Processing kit and processing method

Abstract

The present invention provides a processing kit and processing method with which it is possible to achieve sufficient sensitivity and have little variation in each collected amount of a concentrated liquid of a liquid sample. The processing kit comprises: a water absorbent polymer that absorbs water from a liquid sample containing a test substance and water; a container that accommodates the water absorbent polymer; and a collected liquid container that accommodates a collected liquid containing magnesium chloride and sodium chloride. The collected liquid is added after the liquid sample is inserted into the container.

Description

Processing kit and processing method

[0001] The present invention relates to a processing kit and processing method comprising a container for containing a water-absorbing polymer among a liquid sample including a test substance and water, and a recovery liquid container for containing a recovery liquid containing magnesium chloride and sodium chloride. More particularly, the present invention relates to a processing kit and processing method in which the recovery liquid is added after the liquid sample is placed in the container.

[0002] Conventionally, a technique for concentrating liquid samples, including test substances and water, using superabsorbent polymers (superabsorbent materials) is known (for example, Patent Document 1). The test substance is, for example, a polymer such as an antigen.

[0003] Japanese Patent Application Publication No. 4-355339

[0004] When the present inventors investigated a method for concentrating liquid samples using a superabsorbent polymer, referencing the micro-sample collection device described in Patent Document 1, it became clear that it is difficult to extract the concentrated liquid sample from the micro-sample collection device after the liquid sample has been concentrated. Specifically, from the viewpoint of shortening the concentration time, a larger amount of superabsorbent polymer is preferable, and even when the amount of liquid sample is large, a large amount of superabsorbent polymer is required. If the amount of superabsorbent polymer is large, the amount of concentrated liquid will be small, making it difficult to extract the concentrated liquid. As a result, it may not be possible to obtain a sufficient amount of concentrated liquid from the liquid sample as required. Furthermore, from the viewpoint of shortening the water absorption time, if the saturated sample absorption capacity of the superabsorbent polymer is greater than the amount of liquid sample, water absorption will occur when extracting the concentrated liquid. Therefore, if it takes a long time to extract the concentrated liquid, it may not be possible to obtain the required amount of concentrated liquid from the liquid sample. If the amount of superabsorbent polymer is reduced in order to ensure the amount of concentrated liquid, the concentration ratio of the concentrated liquid will be lower. Moreover, it is also possible to add a small amount of recovered liquid after concentrating the liquid sample to extract the concentrated liquid, but if the amount of recovered liquid is increased, the concentration ratio of the concentrated liquid will be lower. Furthermore, if a recovery solution is added after concentrating a liquid sample, the recovery solution may be absorbed by the superabsorbent polymer, potentially preventing the recovery of high molecular weight substances such as antigens contained in the liquid sample. Thus, the amount of concentrated liquid sample recovered varies from one collection to the next. In other words, the repeatability of concentrated liquid recovery is poor. Additionally, it has become clear that sufficient sensitivity may not always be obtained when applying concentrated liquid samples to immunochromatography.

[0005] The object of the present invention is to provide a processing kit and processing method that have low variability in the amount recovered for each collection of concentrated liquid sample and that provide sufficient sensitivity.

[0006] The above objectives can be achieved with the following configurations. Invention [1] is a processing kit comprising a liquid sample containing a test substance and water, a superabsorbent polymer that absorbs water, a container for containing the superabsorbent polymer, and a recovery liquid container for containing a recovery liquid containing magnesium chloride and sodium chloride, wherein the recovery liquid is added after the liquid sample is placed in the container. Invention [2] is the processing kit according to Invention [1], wherein the magnesium chloride content of the recovery liquid is 10 mg / mL or more and 80 mg / mL or less, and the sodium chloride content of the recovery liquid is 30 mg / mL or more and 200 mg / mL or less. Invention [3] is the processing kit according to Invention [1] or [2], wherein the liquid sample contains biological fluid.

[0007] Invention [4] is a processing method comprising the steps of: placing a liquid sample containing a test substance and water into a container containing a superabsorbent polymer; allowing the water contained in the liquid sample to be absorbed by the superabsorbent polymer to concentrate the liquid sample in the container; adding a recovery solution containing magnesium chloride and sodium chloride to the container; and removing the concentrated liquid sample obtained by concentration in the container from the container. Invention [5] is the processing method according to Invention [4], wherein the magnesium chloride content of the recovery solution is 10 mg / mL or more and 80 mg / mL or less, and the sodium chloride content of the recovery solution is 30 mg / mL or more and 200 mg / mL or less. Invention [6] is the processing method according to Invention [4] or [5], wherein the liquid sample contains biological fluid.

[0008] According to the present invention, it is possible to provide a processing kit and processing method that have small variations in the amount of concentrated liquid sample recovered each time and that provide sufficient sensitivity.

[0009] It is a schematic perspective view showing an example of a container of a processing kit according to an embodiment of the present invention. It is an exploded perspective view showing an example of a container of a processing kit according to an embodiment of the present invention. It is a schematic perspective view showing an example of a recovery liquid container of a processing kit according to an embodiment of the present invention. It is a schematic cross-sectional view showing an example of a cap of a container of a processing kit according to an embodiment of the present invention. It is a schematic perspective view showing another example of a container body of a container of a processing kit according to an embodiment of the present invention. It is a schematic perspective view showing another example of a container body of a container of a processing kit according to an embodiment of the present invention. It is a schematic perspective view showing another example of a container body of a container of a processing kit according to an embodiment of the present invention. It is a schematic diagram for explaining the relationship between the discharge direction of a liquid sample and a flexible wall surface. It is a schematic diagram for explaining the volume changeable amount of a container. It is a schematic cross-sectional view showing one step of an example of a processing method according to an embodiment of the present invention. It is a schematic cross-sectional view showing one step of an example of a processing method according to an embodiment of the present invention. It is a schematic cross-sectional view showing one step of an example of a processing method according to an embodiment of the present invention. It is a schematic cross-sectional view showing one step of an example of a processing method according to an embodiment of the present invention. It is a schematic diagram of one aspect of an insoluble carrier used in the method of the present invention.

[0010] Hereinafter, based on the preferred embodiments shown in the accompanying drawings, the processing kit and the processing method of the present invention will be described in detail. The figures described below are exemplary for explaining the present invention and are simplified for the purpose of explaining the present invention. Therefore, the present invention is not limited to the figures shown below. In the following, "~" indicating a numerical range includes the numerical values described on both sides. For example, ε ｎ is the numerical value ε α ~ the numerical value ε β means that the range of ε ｎ is the range including the numerical value ε α and the numerical value ε β and, in mathematical symbols, ε α ≦ ε ｎ ≦ ε βThe following applies to specific angles, parallelism, and perpendicularity, unless otherwise specified: the generally accepted tolerance range in the relevant technical field is included. Similarly, for specific angles and orthogonality, unless otherwise specified: the generally accepted tolerance range in the relevant technical field is included. For temperature and time, unless otherwise specified: the generally accepted tolerance range in the relevant technical field is included.

[0011] The following provides a detailed explanation of the processing kit and processing method.

[0012] [Processing Kit] Figure 1 is a schematic perspective view showing an example of a container for a processing kit according to an embodiment of the present invention. Figure 2 is an exploded perspective view showing an example of a container for a processing kit according to an embodiment of the present invention. Figure 3 is a schematic perspective view showing an example of a recovery liquid container for a processing kit according to an embodiment of the present invention. Figure 4 is a schematic cross-sectional view showing an example of a cap for a container for a processing kit according to an embodiment of the present invention. The processing kit 10 includes a container 12 shown in Figure 1 and a recovery liquid container 14 shown in Figure 3. More specifically, the processing kit 10 includes a container 12 that contains a water-absorbing polymer 16 and a recovery liquid container 14 (see Figure 3) that contains a recovery liquid 17 containing magnesium chloride and sodium chloride. The water-absorbing polymer 16 absorbs water from a liquid sample containing the test substance and water. The water-absorbing polymer 16 contained in the container is, for example, a superabsorbent polymer (SAP) with high water absorption. Hereinafter, the liquid sample containing the test substance and water will also be referred to as the sample solution. Furthermore, a concentrated liquid sample is a concentrated liquid sample, but both the concentrated liquid sample and the concentrated liquid sample are also simply called a concentrated solution. Unless otherwise specified, the term "concentrated solution" refers to the concentrated liquid sample and the concentrated liquid sample, that is, the concentrated sample solution and the concentrated sample solution.

[0013] The container 12 of the processing kit 10 shown in Figures 1 and 2 is at least partially flexible and has a container body 20 having an opening 21, a cap 22 detachably attached to the opening 21 of the container body 20, and a nozzle cover 24 for the nozzle 22a of the cap 22. The container body 20 of the container 12 has low rigidity and is flexible. The container 12 is used in a concentration process to concentrate a liquid sample in order to obtain a concentrated liquid solution of the liquid sample. The process of obtaining a concentrated liquid solution of a liquid sample from a liquid sample is also called the concentration process.

[0014] The container body 20 of the container 12 shown in Figures 1 and 2 consists of a storage section 20a that houses a water-absorbing polymer 16 (see Figure 1) inside 20h, and a neck section 20b (see Figure 2) having an opening 21 (see Figure 2). In the illustrated example, the storage section 20a is a substantially cylindrical shape with a bottom surface and forms an internal space capable of housing the water-absorbing polymer. The neck section 20b is connected to one of the bottom surfaces, and the opening 21 of the neck section 20b is in communication with the internal space of the storage section 20a. In the example shown in Figures 1 and 2, the storage section 20a has a diameter-reducing section 20c at the end on the neck section 20b side that narrows toward the neck section 20b.

[0015] As shown in Figure 2, the neck portion 20b is a substantially cylindrical portion having an opening 21 that penetrates from one bottom surface to the other bottom surface. In the illustrated example, the neck portion 20b is positioned so that its central axis (central axis of the cylinder) substantially coincides with the central axis (central axis of the cylinder) of the housing portion 20a. Furthermore, the neck portion 20b is provided with a male screw portion 21a on its outer circumferential surface.

[0016] The area of ​​the housing portion 20a in a cross-section perpendicular to the central axis is larger than the area of ​​the neck portion 20b. In the illustrated example, the diameter of the housing portion 20a in a cross-section perpendicular to the central axis is larger than the diameter of the neck portion 20b. Therefore, the area of ​​the housing portion 20a at the connection point between the housing portion 20a and the neck portion 20b is larger than the area of ​​the neck portion 20b. Hereinafter, the bottom surface of the housing portion 20a to which the neck portion 20b is connected will also be referred to as the shoulder portion.

[0017] Furthermore, at least a portion of the housing portion 20a is flexible, allowing the water-absorbing polymer 16 housed in the housing portion 20a to be pressed through the inner wall of the housing portion 20a. In the illustrated example, it is preferable that at least a portion of the circumferential surface of the housing portion 20a is flexible, and the entire housing portion 20a may be flexible.

[0018] In the examples shown in Figures 1 and 2, the cap 22 is a member that closes the opening 21 of the neck 20b of the container body 20. Figure 4 shows a cross-sectional view of the cap 22. As shown in Figures 1, 2 and 4, the cap 22 is a cylindrical member having one bottom surface, and a female threaded portion 22c is provided on its inner circumference. By screwing the female threaded portion 22c onto the male threaded portion 21a of the neck 20b of the container body 20, the cap 22 can be easily attached to and removed from the container body 20.

[0019] The cap 22 also has a nozzle 22a that protrudes outward from its bottom surface, and a through-hole is provided that penetrates the nozzle 22a, which serves as the discharge section 22b. The nozzle 22a is for discharging the concentrated liquid to the outside of the container 12. The concentrated liquid is dispensed to the outside of the container 12 and stored in a recovery container, such as a recovery liquid cup 50 (see Figure 13). The concentrated liquid is used, for example, in tests utilizing antigen-antibody reactions using an immunochromatography kit. The nozzle cover 24 covers the through-hole (discharge section 22b) of the nozzle 22a of the cap 22, thereby blocking the through-hole. By attaching the nozzle cover 24 to the nozzle 22a, for example, when concentrating a liquid sample (sample solution), leakage of the concentrated liquid can be prevented. The shape of the nozzle cover 24 is not particularly limited. The nozzle cover 24 may also be connected to the cap 22.

[0020] The recovery liquid container 14 shown in FIG. 3 is for adding the recovery liquid 17 into the container 12 in a state where the water-absorbent polymer 16 is stored, that is, inside the container body 20, namely, the interior 20h of the container body 20, after a liquid sample is put in. The recovery liquid 17 is added after the liquid sample is put into the container 12. The recovery liquid container 14 has, for example, as shown in FIG. 3, a cylindrical base portion 40 and a cylindrical main body portion 42 extending in one direction with a bottom, and the outer diameter of the base portion 40 is larger than that of the main body portion 42. The recovery liquid 17 is contained in the interior 42a of the main body portion 42, and the opening 40a of the base portion 40 is sealed by a sealing material 44. The recovery liquid 17 is stored in the interior 42a of the main body portion 42 by the sealing material 44. By removing the sealing material 44, the recovery liquid 17 in the main body portion 42 can be made to flow out from the opening 40a to the outside of the recovery liquid container 14. The configuration of the recovery liquid container 14 is not particularly limited as long as it can contain the recovery liquid 17 and can add the recovery liquid 17 into the container 12. The amount of the recovery liquid 17 added by the recovery liquid container 14 is less than that of the liquid sample. Further, the base portion 40 and the main body portion 42 of the recovery liquid container 14 are made of, for example, polyethylene (PE) and polypropylene (PP) or the like. For the sealing material 44, for example, an aluminum foil is used. The sealing material 44 seals the opening 40a of the base portion 40 by heat sealing, for example.

[0021] Further, in the example shown in FIG. 4, as a preferred embodiment, a filter 23 is disposed on the bottom surface side inside the cap 22. The filter 23 allows the concentrated liquid to pass through and does not allow the water-absorbent polymer 16 (see FIG. 1) to pass through. The filter 23 is not particularly limited as long as it allows the concentrated liquid to pass through and does not allow the water-absorbent polymer 16 to pass through. For the filter 23, for example, a membrane filter is used.

[0022] The container 12 having the container body 20 and the cap 22 shown in FIGS. 1 and 2 can remove the cap 22 from the container body 20 and put the water-absorbent polymer 16 before water absorption (see FIG. 1) into the storage portion 20a from the opening 21 of the neck portion 20b. Also, a liquid sample 30 (see FIG. 9) can be put in from the opening 21 of the neck portion 20b. That is, the opening 21 of the container body 20 is an intake portion for taking in the liquid sample.

[0023] After taking in the liquid sample, the cap 22 is attached to the container body 20. At this time, the nozzle cover 24 is attached to the cap 22. After a predetermined time has elapsed, the superabsorbent polymer 16 absorbs the water from the liquid sample, and the liquid sample is concentrated. After that, the concentrated liquid is stirred in the container 12 as needed. Next, using the recovery liquid container 14, the recovery liquid 17 is poured into the container 12 through the opening 21 of the neck 20b, that is, into the inside of the container body 20, and then the concentrated liquid is discharged from the discharge part 22b provided on the nozzle 22a of the cap 22. At this time, the nozzle cover 24 is removed from the cap 22. At least a part of the container body 20 is flexible, and the superabsorbent polymer can be pressed through the inner wall of the container body 20. For this reason, the swollen superabsorbent polymer 16 after water absorption can be pressed through the inner wall of the container body 20, and the concentrated liquid can be removed from the container body 20.

[0024] After the superabsorbent polymer 16 has absorbed the moisture from the liquid sample, the recovered liquid 17 is poured into the recovered liquid container 14 through the opening 21 at the neck 20b. Next, the cap 22 is attached. At this time, the nozzle cover 24 is attached to the cap 22. After that, if necessary, the recovered liquid may be stirred in the container 12 and the concentrated liquid may be removed from the discharge part 22b provided on the nozzle 22a of the cap 22. Alternatively, when stirring the concentrated liquid and stirring the recovered liquid, the container 12 may be sealed with a cap that does not have a nozzle 22a (discharge part 22b) and stirred, and then, when discharging the concentrated liquid, the cap 22 with a nozzle 22a may be attached to remove the concentrated liquid.

[0025] In the examples shown in FIGS. 1 and 2, the area of the accommodating portion 20a at the connection position between the accommodating portion 20a and the neck portion 20b is larger than the area of the neck portion 20b, and the accommodating portion 20a is configured to have a shoulder, but the present invention is not limited thereto. For example, as shown in FIG. 5, the end portion of the accommodating portion 20d of the container body 25 on the neck portion 20b side may have a diameter-reducing portion 44e that reduces in diameter to the same diameter as the neck portion 20b toward the neck portion 20b and is connected to the neck portion 20b. That is, the example shown in FIG. 5 is an example in which the area (diameter) of the accommodating portion 20d at the connection position between the accommodating portion 20d and the neck portion 20b is equal to the area (diameter) of the neck portion 20b, and the accommodating portion 20d does not have a shoulder. Note that FIG. 5 is a schematic perspective view showing another example of the container body of the container of the processing kit according to the embodiment of the present invention.

[0026] In order to prevent the neck portion 20b, to which the cap 22 with the nozzle cover 24 attached is attached, from being deformed when the accommodating portion 20a is pressed to take out the concentrated liquid from the container body 20, the accommodating portion 20a preferably has a shoulder, that is, a configuration in which the area of the accommodating portion 20a at the connection position between the accommodating portion 20a and the neck portion 20b is larger than the area of the neck portion 20b.

[0027] In the examples shown in FIGS. 1 and 2, the neck portion 20b side end portion is configured to have a diameter-reducing portion 20c, but it may have a shape without the diameter-reducing portion 20c. The configuration having the diameter-reducing portion 20c at the end portion on the neck portion 20b side is preferable in that it facilitates the extraction of the concentrated liquid. In the examples shown in FIGS. 1 and 2, the accommodating portion 20a of the container body 20 has a substantially cylindrical shape, but the present invention is not limited thereto. For example, the accommodating portion 20a may have a polygonal cylindrical shape such as a triangular cylindrical shape or a quadrangular cylindrical shape, or an elliptical cylindrical shape.

[0028] Further, the accommodating portion 20a is not limited to a substantially cylindrical shape and can have various shapes. For example, as shown in FIG. 6, the accommodating portion 20f of the container body 25a may have a substantially circular cross-sectional shape on the neck portion 20b side and may be flattened toward the side opposite to the neck portion 20b so that the cross-sectional area becomes smaller. FIG. 6 is a schematic perspective view showing another example of the container body of the container of the processing kit according to the embodiment of the present invention.

[0029] Furthermore, as shown in Figure 7, the container body 25b may have a neck portion 20b with an opening 21 on which the cap 22 can be attached and detached, and a bag-shaped storage portion 26 for containing the superabsorbent polymer. Note that "bag-shaped" refers to a container made of a material that does not have self-supporting properties. Figure 7 is a schematic perspective view showing another example of the container body of the processing kit container in an embodiment of the present invention.

[0030] Furthermore, in the above example, the container bodies 20, 25, 25a, and 25b and the cap 22 are configured to be screwed together by providing a male threaded portion 21a and a female threaded portion 22c, respectively, but the configuration is not limited to this. In addition to screwing, fitting may be used to attach the container bodies 20, 25, 25a, and 25b to the cap 22. One of the container bodies 20, 25, 25a, and 25b and the cap 22 may have a convex portion and the other may have a concave portion, with the concave portion and convex portion being locked together. Known detachable fixing methods can be used as appropriate to attach the container bodies 20, 25, 25a, and 25b to the cap 22.

[0031] Here, "at least a part of the container is flexible" means that the flexible portion of the container is made of a resin material or elastomer material such as polyethylene (PE), polyethylene terephthalate (PET), polystyrene (PS), polypropylene (PP), polyvinyl chloride (PVC), or acrylic resin (PMMA), and has a thickness of 1000 μm or less. In addition, it may be a composite material with a material that has necessary functions such as low moisture permeability, gas barrier properties, light shielding properties, and decorative properties in addition to the above materials. From the viewpoint of high flexibility, relatively high strength, chemical resistance, and cost, the resin material of the flexible portion of the container is preferably either polyethylene (PE) or polypropylene (PP).

[0032] The thickness of the flexible portion of the container is preferably 1000 μm or less, more preferably 800 μm or less, and even more preferably 600 μm or less. The lower limit is not particularly limited, but is preferably 20 μm or more.

[0033] Here, it is preferable that the wall surface of the container parallel to the discharge direction of the concentrated liquid sample (concentrate) at the discharge section is flexible. This point will be explained using Figure 8. Figure 8 is a schematic diagram to explain the relationship between the discharge direction of the liquid sample and the flexible wall surface. Figure 8 conceptually shows a cross-sectional view of the container 28 of the processing kit. In the container 28 shown in Figure 8, a discharge section 22b is provided on the upper surface of the container 28 in the figure. That is, the direction of discharge of the concentrated liquid from the discharge section 22b in the illustrated example is upward in the figure, as indicated by arrow D. Therefore, it is preferable that the wall surface 29 of the container 28 parallel to this arrow D is flexible.

[0034] The container 28 has a flexible wall surface 29 parallel to the discharge direction of the concentrated liquid from the discharge section 22b, which allows the container 28 to be pressed in a direction substantially perpendicular to the discharge direction of the concentrated liquid. This prevents the superabsorbent polymer from being compressed, which would narrow the gaps between the polymers and prevent the concentrated liquid (recovered liquid) from moving. As a result, the concentrated liquid (recovered liquid) can be efficiently spread out, and the removal of the concentrated liquid becomes easier. For example, in the case of the container 12 shown in Figure 1, the discharge direction of the concentrated liquid is upward in the figure, so it is preferable that at least a part of the circumferential surface of the housing section 20a of the container body 20 is flexible, and it is more preferable that the entire circumferential surface is flexible.

[0035] Furthermore, it is preferable that the amount by which the container's volume can be changed is greater than the difference δ between the container's volume V and the volume Vs of the liquid sample contained in the container and the volume Vp of the superabsorbent polymer. This point will be explained using Figure 9. The difference δ is expressed as δ = V - (Vs + Vp). Figure 9 is a schematic diagram for explaining the amount by which the container's volume can be changed, and conceptually shows the container 12 shown in Figures 1 and 2. As shown in Figure 9, if the total volume of the container 28 is V, the total volume of all the superabsorbent polymers 16 in the container 28 before water absorption is Vp, and the volume of the liquid sample 30 to be placed in the container 28 is Vs, then the difference δ (= V - (Vs + Vp)) between the container's volume V and the volume Vp of the superabsorbent polymers 16 and the volume Vs of the liquid sample 30 is the volume Va of the space in the container 28 that is not filled with superabsorbent polymers 16 and / or liquid sample 30.

[0036] Since the sum of the volume of the superabsorbent polymer after water absorption and the volume of the remaining concentrated liquid is approximately equal to the sum of the volume Vp of the superabsorbent polymer 16 and the volume Vs of the liquid sample 30 before water absorption, by making the amount of changeable volume of the container 28 larger than the difference δ (= V - (Vs + Vp)), that is, by making it larger than the volume Va of the empty space in the container 28 before deformation, i.e., the void portion in the container 28, the concentrated liquid can be discharged more reliably even if at least a portion of the air inside the container 28 is discharged when the concentrated liquid is discharged from the container 28.

[0037] Furthermore, it is preferable that the ratio of the surface area S2 of the flexible wall to the total surface area S1 of the container wall is 50% or more. By setting the ratio of the surface area S2 of the flexible wall to the total surface area S1 of the container wall to 50% or more, the amount of volume change possible in the container can be increased, and the concentrated liquid can be discharged more reliably when it is discharged from the container 28.

[0038] As shown in Figure 1, when the container 12 has a container body 20 and a cap 22, it is preferable that at least a part of the circumferential surface of the housing portion 20a of the container body 20 is flexible, and it is more preferable that the entire circumferential surface is flexible. Alternatively, the entire housing portion 20a may be flexible. The neck portion 20b and the cap 22 may be flexible or not, but it is preferable that they are not flexible. When the housing portion 20a and the neck portion 20b shown in Figure 2 are integrally formed from the same material, the housing portion 20a can be made flexible and the neck portion 20b not flexible by making them different thicknesses. Alternatively, the housing portion 20a and the neck portion 20b may be formed from different materials.

[0039] Furthermore, in a direction parallel to the discharge direction of the concentrated liquid, the distance from the tip of the discharge opening of the container to the flexible wall surface (i.e., the pressing area) is preferably 70 mm or less, more preferably 50 mm or less, and even more preferably 30 mm or less. The lower limit is not particularly limited, but is preferably 1 mm or more. This allows for more reliable discharge of the concentrated liquid when it is discharged from the container.

[0040] Furthermore, the ratio of the width in the direction perpendicular to the height direction (the diameter of the cross-section in the case of a cylindrical shape) of the container's containment section for the superabsorbent polymer to the height in the direction of discharge of the concentrated liquid is preferably 8 or less, more preferably 5 or less, and even more preferably 3 or less. The lower limit of the ratio is not particularly limited, but it is preferably 0.5 or more. This allows for more reliable discharge of the concentrated liquid when it is discharged from the container.

[0041] [Example of Processing Method] Figures 10 to 13 are schematic cross-sectional views showing an example of a processing method according to an embodiment of the present invention in order of steps. In Figures 10 to 13, components identical to those in the processing kit 10 shown in Figure 1 are denoted by the same reference numerals, and their detailed descriptions are omitted. As shown in Figure 10, a container 12 containing a water-absorbing polymer 16 is prepared. The water-absorbing polymer 16 is contained inside 12c of the container 12. Although not shown, the container 12 has an intake section for taking in a liquid sample (sample solution) and an outlet section for discharging a concentrated liquid sample (concentrated solution). The water-absorbing polymer 16 has the same configuration as the water-absorbing polymer 16 shown in Figure 1 and is a superabsorbent polymer (SAP) with high water absorption. The water-absorbing polymer 16 will be described later.

[0042] Next, a step is performed in which the liquid sample 30 is placed in the container 12 containing the superabsorbent polymer 16, and then a step is performed in which the water contained in the liquid sample 30 is absorbed by the superabsorbent polymer 16 to concentrate the liquid sample 30 in the container 12. Specifically, as shown in Figure 11, the liquid sample 30 is injected into the interior 12c of the container 12 containing the superabsorbent polymer 16 before water absorption. The method of injecting the liquid sample 30 is the same as in the first example of the processing method described above. The amount of liquid sample 30 injected is appropriately determined according to the capacity of the container 12, the amount of superabsorbent polymer 16, and the target amount of concentrated liquid to be recovered, for example, several tens of mL. When the liquid sample 30 is injected, the superabsorbent polymer 16 absorbs the water contained in the liquid sample 30 and swells. As shown in Figure 12, it becomes a superabsorbent polymer 18 that has absorbed water and swelled. A portion of the liquid sample 30 that was not absorbed by the superabsorbent polymer 18 is concentrated, and the concentrated liquid 34 accumulates inside the container 12c, generating a concentrate 35, which is a concentrated form of the liquid sample 30. After the above-described step of concentrating the liquid sample 30 in the container 12, a predetermined amount of recovery liquid 17 containing magnesium chloride and sodium chloride is then injected from the recovery liquid container 14 into the inside 12c of the container 12.

[0043] Following the step of placing the recovery liquid 17 containing magnesium chloride and sodium chloride into the container 12, the next step is to remove the concentrated liquid 34 of the liquid sample 30 obtained by concentrating it in the container 12 from the container 12. Here, at least a part of the container 12 is flexible, and the superabsorbent polymer 18 can be pressed through the inner wall of the container 12. Therefore, as shown in Figure 13, the swollen superabsorbent polymer 18 after water absorption can be pressed through the inner wall of the container 12, and the concentrated liquid 34 containing the recovery liquid 17 can be removed from the container 12. The concentrated liquid 34 removed from the container 12 is stored in, for example, a recovery liquid cup 50. The recovery liquid cup 50 is, for example, a cylindrical container having a bottom 50b and an opening 50a facing the bottom 50b. The concentrated liquid 34 enters the interior 50c through the opening 50a. The recovery liquid cup 50 is not particularly limited to a cylindrical container.

[0044] The recovered solution 17 containing magnesium chloride and sodium chloride is not easily absorbed by the superabsorbent polymer 16, thus suppressing the reduction in the amount of concentrated solution 34. Therefore, when recovering the concentrated solution 34, variations in the amount of concentrated solution 34 recovered each time can be suppressed. In addition, because the recovered solution 17 is not easily absorbed by the superabsorbent polymer 16, the concentrate 35 can be transferred to the concentrated solution 34. This allows the concentration of the concentrate 35 in the concentrated solution 34 to be maintained, and the predetermined concentration ratio can be maintained. Furthermore, because the recovered solution 17 contains magnesium chloride and sodium chloride, the liquid sample can be further concentrated. This increases the detection sensitivity in tests using antigen-antibody reactions with immunochromatography kits, for example. That is, the sensitivity ratio in the tests using antigen-antibody reactions described above increases. Hereafter, the detection sensitivity in tests using antigen-antibody reactions will also be simply referred to as detection sensitivity. Hereafter, high detection sensitivity will also be referred to as "excellent sensitivity." The ability to extract a concentrated solution, high detection sensitivity, and small variation in the amount of recovered solution can all be collectively described as "the superior effects of the present invention."

[0045] As described above, the inventors' studies have revealed that with conventional processing kits using superabsorbent polymers, it is difficult to remove the concentrated liquid from the processing kit, i.e., the container, after the liquid sample has been concentrated. Specifically, from the viewpoint of shortening the concentration time, a larger amount of superabsorbent polymer is preferable, but if the amount of superabsorbent polymer is large, the amount of concentrated liquid decreases, making it difficult to remove the concentrated liquid. As a result, it may not be possible to obtain a sufficient amount of concentrated liquid from the liquid sample. Also, if the saturated sample absorption capacity of the superabsorbent polymer is greater than the amount of liquid sample, water absorption will occur when removing the concentrated liquid. Therefore, it may not be possible to obtain a sufficient amount of concentrated liquid from the liquid sample. On the other hand, if the amount of superabsorbent polymer is reduced or the amount of recovered liquid is increased in order to secure the amount of concentrated liquid, the concentration ratio of the concentrated liquid will decrease. A low concentration ratio is undesirable because it reduces the amount of test substances such as antigens in the concentrated liquid, which can lead to a decrease in the detection accuracy of antigens, etc.

[0046] In contrast, the container 12 of the present invention is flexible in at least a portion and allows the superabsorbent polymer 18 to be pressed through the inner wall of the container 12. By pressing the superabsorbent polymer 18 through the inner wall of the container 12, the superabsorbent polymer 18 deforms and the internal volume of the container 12 decreases, resulting in a stirring effect that allows some of the unabsorbed liquid sample (concentrate 34) and recovered liquid 17 to spread into the gaps between the superabsorbent polymers 18. This allows for the recovery of more of the concentrate 35 remaining near the superabsorbent polymers 18, and increases the concentration ratio of the concentrate 34. Furthermore, because the container 12 allows the superabsorbent polymer 18 to be pressed through the inner wall, the contents (concentrate 34) can be directly pushed towards the discharge port. Therefore, compared to a configuration in which the concentrate is pushed out only by air pressure from a pump, etc., the concentrate can be easily removed. In addition, even a small amount of concentrate can be efficiently spread and removed from the container 12, thus increasing the concentration ratio of the concentrate 34. Furthermore, it becomes easier to secure the required amount of concentrated liquid 34, and it becomes possible to keep the concentration ratio of the extracted concentrated liquid 34 constant.

[0047] Furthermore, since the container 12 can be deformed by, for example, pressing it with the user's fingers, uneven deformation is likely to occur, allowing it to be deformed into various shapes. Also, because the superabsorbent polymer 18 is movable, it is possible to prevent the superabsorbent polymer 18 from being compressed, which would narrow the gaps between the polymers and prevent the concentrated liquid 34 from moving. As a result, even if there is a large amount of superabsorbent polymer 18, the concentrated liquid 34 can be efficiently dispensed and efficiently removed from the container 12.

[0048] Furthermore, since the concentrated liquid can be dispensed from the container 12 by pressing it with the user's finger, the dispensing operation can be performed easily, and the time required for the collection operation can be shortened.

[0049] As described above, the container 12 takes in a liquid sample 30 and discharges a concentrated liquid 34. Therefore, the container 12 has an intake section for taking in the liquid sample 30 and a discharge section for discharging the concentrated liquid 34. There are no particular limitations on the intake section; various configurations can be used as long as the liquid sample 30 can be taken into the container 12. Similarly, there are no particular limitations on the discharge section; various configurations can be used as long as the concentrated liquid 34 can be discharged from the container 12. Furthermore, the intake section and the discharge section may be shared. However, the intake section is preferably a relatively large opening, in order to easily take in the liquid sample 30 into the container 12 and to allow the superabsorbent polymer before water absorption to be placed inside the container 12, and it is preferable that the opening is larger than the particle size of the superabsorbent polymer before water absorption. On the other hand, the discharge section is preferably a relatively large opening, smaller than the particle size of the absorbent polymer after water absorption, in order to allow the concentrated liquid to be discharged without the absorbent polymer being discharged after water absorption, and to prevent air leakage that would make it difficult to remove the concentrated liquid 34 when the container 12 is pressed to discharge the concentrated liquid 34. The configuration of the processing kit will be described in more detail below.

[0050] [Container Body] The container bodies 20, 25, 25a, and 25b shown in Figures 1, 5, 6, and 7 are made of resin material such as polyethylene (PE), polyethylene terephthalate (PET), polystyrene (PS), polypropylene (PP), polyvinyl chloride (PVC), or acrylic resin (PMMA) in the flexible portion of the container, as described above, and have a thickness of 1000 μm or less. From the viewpoint of high flexibility, relatively high strength, chemical resistance, and cost, the resin material of the flexible portion of the container is preferably either polyethylene (PE) or polypropylene (PP).

[0051] [Superabsorbent Polymer] The superabsorbent polymer contained in the container is, for example, a superabsorbent polymer (SAP). The superabsorbent polymer is in particulate form, and a large number of particulate superabsorbent polymers are contained in the container. The particulate superabsorbent polymer is schematically shown as a spherical shape of uniform size, but the actual particle shape does not have to be spherical, and the particle size does not have to be uniform, but can be non-uniform. The composition of the superabsorbent polymer is not particularly limited, but for reasons that the effects of the present invention are superior, it is preferably a polyacrylic acid-based, polyacrylamide-based, cellulose-based, or polyethylene oxide-based polymer, and more preferably a polyacrylic acid-based or polyacrylamide-based polymer.

[0052] <Swelling Rate> The swelling rate of the superabsorbent polymer described above is not particularly limited, but is preferably greater than 0.2 g / g and less than 800 g / g, more preferably 1.0 g / g or more and 600 g / g or less, even more preferably 10 g / g or more and 500 g / g or less, and particularly preferably 20 g / g or more and 100 g / g or less. Here, the swelling rate is defined as the "mass (g) of water held by 1 g of superabsorbent polymer".

[0053] (Method for measuring swelling rate) The mass of the superabsorbent polymer stored at 25°C and 5% RH (relative humidity) for 10 days is measured, and then immediately immersed in a large amount of distilled water. After 120 minutes, the superabsorbent polymer is removed, the surface water is removed, and the mass is measured again. The swelling rate is then calculated using the following formula: {(Mass after water absorption (g) - Initial mass before water absorption (g)) / Initial mass before water absorption (g)}

[0054] The method for adjusting the swelling rate to the specific range described above is not particularly limited, but examples include changing the type of polymer, changing the molecular weight of the polymer, changing the degree of crosslinking, and changing the particle size.

[0055] <Water absorption rate> The water absorption rate of the superabsorbent polymer described above is not particularly limited, but it is preferably 0.01 g / min or more and 40 g / min or less per gram of superabsorbent polymer, and more preferably 0.02 g / min or more and 40 g / min or less per gram of superabsorbent polymer.

[0056] The water absorption rate mentioned above is measured as follows: A superabsorbent polymer stored at 25°C and 5% RH (relative humidity) for 10 days is weighed (weight M0, in units of g) and immersed in a large amount of distilled water. After time T1 has elapsed, the superabsorbent polymer is removed, the surface water is removed, and the mass is measured (mass M). １ Similarly, measure the mass after time T2 and time T3 from immersion. ２ and M ３ The result is obtained. Here, times T1, T2, and T3 are the same interval, and time T3 is smaller than the water absorption saturation time Ts of the superabsorbent polymer, and is within a range where the amount of water absorbed per unit time can be considered constant. For example, time T3 = Ts / 2 is sufficient.

[0057] The amount of water absorbed is defined as follows: Amount of water absorbed after time T1: ΔM1 = (M １ -M ０ ) / M ０ Amount of water absorbed after time T2: ΔM2 = (M ２ -M ０ ) / M ０ Amount of water absorbed after time T3: ΔM3 = (M ３ -M ０ ) / M ０ Using the water absorption amount defined above, the water absorption rate is determined as follows: Three points are plotted on the X-Y plane with time on the horizontal axis (x = T1, T2, T3; per minute) and water absorption amount on the vertical axis (y = ΔM1, ΔM2, ΔM3; unit g water / g polymer amount). The slope of the linear approximation equation of the water absorption amount with respect to time, using the least squares method, is taken as the water absorption rate per unit time (minute).

[0058] <Particle Size> The particle size of the superabsorbent polymer is preferably 5 mm or less, more preferably 3 mm or less, and even more preferably 1.0 mm or less, for the sake of superior effects of the present invention. The lower limit of the particle size of the superabsorbent polymer is preferably 0.01 mm or more, more preferably 0.1 mm or more, and even more preferably 0.6 mm or more, for the sake of superior effects of the present invention. As described above, the particle size of the superabsorbent polymer is even more preferably 0.6 mm (600 nm) or more and 1 mm or less. The particle size of the superabsorbent polymer is generally not uniform but has a distribution. The diameters of 50 particulate superabsorbent polymer particles are measured using an optical microscope, and the arithmetic mean of the measured diameters of the 50 particles is taken as the particle size of the superabsorbent polymer as described above.

[0059] [Ratio of superabsorbent polymer to liquid sample (amount used)] The ratio (amount used) of the superabsorbent polymer to the liquid sample is not particularly limited, but it is preferably 0.01 to 100 g, and more preferably 0.1 to 1 g, per 1 mL of liquid sample, in order to concentrate the liquid sample to an appropriate concentration and to easily extract the concentrated solution.

[0060] [Binding substance that specifically binds to polymers contained in biological fluids] When testing is performed using a concentrated solution obtained by concentrating a liquid sample, the detection sensitivity is increased. Therefore, it is preferable that the container further contains a binding substance that specifically binds to polymers contained in the biological fluids in the sample solution, as described later. When the container contains the above-mentioned binding substance, for example, the antigen-antibody reaction proceeds simultaneously with the concentration of the sample solution, and a complex of the antigen and labeled antibody in the sample solution is formed in a concentrated state, leading to an improvement in detection sensitivity.

[0061] Examples of the binding substances mentioned above include the first binding substance (particularly antibodies) described later. In other words, in the present invention, it is preferable that the polymer contained in the biological fluid is an antigen and the binding substance is an antibody.

[0062] The aforementioned binding substance is preferably contained in the container as a complex with a labeling substance. An example of such a complex is a labeled antibody. Here, a labeled antibody is an antibody to which a detectable labeling substance is bound, and a labeling substance is, for example, a detectable substance, a substance that can be directly detected, such as a substance that can produce electromagnetic waves such as color, fluorescence, or light, or a substance that can scatter electromagnetic waves such as color, fluorescence, or light, and furthermore, a substance or state that includes an enzyme that forms a light-emitting or chromogenic substance by interacting with a light-emitting precursor or a color-developing precursor.

[0063] The labeled antibody described above is preferably an antibody modified with metal particles that exhibit a vivid color when irradiated with electromagnetic waves such as visible light. The metal particles described above are more preferably gold particles. The labeled antibody described above is preferably an antibody labeled with gold particles, that is, gold particles modified with the antibody (modified gold particles described later). The labeled antibody described above may be contained in the container as a pad (gold colloid holding pad) that holds modified gold colloid particles, which are gold colloid particles modified with the antibody.

[0064] [Casein, Tricin] When testing is performed using a concentrated solution obtained by concentrating a liquid sample, the detection sensitivity is increased. Therefore, it is preferable that the container further contains at least one selected from the group consisting of casein and tricine, and it is more preferable that it contains both casein and tricine. Casein is thought to have the effect of suppressing false positives. In addition, when the pH (hydrogen ion concentration) of a sample solution such as urine is acidic, false positives are likely to occur, but tricine is thought to have the effect of suppressing false positives by adjusting the pH to neutral to alkaline.

[0065] [Liquid Sample (Specimen Solution)] The liquid sample (specimen solution) contains the test substance and water. The liquid sample (specimen solution) may be collected from a living organism, in which case the liquid sample contains biological fluid. Preferably, the liquid sample is an aqueous solution containing polymers contained in biological fluid. The water contained in the liquid sample may be water contained in biological fluid. Specific examples of liquid samples include animal (especially human) body fluids (e.g., blood, serum, plasma, cerebrospinal fluid, tears, sweat, urine, pus, nasal mucus, or sputum), gargle solution, etc. Among these, serum, plasma, urine, and nasal mucus are preferred as samples containing antigens as polymers, and urine is particularly preferred because it can be suitably used in processing kits and processing methods. Furthermore, the liquid sample may be an artificially mimicked biological fluid, for example, artificial urine. When the liquid sample (sample solution) is urine, the concentration of urea in the antigen concentrate obtained in the concentration step is preferably five times or less the concentration of urea in the sample solution, for the reason that the effects of the present invention are superior.

[0066] <Biological Fluids> Biological fluids are biological or bioorganic fluids produced by living organisms. They are any liquids that living organisms possess in some form within their bodies. Any liquid collected from a living organism can be used as biological fluid, such as blood, tissue fluid, body cavity fluid, digestive fluid, urine, saliva, sweat, tears, nasal mucus, semen, lymph, vaginal fluid, amniotic fluid, breast milk, cerebrospinal fluid, synovial fluid, and cell suspensions. Alternatively, biological samples may be those from which cellular components have been pre-disrupted or removed. Among biological fluids, blood, saliva, sweat, tears, and urine are readily available.

[0067] <Macromolecules contained in biological fluids> Examples of macromolecules (especially antigens) contained in biological fluids as described above include, for example, bacteria (e.g., Mycobacterium tuberculosis, lipoarabinomannan (LAM) contained in Mycobacterium tuberculosis), viruses (e.g., influenza virus), and their nucleoproteins, which are macromolecules that are mainly useful for diagnosing diseases and can be detected in biological fluids. LAM is a major antigen in tuberculosis and is a glycolipid that is a major component of the cell membrane and cell wall. The macromolecules contained in biological fluids as described above are preferably antigens, and the antigens are more preferably viruses (especially influenza virus) or LAM, and even more preferably LAM. The molecular weight of the macromolecules contained in biological fluids is preferably 1000 or more, and more preferably 2000 or more. If the molecular weight is a macromolecule useful for diagnosing diseases and has a known structural formula, the theoretical value calculated from the structural formula can be used. If the structural formula is not determined, it can be calculated by comparing it with a substance whose molecular weight is known using electrophoresis, or by liquid chromatography-mass spectrometry (LC-MS).

[0068] <Pretreatment of Liquid Samples> The above-mentioned liquid samples can be used as is, or in the form of a liquid obtained by extracting the antigen using a suitable extraction solvent, or in the form of a diluted solution obtained by diluting the extracted solution with a suitable diluent, or in the form of a concentrated solution obtained by a suitable method. As the extraction solvent, solvents commonly used in immunological analysis methods (e.g., water, physiological saline, or buffer solution) or water-miscible organic solvents that can be used to directly carry out antigen-antibody reactions by dilution with such solvents can also be used.

[0069] [Recovery Solution] The recovery solution contains magnesium chloride and sodium chloride. The sodium chloride and magnesium chloride contained in the recovery solution can increase the detection sensitivity, i.e., the sensitivity ratio mentioned above, and reduce the variability in the amount of concentrated solution recovered. The recovery solution contains a solvent used in conventional immunological analytical methods, or a water-miscible organic solvent that can be used to directly carry out an antigen-antibody reaction by diluting with such a solvent. Solvents used in conventional immunological analytical methods include, for example, water, physiological saline, or buffer solution. The recovery solution may also be given functionality by buffering agents, surfactants, and other additives as needed. The recovery solution is preferably a buffer solution, and more preferably PBS (Phosphate Buffered Salts). A portion of the sample solution may also be used as the recovery solution. The recovery solution may contain a preservative, for example, Proclin 950 (trade name, manufactured by Sigma-Aldrich Japan LLC).

[0070] The magnesium chloride content of the recovered solution is preferably 10 mg / mL (milliliters) or more and 80 mg / mL (milliliters) or less. A magnesium chloride content of 10 mg / mL or more and 80 mg / mL or less in the recovered solution is preferable because it reduces the variation in the amount of concentrated solution recovered and further increases the detection sensitivity, i.e., the sensitivity ratio mentioned above. The sodium chloride content of the recovered solution is preferably 30 mg / mL or more and 200 mg / mL or less in the recovered solution. A sodium chloride content of 30 mg / mL or more and 200 mg / mL or less in the recovered solution is preferable because it suppresses the absorption of the recovered solution by the superabsorbent polymer, reduces the variation in the amount of concentrated solution recovered, and further increases the detection sensitivity, i.e., the sensitivity ratio mentioned above. Note that the magnesium chloride and sodium chloride content of the recovered solution are the mass of the solute salt relative to the amount of solvent in the recovered solution. The magnesium chloride and sodium chloride content of the recovered solution are the values ​​obtained by converting the mass of magnesium chloride and sodium chloride in the recovered solution to mass per milliliter.

[0071] The amount of recovered solution is less than the amount of liquid sample injected into the container, from the viewpoint of concentrating the liquid sample (sample solution). The ratio of the amount of recovered solution to the amount of liquid sample injected into the container (amount of recovered solution / amount of liquid sample (sample solution)) should be less than 100% by volume, preferably 30% or less, more preferably 20% or less, and even more preferably 0.01% or more and 10% or less.

[0072] The salts contained in the recovered solution are required not to inhibit the antigen-antibody reaction in immunochromatography. The effect of salts and other substances in the recovered solution on the antigen-antibody reaction of the immunochromatographic kit was investigated. Four types of recovered solutions were used, each containing sodium chloride, calcium chloride, magnesium chloride, or citric acid as additives. To evaluate the effect on the antigen-antibody reaction, an immunochromatographic kit for detecting Mycobacterium tuberculosis antigen, used to detect lipoarabinomannan antigen as the test substance, as described in International Publication No. 2020 / 045625, was used. The configuration of the immunochromatographic kit was as shown in Figures 1 to 3 of International Publication No. 2020 / 045625. Artificial urine (JIS (Japanese Industrial Standards) T3214) containing 1 mg / mL of BSA (Bovine Serum Albumin) with LAM (lipoarabinomannan) antigen was used as the sample solution. It was diluted to an LAM concentration equivalent to 200 pg / mL. Furthermore, the recovery solution was added to the sample solution to investigate its effects.

[0073] Following the operating procedure for the immunochromatography kit described in the aforementioned International Publication No. 2020 / 045625, 200 μL of the sample solution with the aforementioned recovery solution added was placed in a tube. The sample solution with the aforementioned recovery solution added was then immersed in a pad from the tube and allowed to stand for 40 minutes to react with the gold colloid and antigen. Subsequently, the sample was spotted onto the immunochromatography kit, and its effect on the antigen-antibody reaction was evaluated. The results of the evaluation of the effect on the antigen-antibody reaction are shown in Table 1 below. The effect on the antigen-antibody reaction was evaluated in terms of its effect on the surface and its effect on sensitivity and false positives. The effect on the surface was visually confirmed by observing the surface through the observation window of the immunochromatography kit. The effect on sensitivity and false positives was visually confirmed by increasing the amount of additive and observing the coloration of the test line and the line indicating a positive result on the surface observed through the observation window of the immunochromatography kit. When the test line and the positive line are easily visible, the signal-to-noise ratio (S / N ratio) is higher, sensitivity is higher, and the likelihood of false positives is lower. On the other hand, when the test line and the positive line are difficult to see, the S / N ratio is lower, sensitivity is lower, and the likelihood of false positives is higher. Note that, as described later, the evaluation results for the effect on surface properties of calcium chloride and citric acid were poor, so the effect on sensitivity and false positives was not investigated. For this reason, "-" is written in the "Effect on Sensitivity and False Positives" column in Table 1 below.

[0074] When calcium chloride and citric acid were used as additives in the recovery solution, the surface observed through the observation window of the immunochromatograph kit turned black, making it impossible to determine whether the result was positive or negative. When sodium chloride and magnesium chloride were used as additives in the recovery solution, the effect on the surface observed through the observation window of the immunochromatograph kit was small, therefore sodium chloride and magnesium chloride are used. From the standpoint of the effect on the surface observed through the observation window and the effect on sensitivity and false positives, the recovery solution shall contain sodium chloride and magnesium chloride. The surface refers to the immunochromatographic reaction area (the component holding the various lines) visible through the observation window. Furthermore, regarding the effect on the surface, a small effect on the surface means that there is little discoloration such as blackening, and that the effect of discoloration on determining positive or negative results is small. A small effect on the surface and a small effect on the surface are synonymous.

[0075]

[0076] The concentrated liquid sample (sample solution) obtained using the processing kit and processing method is used in a sample solution testing method for detecting polymers in the sample solution, which is an aqueous solution containing polymers. Polymers in the concentrated solution obtained by concentrating the sample solution can be detected by various known methods. The concentrated solution can be concentrated to a high concentration ratio, reliably extracted, and the variation in the amount of concentrated solution recovered each time is small. Therefore, the detection sensitivity is high, and the repeatability of detection is also high.

[0077] Methods for detecting polymers in concentrated solutions preferably involve antigen-antibody reactions, such as enzyme immunoassay (EIA), solid-phase enzyme immunoassay (ELISA), radioimmunoassay (RIA), fluorescence immunoassay (FIA), Western blotting, and immunochromatography. Among these, the processing kit and processing method are particularly suitable for concentrating liquid samples (sample solutions) for immunochromatography. Specific methods for detecting polymers in concentrated solutions, and the configuration of test kits for detecting polymers in sample solutions (concentrated solutions) containing polymers, are described, for example, in Japanese Patent Application Publication No. 2009-150869 and International Publication No. 2021 / 065300.

[0078] [Immunoassay Method] The immunoassay method using the above-described processing method in the concentration step will be described below. The immunoassay method comprises a concentration step of obtaining an antigen concentrate by concentrating a liquid that may contain an antigen (liquid sample or specimen solution) by mixing it with the above-described superabsorbent polymer, and a detection step of detecting the antigen in the antigen concentrate using an antigen-antibody reaction. The recovered solution used in the concentration step contains magnesium chloride and sodium chloride. As described above, from the viewpoint of sensitivity ratio and variability of recovered amount, it is preferable that the magnesium chloride content of the recovered solution is 10 mg / mL or more and 80 mg / mL or less, and the sodium chloride content of the recovered solution is 30 mg / mL or more and 200 mg / mL or less. The steps of the immunoassay method will be described below.

[0079] [Concentration Process] The concentration process is a process for obtaining an antigen concentrate, and Figures 10 to 13 above show the processing method of an embodiment of the present invention. First, as shown in Figure 11, a liquid that may contain an antigen corresponding to the liquid sample 30 is placed in a container 12 containing a superabsorbent polymer 16. Next, the water contained in the antigen-containing liquid is absorbed by the superabsorbent polymer 16 to concentrate the antigen-containing liquid in the container 12. Then, as shown in Figure 12, a predetermined amount of recovery liquid 17 containing magnesium chloride and sodium chloride is injected from the recovery liquid container 14 into the interior 12c of the container 12. The amount of recovery liquid 17 is less than the amount of the antigen-containing liquid. Here, as described above, at least a part of the container 12 is flexible, and the superabsorbent polymer 18 can be pressed through the inner wall of the container 12. Therefore, as shown in Figure 13, the swollen superabsorbent polymer 18 after water absorption is pressed through the inner wall of the container 12 to remove the concentrate 34 containing the recovery liquid 17 from the container 12 and collect it in the recovery liquid cup 50. The concentrated solution 34 is a concentrated solution of a liquid that may contain an antigen, i.e., an antigen concentrate. For the concentration process, the container 12 with the configuration shown in Figure 1 and the recovery liquid container 14 with the configuration shown in Figure 3 can be used.

[0080] [Detection Step] The detection step is a step in which the antigen in the antigen concentrate obtained in the concentration step described above is detected using an antigen-antibody reaction. The detection step is not particularly limited as long as it uses an antigen-antibody reaction, but examples include enzyme immunoassay (EIA), solid-phase enzyme immunoassay (ELISA), radioimmunoassay (RIA), fluorescence immunoassay (FIA), Western blotting, and immunochromatography. Among these, immunochromatography is preferred because it is more effective than the method of the present invention. In other words, the method of the present invention is preferably immunochromatography. [Antigen Concentrate] The antigen concentrate obtained in the concentration step described above is used in the detection step.

[0081] [Preferred Embodiments] The following describes preferred embodiments when the detection step is immunochromatography. For reasons that the effects of the present invention are superior, the detection step preferably comprises: a spreading step in which a gold particle complex is formed, which is a complex of the antigen in the antigen concentrate and modified gold particles modified with a first binding substance capable of binding to the antigen, and then spread onto an insoluble carrier having a reaction site on which a second binding substance capable of binding to the antigen is immobilized; a capturing step in which the gold particle complex is captured at the reaction site of the insoluble carrier; and a silver amplification step in which the gold particle complex captured in the capturing step is silver amplified. For reasons that the effects of the present invention are superior, it is preferable that at least one of the first binding substance and the second binding substance is a monoclonal antibody, and it is preferable that both the first binding substance and the second binding substance are monoclonal antibodies. The following describes each step of the preferred embodiment.

[0082] [Development Step] The development step involves developing a gold particle complex, which is a composite of the antigen in the antigen concentrate obtained in the concentration step described above and modified gold particles, which are gold particles modified with a first binding substance capable of binding to the antigen, onto an insoluble carrier having a reaction site on which a second binding substance capable of binding to the antigen is immobilized.

[0083] [Gold Particle Composite] As described above, in the development step, a gold particle composite is first formed, which is a composite of the antigen in the antigen concentrate obtained in the concentration step described above and modified gold particles, which are gold particles modified with a first binding substance capable of binding to the antigen. <Modified Gold Particles> Modified gold particles are gold particles modified with a first binding substance capable of binding to the antigen.

[0084] (Gold Particles) The gold particles are not particularly limited. The gold particles act as a catalyst to reduce silver ions in the silver amplification step described later. The particle size of the gold particles is preferably 100 nm or less, more preferably 50 nm or less, even more preferably 30 nm or less, and particularly preferably 15 nm or less, for the sake of superior effects of the present invention. The lower limit of the particle size of the gold particles is not particularly limited, but for the sake of superior effects of the present invention, it is preferably 1 nm or more, more preferably 2 nm or more, and even more preferably 5 nm or more. The particle size can be measured using a commercially available particle size analyzer. Known methods for measuring particle size distribution include optical microscopy, confocal laser microscopy, electron microscopy, atomic force microscopy, static light scattering, laser diffraction, dynamic light scattering, centrifugal sedimentation, electrical pulse measurement, chromatography, and ultrasonic attenuation, and devices corresponding to each principle are commercially available. As a method for measuring particle size, dynamic light scattering can be preferably used due to the particle size range and ease of measurement. Examples of commercially available measuring devices using dynamic light scattering include the NanoTrack UPA (Nikkiso Co., Ltd.), the Dynamic Light Scattering Particle Size Distribution Analyzer LB-550 (Horiba, Ltd.), and the Concentrated Particle Size Analyzer FPAR-1000 (Otsuka Electronics Co., Ltd.). In this invention, the value is determined as the median diameter (d=50) measured at a measurement temperature of 25°C.

[0085] (First binding substance) The first binding substance is not particularly limited as long as it can bind to the antigen, but it is preferably a protein, more preferably an antibody (e.g., a polyclonal antibody or a monoclonal antibody) for the sake of superior effects of the present invention, and even more preferably a monoclonal antibody in terms of achieving higher detection sensitivity. The antibody is not particularly limited, but for example, antiserum prepared from the serum of an animal immunized with the antigen, or an immunoglobulin fraction purified from the antiserum can be used, or monoclonal antibodies obtained by cell fusion using spleen cells of an animal immunized with the antigen, or fragments thereof [e.g., F(ab')2, Fab, Fab', or Fv] can be used. These antibodies can be prepared by conventional methods. An example of the first binding substance when the antigen is LAM is the A194-01 antibody described in International Publication No. 2017 / 139153. All information contained in International Publication 2017 / 139153 relating to the A194-01 antibody is incorporated herein by reference as part of the disclosure herein. Another example of a first conjugate when the antigen is LAM is an antibody having the sequence described as MoAb1 in paragraph

[0080] of International Publication 2013 / 129634. All information contained in International Publication 2013 / 129634 relating to the MoAb1 antibody is incorporated herein by reference as part of the disclosure herein.

[0086] (Method for producing modified gold particles) The method for producing modified gold particles is not particularly limited, and known methods can be used. For example, a chemical bonding method can be used that utilizes the chemical bonding between gold and SH groups, by introducing SH groups into an antibody, and then immobilizing the gold particles with Au-S bonds formed on the Au surface when the antibody approaches the SH bond, which cleaves.

[0087] [Insoluble Carrier] The insoluble carrier is an insoluble carrier having a reaction site (test line) on which a second binding substance capable of binding to an antigen is immobilized. The insoluble carrier may have multiple test lines depending on the type of antigen (for example, a test line for influenza A virus and a test line for influenza B virus). The insoluble carrier may also have a control line downstream of the test line to confirm the development of the gold particle complex. Furthermore, if a reducing agent solution is used in the silver amplification step described later, a chromogenic reagent immobilization line may be provided downstream of the test line to detect the reducing agent solution. A specific embodiment of the insoluble carrier is, for example, a nitrocellulose membrane 100 having, from the upstream side, a gold colloid holding pad 1, a test line 2, a control line 3, and a chromogenic reagent immobilization line 4, as shown in Figure 14. Here, the gold colloid holding pad 1 is a pad that holds gold particles modified with a first binding substance (modified gold particles), the test line 2 is a line on which the second binding substance is immobilized, the control line 3 is a line for confirming the development, and the color development reagent immobilization line 4 is a line for detecting the reducing agent solution described later. Here, upstream and downstream refer to the development of the gold particle complex from the upstream side to the downstream side. More specific embodiments of the insoluble carrier or immunochromatographic kit having the same include, for example, the insoluble carrier and immunochromatographic kit described in Japanese Patent Publication No. 5728453, and all the contents of Japanese Patent Publication No. 5728453 relating to the insoluble carrier and immunochromatographic kit are incorporated herein by reference as part of the disclosure herein.

[0088] <Insoluble Carrier> A porous carrier is preferred as the insoluble carrier. In particular, for reasons that the effects of the present invention are superior, nitrocellulose membranes, cellulose membranes, acetylcellulose membranes, polysulfone membranes, polyethersulfone membranes, nylon membranes, glass fibers, nonwoven fabrics, cloths, or yarns are preferred, and nitrocellulose membranes are more preferred.

[0089] (Second Binding Substance) The second binding substance is not particularly limited as long as it can bind to the antigen. Specific examples and preferred embodiments of the second binding substance are the same as those of the first binding substance described above. The second binding substance may be the same as or different from the first binding substance described above, but it is preferable that it be a different substance for reasons that the effects of the present invention are superior. Furthermore, if the first and second binding substances are antibodies, it is preferable that the antibody that is the first binding substance and the antibody that is the second binding substance be different for reasons that the effects of the present invention are superior. Furthermore, if the first and second binding substances are antibodies, it is preferable that the epitope of the first binding substance (a part of the antigen recognized by the first binding substance) and the epitope of the second binding substance (a part of the antigen recognized by the second binding substance) be different for reasons that the effects of the present invention are superior. The difference in antibody epitopes can be confirmed, for example, by solid-phase enzyme immunoassay (ELISA).

[0090] [Development] There are no particular limitations on the method of developing the gold particle composite onto an insoluble carrier having a test line. For example, one method involves preparing the nitrocellulose membrane 100 (or an immunochromatography kit having the same) shown in Figure 14 above, dropping the antigen concentrate obtained in the concentration step above onto the gold colloid holding pad, and moving it from the upstream side to the downstream side using capillary action as shown in Figure 14.

[0091] [Capture Step] The capture step is a step in which the gold particle complex is captured at the reaction site of the insoluble carrier. As described above, a second binding substance capable of binding to the antigen is immobilized at the reaction site of the insoluble carrier, so the gold particle complex (complex of antigen and modified gold particles) that is deployed on the insoluble carrier in the deployment step is captured at the reaction site (test line) of the insoluble carrier. Note that if the sample solution does not contain the antigen, the gold particle complex will not be formed, and therefore the gold particle complex will not be captured at the reaction site of the insoluble carrier.

[0092] [Silver Amplification Process] The silver amplification process is a process of silver amplification of the gold particle complex captured in the capture process. The silver amplification process is a process in which silver ions are added to the insoluble support after the capture process, thereby forming larger silver particles in the gold particle complex captured at the reaction site of the insoluble support. More specifically, it is a process in which silver ions are reduced using the gold particles of the gold particle complex as a catalyst, and silver particles (for example, with a diameter of 10 μm or more) are formed. As a result, the detection sensitivity of the captured gold particle complex is significantly improved.

[0093] [Preferred Embodiments] The method for imparting silver ions to the insoluble carrier after the capture step is not particularly limited, but a method using the following reducing agent solution and silver amplification solution is preferred for better effects of the present invention. In addition to the reducing agent solution and silver amplification solution, a washing solution may be used to wash away any complexes remaining on the insoluble carrier other than those resulting from specific binding reactions. The reducing solution may also serve as the washing solution.

[0094] <Reducing Agent Solution> The reducing agent solution contains a reducing agent capable of reducing silver ions. Any inorganic or organic material, or a mixture thereof, can be used as the reducing agent capable of reducing silver ions to silver. Examples of inorganic reducing agents include Fe. ２＋ , V ２＋ Ti ３＋ Examples of preferred reducing metal salts and reducing metal complex salts are those whose valence can change with metal ions such as the following. When using inorganic reducing agents, it is necessary to remove or detoxify the oxidized ions by forming a complex or reducing them. For example, Fe ２＋ In systems using citric acid or ethylenediaminetetraacetic acid (EDTA) as a reducing agent, the oxide Fe ３＋ It can form a complex and render harmless. In the present invention, it is preferable to use such an inorganic reducing agent, and in a more preferred embodiment of the present invention, Fe ２＋It is preferable to use a metal salt of as a reducing agent. Furthermore, developing agents used in wet silver halide photographic materials (e.g., methyl gallate, hydroquinone, substituted hydroquinone, 3-pyrazolidones, p-aminophenols, p-phenylenediamines, hindered phenols, amidoximes, azines, catechols, pyrogallols, ascorbic acid (or its derivatives), and leuco dyes), and other materials that are obvious to those skilled in the art, such as the materials described in U.S. Patent No. 6,020,117, can also be used as reducing agents.

[0095] As a reducing agent, ascorbic acid reducing agents are also preferred. Useful ascorbic acid reducing agents include ascorbic acid and its analogs, isomers and derivatives, and for example, D- or L-ascorbic acid and its sugar derivatives (e.g., γ-lactoascorbic acid, glucoascorbic acid, fucoscorbic acid, glucoheptascorbic acid, maltoascorbic acid), sodium salt of ascorbic acid, potassium salt of ascorbic acid, isoascorbic acid (or L-erythroascorbic acid), salts thereof (e.g., alkali metal salts, ammonium salts or salts known in the art), enediol-type ascorbic acid, enaminol-type ascorbic acid, thioenol-type ascorbic acid, etc., with D, L, or D, L-ascorbic acid (and its alkali metal salts) or isoascorbic acid (or its alkali metal salts) being particularly preferred, and sodium salts being preferred salts. Mixtures of these reducing agents can be used as needed. For the best results of the present invention, it is preferable to flow the reducing agent solution such that the angle between the development direction in the development process and the development direction of the reducing agent solution is between 0 and 150 degrees, and more preferably between 0 and 135 degrees. As an example of how to adjust the angle between the development direction in the development process and the development direction of the reducing agent solution, the method described in the examples of Japanese Patent Application Publication No. 2009-150869 can be used.

[0096] <Silver Amplification Solution> The silver amplification solution is a solution containing a compound containing silver ions. Examples of compounds containing silver ions include organic silver salts, inorganic silver salts, or silver complexes. Preferably, the silver ion-containing compound has high solubility in a solvent such as water, and examples include silver nitrate, silver acetate, silver lactate, silver butyrate, and silver thiosulfate. Silver nitrate is particularly preferred. As for the silver complex, a silver complex coordinated to a ligand having a water-soluble group such as a hydroxyl group or a sulfone group is preferred, and examples include silver hydroxythioether. The organic silver salt, inorganic silver salt, or silver complex is contained in the silver amplification solution as silver at a concentration of, for example, 0.001 mol / L to 5 mol / L, preferably at a concentration of 0.005 mol / L to 3 mol / L, and more preferably at a concentration of 0.01 mol / L to 1 mol / L.

[0097] Auxiliaries for the silver amplification solution include buffers, preservatives, such as antioxidants or organic stabilizers, and rate regulators. As buffers, for example, acetic acid, citric acid, sodium hydroxide, or salts thereof, or buffers using tris(hydroxymethyl)aminomethane, or other buffers commonly used in chemical experiments can be used. These buffers can be used as appropriate to adjust the pH of the amplification solution to the optimal level. Alkylamines can also be used as an anti-fogging agent, with dodecylamine being particularly preferred. Furthermore, surfactants can be used to improve the solubility of these auxiliary agents, with C being particularly preferred. ９ H １９ -C ６ H ４ -O-(CH ２ CH ２ O) ５０ H. The silver amplification solution is preferably flowed in the opposite direction to the development step described above, for better effects of the present invention, and more preferably flowed so that the angle between the development direction in the development step and the development direction of the reducing agent solution is 45 to 180 degrees. As a method for adjusting the angle between the development direction in the development step and the development direction of the silver amplification solution, for example, the method described in the example of Japanese Patent Application Publication No. 2009-150869 can be cited.

[0098] The present invention is basically configured as described above. Although the processing kit and processing method of the present invention have been described in detail above, the present invention is not limited to the embodiments described above, and various improvements or modifications may be made without departing from the spirit of the present invention.

[0099] The features of the present invention will be further described in detail below with reference to examples. The materials, reagents, amounts and proportions of substances, and procedures shown in the following examples can be modified as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention is not limited to the following examples. In these examples, the sensitivity ratio and the variability of the recovered concentrated solution were evaluated for Examples 1 to 11 and Comparative Examples 1 to 13.

[0100] [1] Preparation of the concentration jig The container body is made of polyethylene. The thickness of the containment section is 600 μm. Therefore, the containment section is flexible. The containment section contains SAP obtained by classifying Aquaric CA H2 manufactured by Nippon Shokubai Co., Ltd. Specifically, the classification was performed by using two sieves with different mesh sizes, obtaining SAP that passed through the sieve with the larger mesh size but not through the sieve with the smaller mesh size. The sieves used this time were 600 μm and 1000 μm, and the calculated particle size was 800 μm. The amount of SAP to be contained in the containment section (hereinafter also referred to as "SAP amount") was 0.55 g. As described above, since the containment section is flexible, the SAP can be pressed through the inner wall of the container.

[0101] [2] Preparation of sample solution Artificial urine (Catalog No.: 90048945, manufactured by Isekyu Co., Ltd.) was diluted 16 times with ultrapure water (Milli-Q) to which lipoarabinomannan (LAM) (02249-61, manufactured by Nacalai Tesque Co., Ltd.) extracted from Mycobacterium tuberculosis and 0.1% BSA (Bovine serum albumin) (Catalog No.: A7906, manufactured by SIGMA Corporation) were added to prepare an artificial urine sample solution containing 20 pg / mL of LAM.

[0102] [3] Preparation of gold colloid retaining pads 9 mL of a solution containing gold colloid particles (particle size: 50 nm) (product number: EM.GC50, manufactured by BBI) is mixed with 50 mmol / L of KH ２ PO ４The pH was adjusted by adding 1 mL of buffer (pH 8.0). To the pH-adjusted solution, 1 mL of a solution containing 20 μg / mL anti-lipoarabinomannan (LAM) monoclonal antibody (Nacalai Tesque Co., Ltd., product code 05494-84) was added and stirred for 10 minutes. After standing for 10 minutes, 550 μL of an aqueous solution containing 1% by mass polyethylene glycol (PEG (polyethylene glycol); weight-average molecular weight (Mw.): 20000, catalog number: 168-11285, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added and stirred for 10 minutes. Subsequently, 1.1 mL of an aqueous solution of 10% by mass bovine serum albumin (BSA (Bovine serum albumin); Fraction V, catalog number: A-7906, manufactured by SIGMA) was added and stirred for 10 minutes. This solution was centrifuged using a centrifuge (himacCF16RX, manufactured by Eppendorf Himac Technologies, Ltd.) at 8000 x g and 4°C for 30 minutes. The supernatant was removed, leaving 1 mL at the bottom of the container, and the gold colloid particles contained in the remaining 1 mL were redispersed using an ultrasonic cleaner. After this, the particles were dispersed in 20 mL of gold colloid preservation solution (20 mmol / L Tris-HCl buffer (pH 8.2), 0.05% PEG (Mw. 20000), 150 mmol / L NaCl, 1% BSA), and centrifuged again using the same centrifuge under the same conditions. The supernatant was removed, ultrasonically dispersed, and then dispersed in the gold colloid preservation solution to obtain a solution of antibody-modified gold colloid particles (labeled antibody), which are gold colloid particles (particle size: 50 nm) modified with an anti-LAM monoclonal antibody. The obtained solution was adjusted in concentration with buffer, then dropped onto a 5 mm x 30 cm glass fiber pad (Merck GFDX203000), dried in a vacuum dryer for 15 hours, and then cut to obtain a pad (gold colloid-holding pad) (5 mm x 4 mm) containing antibody-modified gold colloid particles (labeled antibody), which are gold colloid particles modified with an anti-LAM monoclonal antibody.

[0103] [4] Immunochromatography [Concentration Step] Add 12 mL of the sample solution described above to the concentration jig (SAP amount: 0.55 g) prepared as described above and mix. In this way, the sample solution and SAP were mixed. After standing for 20 minutes, 1.2 mL of the recovered solution was added to the concentration jig and the container was shaken for 15 seconds to mix. Then, the SAP was pressed through the inner wall of the flexible container of the concentration jig to extract the antigen concentrate from the concentration jig. If no solution was obtained, add another 1.2 mL of the recovered solution to the concentration jig, shake the container for 15 seconds to mix, and then extract the concentrate.

[0104] <Recovered Solution> In Examples 1-11 and Comparative Examples 6-8, the recovered solution was a solution obtained by adding sodium chloride and magnesium chloride to pure water. In Examples 1-11, a preservative (Proclin 950) was added at a concentration of 0.05%. The sodium chloride and magnesium chloride content is shown in Table 2 below. In Comparative Examples 2-4 and 10-12, the recovered solution was a solution obtained by adding sodium chloride to pure water. The sodium chloride content is shown in Table 2 below. In Comparative Examples 5 and 13, the solution was a solution obtained by adding magnesium chloride to pure water. The magnesium chloride content is shown in Table 2 below.

[0105] [Deployment Process] As shown in Figure 14, a nitrocellulose membrane 100 was prepared, having a gold colloid holding pad 1, a test line 2, a control line 3, and a chromogenic reagent immobilization line 4, starting from the upstream side. The gold colloid holding pad 1 is a pad that holds gold colloid modified with anti-LAM monoclonal antibody (modified gold particles), the test line 2 is a line on which anti-LAM monoclonal antibody is immobilized, the control line 3 is a line for confirming deployment, and the chromogenic reagent immobilization line 4 is a line for detecting the reducing solution of the silver amplification process described later. The obtained antigen concentrate was dropped onto the gold colloid holding pad 1. This formed a gold particle complex, which is a complex of LAM in the solution and gold colloid particles modified with anti-LAM monoclonal antibody (modified gold particles). The formed gold particle complex was deployed from the upstream side to the downstream side of the nitrocellulose membrane 100 shown in Figure 14. [Capture Process] The gold particle complex deployed in the deployment process is captured by the test line 2 (see Figure 14).

[0106] [Silver Amplification Process] The silver amplification process was carried out as follows: <Preparation of Reducing Agent Solution> 23.6 mL of 1 mol / L iron nitrate aqueous solution, prepared by dissolving iron(III) nitrate nonahydrate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) in water, and 13.1 g of citric acid (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were dissolved in 290 g of water. After everything was dissolved, 36 ml of nitric acid (10% by mass) was added while stirring with a stirrer, and 60.8 g of ammonium iron(II) sulfate hexahydrate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added to make this the reducing agent solution. <Preparation of Silver Amplification Solution> 8 mL of silver nitrate solution (containing 10 g of silver nitrate) and 24 mL of 1 mol / L iron nitrate aqueous solution were added to 66 g of water. Furthermore, this solution was mixed with 5.9 mL of nitric acid (10% by mass), 0.1 g of dodecylamine (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and surfactant C. １２ H ２５ -C ６ H ４ -O-(CH ２ CH ２ O) ５０ A solution of 0.1 g of H dissolved in 47.6 g of water was mixed with the silver amplification solution.

[0107] <Development of reducing agent solution> On the nitrocellulose membrane 100 (see Figure 14), the reducing agent solution prepared as described above was flowed from the same direction as the development step described above (from further upstream). <Development of silver amplification solution> After the color development reagent immobilization line 4 (see Figure 14) changed color, the silver amplification solution prepared as described above was flowed from the opposite direction to the development direction in the development step (from downstream). In this way, the gold particle composite captured in test line 2 (see Figure 14) was amplified by silver.

[0108] <Calculation of Sensitivity Ratio> The sensitivity ratios for Examples 1-11 and Comparative Examples 1-8 and 13 were calculated based on the signal intensity of the test line (hereinafter referred to as ΔOD). From images captured by a chemiluminescence CCD measuring device (Fujifilm Corporation, LAS-4000), the difference in signal intensity between the test line and the surrounding background was defined as ΔOD from the data obtained by quantifying the intensity using analysis software (Fujifilm Corporation, Multigauge). In order to determine the sensitivity ratio from ΔOD, first, the correlation between ΔOD and the LAM concentration in the sample was evaluated, and a calibration curve between ΔOD and LAM concentration was calculated. Specifically, a and b in the following formula were calculated: (LAM concentration) = a・exp(b・ΔOD) Next, the ΔOD in Examples 1-11 and Comparative Examples 1-8 and 13 were converted to LAM concentration (c) using the above formula. Finally, the sensitivity ratio (ε) is calculated using the LAM concentration (c) of Comparative Example 1 (no concentration, and no addition of sodium chloride or magnesium chloride to the sample). ０ The LAM concentration (c) was calculated from the evaluation results at each level and the following formula: ε = c / c ０

[0109] <Variation in the amount of concentrated solution recovered> To investigate the variation in the amount of concentrated solution recovered, the concentrated solution was recovered 10 times. At each recovery, the amount of concentrated solution that was not absorbed by the superabsorbent polymer was weighed. From the recovered amounts of concentrated solution for the 10 times, the standard deviation (σ) and mean were calculated, and the coefficient of variation was determined. The coefficient of variation is (standard deviation (σ) / mean) × 100 (%). The coefficient of variation was evaluated based on the evaluation criteria below. For comparative examples 1 to 8, which do not have a concentration process, there was no recovery work, so "-" is written in the "Variation in the amount of recovered" column in Table 2 below. The variation in the amount of recovered in Table 2 is the variation in the amount of concentrated solution recovered.

[0110] Examples 1 to 11 and Comparative Examples 1 to 13 will be described below. (Example 1) In Example 1, a concentration process was carried out and the recovered liquid magnesium chloride (MgCl) ２ (Example 1) Example 2 differs from Example 1 in that the sodium chloride content of the recovered solution was 62.5 mg / mL. Otherwise, it was the same as Example 1. (Example 3) Example 3 differs from Example 1 in that the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1.

[0111] (Example 4) Example 4 differs from Example 1 in that the sodium chloride content of the recovered solution was 156.25 mg / mL. Otherwise, it was the same as Example 1. (Example 5) Example 5 differs from Example 1 in that the sodium chloride content of the recovered solution was 187.5 mg / mL. Otherwise, it was the same as Example 1. (Example 6) Example 6 differs from Example 1 in that the sodium chloride content of the recovered solution was 250 mg / mL. Otherwise, it was the same as Example 1.

[0112] (Example 7) Example 7 differs from Example 1 in that the magnesium chloride content of the recovered solution was 15 mg / mL and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1. (Example 8) Example 8 differs from Example 1 in that the magnesium chloride content of the recovered solution was 45 mg / mL and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1. (Example 9) Example 9 differs from Example 1 in that the magnesium chloride content of the recovered solution was 75 mg / mL and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1.

[0113] (Example 10) Example 10 differs from Example 1 in that the magnesium chloride content of the recovered solution was 90 mg / mL and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1. (Example 11) Example 11 differs from Example 1 in that the magnesium chloride content of the recovered solution was 120 mg / mL and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it was the same as Example 1.

[0114] (Comparative Example 1) Comparative Example 1 differs from Example 1 in that the concentration process is not performed and the recovered liquid is not used. Otherwise, it is the same as Example 1. For Comparative Example 1, "-" is written in the "Magnesium Chloride", "Sodium Chloride", and "Variation of Recovery Amount" columns of Table 2. (Comparative Example 2) Comparative Example 2 differs from Example 1 in that the concentration process is not performed and only sodium chloride is added to the recovered liquid. Otherwise, it is the same as Example 1. The sodium chloride content of the recovered liquid was 62.5 mg / mL. For Comparative Example 2, "-" is written in the "Magnesium Chloride" and "Variation of Recovery Amount" columns of Table 2. (Comparative Example 3) Comparative Example 3 differs from Example 1 in that the concentration process is not performed and only sodium chloride is added to the recovered liquid. Otherwise, it is the same as Example 1. The sodium chloride content of the recovered liquid was 125 mg / mL. For Comparative Example 3, "-" is indicated in the "Magnesium Chloride" and "Variation in Recovery Amount" columns of Table 2. (Comparative Example 4) Comparative Example 4 differs from Example 1 in that the concentration process is not performed and only sodium chloride is added to the recovered solution. Otherwise, it is the same as Example 1. The sodium chloride content of the recovered solution was set to 250 mg / mL. For Comparative Example 4, "-" is indicated in the "Magnesium Chloride" and "Variation in Recovery Amount" columns of Table 2.

[0115] (Comparative Example 5) Comparative Example 5 differs from Example 1 in that the concentration process is not performed and only magnesium chloride is added to the recovered solution. Otherwise, it is the same as Example 1. The magnesium chloride content of the recovered solution was 60 mg / mL. For Comparative Example 5, "-" is written in the "Sodium Chloride" and "Variation of Recovered Amount" columns of Table 2. (Comparative Example 6) Comparative Example 6 differs from Example 1 in that the concentration process is not performed and the sodium chloride content of the recovered solution was 62.5 mg / mL. Otherwise, it is the same as Example 1. For Comparative Example 6, "-" is written in the "Variation of Recovered Amount" column of Table 2. (Comparative Example 7) Comparative Example 7 differs from Example 1 in that the concentration process is not performed and the sodium chloride content of the recovered solution was 125 mg / mL. Otherwise, it is the same as Example 1. For Comparative Example 7, "-" is written in the "Variation of Recovered Amount" column of Table 2. (Comparative Example 8) Comparative Example 8 differs from Example 1 in that it does not involve a concentration step and the sodium chloride content of the recovered solution is 250 mg / mL. Otherwise, it is the same as Example 1. For Comparative Example 8, "-" is indicated in the "Variation of recovered amount" column of Table 2.

[0116] (Comparative Example 9) Comparative Example 9 differs from Example 1 in that it does not use a recovered solution. Otherwise, it is the same as Example 1. For Comparative Example 9, "-" is written in the "Magnesium Chloride" and "Sodium Chloride" columns of Table 2. (Comparative Example 10) Comparative Example 10 differs from Example 1 in that only sodium chloride is added to the recovered solution. Otherwise, it is the same as Example 1. The sodium chloride content of the recovered solution was 62.5 mg / mL. For Comparative Example 10, "-" is written in the "Magnesium Chloride" column of Table 2. (Comparative Example 11) Comparative Example 11 differs from Example 1 in that only sodium chloride is added to the recovered solution. Otherwise, it is the same as Example 1. The sodium chloride content of the recovered solution was 125 mg / mL. For Comparative Example 11, "-" is written in the "Magnesium Chloride" column of Table 2. (Comparative Example 12) Comparative Example 12 differs from Example 1 in that only sodium chloride is added to the recovered solution. Otherwise, the procedure was the same as in Example 1. The sodium chloride content of the recovered solution was set to 250 mg / mL. For Comparative Example 12, "-" was written in the "Magnesium Chloride" column of Table 2.

[0117] (Comparative Example 13) Comparative Example 13 differs from Example 1 in that only magnesium chloride was added to the recovered solution. Otherwise, it was the same as Example 1. The magnesium chloride content of the recovered solution was set to 60 mg / mL. For Comparative Example 13, "-" is written in the "Sodium Chloride" column of Table 2. Note that for Comparative Examples 9 to 12, the sensitivity ratio could not be evaluated due to the large variation in the recovered amount. For this reason, for Comparative Examples 9 to 12, "Evaluation not possible due to large variation in recovered amount" is written in the "Sensitivity Ratio" column of Table 2.

[0118]

[0119] As shown in Table 2, Examples 1 to 11 have a higher sensitivity ratio of 2 or more and smaller variation in the amount of concentrated solution recovered compared to Comparative Examples 1 to 13. Note that Comparative Examples 1 to 8 did not have a concentration process, as described above, so the variation in the amount of recovered solution was not evaluated. Also, Comparative Examples 9 to 12 had large variations in the amount of recovered solution, as described above, so the sensitivity ratio could not be evaluated. From Examples 1 to 6, the sensitivity ratio is high when the sodium chloride content is in the range of 30 mg / mL to 200 mg / mL. Furthermore, from Examples 1 to 6, there is a tendency for the variation in the amount of concentrated solution recovered to be smaller when the sodium chloride content is high. From Examples 3, 7 to 11, the sensitivity ratio is high when the magnesium chloride content is in the range of 10 mg / mL to 80 mg / mL. Furthermore, from Examples 3, 7 to 11, there is a tendency for the variation in the amount of concentrated solution recovered to be smaller when the magnesium chloride content is high.

[0120] 10 Processing kit 12 Container 12c Interior 14 Recovery liquid container 16, 18 Superabsorbent polymer 17 Recovery liquid 20 Container body 20a Storage section 20b Neck 20c Reduced diameter section 20d, 20f Storage section 20h, 42a, 50c Interior 21 Opening 21a Male threaded section 22 Cap 22a Nozzle 22b Discharge section 22c Female threaded section 23 Filter 24 Nozzle cover 25, 25a, 25b Container body 26 Storage section 28 Container 29 Wall surface 30 Liquid sample 34 Concentrated liquid 35 Concentrate 40 Base 40a, 50a Opening 42 Main body 44 Sealing material 44e Reduced diameter section 50 Recovery liquid cup 50b Bottom D Arrow

Claims

1. A processing kit comprising a liquid sample containing a test substance and water, a superabsorbent polymer that absorbs the water, a container for containing the superabsorbent polymer, and a recovery liquid container for containing a recovery liquid containing magnesium chloride and sodium chloride, wherein the recovery liquid is added after the liquid sample is placed in the container.

2. The treatment kit according to claim 1, wherein the magnesium chloride content of the recovered liquid is 10 mg / mL or more and 80 mg / mL or less, and the sodium chloride content of the recovered liquid is 30 mg / mL or more and 200 mg / mL or less.

3. The processing kit according to claim 1 or 2, wherein the liquid sample includes biological fluid.

4. A processing method comprising the steps of: placing a liquid sample containing a test substance and water into a container containing a superabsorbent polymer; allowing the superabsorbent polymer to absorb the water contained in the liquid sample and concentrating the liquid sample in the container; placing a recovered liquid containing magnesium chloride and sodium chloride into the container; and removing the concentrated liquid of the liquid sample obtained by concentrating it in the container from the container.

5. The processing method according to claim 4, wherein the magnesium chloride content of the recovered liquid is 10 mg / mL or more and 80 mg / mL or less, and the sodium chloride content of the recovered liquid is 30 mg / mL or more and 200 mg / mL or less.

6. The processing method according to claim 4 or 5, wherein the liquid sample includes biological fluid.

Images