Sensor device

Abstract

To provide a sensor device capable of achieving accurate measurement and efficient agitation of fluid used to measure a test sample, a reaction control reagent, an antigen extraction reagent, and an antibody reagent, etc., in a small size at low cost without using a large-sized machine mechanism, and high-sensitivity measurement.SOLUTION: A sensor device comprises: a sample introduction tank 319 into which a test sample is introduced; a first tank 311 sealed by a first sealing lid 411; a second tank 312 sealed by a second sealing lid 412; a mixing tank 313 for mixing a test sample and an immunological reagent; a first channel 321 connected between the first tank 311 and the second tank 312; a second channel 322 for introducing the test sample introduced into the sample introduction tank 319 into the second tank 312 via the first channel 321; a third channel 323 connected between the second tank 312 and the mixing tank 313; and a fourth channel 324 connected to the mixing tank 313 and introducing a test sample liquid in the mixing tank 313 into a measuring unit 300.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to a sensor device, and more particularly to a sensor device that can be used in an antigen quantification technique that utilizes an immune reaction, for example. [Background technology]

[0002] Antigen quantification techniques commonly used in clinical testing include immunoluminescence, immunoturbidimetry, immunochromatography, etc., such as ELISA (Enzyme-Linked Immunosorbent Assay), CLIA (Chemiluinescense 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, in order to realize antigen testing in the high sensitivity range of pg / mL using immunoluminescence or turbidimetry, (1) a precise optical measurement system is required, and (2) to ensure high sensitivity and quantitation, accurate quantification of test samples and reagents and strict control of reaction time are required, and in many products, complex and expensive mechanical elements such as a dispensing mechanism and a shaking mechanism are indispensable.

[0003] For this reason, such testing methods are not suitable for use as immunochromatography, where medical professionals perform the tests themselves at the bedside or in the 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 technology 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 makes it unnecessary to use the optical system that was necessary for antigen quantification by conventional immunoluminescence, and therefore makes it possible to reduce the size and cost of the measuring device. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent No. 5951527 [Patent Document 2] JP 2022-059154 A Summary of the Invention [Problem to be solved by the invention]

[0006] However, even if the optical system is no longer necessary, the quantification of the antigen to be tested (hereinafter referred to as the "target antigen") contained in the clinical specimen is still performed in the same way as with conventional technology. Therefore, in order to realize the quantification of lower concentrations of the target antigen and ensure high quantitation (small variation in results), it is necessary to strictly control the liquid volumes of the test sample, reaction control reagent, antigen extraction reagent, and antibody reagent, as well as the reaction time, as with conventional antigen quantification devices using immunoluminescence. In addition, as with 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 causes the device to become large and expensive.

[0007] In addition, even if the detection unit can be made smaller and less expensive by using an antigen detection system based on the electric sensing zone method, accurate control of the liquid volume and reaction time, and stirring are still required as in the conventional method. Therefore, even if the antigen detection system based on the electric sensing zone method is used, if a dispensing mechanism or stirring means made of mechanical elements is used as in the conventional method, the measurement device as a whole cannot be made smaller and less expensive.

[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 the use of large mechanical mechanisms, and is small and low-cost, thereby enabling highly sensitive measurements. [Means for solving the problem]

[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 immune reagent 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 as to introduce the test sample introduced into the sample 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 connected to the mixing tank for introducing the test sample liquid in the mixing tank to a measurement section 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 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 liquid, 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 as to introduce a 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 fourth flow path that introduces the mixed liquid into a measurement section for measuring 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 as to introduce a 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.

[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 for mixing the first fluid with 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 stopper provided midway through the third flow path; and a fourth flow path for introducing the mixed liquid into a measurement unit 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. Effect of the Invention

[0012] According to the present invention, it is possible to provide a sensor device that can accurately measure and efficiently mix fluids used in clinical testing, such as test samples, reaction control reagents, antigen extraction reagents, and antibody reagents, without the use of large mechanical mechanisms, and that is small in size and low cost, thereby enabling highly sensitive measurements to be achieved. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1(a) is a schematic diagram explaining the principle of particle measurement by 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 liquid passes through the pore. [Diagram 2]Figure 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 due to particles passing through the pore, Figure 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 due to particles passing through the pore, and Figure 2(c) is an explanatory diagram showing the relationship between the state of particles in chamber 110 and the change in current value due to particles passing through the pore when the concentration of antigen in the sample is higher than in Figure 2(b). [Diagram 3] FIG. 3 is a plan view showing an example of the sensor device according to the first embodiment of the present invention. [Figure 4] 4(a) is a cross-sectional view of the sensor device according to the first embodiment taken along the IVa-IVa cross section of 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 cross section of FIG. 3. [Diagram 5] 5(a) is a cross-sectional view of a sensor device according to the first embodiment taken along the VV section of 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 VV section of FIG. 3. [Figure 6] FIG. 6 is a cross-sectional view of the sensor device according to the first embodiment taken along the line VI-VI of FIG. [Figure 7] 7(a) and 7(b) are flowcharts illustrating an example of the operation of the sensor device and the method of analyzing a sample according to the first embodiment. [Figure 8] FIG. 8 is a flowchart illustrating an example of the operation of the sensor device and the method of analyzing a sample according to the first embodiment. [Figure 9] FIG. 9(a) is an explanatory diagram showing the positional relationship between the first flow path and the second flow path included in the sensor device according to the first embodiment, and FIG. 9(b) is an explanatory diagram showing a comparative example. [Figure 10] FIG. 10 is a plan view illustrating an example of a sensor device according to the second embodiment of the present invention. [Figure 11]11(a) and 11(b) are flowcharts illustrating an example of the operation of the sensor device and the method of analyzing a sample according to the second embodiment. [Figure 12] FIG. 12 is a flowchart illustrating an example of the operation of the sensor device and the method of analyzing a sample according to the second embodiment. [Figure 13] FIG. 13 is a cross-sectional view showing a first modified example of the sensor device according to the first embodiment. [Figure 14] FIG. 14 is a cross-sectional view showing a second modified example of the sensor device according to the first embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The following describes the embodiments of the present invention with reference to the drawings. The embodiments described below 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 structures and arrangements of the components described below.

[0015] The principle of particle measurement by the electrical sensing zone method will be described with reference to Fig. 1(a) to Fig. 1(c). Fig. 1(a) is an example 100 of a cross-sectional structure of a sensor device based on the electrical sensing zone method. A chamber 110 and a chamber 120 communicate with each other through a fine hole 140 opened in a partition wall 141. An electrode 112 is provided on the inner wall of the chamber 110, and an electrode 122 is provided on the inner wall of the chamber 120, and these electrodes 112 and 122 are connected to an amplifier 150, an ammeter 151, and a voltage source 152. An injection port 111 is provided in the chamber 110, and an injection port 121 is provided in the chamber 120. Fig. 1(b) is an example of an electron microscope photograph of a fine hole using a solid pore made of semiconductor Si. In this example, a SiN thin film with a thickness of about 50 nm is used as the partition wall 141. Here, the term chamber refers to a space near the fine hole that is spatially connected to the fine hole. A space through which a liquid flows is commonly called a flow path, and the chamber may be a part of the flow path or a space provided for the purpose of connecting the flow path to the pore.

[0016] In the electrical sensing zone method, for example, the chamber 110 of the sensor device having such a structure is filled with a sample solution in which target particles are suspended in an electrolyte, and the chamber 120 is filled with the electrolyte. Then, the sample solution in the chamber 110 and the electrolyte in the chamber 120 are electrically connected via the pore 140. When a voltage is applied between the electrodes 112 and 122, an ion current flows between the electrodes 112 and 122 through the sample solution, the pore 140, and the electrolyte. Here, when the target particle 101 in the sample solution passes through the pore 140, the movement of the cations and anions that carry the ion current through the pore 140 is temporarily hindered, and as a result, the ion current decreases only while the particle passes through. When this is observed with the ammeter 151, for example, a pulse signal 210 as 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 so long as it is in contact with the sample liquid. The electrode 122 may be located anywhere so long as it is in contact with the electrolyte.

[0017] 2(a) to 2(c) are explanatory diagrams for explaining the principle of antigen quantification by the electric detection zone method. Here, an example is shown in which the chamber 110 is filled with a mixed liquid containing antibody-modified particles 211, 212, 213, 232, and 252, the surfaces of which are modified with antibodies 202 that specifically bind to a detection / quantification target antigen 201 (see FIG. 2(b) hereinafter, also referred to as "target antigen 201"), and the chamber 120 is filled with an electrolyte. Electrodes 112 and 122 are omitted from the illustration. When the mixed liquid filled in the chamber 110 does not contain the target antigen 201, the number of aggregates 219 of antibody-modified particles bound via the target antigen 201 in the mixed liquid is small, as shown in FIG. 2(a). When a voltage is applied between the electrodes 112 and 122 in this state, the antibody-modified particles 211, 212, and 213 in the test sample pass through the pores 140 individually, 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 in which antibody-modified particles specifically aggregate with each other exist, 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 bound to antibody-modified particles in chamber 110 as shown in Figures 2(b) and 2(c). 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 to form larger aggregates 231, 233, 251, 253.

[0019] The waveforms of pulse signals 241, 243, 261, 263 generated when these aggregates 231, 233, 251, 253 pass through the pore 140 change in shape compared to pulse signals 242, 262 generated when single antibody-modified particles 232, 252 pass through the pore 140. This is because when the large antibody-modified particles, aggregates 231, 233, 251, 253, pass through the pore 140, the ion current flowing through the pore 140 is more greatly hindered than when the small antibody-modified particles 232, 252 pass through, and the shapes of the aggregates 231, 233, 251, 253 are significantly different from those of the single antibody-modified particles 232, 252. For example, in FIG. 2(c), the pulse signals generated when the aggregate 251, the single antibody-modified particle 252, and the aggregate 253 pass through the pore 140 are 261, 262, and 263, respectively. This is the reason why the peak current values ​​of the waveforms of pulse signals 261, 263 of agglutinate 251 and further of agglutinate 253 are greater than the waveform of pulse signal 262 of a single antibody-modified particle 252. Therefore, the higher the antigen concentration in the mixed solution, the higher the ratio of pulses with large peak current values ​​(e.g., pulse signals 241, 243, 261, 263 in Figures 2(b) and 2(c)) to the observed pulse signals. In the present invention, the binding / aggregation state of antibody-modified particles is inferred from various characteristics of the pulse shape, not just the peak current value of the pulse signal.

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

[0021] First, the antigen quantification technology based on the electrical detection zone method is highly sensitive. While all conventional techniques use light (light emission or light absorption) to detect macroscopic state changes caused by 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 low concentrations, such as those with an antigen concentration of 1 pg / mL or less.

[0022] Secondly, the antigen quantification technology based on the electric detection zone method requires less liquid volume of the test sample and reagent. This is because the antigen quantification based on the electric detection zone method observes the binding state of the particles one by one. For example, let us consider an example 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^-8-10^-7mL, that is, 10-100pL. For example, the amount of liquid required to measure 1000 pulses is about 10-100nL near the pore 140. Even taking into account the amount of liquid required to fill the flow path, the antigen quantification technology based on the electric detection zone method can significantly reduce the amount of reagent and sample compared to conventional methods, leading to reduced manufacturing costs of reagents and reduced invasiveness when collecting samples.

[0023] In general, in the process of quantifying antigens using immune reactions, a reaction promotion reagent (hereinafter referred to as "R1 reagent") and an antibody reagent (hereinafter referred to as "R2 reagent") containing an antibody that immunologically binds to the target antigen are used. First, the R1 reagent is mixed with the clinical specimen to cause a reaction. Next, the R2 reagent is mixed with the clinical specimen mixed with the R1 reagent to cause an immune reaction, and then measurement is performed. In order to shorten the test time, the immune reaction may be accelerated by mixing the R1 reaction liquid and the R2 reagent and then shaking. Here, blood (including whole blood, plasma, serum, etc.), pus, saliva, and other samples collected from the human body are called "clinical specimens". The samples that are processed to be introduced into the sensor device of the present invention are called "test samples". For example, test samples include samples in which the R1 reagent or R2 reagent is mixed with the clinical specimen, samples in which microorganisms such as viruses and bacteria in a clinical specimen containing pathogens are destroyed and antigens specific to the microorganisms to be quantified are extracted, and the like, but are not limited to these, and any sample that can be introduced into the sensor device of the present invention is called a test sample.

[0024] In the clinical specimen processing during such measurements, in order to improve the quantitative performance, i.e., to suppress the variation from measurement to measurement, it is necessary to control the clinical specimen, the liquid volume of the R1 reagent and the R2 reagent, and also the reaction time of the R1 reagent and the R2 reagent so that they do not vary from measurement to measurement. For this reason, in antigen quantitative tests using conventional technology, the liquid volume and reaction time of the clinical specimen, the R1 reagent and the R2 reagent are 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 are generally large, complex and expensive.

[0025] As described above, antigen quantification by the electrical detection zone method does not require an optical system, and the configuration of the measurement unit can be greatly simplified and miniaturized. However, if a complex dispensing mechanism similar to that of the conventional technology is used for immune reaction control, the characteristics of the simple and compact measurement system of the electrical detection zone method cannot be utilized. The sensor device according to the embodiment of the present invention is provided with a reaction control unit 310 that can easily and accurately control the liquid volume of the test sample, the R1 reagent, and the R2 reagent, the reaction time, and even the promotion of the immune reaction by stirring on a small and inexpensive sensor device. This makes it possible to provide a sensor device that can easily and inexpensively realize highly sensitive antigen detection or quantification without the need for a user to control the reaction time using an automatic machine or the like for the test sample and the R2 reagent.

[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 uses an electrical detection 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 specified volume of the test sample obtained by mixing a clinical specimen with an R1 reagent, mixes the test sample with an R2 reagent, which is a solid-phased immune reagent, and then stirs the mixture for a certain period of time. The measurement unit 300 performs quantitative antigen measurement using the electrical detection zone method.

[0027] The reaction control unit 310 comprises 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 upper part is sealed by a flexible first sealing lid 411, a second tank 312 whose upper part is sealed by a flexible second sealing lid 412, and a mixing tank 313 in which a component that reacts with the object to be measured (target antigen) that may be contained in the first fluid is mixed with the first fluid to prepare a mixed liquid. 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 from any flow path, drainage path, exhaust port, or tank in 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 an electric detection 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] Fig. 4(a) shows an example of a cross-sectional view of a sensor device according to an embodiment of the present invention along the IVa-IVa cross section in Fig. 3, and Fig. 4(b) shows an example of a cross-sectional view of a sensor device according to an embodiment of the present invention along the IVb-IVb stepped cross section in Fig. 3. As shown in Fig. 4(b), the first tank 311 and the second tank 312 are sealed at their tops by a first sealing lid 411 and a second sealing lid 412, which are flexible, respectively. As shown in Fig. 4(a), the top of the mixing tank 313 is sealed by a third sealing lid 413.

[0029] The first sealing lid 411 and the second sealing lid 412 may be formed of a material that is displaceable (deformable) to such an extent that the first sealing lid 411 and the second sealing lid 412 are pressed downward from above and bend, thereby allowing the 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 of an elastic member 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 of a material that is flexible and water-resistant. The first sealing lid 411 and the second sealing lid 412 may also be formed of a material that is transparent to such an extent that a user can see 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 sealing to seal the first tank 311 and the second tank 312, for example, 0.05 to 2 mm, and in one embodiment, 0.1 to 1 mm. The first sealing lid 411 and the second sealing lid 412 may have a laminated structure in which a plurality of materials are laminated. The third sealing lid 413 is formed of 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 constituting the sensor device.

[0031] FIG. 5(a) shows an example of a cross-sectional view of the sample introduction tank 319. The upper part of the sample introduction tank 319 has a first open part 318 that is open toward the outside, and the first fluid introduced into the sample introduction tank 319 is connected to the first flow path 321 via the 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 tank 319 may include a first lid 501 that is provided in the first open part 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 two lids, the first lid 501 and the second lid 502, above the sample introduction tank 319, the opening and closing of the exhaust port 503 can be controlled. This allows accurate measurement of the liquid introduced from the sample introduction tank 319 to the second tank 312.

[0032] The first tank 311 is filled with a gas such as air. When the first sealing lid 411 is pressed downward from the outside in a state in which the first fluid introduced into the sample introduction tank 319 is filled into the second tank 312 and the sample introduction tank 319 is sealed by, for example, the first lid 501 and the second lid 502, 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), the solid-phased immunoreagent 314, for example, the R2 reagent containing an antibody that immunologically binds to a target antigen that may be contained in the test sample, is placed in the mixing tank 313. When the first fluid flows into the mixing tank 313, a test sample liquid in which the test sample is mixed with the immunoreagent 314 is prepared in the mixing tank 313. In this manner, 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, for example, when an external force is applied to a flexible material and the object is bent. The specific configuration of the first sealing lid 411 is not limited as long as it has a configuration capable of pressing the gas in the first tank 311 to such an extent that the gas in the first tank 311 can be pressed 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 pressing down 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 repeating the pressing down and releasing of 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 and is stirred. By repeating the pressing down 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 stirring the test sample liquid.

[0034] When the second sealing lid 412 is pressed downward from the outside while the sample introduction tank 319 is sealed and the first sealing lid 411 is pressed downward in a state where the test sample is mixed with the immunoreagent 314 in the mixing tank 313, the second sealing lid 412 is displaced so that the test sample liquid in the mixing tank 313 is introduced from the mixing tank 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 tank 312 so that the gas in the second tank 312 can be pushed into the mixing tank 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 tank 319 and the mixing with the immunoreagent 314 can be easily controlled by pressing or releasing the pressing of the flexible first sealing lid 411 and second sealing lid 412 downward to displace 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 is preferably provided with 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 in the middle, 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 with two different layers. The first fluid that has flowed into the second tank 312 proceeds through the third flow path 323, but stops before the via hole 333 due to surface tension. Due to such a liquid stopping structure in the third flow path 323, the first fluid that is retained near the inside of the second tank 312 is stopped from proceeding 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 third flow path 323 may have any configuration other than two or more flow paths passing through via hole 333. For example, it may be a bent portion of 270 degrees or more formed in third flow path 323, preferably in the same layer, or it may be a pillar or bump formed so as to protrude from the inner wall surface of third flow path 323. Alternatively, it may be a roughened layer formed on the inner wall surface of third flow path 323 and having projections and recesses 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 in the mixing chamber 313 where the immunoreagent 314 is placed 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 FIG. 6), an upstream flow path electrode liquid contact portion 351 provided on the lower surface 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, a 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 a laminated structure of, for example, five layers as shown in FIG. 6. The upstream flow path reservoir 302 extends in the depth direction from the first layer to the fourth layer, and as shown in FIG. 3, the first layer (top layer) is hollowed out in an elliptical cylindrical shape, and the second to fourth layers below it are hollowed out in a cylindrical shape smaller than the first layer. In this way, the upstream flow path reservoir 302 has a shape in which the cross-sectional area increases from bottom to top, so that even if the amount of liquid introduced from the fourth flow path 324 is too large, the liquid can be held on the upper side of the upstream flow path reservoir 302 so as not to overflow. In addition, the dimensions and shape of the upstream flow path reservoir 302 are determined so that the lower part of the upstream flow path reservoir 302 is small so that the upstream flow path reservoir 302 does not interfere with the upstream flow path connecting flow path 350 and the pore chip 317 arranged adjacent to the upstream flow path reservoir 302, and the measurement unit 300 can be made smaller. 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 has a tapered shape 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 changes, which may cause measurement to become unstable. 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 with reference to the sensor device of FIGS. 3 to 6 and the flow charts of FIGS. 7(a), 7(b) and 8. First, a test sample is introduced into the sample introduction reservoir 319 of FIG. 3 by, for example, dropping (step S101). The test sample moves from the sample introduction reservoir 319 of FIG. 3 through the second flow path 322, the junction 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 does not have an opening connected to the outside, and the air is sealed. On the other hand, the second reservoir 312 is connected to the flow path of the measurement unit 300 having an opening connected to the outside via the third flow path 323, the mixing reservoir 313 and the fourth flow path 324, and can be exhausted. Therefore, in step S102, the test sample flows from the junction 320 not toward first tank 311 but toward second tank 312 only, and second tank 312 is filled with the test sample. Regardless of the amount of test sample introduced into sample introduction tank 319, a certain amount of test sample determined by the volume of second tank 312 is measured in step S102. Also, first tank 311 is in an airtight state.

[0043] After completing step S102, the sample introduction reservoir 319 is sealed (step S103). Next, with the sample introduction reservoir 319 sealed, the test sample is stored in the second reservoir 312 and waits for a predetermined R1 reaction time (step S104). Next, with the test sample held in the second reservoir 312, the first sealing lid 411 on the top of the first reservoir 311 is pressed down (step S105). Since 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 tank 311 is pressed downward by the first sealing lid 411, the gas sealed in the first tank 311 flows into the second tank 312, and the test sample in the second tank 312 is pushed out by the gas and moves into the mixing tank 313 via the third flow path 323 (step S105).

[0044] In general, the progress of immune reactions is highly sensitive to the volume of liquid and the reaction time. The test sample and R2 reagent are optimized so that they can exhibit accuracy and performance only when they are used while strictly adhering to the specified volume of liquid and reaction time. In this embodiment, the test sample and reagent can be measured in a specified amount more accurately by adopting at least one of the first to third techniques described below.

[0045] The first technique is a liquid stopping structure used in step S102 when filling the test sample. For example, as shown in FIG. 4(b), a via hole 333 is provided midway in 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 with respect to the via hole 333. The test sample flowing into the second tank 312 in step S102 stops just before the via hole 333 due to surface tension. Due to such a liquid stopping structure in the third flow path 323, the test sample does not proceed to the mixing tank 313 in step S102, so that the specified amount of the test sample to be filled in the second tank 312 can 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 part 318 of the sample introduction reservoir 319 can be closed in two stages by the first lid 501 and the second lid 502 shown in FIG. 5(b). Specifically, first, after step S102 is completed, the upper part of the sample introduction reservoir 319 is closed with the first lid 501 having the exhaust port 503. Then, the air remaining in the upper part of the sample introduction reservoir 319 is exhausted through the exhaust port 503. Then, the exhaust port 503 is closed with the second lid 502. In this way, it is possible to prevent the air remaining in the sample introduction reservoir 319 from forming bubbles and flowing into the second reservoir 312. Also, if the sample introduction reservoir 319 is closed in one stage in step S103, pressure is applied to the test sample filling the second reservoir 312 during the closing operation, and the test sample may enter the mixing reservoir 313 beyond the liquid stopping structure. This two-stage closing of the sample introduction reservoir 319 works more effectively for accurate metering of the liquid in the second reservoir 312 .

[0047] The third technology is an optimized structure for preventing air from entering from the sample introduction tank 319. In this embodiment, the sample introduction tank 319, the first tank 311 that functions as a pump, and the second tank 312 for measuring the test sample are configured as shown in FIG. 9(a). As a result, at the stage where step S102 is completed, the test sample is in a state where the second flow path 322, the first flow path 321 from the junction 320 to the second tank 312 side, and the second tank 312 are filled with the test sample. Therefore, when the first sealing lid 411 of the first tank 311 is pressed down in step S105, the air in the first tank 311 divides the test sample liquid at the junction 320, and then the test sample in the second tank 312 can be pushed out to the mixing tank 313 side without generating air bubbles. If the sample introduction reservoir 319, the first reservoir 311, and the second reservoir 312 have a configuration as shown in FIG. 9(b), that is, if the sample introduction reservoir 319 and the 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 air 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 with respect to the length direction (fluid flow direction) of the first flow path 321, as shown in FIG. 9(a).

[0048] The test sample moved to the mixing chamber 313 is mixed with the dried antibody-modified particles (immunoreagent 314) already present on the inner wall surface of the mixing chamber 313, and a test sample liquid is formed, which is a mixed liquid in which the antibody-modified particles are 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 this step S106, a reaction occurs in which the antibody-modified particles bind to each other via the target antigen, as illustrated diagrammatically in FIG. 2.

[0049] In order to quantify a more dilute target antigen by promoting an immune reaction in the mixture or to shorten the reaction time required for antigen quantification, it is effective to stir the mixture. A method for promoting an immune reaction will be described with reference to the flowchart of FIG. 7(b). In step S106, the sample introduction tank 319 is sealed and the first sealing lid 411 is pressed down, so that the mixture is in the mixing tank 313. When the first sealing lid 411 is opened from this state, the fourth flow path 324 side is opened, so that the mixture in the mixing tank 313 returns to the second tank 312 via the third flow path 323 (step S111). From this point, the first sealing lid 411 is repeatedly pressed down and opened for a predetermined R2 reaction time, so that the mixture is moved back and forth between the second tank 312 and the mixing tank 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 middle of 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, the mixed liquid may pass through the via hole 333. In this embodiment, an element that obstructs the passage of the mixed liquid is provided in the middle of the third flow path 323, and turbulence is generated when the mixed liquid passes through the third flow path 323, so that the mixed liquid can be mixed more efficiently. The liquid retention structure provided in the middle of the third flow path 323 may be any structure, and may be, for example, a via hole, a pillar, or a fin. Such a liquid retention structure may be provided in the second tank 312 or the mixing tank 313 other than in the second flow path.

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

[0052] In this manner, in the sensor device according to the embodiment of the present invention, a prescribed amount of the test sample mixed with R1 required for the test sample is measured (step S102), the test sample and R1 are mixed and reacted for a prescribed time (step S104), the test sample and R1 are mixed with each other after the reaction between the test sample and R1 (step S106), and the mixture is stirred for a prescribed time for 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 quantifying antigens based on the electric 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 an example shown in FIG. 3, the upper part of the upstream flow reservoir 302 is open, and the intake and exhaust performed in the operations of steps S101 to S106 and S111 to S115 may be performed through the open part of the upstream flow reservoir 302. Next, the reacted mixture that has entered the upstream flow reservoir 302 enters the upstream flow path 303 and reaches the liquid stop tank 357 via the upper part of the fine hole 399 (step S121). The upstream flow electrode liquid contact part 351 is connected to the bottom of the upstream flow reservoir 302, and the reacted mixture is introduced from the fourth flow path 324, so that the upstream flow reservoir 302, the upstream flow path 303, and the upstream flow electrode liquid contact part 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 is not open at the top, the liquid supply control in the reaction control unit 310 can be performed without any problem 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 part 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 is in contact with the reacted mixed liquid in contact with the fine hole 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 part 356, and fills the downstream flow path 308 (step S122). In this state, the reacted mixed liquid filled in the upstream flow path 303 and the electrolyte filled in the downstream flow path 308 come into contact with each other through the pore 399. As a result, the upstream flow path electrode 301 and the downstream flow path electrode 306 are electrically connected through the reacted mixed liquid 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 in FIG. 1(a), the particles in the reacted mixed liquid in the upstream flow path 303 move into the electrolyte in the downstream flow path 308 through the pore 399. Every time the antibody-modified particles pass through the pore 399, a pulse signal 210 as exemplified 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] According to the sensor device of the first embodiment of the present invention, the processes that were previously performed using complex and expensive dispensing and shaking mechanisms, such as measuring a fixed amount of the test sample, stirring, reaction time control, and highly sensitive antigen quantification using the electrical detection zone method, can all be performed within a small sensor measuring just a few centimeters square.

[0056] In the example of Fig. 3, 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 is illustrated. Next, in Fig. 10, 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, is described.

[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 upper part 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 upper part is open to the outside and into which a second fluid is introduced, a first tank 911 whose upper part is sealed by a first sealing lid 811, a second tank 912 whose upper part is sealed by a second sealing lid 812, a third tank 951 whose upper part is sealed by a third sealing lid 831, a fourth tank 952 whose upper part 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 liquid.

[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 measurement unit 900. In this example, the upper part of the upstream flow path reservoir 972 is open, and intake and exhaust are possible from here. In the example of Fig. 10, the third flow path 923 and the seventh flow path 943 are connected via a junction 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 that of the first sealing lid 411 and the second sealing lid 412 in FIG. 3. The cross-sectional structures of the first introduction reservoir 919 and the second introduction reservoir 949 are substantially similar to that of the sample introduction reservoir 319 in FIG. 3. The structures of the first flow path 921 and the fifth flow path 941 are substantially similar to that of the first flow path 321 in FIG. 3, and the structures of the second flow path 922 and the fifth flow path 941 are substantially similar to that of the second flow path 322 in FIG. 3. The structure of the fourth flow path 924 is also substantially similar to that of the fourth flow path 324 in 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. Moreover, 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, it is possible to more appropriately measure the first fluid and the second fluid 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 including 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 side into which the first fluid is introduced, and a test sample may be introduced into the second introduction tank 949 side into which the second fluid is introduced.

[0063] The first open part 918 and the second open part 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 capable of opening or closing the exhaust port 503, as shown in Fig. 5(b). The first open part 918 and the second open part 948 of the first introduction tank 919 and the second introduction tank 949 can be controlled to be closed in two stages by the first lid 501 and the second lid 502, thereby enabling accurate measurement of the liquid. Also, in the sensor device of Fig. 10, it is preferable that a liquid stopper consisting of two flow paths in different layers is provided midway through the third flow path 923, for example, via a via hole 933.

[0064] The measuring unit 900 includes an upstream liquid reservoir 972 connected to the fourth flow path 924, an upstream flow path 971 connected to the upstream liquid 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 fine hole 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 adjust the liquid level in the downstream flow path 981 to a constant level. In this embodiment, the upstream flow path electrode 873 contacts the reacted mixture at the bottom of the upstream flow path liquid reservoir 972. The downstream flow path electrode 986 and the electrolyte filling the downstream flow path are in contact with each other at a liquid contact portion 984.

[0065] When the first sealing lid 811 is pressed down 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, 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 sealing lid 811 and the third sealing lid 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 manner, by repeatedly pressing the first sealing lid 811 and the third sealing lid 831 downward and releasing the 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 with reference to the sensor device of FIG. 10 and the flow charts of FIG. 11(a), FIG. 11(b) and FIG. 12. First, a test sample is introduced as a first fluid into the first introduction tank 919 of FIG. 10 by, for example, dropping (step S1001). The test sample moves from the first introduction tank 919 of FIG. 10 through the second flow path 922 and the first flow path 921 to the second tank 912. When the second tank 912 is filled with the test sample, the movement of the test sample stops (step S1002). At this time, the first tank 911 does not have an opening connected to the outside, and the air is sealed. On the other hand, the second tank 912 is connected to a flow path of the measurement unit 900 having an opening connected to the outside via the third flow path 923, the mixing tank 913 and the fourth flow path 924, and can be exhausted. Therefore, in step S1002, the test sample flows from the junction of first flow path 921 and second flow path 922 only to second tank 912, not to first tank 911, and second tank 912 is filled with the test sample. Regardless of the amount of the test sample introduced into first introduction tank 919, a certain amount of the test sample determined by the volume of second tank 912 is measured by this step S1002.

[0069] Similarly, a reagent such as the R2 reagent is introduced as the second fluid into the second introduction reservoir 949 of FIG. 10 by, for example, dropping (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 according to the volumes of the second reservoir 912 and the fourth reservoir 932, respectively.

[0070] After completing step S1004, the upper portions of the first introduction reservoir 919 and the second introduction reservoir 949 are sealed (step S1005). The progress of immune reactions is highly sensitive to the liquid volume and reaction time. The test sample and the R2 reagent are optimized so that accuracy and performance can be exhibited only when the predetermined liquid volume and reaction time are precisely observed. After completing step S1005, the first introduction reservoir 919 is sealed and the test sample is held in the second reservoir 912 and waits for the predetermined R1 reaction time (step S1006). Next, the first sealing lid 811 on the top of the first reservoir 911 is pressed down with the test sample held in the second reservoir 912, and the third sealing lid 831 on the top of the third reservoir 951 is pressed down preferably at the same time with the R2 sample held in the fourth reservoir 952 (step S1007). In order to appropriately control the reaction time, it is preferable to press down the first sealing lid 811 and the third sealing lid 831 almost simultaneously, but they do not have to be pressed down completely simultaneously. Since the first sealing lid 811 and the third sealing lid 831 are made of a flexible material, by pressing down the first sealing lid 811, a gas such as air sealed in the first tank 911 moves 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 pressing down the second sealing lid 812 and the fourth sealing lid 832 while pressing down the first sealing lid 811 and the third sealing lid 831, the first fluid and the second fluid are introduced into the mixing tank 913, and a mixed liquid, which is a mixture of the first fluid and the second fluid, 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. This allows the test sample liquid measured in the second tank 912 and the R2 reagent measured in the fourth tank 952 to be delivered to the mixing tank 913 without air bubbles when the first sealing lid 811 and the third sealing lid 831 are pressed down.

[0072] The target antigen and the antibody in the R2 reagent undergo an immunological reaction 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 for antigen quantification, it is effective to stir the mixture.

[0073] A method for promoting an immune response according to the second embodiment of the present invention will be described with reference to the flow chart of FIG. 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 down, so that the mixed liquid is in the mixing tank 913. When the first sealing lid 811 and the third sealing lid 831 are opened from this state, the mixed liquid in the mixing tank 913 returns to the fourth tank 952 via the third flow path 923 and the second tank 912 and the third flow path 923 and the seventh flow path 943 via the third flow path 923 (step S1011). From this point, the first sealing lid 811 and the third sealing lid 831 are pressed down and opened, so that 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 stirring the mixed liquid (step S1012). The mixture 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 middle of the third flow path 923. Therefore, when the mixed liquid travels back and forth between the second tank 912 and the fourth tank 952 and the mixing tank 913 in step S1012, the mixed liquid passes through this via hole 933. In this way, an element that obstructs the passage of the mixed liquid is provided in the middle of the third flow path 923, and turbulence is generated when the mixed liquid passes through the third flow path 923, thereby making it possible to perform stirring more efficiently. The liquid retention structure provided in the middle of the third flow path 923 can have a configuration similar to that described for the sensor device in FIG. 3.

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

[0076] In the sensor device according to the second embodiment of the present invention, a specified amount of a test sample, which is a mixture of a clinical specimen and an R1 reagent, is measured (step S1002), a specified amount of an R2 reagent is measured (step S1004), the test sample is held in the second tank for a predetermined reaction time of 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 for an efficient immune reaction (S1012-S1013). Such processing can be easily performed on the small and inexpensive sensor device according to this embodiment, and further, by introducing the reacted mixture obtained as a result of the processing into the measurement unit 900, antigen quantification can be performed with high sensitivity and speed by the electrical detection zone method.

[0077] The process of antigen quantification based on the electric 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 fine hole 999, and reaches the liquid stop tank 974 via the upper part of the fine hole 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 electrode liquid contact part 351 are filled with the reacted mixture. The upper part of the upstream flow exhaust port 975 is open, and even if the upstream flow reservoir 972 is not open to the upper part, the supply control in the reaction control unit 910 can be performed without any problem by the intake and exhaust from the upstream flow exhaust port 975.

[0078] Next, when the electrolyte is introduced into the downstream flow path introduction tank 982, the electrolyte passes through the downstream flow path 981 and contacts the pore 999 and the downstream flow path electrode liquid contact portion 984, filling the downstream flow path 981 (step S1022). The reacted mixture filled in the upstream flow path 971 and the electrolyte filled in the downstream flow path 981 come into contact with each other through the pore 999. This allows electrical conduction between the upstream flow path electrode 873 and the downstream flow path electrode 986 through the reacted mixture 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 mixture in the upstream flow path 971 move into the electrolyte in the downstream flow path 981 through the pore 999. A pulse signal is measured every 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] According to the sensor device of the second embodiment of the present invention, the processes of measuring a fixed amount of the test sample, stirring, reaction time control, and highly sensitive antigen quantification using the electrical detection zone method, which were previously performed using complex and expensive dispensing and shaking mechanisms, can all be performed within a small sensor measuring just a few centimeters square.

[0080] Although the present invention has been described by the above embodiment, the description and drawings forming a part of this disclosure should not be understood as limiting the present invention. The present disclosure is not limited to the above embodiment, and the components can be combined and modified to be embodied without departing from the spirit of the present disclosure.

[0081] For example, in the sensor devices shown in Fig. 3 and Fig. 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 manufactured as separate devices and then integrated by assembling them. Also, a module of the measurement units 300, 900 alone may be manufactured and used in conjunction with a chip or the like that uses an existing electrical detection zone method. In Fig. 10, as in the sensor device of Fig. 3, it is also possible to fill the mixing chamber 913 with the solid phase of the R2 reagent in advance, and introduce any other reagent such as a reaction promoter from the second introduction chamber 949.

[0082] In one example of FIG. 3, the first and second flexible sealing lids 411 and 412 are respectively installed on the top of the first tank 311 and the second tank 312. Similarly, in one example of FIG. 10, the first and second flexible sealing lids 811, 812, the third and fourth tanks 831 and 832 are respectively installed on the top of the first tank 911, the second tank 912, the third tank 951 and the fourth tank 952. However, the first and second flexible sealing lids 411 and 811, the second and third flexible sealing lids 831 and 832 may be arranged not only on the top of each tank but also on the bottom. That is, as shown in FIG. 13, the first and second tanks 311 and 312 may have openings on the lower side, and the first and second sealing lids 411 and 412 may be arranged to close 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 concentrated and arranged 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. With this configuration, 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 can be prevented, and a smaller measuring device can be realized.

[0084] As shown in FIG. 14, the upstream flow path electrode 301 and the downstream flow path electrode 306 may be disposed on the upper surface side of the measuring unit 300, or one may be disposed on the upper surface side and the other on the lower surface side. In cases where the flow path configurations on the lower layer side of the sensor device are close to each other, the sensor device can be made smaller by disposing the upstream flow path electrode 301 on the upper surface side of the sensor device. Similarly, the upstream flow path reservoir 302 is formed to be shifted from each other in the vertical direction of the paper surface 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 shifted to the side closer to the suction pad 30, thereby making it possible to make the sensor device smaller. In this way, this embodiment can include various aspects that can be modified in design within the scope of technical common sense of a person skilled in the art. [Explanation of symbols]

[0085] 101: Particles to be detected 110, 120: Chamber 111, 121: Inlet 112, 122: Electrode 140: pore 141: Bulkhead 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 particles 219, 231, 233, 251, 253: Agglomerates 221, 222, 241, 242, 243: Pulse signal 300: Measurement section 301:Upstream path electrode 302: Upstream liquid reservoir 303:Upstream route 304: Exhaust port (upstream exhaust port) 306: Downstream path electrode 307: Downstream introduction tank 308: Downstream path 310, 910: reaction control section 311, 911: Tank 1 312, 912: 2nd tank 313, 913: Mixing tank 314: Immunoreagents (R2 Reagents) 317: Pore chip 318, 918: 1st opening 319, 919: First introduction tank (sample introduction tank) 320: Junction 321, 921: First flow path 322, 922: Second flow path 323, 923: 3rd flow path 324, 924: 4th channel 333: Beer hall 350: Upstream connection channel 351:Upstream channel electrode wetted part 356: Downstream electrode wetted part 357: Liquid stop tank 399: Pore 411, 811: 1st sealing lid 412, 812: 2nd sealing lid 501: 1st lid 502:Second lid 503: Exhaust port 831:Third sealing lid 832: 4th sealing lid 833: Beer hall 873:Upstream path electrode 900: Measurement section 933: Beer hall 941: 5th channel 942: 6th Channel 943: 7th Channel 948:Second opening 949:Second introduction tank 951:Third tank 952: Tank 4 971:Upstream route 972: Upstream liquid reservoir 974: Liquid stop tank 975: Upstream exhaust port 981: Downstream path 982: Downstream introduction tank 983: Absorbent pad 984: Downstream electrode wetted part 986: Downstream electrode 999: pores

Claims

1. a sample introduction tank having an opening portion that is open to the outside and into which a test sample is introduced; a first tank sealed by a flexible first sealing lid; a second tank sealed by a flexible second sealing lid; a mixing vessel for mixing the test sample with an immune reagent 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 reservoir so as to introduce the test sample to be introduced into the sample introduction reservoir into the second reservoir via the first flow path; a third flow path connected between the second tank and the mixing tank; a fourth flow path connected to the mixing chamber and configured to introduce the test sample liquid in the mixing chamber into a measurement section for measuring the presence or absence or the concentration of the target antigen in the test sample liquid; A sensor device comprising:

2. 2. The sensor device according to claim 1, wherein 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.

3. when the first sealing lid is pressed from the outside in a state in which the test sample introduced into the sample introduction reservoir is filled in 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 in a state in which the test sample is mixed with the immunoreagent in the mixing chamber, the test sample liquid in the mixing chamber returns to the second chamber via the third flow path, The sensor device described in claim 1, characterized in that the first sealing lid is displaced by repeatedly pressing and releasing the pressure on the first sealing lid so that the test sample liquid is stirred by moving back and forth between the second tank and the mixing tank.

4. When the first sealing lid is pressed from the outside in a state in which the test sample introduced into the sample introduction reservoir is filled in the second reservoir and the sample introduction reservoir is sealed, the first sealing lid is displaced so that 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, The sensor device according to claim 1, characterized in that when the second sealing lid is pressed from the outside while the sample introduction tank is sealed and the first sealing lid is pressed, the second sealing lid is displaced so that the test sample liquid in the mixing tank is introduced from the mixing tank to the measurement section via the fourth flow path.

5. The sensor device according to 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 liquid stopping structure is At least two flow paths formed of at least two different layers via via holes; A bent portion formed in the third flow path; a pillar or bump formed so as to protrude from an inner wall surface of the third flow path; A roughened layer formed on the inner wall surface of the third flow path.

6. The sensor device according to claim 5, further comprising:

7. a first introduction tank having a first opening portion that is open to the outside and into which a first fluid is introduced; a second introduction tank having a second opening portion that is open to the outside and into which a second fluid is introduced; a first tank sealed by a flexible first sealing lid; a second tank sealed by a flexible second sealing lid; a third tank sealed by a flexible third 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 measurement target 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 fourth flow path that introduces the mixed liquid into a measurement section for measuring the presence or absence or concentration of the measurement target substance 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; a seventh flow path connected between the fourth tank and the third flow path; A sensor device comprising:

8. 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 into the fourth tank and the second open portion is sealed, when the third sealing lid is pressed from the outside, 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, The sensor device according to claim 7, characterized in that the first sealing lid and the third sealing lid are displaced so that the first fluid and the second fluid flowing into the mixing chamber are mixed.

9. 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 through the third flow path, The sensor device according to claim 7 or 8, characterized in that the first sealing lid and the third sealing lid are displaced such 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 by repeatedly pressing and releasing the first sealing lid and the third sealing lid.

10. 9. The sensor device according to claim 7, wherein 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. 9. The sensor device according to claim 7, wherein the sixth flow path is connected to the fifth flow path so as to merge with the fifth flow path from a side of the fifth flow path.

12. a first introduction tank having a first opening portion that is open to the outside and into which a first fluid is introduced; A first cover provided in the first open portion and having an exhaust port; A second cover capable of opening and closing the exhaust port; a first tank sealed by a flexible first sealing lid; a second tank sealed by a flexible second sealing lid; a mixing tank for mixing the first fluid with a component that reacts with a measurement target 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 stopper portion provided midway through the third flow path; a fourth flow path that introduces the mixed liquid into a measurement section for measuring the presence or absence or concentration of a measurement target that may be contained in the mixed liquid in the mixing tank; A sensor device comprising:

13. The sensor device according to 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 a side of the first flow path.

14. the first fluid comprises a test sample including a clinical specimen; 9. The sensor device according to claim 7, wherein the second fluid contains 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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