Immunochromatography Inspection System

Abstract

In an immunochromatographic test device, according to the present invention, a cartridge comprising: a drop applying region where a drop of a sample is applied; a test region where a color development state changes according to whether the sample is positive or negative; a carrier having at least three color development regions where the color development state changes due to a reaction with a first reagent; and a second reagent holding unit that holds a second reagent which develops in the test region after the first reagent develops in the color development regions, comprises a loaded unit that is loaded in a detachable manner, a detection unit that detects the color development state in the color development regions, a second reagent supplying mechanism for initiating the supply of the second reagent from the second reagent holding unit to the carrier, and a processor that operates the second reagent supplying mechanism on the basis of changes in the color development state in the color development regions.

Description

[[Technical Field]]

[0001] The present disclosure relates to an immunochromatographic test device. [[Background Art]]

[0002] Among immunoassay methods, the immunochromatography method is generally widely used because it is easy to operate and allows testing in a short time.

[0003] WO 2016 / 114122 and WO 2017 / 104143 disclose an immunochromatographic kit using an immunochromatography method. The immunochromatographic kit includes an immunochromatographic carrier to which a sample is supplied. The immunochromatographic carrier has a test region on which an antibody that specifically binds to an antigen, which is the test substance, is immobilized. When a labeled antibody that specifically binds to an antigen is developed on an immunochromatographic carrier together with a sample containing the antigen, the antigen binds to the antibody immobilized on the test region, and the labeling substance is captured via the antigen. When the test region develops color due to the labeling substance captured in this test region, it is determined as positive. When the amount of the labeling substance captured in the test region is very small, the color development is weak, and the result may be determined as false negative. Therefore, WO 2016 / 114122 and WO 2017 / 104143 disclose an amplification technique for amplifying a labeled signal emitted by a labeling substance. The disclosed amplification technique is a silver amplification technique in which gold colloid particles are used as the labeling substance, and silver ions and a silver ion reducing agent are used as reagents for amplification. In silver amplification, an amplification reaction that generates silver particles having a relatively large particle size using gold colloid particles as a catalyst is caused. This amplification reaction amplifies the labeled signal emitted by the gold colloid particles.

[0004] The immunochromatographic kits described in International Publication Nos. 2016 / 114122 and 2017 / 104143 comprise a first amplification solution pod holding a first amplification solution (corresponding to the first reagent) containing a silver ion reducing agent, and a second amplification solution (corresponding to the second reagent) holding a second amplification solution (corresponding to the second reagent) containing silver ions. The immunochromatographic kit also includes operating structures such as buttons for applying pressure to the first amplification solution pod and buttons for applying pressure to the second amplification solution pod. By applying pressure through these operating structures, the first and second amplification solutions can be supplied to the immunochromatographic carrier, thereby inducing an amplification reaction.

[0005] On the other hand, Japanese Patent Publication No. 2012-103150 discloses an analytical device that includes a loading unit for loading a cartridge equivalent to an immunochromatograph kit, and optically analyzes the reaction state of a sample and reagent in the testing area. The analytical device includes a sensor for optically detecting the reaction state and a display unit for displaying the detection results. By using the analytical device, the user can mechanically determine whether a sample is positive or negative simply by loading a cartridge on which the sample has been applied. The cartridge described in Japanese Patent Publication No. 2012-103150 includes an amplification liquid pod, and the analytical device includes an internal mechanism such as a pressing mechanism for pressing the amplification liquid pod, in addition to the sensor and display unit. Therefore, within the analytical device, the internal mechanism presses the amplification liquid pod, supplying the amplification liquid from the amplification liquid pod to the immunochromatograph carrier. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Testing devices that use cartridges equipped with a first amplification solution pod and a second amplification solution pod are also known, such as the immunochromatography kits (equivalent to cartridges) described in International Publication Nos. 2016 / 114122 and International Publication Nos. 2017 / 104143. In conventional testing devices, the timing of supplying the second reagent is controlled using a timer after the supply of the first reagent has started, in order to supply the second reagent after the first reagent has been released. By controlling the timing of supplying the second reagent in this way, the time from the start of supplying the first reagent to the completion of the result becomes constant, regardless of individual differences between cartridges.

[0007] However, there may be individual differences in the development time of the first reagent from cartridge to cartridge; for example, some cartridges may have a long development time for the first reagent, while others may have a short development time. When managing the timing of supplying the second reagent while taking these individual differences into account, it is necessary to set the waiting time until the supply of the second reagent to match the cartridge with the longest development time for the first reagent. In that case, cartridges with a relatively short development time for the first reagent will also occupy the testing equipment for the same amount of time as cartridges with the longest development time for the first reagent. When testing multiple cartridges, it is desirable to minimize the time that each cartridge occupies the testing equipment in order to improve throughput.

[0008] In the analytical apparatus described in Japanese Patent Publication No. 2012-103150, the time from sample application to the cartridge to the determination is controlled. However, in Japanese Patent Publication No. 2012-103150, the determination is made by time control without considering individual differences in cartridges, and therefore, it has the same problems as in International Publication Nos. 2016 / 114122 and International Publication Nos. 2017 / 104143.

[0009] This disclosure is made in view of the above circumstances and aims to provide an immunochromatographic testing device that can shorten the time that a cartridge occupies the immunochromatographic testing device in accordance with individual differences in cartridges, compared to the conventional method which uses a timer to manage the timing of supplying the second reagent. [Means for solving the problem]

[0010] The immunochromatographic testing apparatus of this disclosure includes a carrier having at least three regions: a dotting region on which a sample is dotted; a testing region whose color state changes depending on whether the sample is positive or negative; and a color-developing region whose color state changes due to a reaction with a first reagent; and a loading section in which a cartridge is detachably loaded, the cartridge having a second reagent that is spread in the testing region after the first reagent has been spread in the color-developing region. A detection unit that detects the color development state of the color development area, A second reagent supply mechanism for initiating the supply of the second reagent from the second reagent holding section to the carrier, A processor that determines whether or not there is a change in the color state of the color-developing area based on the change in the color state of the color-developing area, and activates the second reagent supply mechanism, It is equipped with.

[0011] In the immunochromatographic testing apparatus of this disclosure, it is preferable that at least the second reagent among the first reagent and the second reagent is an amplification solution that amplifies the color development in the test area.

[0012] In the immunochromatographic testing apparatus of this disclosure, the first reagent and the second reagent can react with each other to form an amplification solution that amplifies the color development in the test area.

[0013] In the immunochromatographic inspection apparatus of this disclosure, if the processor determines that there is no change in the color state of the color region, it is preferable to determine again whether there is a change in the color state of the color region after a preset first set time has elapsed since the determination.

[0014] In the immunochromatographic inspection apparatus of this disclosure, it is preferable that the processor notify an error if the color development state of the color development area does not change even after a preset second set time has elapsed from a preset time after the cartridge has been loaded, or after a preset number of discrimination repetitions have been repeated.

[0015] In the immunochromatographic inspection apparatus of this disclosure, when the direction toward the inspection area is considered to be the downstream side of the carrier with respect to the spot application area, the carrier of the cartridge is provided downstream of the inspection area and has a control area that indicates by a change in color development that the sample supplied to the carrier from the spot application area has spread to the inspection area. When the detection unit is called a first detection unit, the color development state detected by the first detection unit is called a first color development state, the color development state of the control area is called a second color development state, and the color development state of the inspection area is called a third color development state, the apparatus comprises a second detection unit that detects the second color development state of the control area and a third detection unit that detects the third color development state of the inspection area. Preferably, the processor determines whether or not there has been a change in the third color development state of the inspection area when it has determined that there has been a change in the first color development state of the color development area and a change in the second color development state of the control area.

[0016] In the immunochromatographic testing apparatus of this disclosure, it is preferable that the processor activates the second reagent supply mechanism only when it determines that there is a change in the first color development state of the color development region and no change in the second color development state of the control region.

[0017] In the immunochromatographic testing apparatus of this disclosure, it is preferable that the processor notifies an error if, after activating the second reagent supply mechanism, there is no change in the second color development state of the control area even after a preset third set time has elapsed.

[0018] In the immunochromatographic testing apparatus of this disclosure, it is preferable that the cartridge includes a first reagent holder for holding a first reagent.

[0019] In the immunochromatographic testing apparatus of this disclosure, it is preferable that a cartridge can be loaded in which the supply of the first reagent from the first reagent holding unit to the carrier has already begun.

[0020] In the immunochromatographic test device of the present disclosure, it is preferable to comprise a first reagent supply mechanism for starting the supply of the first reagent from the first reagent holding unit to the carrier with respect to the cartridge loaded in the loading unit.

[0021] In the immunochromatographic test device of the present disclosure, when the direction toward the test region based on the spotting region is defined as the downstream side of the carrier in the cartridge, it is preferable that the color development region is disposed on the downstream side of the test region, and the first reagent holding unit is provided on the upstream side of the spotting region.

[0022] In the immunochromatographic test device of the present disclosure, when the color development region disposed on the downstream side of the test region is defined as the downstream color development region, the carrier of the cartridge has an upstream color development region that is disposed on the upstream side of the test region and between the first reagent holding unit and the test region, and the color development state of the upstream color development region changes through reaction with the first reagent; when the detection unit is defined as a downstream detection unit that detects the color development state of the downstream color development region, the immunochromatographic test device comprises an upstream detection unit that detects the color development state of the upstream color development region, and it is preferable that the processor activates the first reagent supply mechanism when it is determined that there is no change in the color development state of the upstream color development region. [Advantageous Effects of Invention]

[0023] According to the immunochromatographic test device of the present disclosure, compared with the conventional technology that uses a timer for time management of the supply timing of the second reagent, it is possible to shorten the occupation time that the cartridge occupies the immunochromatographic test device according to individual differences between cartridges. [Brief Description of Drawings]

[0024] [Figure 1] It is a perspective view showing the appearance of an immunochromatographic test device. [Figure 2] It is a perspective view of a cartridge. [Figure 3] It is an exploded perspective view of a cartridge. [Figure 4]This diagram shows the positional relationship between the test strip, the multi-functional component, the first reagent holder, and the second reagent holder within the cartridge. [Figure 5] This is an explanatory diagram of the immunochromatography method. [Figure 6] This is a partially fractured side view of the inspection device with a cartridge loaded. [Figure 7] This is a partially broken side view of the testing device with the cartridge loaded and the second reagent supply mechanism activated. [Figure 8] This is a diagram showing the first inspection flow. [Figure 9] This diagram shows the first inspection flow within the inspection device. [Figure 10] This diagram shows the second inspection flow within the inspection device. [Figure 11] This is a side view of inspection strip 1A, an example of a design change. [Figure 12] This figure shows the third inspection flow within the inspection device for the design change example. [Modes for carrying out the invention]

[0025] Embodiments of the immunochromatographic inspection apparatus of this disclosure will be described with reference to the drawings. Figure 1 is a perspective view showing the external appearance of an immunochromatographic inspection apparatus 110 (hereinafter simply referred to as the inspection apparatus 110) according to one embodiment. Figure 2 is an external view of a cartridge 100 mounted in the inspection apparatus 110, and Figure 3 is an exploded perspective view of the cartridge 100. Figure 4 is a diagram showing the positional relationship of the main housing components inside the cartridge 100.

[0026] Cartridge 100 is a single-use type, with one cartridge used for each sample to be tested. As shown in Figure 3, cartridge 100 contains a test strip 1 containing an immunochromatographic carrier 2 (hereinafter referred to as carrier 2). The carrier 2 has a test area L1, and its color development changes depending on whether the sample contains the test substance or not, that is, whether the sample is positive or negative.

[0027] Any sample that may contain the test substance is acceptable, and the sample is not particularly limited. Examples of samples include biological samples, particularly animal (especially human) blood, serum, plasma, cerebrospinal fluid, tears, sweat, urine, pus, nasal secretions, nasal swabs, pharyngeal swabs, nasal aspirates, or other bodily fluids such as sputum, or excretions, organs, tissues, mucous membranes and skin or swabs containing them, or liquid samples containing plants and animals themselves or their dried forms. Examples of test substances include antigens, antibodies, proteins and low-molecular-weight compounds.

[0028] In this example, the testing device 110 is loaded with a cartridge 100 on which a sample has been applied. The testing device 110 then detects the color development of the test area L1 of the loaded cartridge 100 and displays a result indicating whether the sample is positive or negative. When testing multiple samples, one cartridge 100 for each sample is loaded into the testing device 110.

[0029] In the following description, the cartridge 100 is assumed to be loaded into the testing device 110. However, the cartridge 100 in this example has a configuration that allows the user to visually confirm whether the sample is positive or negative without using the testing device 110. Such a cartridge 100 is also called an immunochromatographic testing device or an immunochromatographic testing kit.

[0030] As shown in Figure 1, the inspection device 110 includes a housing 111, which includes a cartridge loading section 112 into which a cartridge 100 is detachably loaded. For example, the front of the housing 111 is provided with an opening for inserting the cartridge 100 into the housing 111 and an opening / closing lid 112a for opening and closing this opening. When loading the cartridge 100, the opening / closing lid 112a is opened, the cartridge 100 is inserted into the housing 111, and once loaded into the cartridge loading section 112, the opening / closing lid 112a is closed. The inspection is performed with the opening / closing lid 112a closed.

[0031] Furthermore, a power switch 113 is provided on the front of the housing 111, and a monitor 119 is provided on the top surface of the housing 111. The monitor 119 displays the judgment results and error messages, etc. The monitor 119 is, for example, a touch panel monitor, and various operation screens are displayed. Through the operation screen, the user can input operation instructions such as inputting a command to start processing and selecting an inspection procedure.

[0032] As shown in Figures 2 and 3, the cartridge 100 includes, for example, a housing 9 composed of a case member 20 and a cover member 10. The housing 9 is made of, for example, a resin material. The case member 20 has an opening at the top and houses the test strip 1, as well as a first reagent holder 40 and a second reagent holder 45 inside. The cover member 10 is attached to the opening of the case member 20, thereby covering the opening of the case member 20. The housing 9 has an overall elongated shape to match the elongated shape of the test strip 1.

[0033] In this example, the upper part of the housing 9, which is formed by the cover member 10, is provided with a dropper port 16, an observation window 18, a first pressure-sensitive area 11, and a second pressure-sensitive area 12. These parts are integrally molded with the cover member 10, for example. The dropper port 16 is an opening for dropping a sample into the housing 9. A boss is erected on the edge of the dropper port 16, facing upwards. The observation window 18 is a window for observing the test area L1 from the outside, and is formed of a transparent material, for example. In this example, the size of the observation window 18 is such that, in addition to the test area L1, the control area L2 and the color development area L3, which will be described later, can also be observed.

[0034] The first pressing section 11 is an operating section operated to supply the first reagent 41 (see Figure 4) from the first reagent holding section 40 to the carrier 2. The second pressing section 12 is an operating section operated to supply the second reagent 46 (see Figure 4) from the second reagent holding section 45 to the carrier 2. The first reagent 41 and the second reagent 46 are amplification solutions for amplifying the color development in the test area L1 when the sample is positive, as will be described later.

[0035] When an external force is applied to the first pressed portion 11, the first pressed portion 11 deforms. For example, if the first pressed portion 11 is shaped like a square pyramid, and an external force is applied from above to the area including the vertex of the square pyramid, the vertex of the square pyramid deforms so that it sinks into the interior of the housing 9. When the first pressed portion 11 deforms in this way, an external force is applied to the first reagent holding portion 40 inside the housing 9. The first reagent holding portion 40 undergoes deformation due to the external force applied through the first pressed portion 11. This deformation causes the first reagent 41 held by the first reagent holding portion 40 to be supplied to the test strip 1.

[0036] Furthermore, it is preferable that the first pressed portion 11 maintains its deformed state after being deformed by pressing. The reason is as follows: As will be described later, the inspection device 110 in this example can be loaded with a cartridge 100 in which the first pressed portion 11 has been pressed in advance by the user. If the first pressed portion 11 is pressed by the user before being loaded into the inspection device 110, it is easier to continue supplying the first reagent 41 if the deformation of the first pressed portion 11 is maintained even after the user releases their hand.

[0037] Similarly, when an external force is applied to the second pressed portion 12, the second pressed portion 12 deforms. In this example, the second pressed portion 12, like the first pressed portion 11, has a square pyramidal shape, and when an external force is applied from above to the region including the vertex of the square pyramid, the vertex of the square pyramid deforms so that it sinks into the interior of the housing 9. When the second pressed portion 12 deforms in this way, an external force is applied to the second reagent holding portion 45 inside the housing 9. The second reagent holding portion 45 undergoes deformation due to the external force applied through the second pressed portion 12. This deformation allows the second reagent 46 held by the second reagent holding portion 45 to be supplied to the test strip 1. In this example, the second pressed portion 12 is provided with a contact portion 12b that contacts the second reagent holding portion 45 (see Figures 6 and 7).

[0038] Furthermore, when the cartridge 100 is inspected with the inspection device 110, the second pressed portion 12, unlike the first pressed portion 11, is pressed by the internal mechanism of the inspection device 110 in either of the inspection flows selectable by the inspection device 110. Therefore, it is sufficient that the second pressed portion 12 can be pressed by the internal mechanism. Of course, if the cartridge 100 is used without the inspection device 110, it is preferable that the second pressed portion 12 can also be pressed by the user.

[0039] As shown in Figures 3 and 4, the case member 20 houses the test strip 1, which includes the carrier 2, along its longitudinal direction. A first reagent holding section 40 is located on one end of the case member 20 in the longitudinal direction (the upstream side as shown in Figure 4). In the case member 20, a first housing section 24 is formed in the area where the first reagent holding section 40 is located, with a recessed shape to match the shape of the first reagent holding section 40. One end of the test strip 1 is positioned above the first reagent holding section 40, which is housed in the first housing section 24.

[0040] The first reagent holder 40 holds the first reagent 41. The first reagent holder 40 is composed of, for example, a container 42 made of a resin material and having an opening on one side, and a sheet member 43 that covers the opening of the container 42 and is breakable. The container 42 is filled with the first reagent 41, and the opening of the container 42 is sealed by the sheet member 43. The first reagent holder 40 is positioned within the first storage section 24 with the sheet member 43 facing upward. The pressing force applied from the first pressed portion 11 is transmitted to the sheet member 43 of the first reagent holder 40 via the end of the test strip 1, causing the sheet member 43 to break (see Figures 6 and 7). As the sheet member 43 breaks, the first reagent 41 is supplied to the test strip 1. In this example, the first pressed portion 11 is provided with a protruding portion 11b that contacts the sheet member 43 (see Figures 6 and 7). The protruding portion 11b has an elongated shape, for example, with its longitudinal direction extending in the width direction of the inspection strip 1, and its tip is pointed toward the sheet member 43, in order to facilitate the breaking of the sheet member 43.

[0041] Furthermore, the cartridge 100 includes a multifunctional member 30 that has the function of housing the second reagent holding section 45. The multifunctional member 30 is located at the other end of the case member 20 (the downstream side shown in Figure 4) and above the test strip 1. The multifunctional member 30 is a member in which a second housing section 32 and a flow path forming section 35 are integrally formed. The second housing section 32 is the part that houses the second reagent holding section 45. The second housing section 32 has a box shape with an open top. As shown in Figure 4, the bottom of the second housing section 32 has a projection 34 for breaking the sheet member 48 of the second reagent holding section 45 (described later) and an opening 33 for allowing the second reagent 46 flowing out from the second reagent holding section 45 to flow towards the test strip 1.

[0042] Furthermore, the channel forming section 35 is provided in connection with the second housing section 32 toward the upstream side. The channel forming section 35 is flat and is positioned in the longitudinal direction of the inspection strip 1 opposite the inspection area L1, etc., and is positioned with a gap between it and the inspection strip 1. The channel forming section 35 forms a channel between itself and the inspection strip 1 that allows the second reagent 46 flowing out from the second housing section 32 to flow toward the inspection area L1, etc. Thus, the channel forming section 35 is positioned between the observation window 18 and the inspection area L1, etc. of the inspection strip 1. For this reason, the channel forming section 35 is made of a transparent material, allowing the inspection area L1, etc. to be observed through the observation window 18.

[0043] The second reagent holder 45 holds the second reagent 46. The second reagent holder 45 is composed of, for example, a container 47 made of a resin material and having an opening on one side, and a sheet member 48 that covers the opening of the container 47 and is breakable. The container 47 is filled with the second reagent 46, and the opening of the container 47 is sealed by the sheet member 48. The second reagent holder 45 is positioned within the second storage section 32 with the sheet member 48 facing downwards. As a result, the sheet member 48 faces the projection 34 within the second storage section 32.

[0044] The pressing force applied from the second pressed portion 12 to the second reagent holding portion 45 acts in a direction that pushes the second reagent holding portion 45 downward, thereby pressing the sheet member 48 against the projection portion 34. When the sheet member 48 is pressed against the projection portion 34, the sheet member 48 is broken (see Figures 6 and 7). When the sheet member 48 is broken, the second reagent 46 is supplied to the test strip 1 through the flow path formed by the opening 33 at the bottom of the second housing portion 32 and the flow path forming portion 35.

[0045] As shown in Figure 4, a gap (clearance) D corresponding to the flow path of the second reagent 46 is formed between the back surface 36 of the flow path forming portion 35 of the multifunctional member 30 and the carrier 2 of the inspection strip 1. The gap D is, for example, in the range of 0.01 mm to 1 mm. The second reagent 46 flows out from the opening 33 at the bottom of the second containment portion 32 toward the carrier 2, and the flowed-out second reagent 46 flows through the flow path formed by the gap D and reaches at least the inspection area L1. The second reagent 46 that reaches the inspection area L1 infiltrates the inspection area L1 from the flow path.

[0046] An absorbent pad 6, described later, is positioned at the downstream end of the inspection strip 1. The case member 20 has a support portion 22 that supports the end of the inspection strip 1, including the absorbent pad 6, at a position opposite to the absorbent pad 6. The second housing portion 32 of the multifunctional member 30 is positioned above the absorbent pad 6. The support portion 22 also supports the multifunctional member 30 via the absorbent pad 6. The case member 20 also has a support portion 21 that supports the central part of the inspection strip 1.

[0047] The inspection strip 1 comprises a carrier 2, a fluid delivery pad 4, and an absorbent pad 6. The carrier 2 is fixed and supported on a back adhesive sheet 7.

[0048] The carrier 2 is a porous, insoluble carrier for spreading the sample, and comprises a test area L1, a control area L2, and a color development area L3. The carrier 2 also includes a label-holding pad 3. The label-holding pad 3 constitutes a dot area where the sample is dotted. When the direction toward the test area L1 is considered the downstream side of the carrier 2 with respect to the dot area, the color development area L3 is located downstream of the test area L1. In this example, the test area L1, the control area L2, and the color development area L3 are each line-shaped regions extending in a direction perpendicular to the direction of sample spreading on the carrier 2.

[0049] The diagram shows the test area L1, control area L2, and color-developing area L3 as lines, but these are not always present. As will be explained in detail later, before developing sample 50 (see Figure 5), the first reagent 41 (see Figure 4), and the second reagent 46 (see Figure 4), the colors of the test area L1 and control area L2 are almost the same as the color of carrier 2 (e.g., white), so at this stage, the test area L1 and control area L2 cannot be clearly seen. The test area L1 appears as a line when sample 50 is developed and the developed sample 50 is positive, as the color intensity increases. This makes the test area L1 visible. The color of the test area L1 is amplified by silver amplification, which will be explained later, so the test area L1 develops to a black color.

[0050] When sample 50 is unfolded, the color intensity of the control region L2 increases, causing it to appear as a line. This makes the control region L2 visible. The color of the control region L2 is also amplified by silver, so the control region L2 also appears black.

[0051] On the other hand, only the color-developing region L3 appears as a dark, almost blackish-green line (hereinafter referred to as dark green) even before the first reagent 41 is developed, and is visible. However, when the first reagent 41 is developed, the dark green color in the color-developing region L3 changes to orange, causing it to appear as an orange line.

[0052] For example, a porous material such as a nitrocellulose membrane can be used as the carrier 2. The back adhesive sheet 7 to which the carrier 2 is fixed is a sheet-like substrate on which the surface to which the carrier 2 is attached is an adhesive surface.

[0053] As shown in Figure 5, a labeling substance 53 is fixed to the labeling pad 3. The labeling substance 53 is modified with a first binding substance 52 that specifically binds to the test substance 51 contained in the sample 50. This labeling pad 3 is fixed on the carrier 2 at a position opposite the dropper port 16 of the cover member 10. Therefore, the sample 50 is dropped onto the labeling pad 3 from the dropper port 16. Thus, the labeling pad 3 corresponds to the application area where the sample 50 is applied.

[0054] The label-holding pad 3 is fixed to approximately the center of the carrier 2 in the longitudinal direction. As the labeling substance 53, for example, gold colloid particles with a diameter of 50 nm (EM.GC50, manufactured by BBI) can be used. Note that the labeling substance 53 is not limited to gold colloid, but can also be metal sulfides that can be used in ordinary chromatography methods, colored particles used in immunoaggregation reactions, etc., and metal colloids are particularly preferred. Examples of metal colloids include gold colloid, silver colloid, platinum colloid, iron colloid, aluminum hydroxide colloid, and composite colloids thereof. Gold colloid is particularly preferred because, at an appropriate particle size, it exhibits a red color and silver colloid exhibits a yellow color, and among these, gold colloid is the most preferred.

[0055] As shown in Figure 5, the test area L1 contains a second binding substance 56 that specifically binds to the test substance 51, thereby capturing the test substance 51. When the test substance 51 is captured in the test area L1 by the binding of the second binding substance 56 to the test substance 51, the first binding substance 52 and the labeling substance 53 bound to the test substance 51 are also captured. If the sample 50 contains the test substance 51, the color intensity of the test area L1 rises above a preset standard as the test substance 51 and the labeling substance 53 are captured in the test area L1. The test area L1 is a region for confirming the presence or absence of the test substance 51 by the labeling signal from the labeling substance 53 captured via the test substance 51.

[0056] The control region L2 contains a third binding substance 58 that specifically binds to the first binding substance 52, and captures the labeled substance 53 via the first binding substance 52. When the sample 50 is applied to the label-holding pad 3, the labeled substance 53 modified with the first binding substance 52 that is not bound to the test substance 51 also spreads within the carrier 2 toward the test region L1 along with the sample 50. The labeled substance 53 that is not bound to the test substance 51 passes through the test region L1 without being captured. The labeled substance 53 that has passed through the test region L1 is captured in the control region L2 via the first binding substance 52, as the first binding substance 52 binds to the third binding substance 58. When the labeled substance 53 is captured in the control region L2, the color intensity of the control region L2 rises to a preset standard or higher. The control region L2 is a region for confirming the completion of the spread of the sample 50 by the label signal from the labeled substance 53 captured via the first binding substance 52. Therefore, the control region L2 is sometimes called the verification region.

[0057] The first binding substance 52 that modifies the labeling substance 53 and specifically binds to the test substance 51 is, for example, an antibody against the antigen if the test substance is an antigen, an antigen against the antibody if the test substance is an antibody, or an aptamer for proteins and low molecular weight compounds if the test substance is a protein and low molecular weight compound, etc., and is a substance that specifically binds to the test substance.

[0058] The second binding substance 56, which is fixed in the test area L1 and specifically binds to the test substance 51, is a substance that specifically binds to the test substance, such as an antibody against the antigen if the test substance is an antigen, an antigen against the antibody if the test substance is an antibody, or an aptamer against the protein and low molecular weight compounds if the test substance is a protein and low molecular weight compounds. The first binding substance 52 and the second binding substance 56 may be the same or they may be different.

[0059] The third binding substance 58 that specifically binds to the first binding substance 52 may be the test substance 51 itself, or it may be a compound that has a site recognized by the first binding substance 52. For example, a compound obtained by binding a derivative of the test substance 51 to a protein may be used.

[0060] For example, if the test substance 51 is influenza A virus or its biomarker, anti-influenza A monoclonal antibody (Anti-Influenza A SPTN-5 7307, manufactured by Medix Biochemica) can be used as the first conjugate 52 and the second conjugate 56, and anti-mouse IgG antibody (anti-mouse IgG(H+L), rabbit F(ab')2, catalog number 566-70621, manufactured by Wako Pure Chemical Industries, Ltd.) can be used as the third conjugate 58.

[0061] The color-developing region L3 contains a substance that reacts with the first reagent 41 to change its color state. The color-developing region L3 indicates that the first reagent 41 has reached that region by reacting with the first reagent 41 to develop color or by changing color. For example, when using a mixed aqueous solution of iron nitrate aqueous solution and citric acid (manufactured by Wako Pure Chemical Industries, Ltd., 038-06925) as the first reagent 41, it is preferable that the color-developing region L3 be composed of a color-developing reagent immobilization line in which bromocresol green (manufactured by Wako Pure Chemical Industries, Ltd.) is immobilized in a line. This is the embodiment of the color-developing region L3 in this example, and as described above, the color-developing region L3 in this example is dark green before reacting with the first reagent 41, and changes to orange when the first reagent 41 reaches the color-developing region L3. Furthermore, the color-developing region L3 is sometimes called the amplification indicator region because the change in color development indicates the timing for the first reagent 41 to be deployed and the supply of the second reagent 46.

[0062] The liquid delivery pad 4 is positioned in contact with one end of the carrier 2 and delivers the first reagent 41 to the carrier 2 from upstream of the point contact area (composed of the label holding pad 3). When the first pressure-sensitive portion 11 is pressed, one end of the liquid delivery pad 4 is immersed in the first reagent holding portion 40. The liquid delivery pad 4 is made of a porous material and absorbs the first reagent 41, and delivers the absorbed first reagent 41 to the carrier 2 by capillary action.

[0063] The absorbent pad 6 is positioned in contact with the other end of the carrier 2 and absorbs the sample 50, the first reagent 41, and the second reagent 46 that are spread on the carrier 2. The absorbent pad 6 is also made of a porous material.

[0064] In this embodiment, the first reagent 41 and the second reagent 46 are amplification solutions that, by reacting together, amplify the color development in the test region L1 and the control region L2. When a metallic labeling substance such as gold colloid is used as the labeling substance 53, as in this example, silver amplification is used as a method to amplify the labeling signal of the labeling substance 53. The first reagent 41 and the second reagent 46 are amplification solutions used for silver amplification as an example, and the reaction of the first reagent 41 and the second reagent 46 with the labeling substance 53 as a catalyst is the amplification reaction. The amplification reaction generates silver particles 60 (see Figure 5) with a particle size relatively larger than that of the labeling substance 53.

[0065] More specifically, in this example, the first reagent 41 is a reducing agent that reduces silver ions, and the second reagent 46 is silver ions. When the first reagent 41, which is a reducing agent, and the second reagent 46, which is silver ions, are brought into contact with the labeled substance 53, silver particles 60 (see Figure 5) are generated, and the generated silver particles 60 are deposited on the labeled substance 53, using the labeled substance 53 as a nucleus. The deposition of silver particles on the labeled substance 53 generates silver particles 60 (see Figure 5) that are larger in particle size than the labeled substance 53. As a result, the labeling signal emitted by the labeled substance 53 is amplified, and consequently, the color development of the labeled substance 53 is amplified in the test area L1 and the control area L2.

[0066] (First reagent) As the reducing agent for the first reagent 41, any inorganic or organic material, or a mixture thereof, can be used, as long as it can reduce the silver ions used as the second reagent 46 to silver. As an inorganic reducing agent, Fe 2+ , V 2+ Or Ti 3+ Examples of preferred reducing metal salts and reducing metal complex salts are those whose valence can change with metal ions such as Fe. 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 Fe as a reducing agent, citric acid or EDTA (ethylenediaminetetraacetic acid) is used to reduce the oxide Fe. 3+ It can form a complex and render harmless. In this system, it is preferable to use such an inorganic reducing agent, and more preferably Fe 2+ A metal salt is preferred.

[0067] 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), as well as other materials obvious to those skilled in the art, such as those described in U.S. Patent No. 6,020,117, can also be used.

[0068] As a reducing agent, ascorbic acid reducing agents are also preferred. Useful ascorbic acid reducing agents include ascorbic acid and its analogues, 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 salts of ascorbic acid, potassium salts 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.

[0069] (Second reagent) The silver ion-containing solution used as the second reagent 46 is preferably one in which a silver ion-containing compound is dissolved in a solvent. As the silver ion-containing compound, an organic silver salt, an inorganic silver salt, or a silver complex can be used. Preferably, an inorganic silver salt or a silver complex is used. As the inorganic silver salt, a silver ion-containing compound with high solubility in a solvent such as water can be used, and examples include silver nitrate, silver acetate, silver lactate, silver butyrate, and silver thiosulfate. Silver nitrate is particularly preferred. As 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 an example is silver hydroxythioether.

[0070] <Immunochromatography> Referring to Figure 5, the immunochromatographic method will be explained. Here, the explanation assumes that sample 50 contains the test substance 51, that is, that sample 50 is positive.

[0071] First, the sample 50 is spot-applied onto the label-holding pad 3, which is the spot-applied area (step S1). The test substance 51 in the sample 50 spot-applied onto the label-holding pad 3 specifically binds to the first binding substance 52 that modifies the label substance 53 contained in the label-holding pad 3. The sample 50 is then spread downstream from the label-holding pad 3 within the carrier 2 by capillary action. A portion of the sample 50 is also spread upstream. Arrow S indicates the spreading of the sample 50.

[0072] Next, the first reagent 41 is supplied (step S2). The first reagent 41 is supplied from the liquid delivery pad 4 side. The first reagent 41 is supplied to the carrier 2 via the liquid delivery pad 4 and spreads downstream.

[0073] Subsequently, the system waits until the first reagent 41 is deployed downstream (steps S3-S4). The "Wait" shown in Figure 5 indicates waiting. The first reagent 41 is gradually deployed downstream, and the sample 50 being deployed from the label-holding pad 3 and the labeled substance 53 modified with the first binding substance 52 are pushed downstream by the first reagent 41 (step S3).

[0074] The test substance 51 in the sample 50, which is deployed downstream and reaches the test area L1, is captured by the second binding substance 56 in the test area L1. That is, the labeled substance 53 is captured in the test area L1 via the test substance 51 and the first binding substance 52. On the other hand, the labeled substance 53 that is not bound to the test substance 51 passes through the test area L1 without being captured and is captured by the third binding substance 58 in the control area L2.

[0075] As the first reagent 41 expands and reaches the color-developing region L3 (step S4), the color-developing region L3 reacts with the first reagent 41 and changes color. In this example, the color-developing region L3 is dark green before reacting with the first reagent 41, and changes to orange after reacting with the first reagent 41.

[0076] After the first reagent 41 has fully expanded, the second reagent 46 is supplied to the carrier 2 (step S5). The second reagent 46 is supplied to the carrier 2 from downstream of the color development region L3 and expands upstream. Here, the first reagent 41 is a first amplification solution containing a reducing agent that reduces silver ions, and the second reagent 46 is a second amplification solution containing silver ions. The reaction between the first and second amplification solutions generates silver particles 60 using the gold colloid particles, which are the labeling substance 53, as a catalyst. This amplifies the label signal (step S6).

[0077] Figures 6 and 7 are partially cutaway side views of the inspection device 110 with the cartridge 100 loaded. The configuration and function of the inspection device 110 will be explained below using Figures 6 and 7.

[0078] The testing device 110 in this example allows selection of two testing flows, a first testing flow and a second testing flow, as shown below. In both the first and second testing flows, the sample 50 must be applied to the carrier 2 of the cartridge 100 before loading. However, if the first testing flow is selected, the supply of the first reagent 41 to the carrier 2 of the cartridge 100 must be started by user operation before loading, whereas if the second testing flow is selected, the supply is started by the internal mechanism of the testing device 110 after loading.

[0079] In other words, the first testing flow is a flow in which testing is performed on a cartridge 100 that has been in a state where the sample 50 has been applied and the supply of the first reagent 41 has been started, before loading. In the case of the first testing flow, after loading, only the supply of the second reagent 46 to the carrier 2 of the first reagent 41 and second reagent 46 is performed by the testing device 110.

[0080] The second testing flow is a flow in which testing is performed on cartridge 100 that has only had the sample 50 applied before loading. In the case of the second testing flow, after loading, both the first reagent 41 and the second reagent 46 are supplied to the carrier 2 by the testing device 110.

[0081] In the following, after describing the configuration of the inspection device 110, the first inspection flow will be described, followed by the second inspection flow.

[0082] (Configuration of inspection device 110) As shown in Figures 6 and 7, the inspection device 110 includes a first reagent supply mechanism 116 and a second reagent supply mechanism 118 as internal mechanisms. The first reagent supply mechanism 116 is a mechanism for initiating the supply of the first reagent 41 from the first reagent holding section 40 to the carrier 2. The first reagent supply mechanism 116 uses an actuator such as a solenoid equipped with an electromagnet and a plunger movable relative to the electromagnet. For example, as the plunger moves, the plunger comes into contact with the first pressed section 11 and presses the first pressed section 11. The first reagent supply mechanism 116 is positioned opposite the first pressed section 11 of the loaded cartridge 100.

[0083] The first reagent supply mechanism 116 is a pressing mechanism that applies external pressure to the first pressed portion 11 of the cartridge 100 by pressing the first pressed portion 11. When pressure is applied to the first pressed portion 11 by the first reagent supply mechanism 116, the first reagent 41 is supplied from the first reagent holding portion 40 to the carrier 2 by the above action. In the first test flow, the first reagent supply mechanism 116 is not used, and is used only in the second test flow.

[0084] The second reagent supply mechanism 118 is a mechanism for initiating the supply of the second reagent 46 from the second reagent holding section 45 to the carrier 2. Similar to the first reagent supply mechanism 116, the second reagent supply mechanism 118 also uses an actuator such as a solenoid. The second reagent supply mechanism 118 is positioned opposite the second pressed section 12 of the loaded cartridge 100. The second reagent supply mechanism 118 is a pressing mechanism that applies external pressure to the second pressed section 12 by pressing the cartridge 100. When pressure is applied to the second pressed section 12 by the second reagent supply mechanism 118, the second reagent 46 is supplied from the second reagent holding section 45 to the carrier 2 by the aforementioned action. The second reagent supply mechanism 118 is used in both the first and second testing flows.

[0085] The inspection device 110 includes, within its housing 111, a loading unit 112, a first reagent supply mechanism 116, and a second reagent supply mechanism 118, as well as a detection unit 114, a processor 120, and a memory 121. In Figure 6, the processor 120 and memory 121 are shown outside the housing 111 of the inspection device 110, but this is a schematic diagram, and they are actually located inside the housing 111.

[0086] The detection unit 114 optically detects the color development state of the inspection area L1, the control area L2, and the color development area L3, and outputs a detection signal representing the color development state to the processor 120. The detection unit 114 is an image sensor such as a CMOS (Complementary Metal Oxide Semiconductor) image sensor or a CCD (Charge Coupled Device) image sensor, and captures an observation area including the inspection area L1, the control area L2, and the color development area L3. The captured image is then output from the detection unit 114 to the processor 120.

[0087] As an example, the detection unit 114 is flanked by light sources 115, such as light-emitting diodes, which illuminate the inspection area L1, control area L2, and color development area L3 during imaging.

[0088] The processor 120 comprehensively controls each part of the inspection device 110. An example of the processor 120 is a CPU (Central Processing Unit) that performs various controls by executing a program. By executing the program, the CPU functions as a control unit having a detection unit control unit 122, a color development state discrimination unit 123, a first reagent supply mechanism control unit 124, a second reagent supply mechanism control unit 125, a display control unit 126, and a timer 128. Memory 121 is an example of memory connected to or built into the CPU as the processor 120. For example, a control program is stored in memory 121. The processor 120 is realized when the CPU executes the control program.

[0089] Memory 121 stores not only the control program but also setting information that is pre-configured for the processor 120 to perform various controls. The setting information includes information necessary for the color state determination unit 123 to determine changes in the color state. Examples of setting information include the pre-configured first setting time t1, the pre-configured second setting time t2, and the pre-configured number of times K in the inspection flow described later.

[0090] The detection unit control unit 122 controls the imaging timing by the detection unit 114.

[0091] The first reagent supply mechanism control unit 124 controls the first reagent supply mechanism 116 to press the first pressed portion 11.

[0092] The second reagent supply mechanism control unit 125 operates the second reagent supply mechanism 118 based on the change in the color development state of the color development area, controlling it to press the second pressed portion 12.

[0093] The color development state determination unit 123 performs color development area determination processing, control area determination processing, and inspection area determination processing based on the detection signal output by the detection unit 114. As described above, the detection unit 114 outputs an image of the observation area including the inspection area L1, control area L2, and color development area L3. The color development state determination unit 123 performs each of the above determination processes based on the image.

[0094] The color development region discrimination process determines, based on the image of the observed region, whether the color of the color development region L3 has changed, for example, whether it has changed from dark green (the color before the reaction with the first reagent 41) to orange. A change in color development state of "Yes" means that the first reagent 41 has spread to the color development region L3.

[0095] Furthermore, "change in color development" includes any of the following: a change from a first color different from the color of the carrier to a second color (i.e., discoloration); a change in the color of the carrier due to the development of a different color from the carrier (i.e., color development); or a change in the density of the color (i.e., density change).

[0096] When the color development state determination unit 123 determines that the color development state of the color development region L3 has changed, the processor 120 activates the second reagent supply mechanism 118 via the second reagent supply mechanism control unit 125.

[0097] The control region discrimination process determines whether or not there has been a change in the coloration state of the control region L2 based on the detection signal output by the detection unit 114. In this example, a line appears in the control region L2 when the labeling substance 53 is captured in the control region, or when it is amplified by silver after being captured. Therefore, the process determines whether or not a line appears in the control region L2. The coloration state discrimination unit 123 determines that the coloration state of the control region L2 has changed, that is, that the control region L2 is present, and then executes the inspection region discrimination process in the next step.

[0098] The inspection area discrimination process determines whether or not there is a change in the color development state of the inspection area L1 based on the detection signal output by the detection unit 114. In this example, a line appears in the inspection area L1 when the labeling substance 53 is captured in the inspection area L1, or when it is amplified by silver after being captured. Therefore, the process determines whether or not a line appears in the inspection area L1.

[0099] If the color state determination unit 123 determines that there is a change in the color state of the test area L1, the processor 120 displays the test result as "positive" on the monitor 119 via the display control unit 126. If the processor 120 determines that there is no change in the color state of the test area L1, it displays the test result as "negative" on the monitor 119 via the display control unit 126.

[0100] The procedure for immunochromatographic testing using the testing device 110 of this embodiment will be described with reference to Figures 8 and 9. Here, we will describe a first testing flow in which the user supplies the first reagent 41 and the testing device 110 supplies the second reagent 46.

[0101] (First Inspection Flow) Figure 8 shows the first inspection flow. First, the user drops the sample 50 onto the application area of ​​the carrier 2 from the dropper port 16 of the cartridge 100 (step S11).

[0102] Next, the user presses the first pressure-sensitive portion 11 of the cartridge 100 to start supplying the first reagent 41 (step S12).

[0103] Subsequently, the user loads the cartridge 100 into the loading section 112 of the inspection device 110, which is powered on (step S13).

[0104] An inspection of the loaded cartridge 100 is performed inside the inspection device 110 (step S14).

[0105] The time required for the first reagent 41 to fully unfurl the carrier 2 after the supply of the first reagent 41 begins varies for each cartridge, but is generally about 5 to 10 minutes. The time required from when the user presses the first pressure-sensitive part 11 until it is loaded into the loading part 112 can be determined according to the user's convenience.

[0106] Figure 9 shows a detailed inspection flow of the inspection execution (process S14) using the inspection device 110 shown in Figure 8. When the cartridge 100 is loaded into the inspection device 110, the inspection in the inspection device 110 (step S14 in Figure 8) begins.

[0107] As shown in Figure 9, in the inspection device 110, first, n is set to 1 in the processor 120 (step S20). Here, n is a parameter for the number of times the color development region L3 discrimination process is executed. The processor 120 determines whether or not the color state of the color development region L3 has changed (specifically, a change from dark green to orange) (step S21). Specifically, the processor 120 illuminates the observation area by turning on the light source 115 and operates the detection unit 114 to cause the detection unit 114 to take an image. The processor 120 then acquires the image from the detection unit 114 and determines the change in the color state of the color development region L3 from the acquired image. If it is determined that there has been a change in the color state of the color development region L3, that is, if the color development region L3 has changed from dark green to orange, it means that the first reagent 41 has reached the color development region L3 and the inspection area L1 and control area L2 upstream of it.

[0108] When the color development state of the color development area L3 has changed (step S21: Yes), the processor 120 determines whether or not a line has appeared in the control area L2 (step S22). In step S22, the processor 120 determines a change in the color development state of the control area L2 from the captured image. For example, the processor 120 determines whether or not the color development density of the control area L2 has reached a density equal to or higher than a preset reference, and when the density is equal to or higher than the reference, determines that the control area L2 has appeared. When it is determined that the control area L2 has appeared, this means that the specimen 50 has reached the control area L2 and the test area L1 on the upstream side thereof, and has undergone silver amplification, that is, the second reagent 46 has already been supplied.

[0109] On the other hand, when the color development state of the color development area L3 has not changed (step S21: No), it is determined that it is within the second set time t2 and the number of times n that a change in the color development state of the color development area L3 has been determined is less than K times (n<K) (step S23).

[0110] Here, when the second set time t2 has been exceeded, or n is not less than K (step S23: No), the processor 120 notifies an error (step S26) and ends the test flow. For example, the error notification is performed by displaying an error message on the monitor 119. Note that, as a method for notifying an error, in addition to displaying an error message on the monitor 119, the error message may be notified by voice.

[0111] On the other hand, when it is within the second set time t2 and n<K (step S23: Yes), the processor 120 waits until the first set time t1 elapses (step S24). In step S24 of FIG. 9, this is indicated as "t1 wait". The first set time t1 is, for example, about 30 seconds, and the second set time t2 is preset to, for example, 20 minutes or the like. Thereafter, n is incremented by 1 (indicated as n=n+1 in FIG. 9) (step S25), and the process returns to step S21 of determining again whether or not the color development state of the color development area L3 has changed.

[0112] In determining whether or not control region L2 is expressed (step S22), if control region L2 is expressed (step S22: Yes), the test result can be determined as is because it has already been amplified. Therefore, the processor 120 determines the test result by determining whether or not test region L1 is expressed without amplification (step S30), and terminates the test flow.

[0113] In determining the test result, the processor 120, for example, determines whether the color intensity of the line-shaped test area L1 has reached a predetermined standard or higher. If the concentration is above the standard, it determines that the test area L1 is present. If it is determined that the test area L1 is present, it means that the sample 50 is positive. If it is determined that the test area L1 is not present, it means that the sample 50 is negative. In this way, the processor 120 determines whether the sample 50 is positive or negative based on the presence or absence of the test area L1.

[0114] On the other hand, in the determination of whether or not control region L2 is expressed (step S22), if it is determined that control region L2 is not expressed (step S22: No), it is necessary to amplify the color development.

[0115] Therefore, the processor 120 activates the second reagent supply mechanism 118 to start supplying the second reagent 46 (step S27). In this way, the processor 120 activates the second reagent supply mechanism 118 only when it determines that there is a change in the color development state of the color development region L3 and no change in the color development state of the control region L2. In this embodiment, the processor 120 uses the second reagent supply mechanism 118 to press the second pressed portion 12 of the cartridge 100. When the second pressed portion 12 is pressed, it deforms so that it sinks towards the second reagent holding portion 45. Due to this deformation, the sheet member 48 of the second reagent holding portion 45 is pressed against the projection 34 and breaks, and the second reagent 46 is supplied onto the carrier 2. After that, the system waits for the second reagent 46 to unfold until a preset third set time t3 has elapsed (step S28). The third setting time t3 is set to, for example, about 3 minutes.

[0116] After the third set time t3 has elapsed, the processor 120 again determines whether or not the control area L2 has appeared (step S29).

[0117] In determining whether the control region L2 is present in step S29, the processor 120, if the control region L2 is present (step S29: Yes), performs an inspection result determination by determining whether the inspection region L1 is present (step S30), and terminates the inspection flow.

[0118] If the processor 120, which performed the test result determination, determines that expression in the test region L1 is present, it displays the test result as "positive" on the monitor 119. If it determines that there is no expression in the test region L1, it displays the test result as "negative" on the monitor 119.

[0119] On the other hand, in determining whether or not the control region L2 is expressed in step S29, if the control region L2 is not expressed (step S29: No), an error is reported (step S26) and the test flow is terminated. Note that if the control region L2 is not expressed after the second reagent 46 is deployed, there is a possibility that the sample 50 has not been applied. The test flow within the testing device 110 is as described above.

[0120] As previously described, the inspection device 110 of this embodiment includes a detection unit 114 for detecting the color development state of the color development region L3 of the carrier 2, and a second reagent supply mechanism 118 for initiating the supply of the second reagent 46 from the second reagent holding unit 45 to the carrier 2. The processor 120 then activates the second reagent supply mechanism 118 based on the change in the color development state of the color development region L3. In other words, the inspection device 110 of this disclosure includes a processor 120 that controls the timing of activating the second reagent supply mechanism 118 based on the change in the color development state of the color development region L3 due to the reaction with the first reagent 41. Therefore, even if the development time from when the first reagent 41 is supplied to the carrier 2 until the first reagent 41 reaches the color development region L3 varies due to individual differences in the cartridge 100, the supply of the second reagent 46 to the carrier 2 can be started according to the development time for each cartridge.

[0121] Conventionally, when the supply timing of the second reagent 46 is managed using a timer, it is necessary to ensure the maximum deployment time of the first reagent 41, taking into account individual differences in the cartridge 100. However, according to the technology of this disclosure, it is not necessary to wait for the maximum deployment time. Therefore, compared to conventional methods in which the supply timing of the second reagent 46 is managed using a timer, it is possible to shorten the time that the cartridge 100 occupies the testing device 110 according to individual differences in the cartridge 100.

[0122] In this disclosure, the second reagent supply mechanism 118 is exemplified as an actuator such as a solenoid, but any mechanism capable of initiating the supply of the second reagent 46 from the second reagent holding section 45 in the cartridge 100 is acceptable, depending on the configuration of the second reagent holding section 45 in the cartridge 100. For example, if the second reagent holding section 45 is equipped with a shutter and the supply of the second reagent 46 is initiated by opening the shutter, then the second reagent supply mechanism 118 can be any mechanism for opening the shutter.

[0123] In the above embodiment, the first reagent 41 is the first amplification solution and the second reagent 46 is the second amplification solution, but the first reagent 41 and the second reagent 46 are not limited to this combination. The first reagent 41 may be the developing solution and the second reagent 46 may be the washing solution, or the first reagent 41 may be the developing solution or washing solution and the second reagent 46 may be the amplification solution, etc.

[0124] However, it is preferable that the second reagent 46 is an amplification solution that amplifies the color development. If the second reagent 46 is an amplification solution that amplifies the color development in the test area L1, the color development in the test area L1 is amplified, which can improve the accuracy of the determination.

[0125] Furthermore, as in this embodiment, it is preferable that the first reagent 41 and the second reagent 46 are amplifying solutions that amplify the color development in the test area L1. When the first reagent 41 and the second reagent 46 are amplifying solutions that amplify the color development in the test area L1, the color development in the test area L1 is amplified, which can improve the accuracy of the determination.

[0126] As previously described, in the first inspection flow, if the processor 120 determines that there is no change in the color development state, it determines whether there is a change in the color development state again after a predetermined first set time t1 has elapsed. This ensures that even if the development time for the first reagent 41 to reach the color development region L3 varies due to individual differences in the cartridges 100, reliable testing can be performed according to the development state of each cartridge 100.

[0127] Further, in the first inspection flow, if the color development state does not change even after a preset second set time t2 has elapsed from a preset time point after the cartridge 100 is loaded, or even after repeating the determination a preset number of times, the processor 120 notifies an error. According to this configuration, when the color development state does not change even after the second set time has elapsed or after repeating the determination a preset number of times, an error is notified, thereby preventing the apparatus from being occupied indefinitely when the first reagent fails to develop or does not develop at all.

[0128] Further, the preset time point at which counting of the second set time t2 is started may be a time point when the cartridge is loaded or a time point when determination of the preset number of times is completed. Further, the preset number of times K may be appropriately set to 2 or more. In the present embodiment, it is determined in step S23 whether the time is within t2 and the number of times is less than the preset number of times (n<K), but only one of them may be determined. That is, it may be configured to determine only whether the time is within t2 to decide whether to determine the color development state again or to notify an error. Further, it may be configured to determine only whether n<K to decide whether to determine the color development state again or to notify an error.

[0129] In this embodiment, the detection unit 114 detects all color development states in the inspection area L1, control area L2, and color development area L3. However, a separate detection unit may be provided for each area. That is, if the color development state of the color development area L3 is the first color development state, the color development state of the control area L2 is the second color development state, and the color development state of the inspection area L1 is the third color development state, then a first detection unit for detecting the first color development state, a second detection unit for detecting the second color development state, and a third detection unit for detecting the third color development state may be provided. However, as in this embodiment, by using one detection unit 114 to serve as the first, second, and third detection units, and enabling the detection unit 114 to detect the color development states of the three areas, the configuration can be simplified compared to providing separate detection units, and costs can be reduced. In addition, since only one detection unit 114 is needed, space can be saved. Furthermore, the changes in the color development state of the inspection area L1 and the control area L2 may be configured to be visually confirmed by the user.

[0130] In this first test flow, the processor 120 determines whether there is a change in the third color state of the test area L1 when it determines that there is a change in the first color state of the color development area L3 and a change in the second color state of the control area L2. The processor 120 then activates the second reagent supply mechanism 118 only when it determines that there is a change in the first color state of the color development area L3 and no change in the second color state of the control area L2. As a result, if there is a change in the second color state and the deployment of the second reagent 46 is unnecessary, the second reagent 46 is not deployed, and the presence or absence of a change in the third color state of the test area L1 can be determined, thereby reducing the time the device is occupied.

[0131] Furthermore, as previously described, the processor 120 activates the second reagent supply mechanism 118 and, if there is no change in the second color development state of the control area L2 even after a preset third setting time t3 has elapsed, it notifies an error. This prevents the device from being unnecessarily occupied in the event of a malfunction in the second reagent 46.

[0132] In this embodiment, the cartridge 100 is equipped with a first reagent holding section 40 that holds the first reagent 41, and the testing device 110 can be loaded with the cartridge 100 in a state where the supply of the first reagent 41 from the first reagent holding section 40 to the carrier 2 has started. Therefore, it is possible to start the supply of the first reagent to the carrier by, for example, an operation on the cartridge 100 by the user before loading the cartridge 100, thus reducing the occupation time of the testing device 110 compared to the case where the first reagent 41 is supplied to the carrier 2 after the cartridge 100 is loaded.

[0133] In the first testing flow described above, the supply of the first reagent 41 is performed by the user pressing the first pressure-sensitive part 11, so the testing device 110 does not need to be equipped with the first reagent supply mechanism 116.

[0134] (Second Inspection Flow) Figure 10 shows the second inspection flow. Only the differences from the first inspection flow are explained in detail; the same process as in the first inspection flow is given the same process number and detailed explanations are omitted.

[0135] In the first testing flow described above, the user supplies the first reagent 41 and then loads the cartridge 100 into the loading section 112 of the testing device 110. However, in the second testing flow, after the user applies the sample 50, the cartridge 100 is loaded into the loading section 112 without supplying the first reagent 41.

[0136] The inspection in the inspection device 110 is started when a cartridge 100 is loaded in which the first reagent 41 is not supplied.

[0137] The processor 120 first activates the first reagent supply mechanism 116 to start supplying the first reagent 41 to the carrier 2 (step S19). The processor 120 lowers the first reagent supply mechanism 116 to cause it to press the first pressed portion 11 of the cartridge 100. The first pressed portion 11 is deformed by the pressing, and the protrusion 11b inside the first pressed portion 11 pushes the liquid-feeding pad 4 into the first reagent holding portion 40. The liquid-feeding pad 4 breaks through the sheet member 43 of the first reagent holding portion 40 and is immersed in the first reagent 41, whereby the first reagent 41 is supplied from the liquid-feeding pad 4 to the carrier 2.

[0138] The subsequent flow is substantially the same as the first inspection flow. However, in the present configuration, when there is no change in the color development state of the color development region L3 (step S21: No), if the number of times n that the change in the color development state of the color development region L3 has been determined is less than K times (n<K) (step S23A: Yes), the processor waits for a first set time t1 (step S24); if n is not less than K (step S23A: No), an error is notified (step S26). That is, unlike the case of the first inspection flow, the present configuration is configured to proceed to the next step only based on the number of times the change in the color development state of the color development region L3 has been determined, without determining whether or not a second set time t2 has been exceeded.

[0139] As described above, since the inspection device 110 includes the first reagent supply mechanism 116 for starting supply of the first reagent 41 from the first reagent holding portion 40 to the carrier 2, supply of the first reagent 41 can also be performed in the inspection device 110 as in the second inspection flow, and operations other than sample spotting can also be automated. Even in the second inspection flow in which the first reagent 41 is also supplied by the inspection device 110, according to the present inspection device 110, supply of the second reagent 46 to the carrier 2 can be started according to the development time for each cartridge. Therefore, as in the case of the first inspection flow, compared with the conventional technique in which the supply timing of the second reagent 46 is time-managed using a timer, the occupation time during which the cartridge 100 occupies the inspection device 110 can be shortened according to individual differences between the cartridges 100.

[0140] <Modified Example> Figure 11 is a side view of a modified inspection strip 1A. Components identical to those in the inspection strip 1 described above are denoted by the same reference numerals, and detailed explanations are omitted.

[0141] The carrier 2A of the test strip 1A includes an upstream color-developing region L4 located upstream of the test area L1 and between the first reagent holding portion 40 and the test area L1. In this example, the color-developing region L3 located downstream of the test area L1 is referred to as the downstream color-developing region L3. The upstream color-developing region L4, like the downstream color-developing region L3, changes color state due to reaction with the first reagent 41. Therefore, the first reagent 41 reaches the upstream color-developing region L4 at an earlier stage than the time it takes for the first reagent 41 to spread to the downstream color-developing region L3 after the supply of the first reagent 41 is started, making it possible to confirm early whether the supply of the first reagent 41 has started.

[0142] Furthermore, in the housing containing the above-mentioned inspection strip 1A, the portion facing the upstream color-developing region L4 should be made of a transparent material that allows the upstream color-developing region L4 to be visible.

[0143] Furthermore, when using a cartridge equipped with the above-mentioned test strip 1A, it is preferable that the test device 110 includes an upstream detection unit for detecting the color development state of the upstream color development region L4, in addition to the detection unit 114 for detecting the color development state of the downstream color development region L3. Similar to the downstream detection unit 114, an image sensor can be used as the upstream detection unit. In this case, it is preferable that the processor 120 is configured to activate the first reagent supply mechanism 116 when it determines that there is no change in the color development state of the upstream color development region L4.

[0144] Here, we will describe a third inspection flow using an inspection device 110 equipped with an upstream detection unit and a cartridge containing the above-mentioned inspection strip 1A.

[0145] (Third Examination Flow) Figure 12 shows the third inspection flow. Only the differences from the first inspection flow will be explained in detail; the same process as in the first inspection flow will be given the same process code and detailed explanations will be omitted.

[0146] In this third inspection flow, the first step is to determine whether or not to activate the first reagent supply mechanism 116 (step S18). This is a step to determine whether or not the first reagent 41 is being supplied appropriately. The processor 120 has the upstream detection unit detect the color development state of the upstream color development region L4, and determines whether or not there is a change in the color development state based on the color development state detected by the upstream detection unit. If the processor 120 determines that there is no change in the color development state, it determines that it is necessary to activate the first reagent supply mechanism 116 (step S18: Yes), and activates the first reagent supply mechanism 116 (step S19). On the other hand, if the processor 120 determines that there is a change in the color development state, it determines that it is unnecessary to activate the first reagent supply mechanism 116 (step S18: No).

[0147] Subsequently, the processor 120 determines whether or not the color state of the downstream color-developing region L3 has changed (step S21). The subsequent flow is almost the same as the first inspection flow. However, if the color state of the downstream color-developing region L3 has not changed (step S21: No), and if it is within the second set time t2 (step S23B: Yes), the processor waits for the first set time t1 (step S24), and if it exceeds the second set time t2 (step S23B: No), an error is reported (step S26). In other words, unlike the first inspection flow, the system is configured to proceed to the next step based on whether or not it is within the second set time t2, without counting the number of times the change in the color state of the color-developing region L3 has been determined.

[0148] Furthermore, in this third test flow, if a change in the color development state of the downstream color development region L3 is detected (step S21: Yes), the second reagent supply mechanism 118 is activated without confirming the expression of the control region (step S27). Thus, if a change in the color development state of the color development region L3 is detected, the second reagent supply mechanism 118 may be activated and the second reagent 46 supplied to the carrier 2 without confirming the presence or absence of expression of the control region.

[0149] The inspection device in this design modification example, like the inspection device 110 described above, is equipped with a processor 120 that controls the timing of activating the second reagent supply mechanism 118 based on the change in the color development state of the color development region L3 due to the reaction with the first reagent 41. Therefore, the same effects as the inspection device 110 can be obtained. In other words, compared to the conventional method in which the timing of supplying the second reagent is managed using a timer, it is possible to shorten the time that the cartridge 100 occupies the inspection device 110 according to individual differences in the cartridge 100.

[0150] The carrier 2A in the cartridge of the redesigned example includes an upstream color development region L4 located upstream of the test area L1 and between the first reagent holding section 40 and the test area L1. The upstream color development region L4 changes color state due to reaction with the first reagent 41. The testing device 110 of the redesigned example also includes an upstream detection section that detects the color development state of the upstream color development region L4. The processor 120 activates the first reagent supply mechanism 116 when it determines that there is no change in the color development state of the upstream color development region L4. Therefore, even if the user forgets to start the supply of the first reagent 41 or the operation is insufficient, the supply of the first reagent 41 can be reliably performed.

[0151] In the above design modification example, the first reagent supply mechanism 116 is activated or not by determining the change in the color development state of the upstream color development region L4. However, the determination of whether or not to activate the first reagent supply mechanism 116 is not limited to this embodiment. In the inspection device 110, the processor 120 may be configured to determine whether or not to activate the first reagent supply mechanism depending on whether or not the first pressed portion 11 of the cartridge 100 is in a pressed state. Specifically, the inspection device 110 only needs to have a detection unit that detects whether or not the first pressed portion 11 of the cartridge 100 is in a pressed state. An example of a detection unit is an image sensor that captures the first pressed portion 11. Then, the processor 120 of the inspection device 110 determines, based on the image of the first pressed part 11 captured by the image sensor, that if the first pressed part 11 is in a pressed state, it does not need to activate the first reagent supply mechanism 116, and if it determines that the first pressed part 11 is not in a pressed state, it does need to activate the first reagent supply mechanism 116.

[0152] In the above embodiment, the hardware structure of the processor 120 and the processing unit (Processing Unit) that executes various processes, such as the detection unit control unit 122, the color development state discrimination unit 123, the first reagent supply mechanism control unit 124, and the second reagent supply mechanism control unit 125, can be any of the following types of processors. As mentioned above, the types of processors include a CPU, which is a general-purpose processor that executes software and functions as various processing units, as well as programmable logic devices (PLDs), such as FPGAs (Field Programmable Gate Arrays), which are processors whose circuit configuration can be changed after manufacturing, and dedicated electrical circuits, such as ASICs (Application Specific Integrated Circuits), which are processors with circuit configurations specifically designed to execute specific processes.

[0153] A single processing unit may consist of one of these various processors, or it may consist of a combination of two or more processors of the same or different types (for example, a combination of multiple FPGAs, and / or a combination of a CPU and an FPGA). Alternatively, multiple processing units may be composed of a single processor.

[0154] One example of configuring multiple processing units with a single processor is a configuration where one or more CPUs and software are combined to form a single processor, and this processor functions as multiple processing units. Secondly, there is a configuration using a processor that realizes the functions of the entire system, including multiple processing units, on a single IC (Integrated Circuit) chip, as exemplified by System-on-a-Chip (SoC). Thus, various processing units are configured, in terms of hardware structure, using one or more of the above-mentioned various processors.

[0155] Furthermore, the hardware structure of these various processors can more specifically utilize electrical circuits, which are combinations of circuit elements such as semiconductor devices.

[0156] Based on the above description, the following technologies can be understood. [Additional note 1] A method for operating an immunochromatographic testing apparatus comprising: a carrier having at least three regions, including a dotting region where a sample is dotted, a test region whose color development state changes depending on whether the sample is positive or negative, and a color development region whose color development state changes due to a reaction with a first reagent; a second reagent holding portion that holds a second reagent which is spread in the test region after the first reagent has been spread in the color development region; a loading portion into which a cartridge is detachably loaded; a detection portion for detecting the color development state of the color development region; a second reagent supply mechanism for initiating the supply of the second reagent from the second reagent holding portion to the carrier; and a processor, wherein the apparatus is operated by: The aforementioned processor, Based on the change in the color state of the colored area detected by the detection unit, it is determined whether or not there has been a change in the color state of the colored area. A method for operating an immunochromatographic testing device to activate the second reagent supply mechanism.

[0157] The disclosure of Japanese Patent Application No. 2021-050776, filed on 24 March 2021, is incorporated herein by reference in its entirety. All documents, patent applications, and technical standards described herein are incorporated by reference to the same extent as if each individual document, patent application, and technical standard were specifically and individually noted to be incorporated by reference.

Claims

1. An immunochromatographic inspection device, A loading unit in which a cartridge is detachably loaded comprises a carrier having at least three regions: a dotting region where a sample is dotted, a test region whose color state changes depending on whether the sample is positive or negative, and a color-developing region whose color state changes due to a reaction with a first reagent; and a second reagent holding portion that holds a second reagent which is spread in the test region after the first reagent has been spread in the color-developing region. A detection unit for detecting the color development state of the color development region, A second reagent supply mechanism for initiating the supply of the second reagent from the second reagent holding section to the carrier, A processor that controls the operation of the second reagent supply mechanism based on the change in the color development state of the color development region, Equipped with, An immunochromatographic testing apparatus wherein the processor activates the second reagent supply mechanism on the condition that there is a change in the color development state of the color development region where the first reagent is deployed, and does not activate the second reagent supply mechanism if there is no change in the color development state.

2. The immunochromatographic testing apparatus according to claim 1, wherein at least the second reagent among the first reagent and the second reagent is an amplifying solution that amplifies the color development in the test area.

3. The immunochromatographic testing apparatus according to claim 2, wherein the first reagent and the second reagent are amplifying solutions that, upon reaction, amplify the color development in the test area.

4. The immunochromatographic inspection apparatus according to any one of claims 1 to 3, wherein the processor determines whether or not there is a change in the color state of the color region, and if it determines that there is no change in the color state of the color region, after a preset first set time has elapsed since the determination, it determines again whether or not there is a change in the color state of the color region.

5. The immunochromatograph inspection apparatus according to any one of claims 1 to 4, wherein the processor notifies an error if the color state of the color region does not change even after a preset second set time has elapsed from a preset time after the cartridge has been loaded, or after the determination has been repeated a preset number of times.

6. When the direction toward the inspection area is defined as the downstream side of the carrier with respect to the point application area, the carrier of the cartridge is provided downstream of the inspection area and has a control area that indicates, by a change in color development, that the sample supplied to the carrier from the point application area has spread to the inspection area. When the detection unit is referred to as the first detection unit, the color development state detected by the first detection unit as the first color development state, the color development state of the control area as the second color development state, and the color development state of the inspection area as the third color development state, A second detection unit for detecting the second color development state of the control region, The system includes a third detection unit that detects the third color development state of the inspection area, The immunochromatographic inspection apparatus according to any one of claims 1 to 5, wherein the processor determines whether there is a change in the third color state of the inspection area when it determines that there is a change in the first color state of the color region and there is a change in the second color state of the control area.

7. The immunochromatographic inspection apparatus according to claim 6, wherein the processor activates the second reagent supply mechanism only when it determines that there is a change in the first color development state of the color development region and no change in the second color development state of the control region.

8. The immunochromatographic inspection apparatus according to claim 6 or 7, wherein the processor notifies an error if, after activating the second reagent supply mechanism, a preset third set time has elapsed and there has been no change in the second color development state of the control area.

9. The immunochromatographic testing apparatus according to any one of claims 1 to 8, wherein the cartridge comprises a first reagent holding section for holding the first reagent.

10. The immunochromatographic inspection apparatus according to claim 9, wherein the cartridge can be loaded in a state in which the supply of the first reagent from the first reagent holding unit to the carrier has started.

11. The immunochromatographic testing apparatus according to claim 9 or 10, further comprising a first reagent supply mechanism for initiating the supply of the first reagent from the first reagent holding unit to the carrier with respect to the cartridge which is loaded in the loading unit.

12. The immunochromatographic testing apparatus according to claim 11, wherein in the cartridge, when the direction toward the testing area is taken with respect to the dot area as the downstream side of the carrier, the color development area is located downstream of the testing area, and the first reagent holding section is provided upstream of the dot area.

13. When the color-developing region located downstream of the inspection region is defined as the downstream color-developing region, the carrier of the cartridge has an upstream color-developing region located upstream of the inspection region and between the first reagent holding portion and the inspection region, wherein the color-developing state changes due to reaction with the first reagent. When the detection unit is a downstream detection unit that detects the color development state of the downstream color development region, it is also provided with an upstream detection unit that detects the color development state of the upstream color development region. The immunochromatographic inspection apparatus according to claim 11 or 12, wherein the processor activates the first reagent supply mechanism when it determines that there is no change in the color development state of the upstream color development region.

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