix the solution.

[0172] (7f) The control unit 106 and its method described in this disclosure can be implemented by a dedicated computer, which is provided by a processor and memory configured to perform one or more functions embodied in a computer program. Alternatively, the control unit 106 and its method described in this disclosure can be implemented by a dedicated computer provided by a processor configured with one or more dedicated hardware logic circuits. Alternatively, the control unit 106 and its method described in this disclosure can also be implemented by one or more dedicated computers configured by a combination of a processor and memory configured to perform one or more functions and a processor configured with one or more hardware logic circuits. In addition, the computer program can be stored as instructions to be executed by the computer on a computer-readable non-transitory tangible recording medium. In the method of implementing the functions of each part included in the control unit 106, it is not necessary to include software, and all its functions can also be implemented using one or more hardware components.

[0173] (7g) Multiple functions of one component in the above embodiments can be achieved through multiple components, or one function of one component can be achieved through multiple components. Alternatively, multiple functions of multiple components can be achieved through one component, or one function achieved by multiple components can be achieved through one component. Furthermore, a portion of the structure in the above embodiments may be omitted. Additionally, at least a portion of the structure in the above embodiments may be added to or replaced relative to the structures of other above embodiments.

[0174] (7h) In addition to the detection device 100 described above, this disclosure can also be implemented in various ways, such as a system that includes the detection device 100 as a component, a program that enables a computer to function as the control unit 106 of the detection device 100, a non-transitory physical recording medium such as a semiconductor memory that records the program, a detection method, and a method for detecting objects in a specimen.

[0175] [7. The correspondence between statements] In this embodiment, the second flow path 7 corresponds to the moving flow path. The first operating unit 11 and the ion concentration changing substance supply passage 11A correspond to the changing substance supply unit. The first operating unit 11 corresponds to the changing substance containing unit. The second operating unit 12, the third operating unit 13, the sixth operating unit 16, the bead containing unit 61, the first mixing unit 71, the second mixing unit 73, the adjustment unit 75, the adapter-enzyme containing unit 712, and the substrate solution containing unit 714 correspond to the mixing unit. The sixth operating unit 16 and the bead containing unit 61 correspond to the bead containing unit. The second operating unit 12 and the adapter-enzyme containing unit 712 correspond to the enzyme containing unit. The third operating unit 13 and the substrate solution containing unit 714 correspond to the substrate containing unit. The fourth operating unit 14 and the fifth operating unit 15 correspond to the washing unit. The first flow path 6 corresponds to the driving flow path. S20, 40, and 60 in the process of this disclosure correspond to the sample supply unit. S80 in the process of this disclosure corresponds to the ion concentration changing substance addition unit. Furthermore, in the process of this disclosure, S90 corresponds to the measuring section. Additionally, in the process of this disclosure, S20 corresponds to the bead supply section. Additionally, in the process of this disclosure, S40 corresponds to the adapter-enzyme fusion body supply section. Additionally, in the process of this disclosure, S60 corresponds to the substrate supply section. Additionally, in the process of this disclosure, S60 and S70 correspond to the reaction section.

[0176] [The technical concepts disclosed in this specification] [Project 1] A detector (1, 1A, 1B, 1C, 1D), have: Sensor housing (3) houses a sensor used for measuring ions; Specimen input section (2), for inputting specimens; The moving flow path (7) connects the sample delivery unit and the sensor receiving unit, and supplies the sample to the sensor receiving unit; and The variable substance supply unit (11, 11A) supplies the sensor housing unit with an ion concentration changing substance (C) that causes a change in the ion concentration of the sample.

[0177] [Project 2] According to the detector described in Project 1 The variable substance supply unit includes a variable substance receiving section (11) for receiving the variable substance with varying ion concentration, and has a structure (11) for pushing the variable substance with varying ion concentration toward the sensor receiving section by pressing the variable substance receiving section.

[0178] [Project 3] According to the detector described in Project 1 The variable substance supply unit has an ion concentration variable substance supply passage (11A) connected to the sensor housing unit, and has a structure (11B) for drawing in external air to the sensor housing unit.

[0179] [Project 4] The detector according to any one of Items 1 to 3 At least one of the sample delivery section and the sensor receiving section contains a buffer solution. The buffer solution and the sample form a mixture. The ion concentration-changing substance has the property of causing the ion concentration of the sample contained in the mixture to change to outside the buffer range of the buffer solution.

[0180] [Project 5] The detector according to any one of Items 1 to 4 It also includes a mixing section (12, 13, 16, 61, 71, 73, 75, 712, 714) for mixing the enzyme and substrate with the sample before the sample is fed from the sample feeding section to the sensor receiving section.

[0181] [Project 6] The detector described in Project 5 The enzyme is configured as a adapter-enzyme fusion body fused with a adapter, the adapter having the property of binding to the specimen.

[0182] [Project 7] The detector described in Project 6 The mixing section has a configuration (16, 61) for mixing the beads with the sample before the sample, which is inserted from the sample insertion section, reaches the sensor receiving section.

[0183] [Project 8] According to the detector described in Project 7 The mixing section includes a bead receiving section (16, 61) for receiving the beads, an enzyme receiving section (12, 712) for receiving the linker-enzyme fusion, and a substrate receiving section (13, 714) for receiving the substrate. At least one of the bead receiving section, the enzyme receiving section, and the substrate receiving section is disposed on the moving flow path.

[0184] [Project 9] According to the detector described in Project 7 The mixing section includes a bead-containing section for containing the beads, an enzyme-containing section for containing the linker-enzyme fusion, and a substrate-containing section for containing the substrate. At least one of the bead receiving section, the enzyme receiving section, and the substrate receiving section supplies at least one of the beads, the adapter-enzyme fusion, and the substrate to the sample delivery section and a certain part of the moving flow path.

[0185] [Project 10] The detector according to item 8 or item 9 The sensor housing and the substrate housing are integrally formed.

[0186] [Project 11] The detector according to any one of Items 1 to 10, It also includes a drive flow path (6) for applying fluid pressure to the specimen insertion section to move the specimen from the specimen insertion section toward the sensor receiving section.

Claims

1. A detector (1, 1A, 1B, 1C, 1D), have: Sensor housing (3) houses a sensor used for measuring ions; Specimen input section (2), for inputting specimens; The moving flow path (7) connects the sample delivery unit and the sensor receiving unit, and supplies the sample to the sensor receiving unit; and The variable substance supply unit (11, 11A) supplies the sensor housing unit with an ion concentration changing substance (C) that causes a change in the ion concentration of the sample.

2. The detector according to claim 1, The variable substance supply unit includes a variable substance receiving section (11) for receiving the variable substance with varying ion concentration, and has a structure (11) for pushing the variable substance with varying ion concentration toward the sensor receiving section by pressing the variable substance receiving section.

3. The detector according to claim 1, The variable substance supply unit has an ion concentration variable substance supply passage (11A) connected to the sensor housing unit, and has a structure (11B) for drawing in external air to the sensor housing unit.

4. The detector according to any one of claims 1 to 3, At least one of the sample delivery section and the sensor receiving section contains a buffer solution. The buffer solution and the sample form a mixture. The ion concentration-changing substance has the property of causing the ion concentration of the sample contained in the mixture to change to outside the buffer range of the buffer solution.

5. The detector according to any one of claims 1 to 3, It also includes a mixing section (12, 13, 16, 61, 71, 73, 75, 712, 714) for mixing the enzyme and substrate with the sample before the sample is fed from the sample feeding section to the sensor receiving section.

6. The detector according to claim 5, The enzyme is configured as a adapter-enzyme fusion body fused with a adapter, the adapter having the property of binding to the specimen.

7. The detector according to claim 6, The mixing section has a configuration (16, 61) for mixing the beads with the sample before the sample, which is inserted from the sample insertion section, reaches the sensor receiving section.

8. The detector according to claim 7, The mixing section includes a bead receiving section (16, 61) for receiving the beads, an enzyme receiving section (12, 712) for receiving the linker-enzyme fusion, and a substrate receiving section (13, 714) for receiving the substrate. At least one of the bead receiving section, the enzyme receiving section, and the substrate receiving section is disposed on the moving flow path.

9. The detector according to claim 7, The mixing section includes a bead-containing section for containing the beads, an enzyme-containing section for containing the linker-enzyme fusion, and a substrate-containing section for containing the substrate. At least one of the bead receiving section, the enzyme receiving section, and the substrate receiving section supplies at least one of the beads, the adapter-enzyme fusion, and the substrate to the sample delivery section and a certain part of the moving flow path.

10. The detector according to claim 9, The sensor housing and the substrate housing are integrally formed.

11. The detector according to any one of claims 1 to 3, It also includes a drive flow path (6) for applying fluid pressure to the specimen insertion section to move the specimen from the specimen insertion section toward the sensor receiving section.

12. A detection device (100), The detection device includes a detector and a control unit (106). The detector has the following features: Sensor housing, which houses the sensor used for measuring ions; Specimen input department, for the input of specimens; A moving flow path connects the sample delivery unit to the sensor receiving unit, supplying the sample to the sensor receiving unit; and The variable substance supply unit supplies an ion concentration changing substance, which causes a change in the ion concentration of the sample, to the sensor housing. The control unit includes: The sample supply unit (S20, S40, S60) supplies the sample to the sensor receiving unit. The ion concentration changing substance supply unit (S80) supplies the ion concentration changing substance contained in the changing substance storage unit to the sensor storage unit; and The measuring unit (S90) measures the ion concentration in the sample.

13. A detection device, The detection device includes a detector and a control unit. The detector has the following features: Sensor housing, which houses the sensor used for measuring ions; Specimen input area, for the input of specimens; A moving flow path connects the sample delivery unit to the sensor receiving unit, and supplies the sample to the sensor receiving unit; The variable substance supply unit supplies an ion concentration changing substance, which causes a change in the ion concentration of the sample, to the sensor housing unit; and A mixing section is used to mix the beads, adapter-enzyme fusion compound, and substrate with the sample before the sample, inserted from the sample input section, reaches the sensor receiving section. The control unit includes: The bead supply unit (S20) supplies the beads to the inserted specimen, generating a first conjugate (A) formed by combining the specimen and the beads; The adapter-enzyme fusion unit (S40) supplies the adapter-enzyme fusion to the first conjugate to generate a second conjugate (B) formed by combining the first conjugate and the adapter-enzyme fusion; Substrate supply unit (S60) supplies the substrate to the second assembly; The reaction section (S60, S70) supplies the second conjugate and the substrate to the sensor housing section, causing the enzyme contained in the second conjugate to produce a chemical reaction that consumes the substrate. The ion concentration changing substance supply unit (S80) supplies the ion concentration changing substance contained in the changing substance storage unit to the sensor storage unit; and The measuring unit (S90) measures the ion concentration in the sample.

14. A detection method, using a detector, The detector has the following features: Sensor housing, which houses the sensor used for measuring ions; Specimen input department, for the input of specimens; A moving flow path connects the sample delivery unit to the sensor receiving unit, and supplies the sample to the sensor receiving unit; as well as The variable substance supply unit supplies an ion concentration changing substance, which causes a change in the ion concentration of the sample, to the sensor housing. The detection method comprises the following steps: The sample is inserted into the sample insertion unit; The sample is supplied to the sensor receiving section; The ion concentration-changing substance contained in the changing substance containment section is supplied to the sensor containment section; as well as The ion concentration in the sample was measured.

15. A detection method, using a detector, The detector has the following features: Sensor housing, which houses the sensor used for measuring ions; Specimen input department, for the input of specimens; A moving flow path connects the sample delivery unit to the sensor receiving unit, and supplies the sample to the sensor receiving unit; The variable substance supply unit supplies an ion concentration changing substance, which causes a change in the ion concentration of the sample, to the sensor housing unit; and A mixing section is used to mix the beads, adapter-enzyme fusion compound, and substrate with the sample before the sample, inserted from the sample input section, reaches the sensor receiving section. The detection method comprises the following steps: The sample is inserted into the sample insertion unit; The beads are supplied to the injected sample to prepare a first conjugate formed by combining the sample and the beads; The adapter-enzyme fusion compound is supplied to the first conjugate to prepare a second conjugate formed by combining the first conjugate and the adapter-enzyme fusion compound; The substrate is supplied to the second conjugate; The second conjugate and the substrate are supplied to the sensor housing, causing the enzyme contained in the second conjugate to produce a chemical reaction that consumes the substrate; The ion concentration-changing substance contained in the changing substance containment section is supplied to the sensor containment section; and The ion concentration in the sample was measured.

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