Sensor device

Abstract

Provided is a sensor device capable of performing accurate weighing and efficient stirring of a fluid used for measurement of a test sample, a reaction control reagent, an antigen extraction reagent, an antibody reagent, etc., in a small size and at a low cost without using a large-sized mechanical mechanism, and capable of realizing high-sensitivity measurement. The sensor device comprises a sample introduction tank 319 into which a test sample is introduced, a first tank 311 sealed by a first seal lid 411, a second tank 312 sealed by a second seal lid 412, a mixing tank 313 that mixes the test sample and an immunoreagent, a first flow path 321 connected between the first tank 311 and the second tank 312, a second flow path 322 that introduces the test sample introduced into the sample introduction tank 319 to the second tank 312 via the first flow path 321, a third flow path 323 connected between the second tank 312 and the mixing tank 313, and a fourth flow path 324 that is connected to the mixing tank 313 and introduces the test sample liquid in the mixing tank 313 into a measurement unit 300.

Description

Sensor Device

[0001] The present invention relates to a sensor device, and more particularly to a sensor device that can be used in antigen quantification techniques that utilize immune reactions.

[0002] Commonly used antigen quantification techniques in clinical testing include immunoluminescence, immunoturbidimetry, immunochromatography, etc., such as ELISA (Enzyme-Linked Immunosorbent Assay), CLIA (Chemiluminescence Immunoassay), and CLEIA (Chemiluminescent Enzyme Immunoassay). Except for immunochromatography, which is widely used for antigen testing in the low-sensitivity range of antigen concentrations of ng / mL or more, immunoluminescence and turbidimetry for antigen testing in the high-sensitivity range of pg / mL require (1) a precise optical measurement system, and (2) accurate quantification of test samples and reagents and strict control of reaction time are required to ensure high sensitivity and quantitative performance, and many products require complex and expensive mechanical elements such as a dispensing mechanism and a shaking mechanism.

[0003] For this reason, such testing methods are not suitable for use as immunochromatography, where medical professionals can perform the test themselves at the bedside or in an examination room and obtain test results on the spot. In other words, conventional technology has not been able to provide a testing method that combines the high sensitivity required for many antigen tests with the simplicity and speed required to obtain results immediately in the medical field.

[0004] A technique has been disclosed that enables highly sensitive antigen quantification by electrical measurement by combining the electrical detection zone method with immune agglutination (see Patent Documents 1 and 2). This eliminates the need for an optical system, which was necessary for antigen quantification by conventional immunoluminescence, and therefore allows for the miniaturization and lower cost of the measuring device.

[0005] Japanese Patent No. 5951527 Japanese Patent Application Laid-Open No. 2022-059154

[0006] However, even if an optical system is no longer required, quantifying the antigen to be tested (hereinafter referred to as "target antigen") contained in a clinical sample remains the same as with conventional technology. Therefore, to achieve quantification of lower concentrations of target antigen and ensure high quantitation accuracy (low variability in results), it is necessary to strictly control the volume of each of the test sample, reaction control reagent, antigen extraction reagent, and antibody reagent, as well as the reaction time, just as with conventional immunoluminescence antigen quantification devices. In addition, just like conventional devices, some kind of stirring mechanism may be used to promote the antigen-antibody reaction in order to reduce the time required for the immune reaction. These are relatively complex mechanical mechanisms, which cause the device to be large and expensive.

[0007] Furthermore, even if the use of an antigen detection system based on the electrical sensing zone method makes it possible to reduce the size and cost of the detection unit, accurate control of the liquid volume and reaction time, and stirring are still required, as in the conventional method. Therefore, even in an antigen detection system based on the electrical sensing zone method, if a dispensing mechanism or stirring means made of mechanical elements is used as in the conventional method, the overall measurement device cannot be made small and low-cost.

[0008] The present invention has been made in consideration of the above circumstances, and provides a sensor device that can accurately measure and efficiently mix fluids used in clinical tests, such as test samples, reaction control reagents, antigen extraction reagents, and antibody reagents, without using large mechanical mechanisms, and that is small in size and low in cost, and that can achieve highly sensitive measurements.

[0009] According to one embodiment of the present invention, there is provided a sensor device comprising: a sample introduction tank having an opening open to the outside and into which a test sample is introduced; a first tank sealed by a first flexible sealing lid; a second tank sealed by a second flexible sealing lid; a mixing tank in which the test sample is mixed with an immunoreagent containing an antibody that immunologically binds to a target antigen that may be contained in the test sample to prepare a test sample liquid; a first flow path connected between the first tank and the second tank; a second flow path connected between the first flow path and the sample introduction tank so that the test sample introduced into the sample introduction tank is introduced into the second tank via the first flow path; a third flow path connected between the second tank and the mixing tank; and a fourth flow path connected to the mixing tank and which introduces the test sample liquid in the mixing tank to a measurement unit for measuring the presence or concentration of the target antigen in the test sample liquid.

[0010] According to another embodiment of the present invention, there is provided a measuring device comprising: a first introduction tank having a first open part open to the outside and into which a first fluid is introduced; a second introduction tank having a second open part open to the outside and into which a second fluid is introduced; a first tank sealed by a first flexible sealing lid; a second tank sealed by a second flexible sealing lid; a third tank sealed by a third flexible sealing lid; a fourth tank sealed by a fourth flexible sealing lid; a mixing tank for mixing the first fluid with a second fluid containing a component that reacts with a measurement target that may be contained in the first fluid to prepare a mixed solution; and a first flow path connected between the first tank and the second tank. Provided is a sensor device comprising: a second flow path connected between the first flow path and the first introduction tank so that a first fluid introduced into the first introduction tank is introduced into the second tank via the first flow path; a third flow path connected between the second tank and the mixing tank; a fourth flow path that introduces the mixed liquid into a measuring section that measures the presence or concentration of a measurement target that may be contained in the mixed liquid in the mixing tank; a fifth flow path connected between the third tank and the fourth tank; a sixth flow path connected between the fifth flow path and the second introduction tank so that a second fluid introduced into the second introduction tank is introduced into the fourth tank via the fifth flow path; and a seventh flow path connected between the fourth tank and the third flow path.

[0011] According to yet another embodiment of the present invention, there is provided a sensor device comprising: a first introduction tank having a first open part open to the outside and into which a first fluid is introduced; a first lid provided in the first open part and having an exhaust port; a second lid capable of opening and closing the exhaust port; a first tank sealed by a first flexible sealing lid; a second tank sealed by a second flexible sealing lid; a mixing tank in which the first fluid and a component that reacts with a substance to be measured that may be contained in the first fluid are mixed to prepare a mixed solution; a first flow path connected between the first tank and the second tank; a second flow path connected between the first flow path and the first introduction tank so that the first fluid introduced into the first introduction tank is introduced into the second tank via the first flow path; a third flow path connected between the second tank and the mixing tank; a liquid stopping unit installed midway through the third flow path; and a fourth flow path that introduces the mixed solution into a measuring unit that measures the presence or concentration of a substance to be measured that may be contained in the mixed solution in the mixing tank.

[0012] According to the present invention, a sensor device can be provided that can accurately measure and efficiently mix fluids used in clinical tests, such as test samples, reaction control reagents, antigen extraction reagents, and antibody reagents, without using large mechanical mechanisms, and that is small in size and low in cost, thereby enabling highly sensitive measurements.

[0013] Fig. 1(a) is a schematic diagram illustrating the principle of particle measurement using the electrical sensing zone method, Fig. 1(b) is an electron microscope photograph of a pore using a solid pore made of semiconductor Si, and Fig. 1(c) is a waveform diagram showing an example of a pulse signal measured when a target particle in a sample solution passes through the pore. Fig. 2(a) is an explanatory diagram showing the relationship between the state of particles in chamber 110 when the sample does not contain an antigen and the change in current value caused by the particles passing through the pore. Fig. 2(b) is an explanatory diagram showing the relationship between the state of particles in chamber 110 when the concentration of antigen in the sample is low and the change in current value caused by the particles passing through the pore. Fig. 2(c) is an explanatory diagram showing the relationship between the state of particles in chamber 110 when the concentration of antigen in the sample is higher than that in Fig. 2(b) and the change in current value caused by the particles passing through the pore. Fig. 3 is a plan view showing an example of a sensor device according to a first embodiment of the present invention. FIG. 4(a) is a cross-sectional view of the sensor device according to the first embodiment taken along the IVa-IVa section in FIG. 3 , and FIG. 4(b) is a cross-sectional view of the sensor device according to the first embodiment taken along the IVb-IVb step section in FIG. 3 . FIG. 5(a) is a cross-sectional view of the sensor device according to the first embodiment taken along the V-V section in FIG. 3 , and FIG. 5(b) is a cross-sectional view of a modified example of the sensor device according to the first embodiment taken along the V-V section in FIG. 3 . FIG. 6 is a cross-sectional view of the sensor device according to the first embodiment taken along the VI-VI section in FIG. 3 . FIGS. 7(a) and 7(b) are flowcharts illustrating an example of the operation of the sensor device according to the first embodiment and an example of a sample analysis method. FIG. 8 is a flowchart illustrating an example of the operation of the sensor device according to the first embodiment and an example of a sample analysis method. FIG. 9(a) is an explanatory diagram illustrating the positional relationship between the first flow path and the second flow path provided in the sensor device according to the first embodiment, and FIG. 9(b) is an explanatory diagram illustrating a comparative example. FIG. 10 is a plan view illustrating an example of a sensor device according to a second embodiment of the present invention. 11(a) and 11(b) are flowcharts illustrating an example of the operation of the sensor device and the sample analysis method according to the second embodiment. FIG. 12 is a flowchart illustrating an example of the operation of the sensor device and the sample analysis method according to the second embodiment.13 and 14 are cross-sectional views showing a first and second modified example of the sensor device according to the first embodiment.

[0014] The following description of the preferred embodiments of the present invention will be given with reference to the accompanying drawings. The preferred embodiments are merely examples of devices and methods for embodying the technical concept of the present invention, and the technical concept of the present invention is not limited to the following in terms of the structure, arrangement, etc. of the components.

[0015] The principle of particle measurement using the electrical sensing zone method is explained using Figures 1(a) to 1(c). Figure 1(a) shows an example 100 of the cross-sectional structure of a sensor device based on the electrical sensing zone method. Chambers 110 and 120 are connected to each other via a pore 140 opened in a partition wall 141. An electrode 112 is provided on the inner wall of chamber 110, and an electrode 122 is provided on the inner wall of chamber 120. These electrodes 112 and 122 are connected to an amplifier 150, an ammeter 151, and a voltage source 152. Chamber 110 is provided with an injection port 111, and chamber 120 is provided with an injection port 121. Figure 1(b) is an example of an electron microscope photograph of a pore portion using a solid-state pore made of semiconductor silicon. In this example, a SiN thin film with a thickness of approximately 50 nm is used as the partition wall 141. Here, the term "chamber" refers to the space spatially connected to the pore and surrounding the pore. A space through which a liquid flows is commonly called a flow channel, and the chamber may be a part of the flow channel or a space provided for the purpose of connecting the flow channel to the pore.

[0016] In the electrical sensing zone method, for example, chamber 110 of a sensor device having such a structure is filled with a sample solution in which target particles are suspended in an electrolyte, and chamber 120 is filled with the electrolyte. Then, the sample solution in chamber 110 and the electrolyte solution in chamber 120 are electrically connected via pore 140. When a voltage is applied between electrodes 112 and 122, an ionic current flows between electrodes 112 and 122 through the sample solution, pore 140, and electrolyte. When target particles 101 in the sample solution pass through pore 140, the movement of cations and anions that carry the ionic current through pore 140 is temporarily hindered, resulting in a decrease in the ionic current only while the particles are passing through. When this is observed with ammeter 151, a pulse signal 210, such as that shown in FIG. 1(c), is observed. Since the size, shape, and surface charge distribution of the particle are reflected in this pulse signal 210, it is possible to detect, identify, or quantify the particles to be measured by analyzing this pulse signal 210. In this example, the electrode 112 may be located anywhere as long as it is in contact with the sample liquid. The electrode 122 may be located anywhere as long as it is in contact with the electrolyte.

[0017] 2(a) to 2(c) are explanatory diagrams illustrating the principle of antigen quantification using the electrical detection zone method. Here, the example shows a state in which chamber 110 is filled with a liquid mixture containing antibody-modified particles 211, 212, 213, 232, and 252, the surfaces of which are modified with antibodies 202 that specifically bind to a target antigen 201 to be detected / quantified (see FIG. 2(b) ; hereinafter, also referred to as "target antigen 201"), and chamber 120 is filled with an electrolyte solution. Electrodes 112 and 122 are not shown. When the target antigen 201 is not contained in the liquid mixture filled in chamber 110, the number of antibody-modified particle aggregates 219 bound via the target antigen 201 in the liquid mixture is small, as shown in FIG. 2(a). When a voltage is applied between electrodes 112 and 122 in this state, antibody-modified particles 211, 212, and 213 in the test sample individually pass through pore 140, as shown in FIG. 2(a). 2(a) shows examples of pulse signals 221, 222, and 223 when antibody-modified particles 211, 212, and 213 pass through pore 140. Even when target antigen 201 is not contained in the mixed liquid, aggregates 219 formed by nonspecific aggregation of antibody-modified particles are present, but the number of antibody-modified particle aggregates 219 is smaller than in the examples of FIG. 2(b) and FIG. 2(c).

[0018] When target antigen 201 is present in the test sample liquid, the target antigen 201 is present in the mixed liquid in a state bound to antibody-modified particles as shown in Figures 2(b) and 2(c) in chamber 110. Figure 2(b) shows a case where the antigen concentration of target antigen 201 is low, while Figure 2(c) shows a case where the antigen concentration of target antigen 201 is high. The higher the antigen concentration, the more antibody-modified particles bind to each other, forming larger aggregates 231, 233, 251, and 253.

[0019] The waveforms of pulse signals 241, 243, 261, and 263 generated when these agglomerates 231, 233, 251, and 253 pass through pore 140 have different shapes compared to pulse signals 242 and 262 generated when single antibody-modified particles 232 and 252 pass through pore 140. This is because when large antibody-modified particles, agglomerates 231, 233, 251, and 253, pass through pore 140, the ionic current flowing through pore 140 is more strongly impeded than when small antibody-modified particles 232 and 252 pass through, and the shapes of agglomerates 231, 233, 251, and 253 are significantly different from those of single antibody-modified particles 232 and 252. For example, in Figure 2(c), the pulse signals generated when agglomerate 251, single antibody-modified particle 252, and agglomerate 253 pass through pore 140 are 261, 262, and 263, respectively. This is why the peak current values ​​of the waveforms of pulse signals 261 and 263 for agglutinate 251 and agglutinate 253 are larger than those of pulse signal 262 for single antibody-modified particle 252. Therefore, the higher the antigen concentration in the mixed solution, the higher the proportion of pulses with large peak current values ​​(e.g., pulse signals 241, 243, 261, 263 in Figures 2(b) and 2(c)) in the observed pulse signals. In the present invention, the binding / aggregation state of antibody-modified particles is inferred not only from the peak current value of the pulse signal but also from various characteristics of the pulse shape.

[0020] The antigen quantification technique based on the electrical sensing zone method shown in Figures 2(a) to 2(c) has the following two advantages over conventional optical detection techniques such as immunoluminescence, immunochromatography, and immunoturbidimetry.

[0021] First, antigen quantification techniques based on the electrical detection zone method are highly sensitive. While all conventional techniques use light (emission or absorption) to detect macroscopic state changes due to immune reactions, the method of the present invention based on the electrical detection zone method counts the bonds between antibody-modified particles one by one, making it possible to detect and quantify antigens at particularly dilute concentrations, such as those below 1 pg / mL.

[0022] Second, antigen quantification technology based on the electrical sensing zone method requires less liquid volume for the test sample and reagents. This is because antigen quantification based on the electrical sensing zone method observes the binding state of particles one by one. For example, consider the case where antibody-modified particles with a concentration of 10^9 particles / mL are diluted 10-100 times and measured. In this case, the volume per particle is 10^-8 to 10^-7 mL, or 10-100 pL. For example, the volume of liquid required to measure 1,000 pulses is approximately 10-100 nL near the pore 140. Even taking into account the volume of liquid required to fill the flow path, antigen quantification technology based on the electrical sensing zone method can significantly reduce the amount of reagent and sample required compared to conventional methods, leading to reduced reagent manufacturing costs and less invasiveness during sample collection.

[0023] Generally, antigen quantification using an immune reaction involves the use of a reaction-promoting reagent (hereinafter referred to as the "R1 reagent") and an antibody reagent (hereinafter referred to as the "R2 reagent") containing an antibody that immunologically binds to the target antigen. First, the R1 reagent is mixed with a clinical specimen to allow for reaction. Next, the R2 reagent is mixed with the clinical specimen mixed with the R1 reagent to induce an immune reaction, followed by measurement. To shorten the test time, the immune reaction may be accelerated by, for example, shaking the R1 reaction solution and R2 reagent after mixing. Here, samples collected from the human body, such as blood (including whole blood, plasma, serum, etc.), pus, and saliva, are referred to as "clinical specimens." These samples, once processed to a state suitable for introduction into the sensor device of the present invention, are referred to as "test samples." Examples of test samples include, but are not limited to, samples prepared by mixing the R1 reagent or R2 reagent with a clinical specimen, or samples prepared by destroying microorganisms such as viruses and bacteria in a pathogen-containing clinical specimen to extract antigens specific to the microorganisms to be quantified. The term "test sample" refers to any sample in a state suitable for introduction into the sensor device of the present invention.

[0024] In such clinical sample processing during measurement, to improve quantitation, i.e., to reduce variability between measurements, it is necessary to control the liquid volumes of the clinical sample, R1 reagent, and R2 reagent, as well as the reaction times of the R1 reagent and R2 reagent, so as to prevent variability between measurements. For this reason, in antigen quantitative tests using conventional technology, the liquid volumes and reaction times of the clinical sample, R1 reagent, and R2 reagent have been controlled by a complex dispensing mechanism of an automated machine before being injected into the sensor. For this reason, devices for highly sensitive antigen quantitative tests using conventional technology have generally been large, complex, and expensive.

[0025] As described above, antigen quantification using the electrical sensing zone method does not require an optical system, allowing for significantly simplified and compact measurement unit configuration. However, if a complex dispensing mechanism similar to that of conventional technology were used to control the immune reaction, the advantages of the electrical sensing zone method's simple and compact measurement system would not be fully utilized. The sensor device according to the embodiment of the present invention includes a reaction control unit 310 that can easily and accurately control the volume of the test sample, R1 reagent, and R2 reagent, the reaction time, and even the promotion of the immune reaction through stirring, all on a small, inexpensive sensor device. This eliminates the need for users to control the reaction time of the test sample and R2 reagent using an automated device, etc., and provides a sensor device that can achieve highly sensitive antigen detection or quantification easily and at low cost.

[0026] 3 is a plan view showing an example of a sensor device according to a first embodiment of the present invention. The sensor device according to the embodiment of the present invention includes a measurement unit 300 that utilizes the electrical sensing zone method and a reaction control unit 310 that can control the reaction of a first fluid (test sample) introduced into the measurement unit 300. The reaction control unit 310 accurately measures and collects a predetermined volume of the test sample after mixing the clinical specimen with the R1 reagent, mixes it with the R2 reagent, which is a solid-phase immunoreagent, and then stirs it for a certain period of time. The measurement unit 300 performs antigen quantitative measurement using the electrical sensing zone method.

[0027] The reaction control unit 310 includes a first introduction tank 319 (hereinafter also referred to as "sample introduction tank 319") into which a first fluid is introduced, a first tank 311 whose top is sealed by a flexible first sealing lid 411, a second tank 312 whose top is sealed by a flexible second sealing lid 412, and a mixing tank 313 that mixes the first fluid with a component that reacts with the object to be measured (target antigen) that may be contained in the first fluid to prepare a mixed solution. The reaction control unit 310 further includes a first flow path 321 connected between the first tank 311 and the second tank 312, a second flow path 322 connected between the first flow path 321 and the first introduction tank 319 so as to introduce the first fluid introduced into the first introduction tank 319 into the second tank 312 via the first flow path 321, a third flow path 323 connected between the second tank 312 and the mixing tank 313, and a fourth flow path 324 connected to the mixing tank 313 and introducing the mixed liquid in the mixing tank 313 into the measurement unit 300. The measurement unit 300 includes an opening (e.g., exhaust ports 3033, 304) that leads to the outside of the sensor device any flow path, drainage path, exhaust port, or tank within the measurement unit 300 for observing the target antigen or analyte that may be contained in the test sample liquid or mixed liquid flowing in from the fourth flow path 324 using the electrical sensing zone method. This allows the test sample liquid or the mixture liquid to be introduced appropriately from the reaction control unit 310 to the measurement unit 300.

[0028] Figure 4(a) shows an example of a cross-sectional view of a sensor device according to an embodiment of the present invention taken along the IVa-IVa cross section in Figure 3, and Figure 4(b) shows an example of a cross-sectional view of a sensor device according to an embodiment of the present invention taken along the IVb-IVb stepped cross section in Figure 3. As shown in Figure 4(b), the tops of first tank 311 and second tank 312 are sealed with flexible first sealing lid 411 and second sealing lid 412, respectively. As shown in Figure 4(a), the top of mixing tank 313 is sealed with third sealing lid 413.

[0029] The first sealing lid 411 and the second sealing lid 412 may be formed from a material that is sufficiently deformable (displaceable) when pressed downward, thereby allowing gas in the first tank 311 to be introduced into the second tank 312. For example, the first sealing lid 411 and the second sealing lid 412 may be formed from an elastic material such as silicone rubber, butyl rubber, ethylene-propylene rubber, ethylene-propylene-diene rubber, urethane rubber, fluororubber, acrylic rubber, butadiene rubber, styrene-butadiene rubber, acrylonitrile-butadiene rubber, chloroprene rubber, isoprene rubber, natural rubber, and / or thermoplastic elastomer. The first sealing lid 411 and the second sealing lid 412 are formed from a material that is flexible and water-resistant. The first sealing lid 411 and the second sealing lid 412 may also be formed from a material that is transparent enough to allow a user to view the inside of the sensor device from above.

[0030] The thickness of the first sealing lid 411 and the second sealing lid 412 is not particularly limited, but may be a thickness that ensures sufficient airtightness to seal the first tank 311 and the second tank 312, for example, 0.05 to 2 mm, or in one embodiment, 0.1 to 1 mm. The first sealing lid 411 and the second sealing lid 412 may have a layered structure in which multiple materials are layered. The third sealing lid 413 is formed from a material that is not flexible so as to keep the volume of the mixing tank 313 constant. The third sealing lid 413 is made of a material similar to that of the base material that constitutes the sensor device.

[0031] FIG. 5( a) shows an example of a cross-sectional view of the sample introduction reservoir 319. The top of the sample introduction reservoir 319 has a first open section 318 that is open to the outside, and a first fluid introduced into the sample introduction reservoir 319 is connected to a first flow path 321 via a second flow path 322. A test sample that may contain a target antigen to be measured by the measurement unit 300 is used as the first fluid. As shown in FIG. 5( b), the sample introduction reservoir 319 may include a first lid 501 that is provided in the first open section 318 and has an exhaust port 503, and a second lid 502 that is provided on the first lid 501 and can open or close the exhaust port 503. As shown in FIG. 5( b), by providing the first lid 501 and the second lid 502 above the sample introduction reservoir 319, the opening and closing of the exhaust port 503 can be controlled. This allows for accurate measurement of the liquid introduced from the sample introduction reservoir 319 to the second reservoir 312.

[0032] The first tank 311 is filled with a gas such as air. When the first fluid introduced into the sample introduction tank 319 fills the second tank 312 and the sample introduction tank 319 is sealed, for example, by the first lid 501 and the second lid 502, and the first sealing lid 411 is pressed downward (depressed) from the outside, the gas in the first tank 311 flows into the second tank 312 via the first flow path 321. When the gas flows into the second tank 312, the first fluid in the second tank 312 is pushed out, and the first fluid flows into the mixing tank 313 via the third flow path 323. In the example of FIG. 4( a ), a solid-phase immunoreagent 314, for example, an R2 reagent containing an antibody that immunologically binds to a target antigen that may be contained in the test sample, is disposed in the mixing tank 313. When the first fluid flows into the mixing tank 313, the test sample is mixed with the immunoreagent 314 to produce a test sample liquid in the mixing tank 313. In this way, the first sealing lid 411 is displaced from above to below the first tank 311 by an operator's finger or a pressing member (not shown) so as to push out the air in the first tank 311, thereby transferring the first fluid from the second tank 312 to the mixing tank 313. In this embodiment, the term "displacement" refers to a partial or total change in the position of a member to which an external force is applied from its initial position, such as when an external force is applied to a flexible material and the object bends. The specific configuration of the first sealing lid 411 is not limited as long as it is capable of pressing the gas in the first tank 311 to an extent that the gas in the first tank 311 can be forced into the second tank 312.

[0033] When the first sealing lid 411 is pressed down and the test sample is mixed with the immunoreagent 314 in the mixing chamber 313, if the downward pressing of the first sealing lid 411 is released, the test sample liquid in the mixing chamber 313 returns to the second chamber 312 via the third flow path 323. By repeatedly pressing down and releasing the first sealing lid 411 in this manner, the test sample liquid moves back and forth between the second chamber 312 and the mixing chamber 313, thereby being agitated. By repeating the downward pressing and releasing of the first sealing lid 411 in this manner for a predetermined reaction time, the first sealing lid 411 is displaced up and down above the first chamber 311, thereby agitating the test sample liquid.

[0034] When the second sealing lid 412 is pressed downward from the outside while the sample introduction reservoir 319 is sealed and the first sealing lid 411 is pressed downward in a state in which the test sample is mixed with the immunoreagent 314 in the mixing reservoir 313, the second sealing lid 412 is displaced so that the test sample liquid in the mixing reservoir 313 is introduced from the mixing reservoir 313 to the measurement unit 300 via the fourth flow path 324. The specific configuration of the second sealing lid 412 is not limited as long as it has a configuration that can press the first fluid in the second reservoir 312 so as to push the gas in the second reservoir 312 into the mixing reservoir 313. According to the sensor device according to the first embodiment of the present invention, the transfer of the first fluid introduced into the sample introduction reservoir 319 and the mixing of the first fluid with the immunoreagent 314 can be easily controlled by pressing or releasing the downward pressing of the flexible first sealing lid 411 and second sealing lid 412 and displacing the first sealing lid 411 and second sealing lid 412.

[0035] As shown in FIG. 3, the first flow path 321 linearly connects the first tank 311 and the second tank 312, and the second flow path 322 is preferably connected to the first flow path 321 so as to merge with the first flow path 321 from the side of the first flow path 321, i.e., from a direction intersecting the extension direction of the first flow path 321.

[0036] The third flow path 323 preferably includes a liquid stopping structure for stopping the progress of the first fluid from the second tank 312 to the mixing tank 313. As an example of the liquid stopping structure, the third flow path 323 connecting the second tank 312 and the mixing tank 313 has a via hole 333 therethrough, and the third flow path 323 on the second tank 312 side and the mixing tank 313 side with respect to the via hole 333 are configured as two different layers. The first fluid that has flowed into the second tank 312 proceeds through the third flow path 323 but stops just before the via hole 333 due to surface tension. Due to this liquid stopping structure in the third flow path 323, the first fluid that has accumulated near the second tank 312 is stopped from progressing from the second tank 312 to the mixing tank 313 until the second sealing lid 412 is pressed down.

[0037] The liquid stopping structure in the third flow path 323 may have any configuration other than two or more flow paths passing through the via hole 333. For example, it may be a bent portion of 270 degrees or more formed in the third flow path 323, preferably in the same layer, or may be a pillar or bump formed so as to protrude from the inner wall surface of the third flow path 323. Alternatively, it may be a roughened layer formed on the inner wall surface of the third flow path 323 and having projections and depressions formed by surface roughening treatment or the like.

[0038] The immunoreagent 314 contained in the mixing chamber 313 is, for example, granular antibody-modified particles immobilized thereon. The location of the immunoreagent 314 in the mixing chamber 313 is not particularly limited.

[0039] The measuring unit 300 includes an upstream flow path liquid reservoir 302 connected to the fourth flow path 324, an upstream flow path connecting flow path 350 connected to the bottom of the upstream flow path liquid reservoir 302 (see Figure 6), an upstream flow path electrode liquid contact portion 351 provided on the underside of the upstream flow path connecting flow path 350 and to which the upstream flow path electrode 301 is connected, an exhaust port 3033 connected to the upstream flow path connecting flow path 350, an upstream flow path 303 connected to the upstream flow path liquid reservoir 302, a liquid stop tank 357 connected to the upstream flow path 303, an upstream flow path exhaust port 304 connected to the liquid stop tank 357, a downstream flow path 308 connected below the upstream flow path 303 via a fine hole 399 provided in the upstream flow path 303, a downstream flow path introduction tank 307 into which an electrolyte is introduced, the downstream flow path 308 connected to the downstream flow path introduction tank 307 and the fine hole 399, a downstream flow path electrode liquid contact portion 356 connected to the downstream flow path 308 and to which the downstream flow path electrode 306 is connected, and an absorbent pad 309.

[0040] The sensor device of FIG. 3 may have, for example, a five-layer stacked structure as shown in FIG. 6 . The upstream channel reservoir 302 extends in the depth direction from the first layer to the fourth layer. As shown in FIG. 3 , the first layer (the top layer) is hollowed out in the shape of an elliptical cylinder, and the second to fourth layers below it are hollowed out in the shape of smaller cylinders. By having the upstream channel reservoir 302 thus have a shape in which the cross-sectional area increases from bottom to top, the liquid can be retained above the upstream channel reservoir 302 without overflowing even when the amount of liquid introduced from the fourth channel 324 is excessive. Furthermore, the dimensions and shape of the upstream channel reservoir 302 can be determined so that the area below the upstream channel reservoir 302 is small so that the upstream channel reservoir 302 does not interfere with the upstream channel connection channel 350 or the pore chip 317 arranged adjacent to the upstream channel reservoir 302. This allows the measurement unit 300 to be further miniaturized. The upstream flow path liquid reservoir 302 may have a cross-sectional area that differs for each layer in relation to the flow paths adjacent to the upstream flow path liquid reservoir 302, or may have a continuous inner wall surface that tapers from the lower layer to the upper layer.

[0041] An absorbent pad 309 is connected to the downstream flow path 308 to absorb the liquid introduced into the downstream flow path 308 and adjust the amount of liquid. If the level of the liquid introduced into the downstream flow path 308 is not constant, the head pressure applied to the pores 399 will change, which may cause unstable measurement. If a large amount of liquid is introduced into the downstream flow path 308, the absorbent pad 309 absorbs the excess liquid, so that the liquid level in the downstream flow path 308 is always stable regardless of the amount of liquid introduced into the downstream flow path 308. This suppresses changes in the head pressure applied to the pores 399, allowing particles to pass through the pores stably.

[0042] The operation of the sensor device according to the first embodiment of the present invention will be described using the sensor devices shown in FIGS. 3 to 6 and the flowcharts shown in FIGS. 7A, 7B, and 8. First, a test sample is introduced into the sample introduction reservoir 319 shown in FIG. 3, for example, by dripping (step S101). The test sample moves from the sample introduction reservoir 319 shown in FIG. 3 through the second flow path 322, the confluence 320, and then the first flow path 321 to the second reservoir 312. When the second reservoir 312 is filled with the test sample, the movement of the test sample stops (step S102). At this time, the first reservoir 311 has no opening connected to the outside and is airtight. Meanwhile, the second reservoir 312 is connected to the flow path of the measurement unit 300, which has an opening connected to the outside, via the third flow path 323, the mixing reservoir 313, and the fourth flow path 324, allowing for exhaust of air. Therefore, in step S102, the test sample flows from the confluence 320 only to the second tank 312, not to the first tank 311, and the second tank 312 is filled with the test sample. Regardless of the amount of test sample introduced into the sample introduction tank 319, a certain amount of test sample determined by the volume of the second tank 312 is measured in step S102. Furthermore, the first tank 311 is in an airtight state.

[0043] After step S102 is completed, the sample introduction reservoir 319 is sealed (step S103). Next, with the sample introduction reservoir 319 sealed, the test sample is placed in the second reservoir 312 and waits for the predetermined R1 reaction time (step S104). Next, with the test sample held in the second reservoir 312, the first sealing lid 411 on top of the first reservoir 311 is pressed down (step S105). Because the first sealing lid 411 is made of a flexible material, pressing down the first sealing lid 411 moves gas, such as air, sealed in the first reservoir 311 to the second reservoir 312 via the first flow path 321. When the first reservoir 311 is pressed down by the first sealing lid 411, the gas sealed in the first reservoir 311 flows into the second reservoir 312, and the test sample in the second reservoir 312 is pushed out by the gas and moves into the mixing reservoir 313 via the third flow path 323 (step S105).

[0044] In general, the progress of immune reactions is highly sensitive to the liquid volume and reaction time. Test samples and R2 reagents are optimized to ensure accuracy and performance only when the specified liquid volume and reaction time are strictly adhered to. In this embodiment, the specified amounts of test samples and reagents can be measured more accurately by employing at least one of the following three techniques:

[0045] The first technique is a liquid stop structure used in step S102 when filling the test sample. For example, as shown in FIG. 4B, a via hole 333 is provided midway along the third flow path 323 connecting the second tank 312 and the mixing tank 313. The third flow path 323 on the second tank 312 side and the third flow path 323 on the mixing tank 313 side are formed in different layers relative to the via hole 333. In step S102, the test sample flowing into the second tank 312 stops just before the via hole 333 due to surface tension. This liquid stop structure in the third flow path 323 prevents the test sample from proceeding into the mixing tank 313 in step S102, allowing the specified amount of test sample to be filled into the second tank 312 to be accurately measured.

[0046] The second technique is a two-stage sealing process that can be performed in step S103. In this embodiment, the first open portion 318 of the sample introduction reservoir 319 can be closed in two stages using the first lid 501 and second lid 502 shown in FIG. 5B. Specifically, after step S102 is completed, the top of the sample introduction reservoir 319 is first closed with the first lid 501 having an exhaust port 503. Air remaining in the top of the sample introduction reservoir 319 is then exhausted through the exhaust port 503. The exhaust port 503 is then closed with the second lid 502. This prevents air remaining in the sample introduction reservoir 319 from forming bubbles and flowing into the second reservoir 312. Furthermore, if the sample introduction reservoir 319 were closed in one stage in step S103, pressure would be applied to the test sample filling the second reservoir 312 during the closing operation, which could cause the test sample to overflow the liquid stopping structure and enter the mixing reservoir 313. This two-stage closing of the sample introduction reservoir 319 functions more effectively for accurate measurement of the liquid in the second reservoir 312 .

[0047] The third technology is an optimized structure for preventing air from entering through the sample introduction reservoir 319. In this embodiment, the sample introduction reservoir 319, the first reservoir 311 functioning as a pump, and the second reservoir 312 for measuring the test sample are configured as shown in FIG. 9A . As a result, at the completion of step S102, the test sample is filled in the second flow path 322, the first flow path 321 from the confluence 320 to the second reservoir 312, and the second reservoir 312. Therefore, when the first sealing lid 411 of the first reservoir 311 is pressed down in step S105, the air in the first reservoir 311 is separated from the test sample liquid at the confluence 320, and the test sample in the second reservoir 312 can be pushed toward the mixing reservoir 313 without generating bubbles. If the sample introduction reservoir 319, first reservoir 311, and second reservoir 312 have a configuration as shown in FIG. 9( b), i.e., if the sample introduction reservoir 319 and second reservoir 312 are connected by a flow path 721 and the first reservoir 311 is connected from the side of the flow path 721 via a flow path 722, in step S105, air from the first reservoir 311 may enter both the sample introduction reservoir 319 and the second reservoir 312 connected to the first flow path 321, causing bubbles to form in the second reservoir 312. In this case, even if the first reservoir 311 is pressed down, liquid may not be properly delivered to the mixing reservoir 313. In this embodiment, this problem can be avoided by using a configuration in which the second flow path 322 joins the first flow path 321 from the side relative to the length direction of the first flow path 321 (the direction of fluid flow), as shown in FIG. 9( a).

[0048] The test sample transferred to the mixing chamber 313 is mixed with dried antibody-modified particles (immunoreagent 314) already present on the inner wall surface of the mixing chamber 313, forming a test sample liquid, which is a mixture of the antibody-modified particles dispersed in the test sample (step S106). The antibody-modified particles can be beads having antibodies bound to their surfaces that immunologically bind to the target antigen. If the test sample contains the target antigen in step S106, a reaction occurs in which the antibody-modified particles bind to each other via the target antigen, as schematically illustrated in FIG. 2.

[0049] Stirring the mixture is effective for promoting immune reactions in the mixture, allowing for quantification of more dilute target antigens, or for shortening the reaction time required for antigen quantification. A method for promoting immune reactions will be described using the flowchart in Figure 7(b). In step S106, the sample introduction reservoir 319 is sealed and the first sealing lid 411 is pressed down, resulting in the mixture being in the mixing reservoir 313. When the first sealing lid 411 is opened from this state, the fourth flow path 324 is opened, and the mixture in the mixing reservoir 313 returns to the second reservoir 312 via the third flow path 323 (step S111). From this point, the first sealing lid 411 is repeatedly pressed down and opened over a predetermined R2 reaction time, causing the mixture to move back and forth between the second reservoir 312 and the mixing reservoir 313, thereby stirring the mixture (step S112). The mixture is stirred for the R2 reaction time, and if the R2 reaction time has elapsed in step S113, the process proceeds to step S114.

[0050] In one embodiment of the present invention shown in FIG. 3 , a via hole 333 is provided in the third flow path 323. Therefore, when the mixed liquid travels back and forth between the second tank 312 and the mixing tank 313 in step S112, it may pass through this via hole 333. In this embodiment, an element that obstructs the passage of the mixed liquid is provided in the third flow path 323, thereby generating turbulence as the mixed liquid passes through the third flow path 323, thereby enabling more efficient mixing. The liquid retention structure provided in the third flow path 323 may be any structure, such as a via hole, a pillar, or a fin. Furthermore, such a liquid retention structure may be provided not only in the second flow path but also in the second tank 312 or the mixing tank 313.

[0051] The test liquid, which is the mixture after the R2 reaction time has elapsed after stirring the mixture by the method of step S112, will be referred to below as the “reacted mixture.” When the first sealing lid 411 is pressed down and the reacted mixture is in the mixing tank 313, and the second sealing lid 412 is further pressed down (step S114), the reacted mixture moves to the measurement unit 300 via the fourth flow path 324 (step S115).

[0052] In this way, the sensor device according to the embodiment of the present invention measures out a specified amount of test sample mixed with R1 required for the test sample (step S102), mixes the test sample with R1 and then reacts for a specified time (step S104), mixes the test sample with R1 after the reaction with R1, mixes with R2 reagent (step S106), and then stirs the mixture for a specified time to ensure an efficient immune reaction (step S112). Such processing can be easily performed on the small and inexpensive sensor device according to the present embodiment.

[0053] Next, the process of antigen quantification based on the electrical detection zone method in the measurement unit 300 will be described with reference to FIG. 8 . In step S121, the reacted mixture is sent from the fourth flow path 324 to the upstream flow reservoir 302. In the example shown in FIG. 3 , the upper portion of the upstream flow reservoir 302 is open, and the intake and exhaust performed in steps S101 to S106 and S111 to S115 may be performed through the open portion of the upstream flow reservoir 302. Next, the reacted mixture that has entered the upstream flow reservoir 302 flows into the upstream flow path 303 and reaches the liquid stop tank 357 via the upper portion of the pore 399 (step S121). The upstream flow electrode liquid contact portion 351 is connected to the bottom of the upstream flow reservoir 302. When the reacted mixture is introduced from the fourth flow path 324, the upstream flow reservoir 302, the upstream flow path 303, and the upstream flow electrode liquid contact portion 351 are filled with the reacted mixture. The upper part of the upstream flow path exhaust port 304 is open, and even if the upstream flow path liquid reservoir 302 does not have an open structure at the top, the liquid supply control in the reaction control unit 310 can be performed without any problems by the intake and exhaust from the upstream flow path exhaust port 304. In this embodiment, the upstream flow path electrode liquid contact part 351 is connected to the bottom of the upstream flow path connecting flow path 350 due to the installation position of the upstream flow path electrode 301, but it may be located anywhere as long as it comes into contact with the reacted mixed liquid in contact with the pore 399.

[0054] Next, when the electrolyte is introduced into the downstream flow path introduction tank 307, the electrolyte passes through the downstream flow path 308 and contacts the lower part of the pore 399, the downstream flow path electrode liquid contact portion 356, thereby filling the downstream flow path 308 (step S122). In this state, the reacted mixed solution filled in the upstream flow path 303 and the electrolyte solution filled in the downstream flow path 308 come into contact through the pore 399. This establishes electrical conductivity between the upstream flow path electrode 301 and the downstream flow path electrode 306 through the reacted mixed solution and the electrolyte (step S123). In this state, when a voltage is applied between the upstream flow path electrode 301 and the downstream flow path electrode 306 (step S124), as described with reference to FIG. 1(a), particles in the reacted mixed solution in the upstream flow path 303 move through the pore 399 into the electrolyte solution in the downstream flow path 308. Each time an antibody-modified particle passes through the pore 399, a pulse signal 210, as illustrated in FIG. 1(c), is observed (step S125). In this embodiment, the shape of this pulse signal is analyzed to estimate the concentration of the target antigen in the reacted mixture (step S126).

[0055] The sensor device according to the first embodiment of the present invention can perform all of the steps that were previously performed using complex and expensive dispensing and shaking mechanisms, including measuring a fixed amount of test sample, stirring, reaction time control, and highly sensitive antigen quantification using the electrical detection zone method, all within a small sensor measuring just a few centimeters square.

[0056] 3 illustrates, as one embodiment of the present invention, a sensor device in which dried R2 reagent 314 is fixed in advance in a mixing tank 313. Next, Fig. 10 illustrates an example of a sensor device in which a liquid reagent is used as the immune reagent, that is, an R2 reagent in which the liquid solvent contains an antibody or in which antibody-modified particles are dispersed.

[0057] 10 is an explanatory diagram showing a sensor device according to the second embodiment. The sensor device includes a first introduction tank 919 having a first open part 918 whose top is open to the outside and into which a first fluid is introduced, a second introduction tank 949 having a second open part 948 whose top is open to the outside and into which a second fluid is introduced, a first tank 911 whose top is sealed by a first sealing lid 811, a second tank 912 whose top is sealed by a second sealing lid 812, a third tank 951 whose top is sealed by a third sealing lid 831, a fourth tank 952 whose top is sealed by a fourth sealing lid 832, and a mixing tank 913 in which the first fluid and the second fluid are mixed to prepare a mixed solution.

[0058] The sensor device further includes a first flow path 921 connected between the first tank 911 and the second tank 912, a second flow path 922 connected between the first flow path 921 and the first introduction tank 919 and introducing the first fluid introduced into the first introduction tank 919 into the second tank 912 via the first flow path 921, a third flow path 923 connected between the second tank 912 and the mixing tank 913, and a fourth flow path 924 connected to the mixing tank 913 and introducing the mixed liquid in the mixing tank 913 into the measuring section 900. The sensor device further includes a fifth flow path 941 connected between the third tank 951 and the fourth tank 952, a sixth flow path 942 connected between the fifth flow path 941 and the second introduction tank 949 so as to introduce the second fluid introduced into the second introduction tank 949 into the fourth tank 952 via the fifth flow path 941, and a seventh flow path 943 connected between the fourth tank 952 and the third flow path 923.

[0059] 10 , the fourth flow path 924 is connected to an upstream flow path reservoir 972 of the measuring unit 900. In this example, the upper part of the upstream flow path reservoir 972 is open, allowing intake and exhaust of air from there. In the example of Fig. 10 , the third flow path 923 and the seventh flow path 943 are connected via a confluence such as a via hole 933 formed in the third flow path 923.

[0060] The first sealing lid 811, the second sealing lid 812, the third sealing lid 831, and the fourth sealing lid 832 are each flexible and have a configuration substantially similar to the first sealing lid 411 and the second sealing lid 412 of Fig. 3. The cross-sectional structures of the first introduction reservoir 919 and the second introduction reservoir 949 are substantially similar to the cross-sectional structure of the sample introduction reservoir 319 of Fig. 3. The structures of the first flow path 921 and the fifth flow path 941 are substantially similar to the structure of the first flow path 321 of Fig. 3, and the structures of the second flow path 922 and the fifth flow path 941 are substantially similar to the structure of the second flow path 322 of Fig. 3. The structure of the fourth flow path 924 is also substantially similar to the structure of the fourth flow path 324 of Fig. 3.

[0061] The second flow path 922 is preferably connected to the first flow path 921 so as to merge with the first flow path 921 from the side of the first flow path 921. Furthermore, the sixth flow path 942 is preferably connected to the fifth flow path 941 so as to merge with the fifth flow path 941 from the side of the fifth flow path 941. By employing such a configuration, the first fluid and the second fluid can be more appropriately measured while preventing air from entering the second tank 912 and the fourth tank 952.

[0062] The first fluid may be a test sample including a clinical specimen. The second fluid may be any reagent used in clinical testing, such as a reagent containing an antibody or an antibody reaction reagent that promotes an antibody reaction, for example, a reaction control reagent, an antigen extraction reagent, or an antibody reagent. The first and second fluids may be selected in various ways depending on the test. For example, a reagent may be introduced into the first introduction tank 919, which introduces the first fluid, and a test sample may be introduced into the second introduction tank 949, which introduces the second fluid.

[0063] 5(b), the first open portion 918 and the second open portion 948 of the first introduction tank 919 and the second introduction tank 949 are preferably provided with a first lid 501 having an exhaust port 503 and a second lid 502 that can open or close the exhaust port 503. The first lid 501 and the second lid 502 allow the first open portion 918 and the second open portion 948 of the first introduction tank 919 and the second introduction tank 949 to be closed and controlled in two stages, thereby enabling accurate measurement of the liquid. Also, in the sensor device of FIG. 10, it is preferable to provide a liquid stopper consisting of two flow paths in different layers, for example, via a via hole 933, midway along the third flow path 923.

[0064] The measurement unit 900 includes an upstream flow path reservoir 972 connected to the fourth flow path 924, an upstream flow path 971 connected to the upstream flow path reservoir 972, a liquid stop tank 974 connected to the upstream flow path 971, an upstream flow path exhaust port 975 connected to the liquid stop tank 357, a downstream flow path introduction tank 982 into which the electrolyte is introduced, a downstream flow path 981 connected to the upstream flow path 971 via a pore 999, a downstream flow path electrode 986 connected to the downstream flow path 981, and an absorption pad 983 connected to the downstream flow path 981 and absorbing excess electrolyte flowing through the downstream flow path 981 to maintain a constant liquid level in the downstream flow path 981. In this embodiment, the upstream flow path electrode 973 contacts the reacted mixed liquid at the bottom of the upstream flow path reservoir 972. The downstream flow path electrode 986 and the electrolyte filling the downstream flow path contact each other at a liquid contact portion 984.

[0065] When the first sealing lid 811 is pressed downward in a state in which the second tank 912 is filled with the first fluid introduced into the first introduction tank 919 and the first open portion 918 is sealed, the gas in the first tank 911 flows into the second tank 912 via the first flow path 921, and the first fluid in the second tank 912 flows into the mixing tank 913 via the third flow path 923. When the fourth tank 952 is filled with the second fluid introduced into the second introduction tank 949 and the second open portion 948 is sealed, and the third sealing lid 831 is pressed downward, the gas in the third tank 951 flows into the fourth tank 952 via the fifth flow path 941, and the fluid in the fourth tank 952 flows into the mixing tank 913 via the seventh flow path 943 and the third flow path 923. As a result, the first and third sealing lids 811 and 831 are displaced so that the first and second fluids that have flowed into the mixing chamber 913 are mixed.

[0066] Furthermore, when the first open portion 918 and the second open portion 948 are sealed, a mixed liquid of the first fluid and the second fluid is contained in the mixing tank 913, and the first sealing lid 811 and the third sealing lid 831 are pressed downward, and the downward pressing of the first sealing lid 811 and the third sealing lid 831 is released, the mixed liquid in the mixing tank 913 returns to the second tank 912 and the fourth tank 952 via the third flow path 923. In this way, by repeatedly pressing the first sealing lid 811 and the third sealing lid 831 downward and releasing the downward pressing, the first sealing lid 811 and the third sealing lid 831 are displaced so that the mixed liquid is stirred while reciprocating between the second tank 912 and the mixing tank 913 and between the fourth tank 952 and the mixing tank 913.

[0067] Furthermore, when the first open portion 918 and the second open portion 948 are sealed, a mixed liquid of the first fluid and the second fluid is contained in the mixing tank 913, and the first sealing lid 811 and the third sealing lid 831 are pressed downward, and at least one of the second sealing lid 812 and the fourth sealing lid 832 is pressed downward, the mixed liquid in the mixing tank 913 is introduced from the mixing tank 913 to the measuring section 900 via the fourth flow path 924.

[0068] The operation of the sensor device according to the second embodiment of the present invention will be described using the sensor device shown in FIG. 10 and the flowcharts shown in FIGS. 11(a), 11(b), and 12. First, a test sample is introduced as a first fluid into the first introduction reservoir 919 shown in FIG. 10, for example, by dripping (step S1001). The test sample moves from the first introduction reservoir 919 shown in FIG. 10 through the second flow path 922 and the first flow path 921 to the second reservoir 912. When the second reservoir 912 is filled with the test sample, the movement of the test sample stops (step S1002). At this time, the first reservoir 911 has no opening connected to the outside and is airtight. Meanwhile, the second reservoir 912 is connected to the flow path of the measurement unit 900, which has an opening connected to the outside, via the third flow path 923, the mixing reservoir 913, and the fourth flow path 924, allowing for exhaust. Therefore, in step S1002, the test sample flows from the confluence of first flow path 921 and second flow path 922 only toward second tank 912, not toward first tank 911, and second tank 912 is filled with the test sample. Regardless of the amount of test sample introduced into first introduction tank 919, a constant amount of test sample determined by the volume of second tank 912 is measured in step S1002.

[0069] Similarly, a reagent such as R2 reagent is introduced as a second fluid into the second introduction reservoir 949 of FIG. 10 by, for example, dripping (step S1003). The R2 reagent moves from the second introduction reservoir 949 of FIG. 10 through the sixth flow path 942 and the fifth flow path 941 to the fourth reservoir 952. When the fourth reservoir 952 is filled with the reagent, the movement of the reagent stops (step S1004). In this manner, in this embodiment, the liquid volumes of the test sample liquid and the R2 reagent can be accurately measured. The volumes of the test sample liquid and the R2 reagent can be freely set by the volumes of the second reservoir 912 and the fourth reservoir 932, respectively.

[0070] After completing step S1004, the tops of the first introduction reservoir 919 and the second introduction reservoir 949 are sealed (step S1005). The progress of the immune reaction is highly sensitive to the liquid volume and reaction time. The test sample and R2 reagent are optimized to achieve accuracy and performance only when the specified liquid volume and reaction time are precisely adhered to. After completing step S1005, the first introduction reservoir 919 is sealed, and the test sample is placed in the second reservoir 912 and waits for the specified R1 reaction time (step S1006). Next, with the test sample held in the second reservoir 912, the first sealing lid 811 on top of the first reservoir 911 is pressed down, and with the R2 sample held in the fourth reservoir 952, the third sealing lid 831 on top of the third reservoir 951 is pressed down, preferably simultaneously (step S1007). Note that, to appropriately control the reaction time, it is preferable to push down the first sealing lid 811 and the third sealing lid 831 almost simultaneously, but they do not have to be pushed down exactly simultaneously. Because the first sealing lid 811 and the third sealing lid 831 are made of a flexible material, pushing down the first sealing lid 811 moves gas such as air sealed in the first tank 911 to the second tank 912 via the first flow path 921. The R2 sample sealed in the third tank 951 moves to the fourth tank 952 via the fifth flow path 941. Furthermore, by pushing down the second sealing lid 812 and the fourth sealing lid 832 while the first sealing lid 811 and the third sealing lid 831 are pressed down, the first fluid and the second fluid are introduced into the mixing tank 913, and a mixture of the first and second fluids is prepared in the mixing tank 913 (step S1007).

[0071] As in the description of step S105, in this embodiment, the second flow path 922 joins from the side of the first flow path 921, and the sixth flow path 942 joins from the side of the fifth flow path 941. As a result, when the first sealing lid 811 and the third sealing lid 831 are pressed down, the test sample liquid measured in the second tank 912 and the R2 reagent measured in the fourth tank 952 can be sent to the mixing tank 913 without generating air bubbles.

[0072] An immunological reaction occurs between the target antigen and the antibody in the R2 reagent in the mixture prepared in the mixing tank 913. In order to promote this immune reaction and quantify a more dilute target antigen, or to shorten the reaction time and the time required to quantify the antigen, it is effective to stir the mixture.

[0073] A method for promoting an immune response according to a second embodiment of the present invention will be described using the flowchart in Figure 11(b). In step S1007, the first introduction tank 919 and the second introduction tank 949 are sealed, and the first sealing lid 811 and the third sealing lid 831 are pressed downward, resulting in the mixed liquid being in the mixing tank 913. When the first sealing lid 811 and the third sealing lid 831 are opened from this state, the fourth flow path 924 is opened, and the mixed liquid in the mixing tank 913 returns to the fourth tank 952 via the third flow path 923, the second tank 912, the third flow path 923, and the seventh flow path 943 (step S1011). From this point, the first sealing lid 811 and the third sealing lid 831 are pressed downward and opened, and the mixed liquid is moved back and forth between the second tank 912 and the mixing tank 913 and between the fourth tank 952 and the mixing tank 913, thereby agitating the mixed liquid (step S1012). The mixed solution is stirred for the R2 reaction time, and if the R2 reaction time has elapsed in step S1013, the process proceeds to step S1014.

[0074] 10, a via hole 933 is provided in the third flow path 923. Therefore, in step S1012, the mixed liquid passes through this via hole 933 when traveling back and forth between the second tank 912 and the fourth tank 952 and the mixing tank 913. In this way, by providing an element that obstructs the passage of the mixed liquid in the third flow path 923 and generating turbulence when the mixed liquid passes through the third flow path 923, more efficient stirring can be achieved. The liquid retention structure provided in the third flow path 923 can have a configuration similar to that described for the sensor device of FIG. 3.

[0075] With the first and third sealing lids 811 and 831 pressed down and the reacted mixture in the mixing tank 913, at least one of the second and fourth sealing lids 812 and 832 is pressed down, preferably both, or more preferably both simultaneously (step S1014). Then, the reacted mixture moves to the measuring unit 900 via the fourth flow path 924 (step S1015).

[0076] In the sensor device according to the second embodiment of the present invention, a predetermined amount of test sample, which is a mixture of a clinical specimen and R1 reagent, is measured out (step S1002), a predetermined amount of R2 reagent is measured out (step S1004), the test sample is held in the second tank for a predetermined reaction time for the R1 reagent (step S1006), and then this is mixed with the R2 reagent held in the fourth tank (S1007). Furthermore, the mixture is stirred for a predetermined time to ensure an efficient immune reaction (S1012-S1013). This process can be easily performed on the compact and inexpensive sensor device according to this embodiment, and the reacted mixture obtained as a result of the process can be introduced into the measurement unit 900, enabling highly sensitive and rapid antigen quantification using the electrical detection zone method.

[0077] The process of antigen quantification based on the electrical detection zone method in the measurement unit 900 of FIG. 10 will be described with reference to FIG. 12 . The reacted mixture is sent from the fourth flow path 924 to the upstream flow reservoir 972. Next, the reacted mixture that has entered the upstream flow reservoir 972 enters the upstream flow path 971, reaches the pore 999, and reaches the liquid stop tank 974 via the upper part of the pore 999 (step S1021). By introducing the reacted mixture from the fourth flow path 924, the upstream flow reservoir 972, the upstream flow path 971, and the upstream flow electrode liquid contact portion 351 are filled with the reacted mixture. The upstream flow path exhaust port 975 is open at the top. Therefore, even if the upstream flow path reservoir 972 does not have an open structure at the top, the intake and exhaust from the upstream flow path exhaust port 975 allows the reaction control unit 910 to smoothly control the liquid supply.

[0078] Next, when the electrolyte is introduced into the downstream flow path introduction tank 982, the electrolyte passes through the downstream flow path 981, contacts the pore 999 and the downstream flow path electrode liquid contact portion 984, and fills the downstream flow path 981 (step S1022). The reacted mixed solution filled in the upstream flow path 971 and the electrolyte solution filled in the downstream flow path 981 come into contact through the pore 999. This brings the upstream flow path electrode 873 and the downstream flow path electrode 986 into electrical conduction through the reacted mixed solution and the electrolyte (step S1023). In this state, when a bias voltage is applied between the upstream flow path electrode 873 and the downstream flow path electrode 986 (step S1024), the particles in the reacted mixed solution in the upstream flow path 971 move through the pore 999 into the electrolyte solution in the downstream flow path 981. A pulse signal is measured each time the antibody-modified particles pass through the pore 999 (step S1025). By analyzing the shape of this pulse signal, the concentration of the target antigen in the reacted mixture is estimated (step S1026).

[0079] The sensor device according to the second embodiment of the present invention can perform all of the steps that were previously performed using complex and expensive dispensing and shaking mechanisms, including measuring a fixed amount of test sample, stirring, reaction time control, and highly sensitive antigen quantification using the electrical detection zone method, all within a small sensor measuring just a few centimeters square.

[0080] Although the present invention has been described with reference to the above-described embodiments, the descriptions and drawings that form part of this disclosure should not be understood to limit the present invention. The present disclosure is not limited to the above-described embodiments, and components can be combined and modified to be embodied within the scope of the gist of the present disclosure.

[0081] For example, in the sensor devices shown in Figures 3 and 10, the measurement units 300, 900 and the reaction control units 310, 910 are mounted on the same device, but the measurement units 300, 900 and the reaction control units 310, 910 may be fabricated as separate devices and then integrated by assembly. Alternatively, a module for each of the measurement units 300, 900 may be fabricated and used in conjunction with an existing chip or the like that utilizes an electrical sensing zone method. In Figure 10, as with the sensor device of Figure 3, it is also possible to pre-fill the mixing chamber 913 with a solid-phased R2 reagent, and then introduce any other reagent, such as a reaction accelerator, from the second introduction reservoir 949.

[0082] In the example shown in Fig. 3, the first and second flexible sealing lids 411 and 412 are installed on the tops of the first and second tanks 311 and 312, respectively. Similarly, in the example shown in Fig. 10, the first and second flexible sealing lids 811, 812, 831, and 832 are installed on the tops of the first and second tanks 911, 912, 951, and 952, respectively. However, the first and second sealing lids 411, 811, 412, 812, 831, and 832 may be installed not only on the tops but also on the bottoms of the tanks. That is, as shown in Fig. 13, the first and second tanks 311 and 312 may have openings at the bottom, and the first and second sealing lids 411 and 412 may be installed to seal the openings of the first and second tanks 311 and 312. For example, when sending air in the first tank 311 to the second tank 312, the first sealing lid 411 may be displaced by pushing (pressing) it upward from the outside.

[0083] 13 , the pressing mechanisms (first sealing lid 411, second sealing lid 412) for controlling the introduction of the test sample into the mixing chamber 313 and the stirring of the test sample are arranged together on one side of the sensor device, while the introduction mechanism for the test sample and reagent into the sensor device (sample introduction chamber 319) is arranged on the other side of the sensor device from the viewpoint of ease of dropping the test sample and reagent, etc. This configuration prevents interference between the pressing mechanisms of the first sealing lid 411 and second sealing lid 412 and the introduction mechanisms for the test sample and reagent, thereby realizing a more compact measuring device.

[0084] As shown in FIG. 14 , the upstream path electrode 301 and the downstream path electrode 306 may be disposed on the upper surface of the measurement unit 300, or one may be disposed on the upper surface and the other on the lower surface. In cases where the flow path configurations on the lower layer of the sensor device are close to each other, disposing the upstream path electrode 301 on the upper surface of the sensor device allows for a more compact sensor device. Similarly, the upstream path reservoir 302 may be offset from each other in the vertical direction of the page of FIG. 14 (the longitudinal direction of the sensor chip) so that the upper layer portion is disposed closer to the fourth flow path 324 and the lower layer portion is disposed closer to the suction pad 30. As described above, this embodiment may include various aspects that can be modified in design within the scope of common technical knowledge of a person skilled in the art.

[0085] 101: Particle to be detected 110, 120: Chamber 111, 121: Inlet 112, 122: Electrode 140: Pore 141: Partition 150: Amplifier 151: Ammeter 152: Voltage source 201: Antigen to be detected / quantified 202: Antibody 210, 261, 262, 263: Pulse signal 211, 212, 213, 232, 252: Antibody-modified particle 219, 231, 233, 251, 253: Aggregate 221, 222, 241, 242, 243: Pulse signal 300: Measurement unit 301: Upstream flow path electrode 302: Upstream flow path liquid reservoir 303: Upstream flow path 304: Exhaust port (upstream flow path exhaust port) 306: Downstream flow path electrode 307: Downstream flow path introduction tank 308: Downstream flow path 310, 910: Reaction control unit 311, 911: First tank 312, 912: Second tank 313, 913: Mixing tank 314: Immunoreagent (R2 reagent) 317: Pore tip 318, 918: First open part 319, 919: First introduction tank (sample introduction tank) 320: Confluence part 321, 921: First flow path 322, 922: Second flow path 323, 923: Third flow path 324, 924: Fourth flow path 333: Via hole 350: Upstream flow path connecting flow path 351: Upstream flow path electrode liquid contact part 356: Downstream flow path electrode liquid contact part 357: Liquid stopping tank 399: Pore 411, 811: First sealing lid 412, 812: Second sealing lid 501: First lid 502: Second lid 503: Exhaust port 831: Third sealing lid 832: Fourth sealing lid 833: Via hole 873: Upstream path electrode 900: Measurement section 933: Via hole 941: Fifth path 942: Sixth path 943: Seventh path 948: Second open section 949: Second introduction reservoir 951: Third reservoir 952: Fourth reservoir 971: Upstream path 972: Upstream path liquid reservoir 974: Liquid stopping reservoir 975: Upstream path exhaust port 981: Downstream path 982: Downstream path introduction reservoir 983: Absorbent pad 984: Downstream path electrode liquid contact section 986: Downstream path electrode 999: Pore

Claims

a mixing chamber in which the test sample is mixed with an immunoreagent containing an antibody that immunologically binds to a target antigen that may be contained in the test sample to prepare a test sample liquid; a first flow path connected between the first chamber and the second chamber; a second flow path connected between the first flow path and the sample introduction chamber so as to introduce the test sample introduced into the sample introduction chamber into the second chamber via the first flow path; a third flow path connected between the second chamber and the mixing chamber; and a fourth flow path connected to the mixing chamber for introducing the test sample liquid in the mixing chamber to a measurement section for measuring the presence or concentration of the target antigen in the test sample liquid.

2. The sensor device described in claim 1, characterized in that when the first sealing lid is pressed from the outside while the test sample introduced into the sample introduction tank is filling the second tank and the sample introduction tank is sealed, the first sealing lid is displaced so that gas in the first tank flows into the second tank via the first flow path and the test sample in the second tank flows into the mixing tank via the third flow path, thereby mixing the test sample with the immunoreagent in the mixing tank.

3. The sensor device described in claim 1, characterized in that when the first sealing lid is pressed from the outside while the test sample introduced into the sample introduction reservoir is filling the second reservoir and the sample introduction reservoir is sealed, the gas in the first reservoir flows into the second reservoir via the first flow path, and the test sample in the second reservoir flows into the mixing reservoir via the third flow path, thereby mixing the test sample with the immunoreagent in the mixing reservoir; when the pressure on the first sealing lid is released while the test sample is mixed with the immunoreagent in the mixing reservoir, the test sample liquid in the mixing reservoir returns to the second reservoir via the third flow path; and by repeatedly pressing and releasing the first sealing lid, the first sealing lid is displaced so that the test sample liquid is stirred as it moves back and forth between the second reservoir and the mixing reservoir.

4. The sensor device described in claim 1, characterized in that when the first sealing lid is pressed from the outside while the test sample introduced into the sample introduction reservoir is filling the second reservoir and the sample introduction reservoir is sealed, the first sealing lid is displaced so that gas in the first reservoir flows into the second reservoir via the first flow path and the test sample in the second reservoir flows into the mixing reservoir via the third flow path, thereby mixing the test sample with the immunoreagent in the mixing reservoir; and when the second sealing lid is pressed from the outside while the sample introduction reservoir is sealed and the first sealing lid is pressed, the second sealing lid is displaced so that the test sample liquid in the mixing reservoir is introduced from the mixing reservoir via the fourth flow path to the measurement section.

5. A sensor device described in any one of claims 1 to 4, characterized in that the third flow path is provided with a liquid stopping structure for stopping the progress of the test sample from the second tank to the mixing tank.

6. The sensor device described in claim 5, characterized in that the liquid stopping structure includes any of: at least two flow paths composed of at least two different layers passing through a via hole; a bent portion formed in the third flow path; a pillar or bump formed so as to protrude from the inner wall surface of the third flow path; and a roughened layer formed on the inner wall surface of the third flow path.

7. A first introduction tank having a first open part open to the outside and into which a first fluid is introduced; a second introduction tank having a second open part open to the outside and into which a second fluid is introduced; a first tank sealed by a first flexible sealing lid; a second tank sealed by a second flexible sealing lid; a third tank sealed by a third flexible sealing lid; a fourth tank sealed by a fourth flexible sealing lid; a mixing tank for mixing the first fluid with the second fluid containing a component that reacts with a substance to be measured that may be contained in the first fluid to prepare a mixed liquid; a first flow path connected between the first tank and the second tank; a second flow path connected between the first flow path and the first introduction tank so as to introduce the first fluid introduced into the first introduction tank into the second tank via the first flow path; a third flow path connected between the second tank and the mixing tank; and a fourth flow path for introducing the mixed liquid into a measuring section for measuring the presence or absence or concentration of the substance to be measured that may be contained in the mixed liquid in the mixing tank. a fifth flow path connected between the third tank and the fourth tank; a sixth flow path connected between the fifth flow path and the second introduction tank so as to introduce the second fluid introduced into the second introduction tank into the fourth tank via the fifth flow path; and a seventh flow path connected between the fourth tank and the third flow path.

8. The sensor device according to claim 7, characterized in that when the first sealing lid is pressed from the outside in a state in which the first fluid introduced into the first introduction tank is filled in the second tank and the first open portion is sealed, the gas in the first tank flows into the second tank via the first flow path and the first fluid in the second tank flows into the mixing tank via the third flow path; when the second fluid introduced into the second introduction tank is filled in the fourth tank and the second open portion is sealed, the gas in the third tank flows into the fourth tank via the fifth flow path and the fluid in the fourth tank flows into the mixing tank via the seventh flow path and the third flow path, and the first and third sealing lids are displaced so that the first and second fluids that have flowed into the mixing tank are mixed.

9. The sensor device described in claim 7 or 8, characterized in that when the first open portion and the second open portion are sealed, a mixed liquid of the first fluid and the second fluid is contained in the mixing tank, and the first sealing lid and the third sealing lid are pressed, when the pressure on the first sealing lid and the third sealing lid is released, the mixed liquid in the mixing tank returns to the second tank and the fourth tank via the third flow path, and the first sealing lid and the third sealing lid are displaced by repeatedly pressing and releasing the pressure on the first sealing lid and the third sealing lid so that the mixed liquid is stirred by reciprocating between the second tank and the mixing tank and between the fourth tank and the mixing tank.

10. A sensor device as described in claim 7 or 8, characterized in that when the second sealing lid and the fourth sealing lid are pressed from the outside while the first open portion and the second open portion are sealed, a mixed liquid of the first fluid and the second fluid is contained in the mixing tank, and the first sealing lid and the third sealing lid are pressed, at least one of the second sealing lid and the fourth sealing lid is displaced so that the mixed liquid in the mixing tank is introduced from the mixing tank through the fourth flow path to the measuring section.

11. The sensor device according to claim 7 or 8, characterized in that the sixth flow path is connected to the fifth flow path so as to merge with the fifth flow path from the side of the fifth flow path.

12. A sensor device comprising: a first introduction tank having a first open part open to the outside and for introducing a first fluid; a first lid provided in the first open part and having an exhaust port; a second lid capable of opening and closing the exhaust port; a first tank sealed by a first flexible sealing lid; a second tank sealed by a second flexible sealing lid; a mixing tank for mixing the first fluid and a component that reacts with a substance to be measured that may be contained in the first fluid to prepare a mixed liquid; a first flow path connected between the first tank and the second tank; a second flow path connected between the first flow path and the first introduction tank so as to introduce the first fluid introduced into the first introduction tank into the second tank via the first flow path; a third flow path connected between the second tank and the mixing tank; a liquid stopping section provided midway through the third flow path; and a fourth flow path for introducing the mixed liquid into a measuring section for measuring the presence or absence or concentration of a substance to be measured that may be contained in the mixed liquid in the mixing tank.

13. A sensor device described in any one of claims 1 to 4, 7, 8 and 12, characterized in that the second flow path is connected to the first flow path so as to merge with the first flow path from the side of the first flow path.

14. A sensor device as described in claim 7 or 8, characterized in that the first fluid includes a test sample including a clinical specimen, and the second fluid includes a reagent including an antibody or an antibody reaction reagent that promotes an antibody reaction.

15. A sensor device as described in any one of claims 1, 7, 8 and 12, characterized in that the measurement section is provided with a flow path and a pore for observing a target antigen or analyte that may be contained in the test sample liquid or mixed liquid flowing in from the fourth flow path using an electrical detection zone method.

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