Reaction plate

Abstract

The present application provides a reaction plate which is incorporated in a biochemical reaction substrate used in an allergy test or the like, 1) even in the case where the number of times of washing of a reaction area is reduced, stable examination can be performed by performing reliable washing, and / or 2) by making the detection sensitivity uniform at the outer peripheral portion and the inner peripheral portion of the reaction area, the same examination can be performed at the outer peripheral portion and the central portion. A reaction plate which is assembled to a biochemical reaction substrate, wherein the reaction plate has: a reaction area in which a specific binding substance which specifically reacts with a target substance in a test object is immobilized; a flow-out prevention wall which is formed in a manner of surrounding the reaction area, and a portion where the reaction area and the flow-out prevention wall abut each other is formed with a fillet R which is greater than 0.10 mm.

Description

Technical Field

[0001] This invention relates to reaction plates, which are loaded with a biochemical reaction matrix, for determining the amount of a sample substance having physiological activity based on biochemical reaction treatments such as immunoassays. Background Technology

[0002] The examination of the presence and quantity of substances in the body is crucial for understanding health status and determining treatment methods. Furthermore, in today's diverse living environment, the examination of these biologically related substances requires a variety of tests that are rapid and highly sensitive. For example, in treating allergic diseases, it is important to first understand what kind of allergy one has. In recent years, the causes of allergies have become increasingly diverse, and determining the cause is necessary to receive appropriate medication and other treatments.

[0003] Various methods are known for allergy testing. For example, as described in Patent Document 1, a common method is to quantify IgE antibodies against a specific allergen in a blood sample collected from a subject using a solid-phase sandwich immunoassay. In this method, for example, a substance is prepared to block the protein adsorption sites other than the ligand-capturing antibody adsorption sites with a blocking agent such as casein, by adsorbing ligand-capturing antibodies onto a solid-phase carrier such as a glass filter. On the other hand, a substance containing a ligand-bound specific allergen such as dust mites or pollen is prepared and mixed with the blood sample to form a complex of the ligand-bound specific allergen and the IgE antibody against the specific allergen in the blood sample. Then, a mixture containing this complex is added to the substance containing the ligand-capturing antibody adsorbed on the solid-phase carrier, causing the ligand portion of the complex to bind to the ligand-capturing antibody. Next, an anti-IgE antibody labeled with an enzyme or the like is added, causing the IgE antibody portion of the complex to bind to the labeled anti-IgE antibody. Next, excess labeled anti-IgE antibodies that did not bind to the complex were removed, and a colorimetric reaction corresponding to the type of labeling was performed to detect labeled anti-IgE antibodies that bound to IgE antibodies. The results were compared with a standard curve prepared in advance using labeled IgE antibodies to quantify IgE antibodies against specific allergens in the blood sample.

[0004] To perform the above-mentioned examination, methods and apparatus for detecting the presence or absence of allergies are known, which utilize a porous filter formed by combining a specific allergen with a bioreaction matrix. Furthermore, as an example of other methods and apparatuses for detecting the presence or absence of allergies using a bioreaction matrix, a biochemical reaction matrix described in Patent Document 2 is known. This matrix comprises a reaction plate, an absorbent, a receiving container, and a cover. The receiving container includes a reaction plate receiving portion for receiving the reaction plate, an absorbent receiving portion for receiving the absorbent, and a heat-insulating portion. The cover is assembled to the receiving container such that it covers at least a portion of the reaction plate and the absorbent contained within the receiving container. The reaction plate has: a reaction zone for immobilizing a specific binding substance that specifically reacts with the analyte in the sample; a flow path portion connecting the absorbent to the reaction zone; and an injection port for injecting the sample, etc., into the reaction plate.

[0005] Existing technical documents

[0006] Patent documents

[0007] Patent Document 1: (Japan) Publication No. 2011-133285

[0008] Patent Document 2: International Publication No. WO2019 / 151468 Summary of the Invention

[0009] The technical problem that the invention aims to solve

[0010] For example, in the inspection using the reaction plate described in Patent Document 2, there is a reaction zone and a flow path. The reaction zone is used to immobilize a specific binding substance (e.g., an antigen (allergen)) that specifically reacts with the test substance in the sample. The flow path connects the absorber and the reaction zone. The cover has an injection hole for injecting the sample into the reaction plate. Therefore, the number of inspection steps can be reduced (e.g., the step of absorbing cleaning fluid from the reaction zone to drain the cleaning fluid can be reduced), and the inspection can be performed easily.

[0011] Using such a reaction plate, the following steps are performed within the analytical apparatus: (1) Specimen dilution step: The specimen (which can be diluted, hereinafter also referred to as "diluted specimen") is dispensed into the reaction zone of the reaction plate of the biochemical reaction matrix and binds to IgE immobilized in the reaction zone; (2) Specimen washing step: The reaction zone is washed with a washing solution to remove unreacted specimen components; (3) Labeled antibody addition step: Anti-IgE antibodies labeled with enzymes such as ALP (alkaline phosphatase) (so-called labeled secondary antibodies, hereinafter also referred to as "labeled antibodies" in this specification) are added to bind IgE antibodies to the labeled antibodies; (4) Labeled antibody washing step: The unreacted labeled antibodies are washed with a washing solution; (5) Inspection step: A chemiluminescent matrix is ​​added, and the chemiluminescence intensity is captured by a camera. In particular, in the (4) labeled antibody washing step, multiple (e.g., about 10) washes are performed to wash the labeled antibodies. The more times the labeled antibody washing step is performed, the longer the washing step takes, which makes it impossible to shorten the inspection time. It should be noted that “(4) Labeled antibody washing process” refers to a series of processes starting from the state where the labeled antibody added in process (3) is removed from the reaction area (drainage), then the washing solution is injected into the reaction area, the washing solution is stirred, and the washing solution is removed (drainage). “Number of times the labeled antibody washing process” refers to the number of times this series of processes is performed.

[0012] Therefore, the inventors reconsidered the number of washes required for the labeled antibody and confirmed whether a reduction in the number of washes affected the test results. The results of this confirmation are as follows: Figure 10 As shown in (b), unwanted luminescence was detected in the reaction zone surrounding the effluent prevention wall that divides the reaction zone. This is because unwashed labeled antibody remains in the reaction zone surrounding the effluent prevention wall, and this unwashed labeled antibody emits light. Furthermore, it was determined that the presence of such unwanted luminescence makes accurate inspection impossible or necessitates increasing the number of washing steps.

[0013] Furthermore, in the reaction zone of the reaction plate described in Patent Document 2, a polymer or other immobilizing agent is pre-coated on its surface. Specific binding substances (e.g., antigens (allergens)) that specifically react with the test substance in the sample are immobilized in the reaction zone by this immobilizing agent. However, it has been clarified that when the immobilizing agent is not coated with a uniform thickness across the entire surface of the reaction zone, the amount and density of the specific binding substances immobilized in the reaction zone vary depending on the location of immobilization. Even with the same specific binding substance, there is a problem that different test results may occur depending on the location.

[0014] For example, in the reaction plate described in Patent Document 2, the reaction zone is surrounded by an outflow prevention wall at its outer edge. Therefore, when the immobilizer is coated on the reaction plate, the immobilizer is attracted by the surface tension of the inner wall of the outflow prevention wall, resulting in a thicker coating of the immobilizer in the reaction zone around the outflow prevention wall. Sometimes, the thickness of the coating in the reaction zone around the outflow prevention wall is greater than that in the center of the reaction zone. Consequently, the detection sensitivity changes between the center of the reaction zone and the outer periphery around the outflow prevention wall, sometimes leading to different test results. Therefore, to achieve stable testing, it is considered to use only the center of the reaction zone. However, in this case, the following problem exists: because the area of ​​the reaction zone is larger, the amount of sample (sample diluent), antibody, and washing solution used increases, resulting in a longer washing process time, making it impossible to shorten the testing time.

[0015] Therefore, the present invention was made to solve such problems, and its object is to: 1) provide a reaction plate that can be reliably inspected by performing cleaning even when the number of cleanings of the reaction zone is reduced; and / or 2) provide a reaction plate that can perform the same inspection in the outer periphery and the center of the reaction zone by making the detection sensitivity consistent in the outer periphery and the center.

[0016] Technical solutions for solving technical problems

[0017] The reaction plate of the present invention, which solves the above-mentioned problems, is assembled on a biochemical reaction substrate. The reaction plate is characterized in that it has: a reaction zone for immobilizing a specific binding substance that reacts specifically with the analyte in the sample; and an outflow prevention wall formed to surround the reaction zone, wherein the portion of the reaction zone that abuts the outflow prevention wall forms a rounded corner, wherein the rounded corner R is greater than 0.10 mm.

[0018] Furthermore, in the reaction plate of the present invention, it is preferable that the fillet radius R is greater than 0.12 mm.

[0019] Furthermore, in the reaction plate of the present invention, it is preferable that the angle between the outflow prevention wall and the reaction zone is 91 degrees or more.

[0020] Furthermore, in the reaction plate of the present invention, it is preferable that the angle between the outflow prevention wall and the reaction zone is 95 degrees to 130 degrees.

[0021] Furthermore, in the reaction plate of the present invention, it is preferable that the reaction plate has a flow path portion connected to the reaction zone, which prevents the sample or the like from flowing out when the sample or the like is stirred in the reaction zone, and discharges the sample or the like after stirring.

[0022] Furthermore, in the reaction plate of the present invention, it is preferable that the flow path portion has an inclined surface formed in a manner that rises from the reaction zone.

[0023] Furthermore, in the reaction plate of the present invention, it is preferable that the flow path portion extends from the reaction zone to the end of the reaction plate, and the length of the end of the flow path portion in the width direction is narrower than the length of the reaction zone in the width direction.

[0024] Furthermore, in the reaction plate of the present invention, it is preferable that the flow path portion has a flow path wall that is continuous with the outflow prevention wall, and the flow path wall is formed in a streamline shape along the flow path of the flow path portion.

[0025] Furthermore, in the reaction plate of the present invention, it is preferable that the outflow prevention wall has an opening that opens toward any end of the reaction plate, and the flow path is formed between the opening and the end.

[0026] Furthermore, in the reaction plate of the present invention, it is preferable that the flow path wall of the flow path portion is formed to be curved in a streamlined shape along the flow path in such a way that the width of the flow path narrows from the reaction zone toward the end, and the width of the flow path is widened at the end side.

[0027] Furthermore, in the reaction plate of the present invention, it is preferable that the surface of the reaction zone has a coating that fixes the specific binding substance to the reaction zone.

[0028] Furthermore, in the manufacturing method of the reaction plate of the present invention, the reaction plate is assembled on a biochemical reaction substrate, characterized in that the reaction plate has: a reaction zone for immobilizing a specific binding substance that specifically reacts with the analyte in the sample; and an outflow prevention wall formed to surround the reaction zone, wherein the portion of the reaction zone that abuts the outflow prevention wall forms a rounded corner R, the rounded corner R being greater than 0.10 mm. In the reaction plate, an immobilizing agent is applied to the reaction zone by rotating the reaction plate, the immobilizing agent being used to immobilize the specific binding substance that specifically reacts with the analyte in the sample in the reaction zone.

[0029] Invention Effects

[0030] According to the present invention, the reaction zone of the reaction plate and the portion abutting the outflow prevention wall form a predetermined rounded corner R. Therefore, even with a reduction in the number of cleaning cycles during the cleaning process, reliable cleaning can be performed, reducing liquid residue in the reaction zone around the outflow prevention wall and improving inspection accuracy. Furthermore, according to the present invention, the coating of the reaction plate that fixes the specific binding substance to the reaction zone is formed with a uniform thickness. Therefore, the detection sensitivity remains constant at both the outer periphery and the center of the reaction zone, improving inspection accuracy. Moreover, according to the present invention, it is preferable that the flow path of the flow path communicating with the reaction zone is streamlined, making cleaning during the cleaning process more reliable and improving inspection accuracy. Additionally, since the end side of the flow path is formed with an outwardly widening diameter, liquid outflow from the flow path can be prevented even if the absorbency of the absorbent is reduced. Attached Figure Description

[0031] Figure 1 This is a perspective view of an analytical apparatus for inspecting a reaction plate according to an embodiment of the present invention.

[0032] Figure 2 This is a perspective view of a biochemical reaction substrate assembled with a reaction plate according to an embodiment of the present invention.

[0033] Figure 3 This is an exploded view of a biochemical reaction substrate assembled with a reaction plate according to an embodiment of the present invention.

[0034] Figure 4 This is a perspective view of the reaction plate according to an embodiment of the present invention.

[0035] Figure 5 yes Figure 4 AA section view.

[0036] Figure 6 yes Figure 5 Enlarged view of part B in the image.

[0037] Figure 7 This is a perspective view showing a modified example of the reaction plate according to an embodiment of the present invention.

[0038] Figure 8 This is a top view showing a modified example of the reaction plate according to an embodiment of the present invention.

[0039] Figure 9 It is a top view used to illustrate the configuration of antigens (allergens) in the reaction zone of a reaction plate.

[0040] Figure 10 The figures illustrate the test results of the inspection. (a) is a figure illustrating the test results of the inspection using the reaction plate according to the embodiment of the present invention, and (b) is a figure illustrating the test results of the conventional reaction plate. Detailed Implementation

[0041] The reaction plate of the present invention will now be described with reference to the accompanying drawings. Furthermore, the following embodiments do not limit the invention to any particular technical solution, and not all combinations of features described in the embodiments are necessary for the solution of the invention.

[0042] Figure 1 This is a perspective view of an analytical apparatus for inspecting a reaction plate according to an embodiment of the present invention; Figure 2 This is a perspective view of a biochemical reaction substrate assembled with a reaction plate according to an embodiment of the present invention; Figure 3 This is an exploded view of a biochemical reaction substrate assembled with a reaction plate according to an embodiment of the present invention; Figure 4 This is a perspective view of the reaction plate according to an embodiment of the present invention; Figure 5 yes Figure 4 AA section view in the middle; Figure 6 yes Figure 5 Enlarged view of part B in the image; Figure 7 This is a perspective view showing a modified example of the reaction plate according to an embodiment of the present invention; Figure 8 This is a top view showing a modified example of the reaction plate according to an embodiment of the present invention. Figure 9 This is a top view used to illustrate the configuration of the antigens (allergens) in the reaction zone of a reaction plate. Figure 10 The figures illustrate the test results of the inspection. (a) is a figure illustrating the test results of the inspection using the reaction plate according to the embodiment of the present invention, and (b) is a figure illustrating the test results of the conventional reaction plate.

[0043] like Figure 1 As shown, in this embodiment, the reaction plate 10 is assembled onto the biochemical reaction substrate 5 and, together with the reagent kit 6 containing the sample and washing solution, is inserted into the insertion section 3 of the analytical apparatus 1 for testing. The analytical apparatus 1 includes a housing 4, which has an operation panel 2 and an insertion section 3 for mounting the reagent kit 6 and the biochemical reaction substrate 5. The insertion section 3 can be opened and closed freely through a mounting door (not shown).

[0044] The interior of the analytical apparatus 1 is divided into a mounting area, a barcode reading area, a dispensing area, a stirring area, a detection area, and a draining area. If these areas are arranged in a straight line, their regional efficiency is improved, allowing for miniaturization of the analytical apparatus 1. Additionally, a guide unit (e.g., a moving stage) can be installed to guide the biochemical reaction substrate 5 from the mounting area to the draining area. Furthermore, for example, overlapping portions can be created in these areas within the stirring and dispensing areas, with adjacent areas overlapping each other.

[0045] like Figure 2As shown, the reaction plate 10 of this embodiment can be assembled into the biochemical reaction substrate 5 for use. The biochemical reaction substrate 5 only needs to be able to house the reaction plate 10 of this embodiment internally, and may also have a cover 30 and / or a housing container 40. The cover 30 has a generally circular injection hole 31, which, when the reaction plate 10 of this embodiment is housed, is configured to be positioned corresponding to the reaction zone 11 of the reaction plate 10. Additionally, a concave heat-insulating portion 43 may be formed at the bottom of the housing container 40, which can be configured to contact the heater of the analytical apparatus described later to heat the reaction zone 11. Furthermore, an uneven anti-slip unit 35 may be formed on the outer periphery of the cover 30's sidewall, allowing the inspector to reliably hold the biochemical reaction substrate 5.

[0046] like Figure 3 As shown, the storage container 40 has a reaction plate storage section 41 for storing the reaction plate 10 of this embodiment, and may also include an absorbent storage section 42 for storing an absorbent 20 made of a porous material such as a sponge that can fully absorb liquid. Furthermore, when the reaction plate 10 and absorbent 20 of this embodiment are stored in the storage container 40, the front end of the flow path section 12 of the reaction plate 10 is arranged to abut against the absorbent 20.

[0047] like Figure 4 As shown, the reaction plate 10 of this embodiment only needs to be able to fit into the reaction plate receiving portion 41 formed in the receiving container 40 without obstruction, for example, it has a plate-shaped base portion 14 that can fit into the reaction plate receiving portion 41. Furthermore, the reaction plate 10 has an outflow prevention wall 13, which can be erected from the base portion 14. The outflow prevention wall 13 is, for example, a generally annular wall, and the interior of the outflow prevention wall 13 is divided into a reaction zone 11. In addition, the reaction plate 10 has a flow path communicating with the reaction zone 11, and the flow path includes a slope 15, which constitutes at least a portion of the flow path portion 12. Furthermore, the outflow prevention wall 13 only needs to be shaped so that it will not stagnate during drainage; for example, it can be formed as a circle or an ellipse, and the reaction zone 11 inside the outflow prevention wall 13 can also be formed as a circle or an ellipse.

[0048] like Figures 3 to 5 As shown, a ramp 15 forming a flow path is provided from the reaction zone 11 to a position opposite the absorber 20. The flow path is divided by a pair of flow path walls 12X and 12Y extending from the outer periphery of the outflow prevention wall 13. Preferably, the pair of flow path walls 12X and 12Y are curved in a streamlined shape along the flow path in such a way that the width of the flow path (including the ramp 15) (i.e., the spacing between the pair of flow path walls 12X and 12Y) changes continuously. For example, as Figure 4 and Figure 5 As shown, the gap between a pair of flow path walls 12X and 12Y is formed such that it gradually narrows and then gradually widens from the reaction zone 11 toward the end side 15' of the inclined surface 15. These flow path walls 12X and 12Y define a flow path (including the inclined surface 15) for discharging samples, etc., dispensed into the reaction zone 11, and the discharged liquid is absorbed by the absorbent body 20. That is, the pair of flow path walls 12X and 12Y have an inclined surface 15 formed in a manner that rises from the reaction zone 11, so that by tilting the biochemical reaction substrate 5, samples, etc., can be discharged from the reaction zone 11 through the flow path toward the absorbent body 20 without waste.

[0049] The flow path walls 12X and 12Y are continuously formed with the outflow prevention wall 13. Preferably, the flow path walls 12X and 12Y are curved from the outflow prevention wall 13, and preferably without sharp corners. If there is a portion deemed a "seam" where the inner wall of the flow path walls 12X and 12Y meets the inner wall of the outflow prevention wall 13, for example, if there is a protruding shape forming an angle, a concave shape, or a shape with different heights, the liquid to be discharged is likely to remain in the seam, and the liquid cannot be completely discharged and is likely to remain in the seam. As a result, this becomes a major reason for increasing the number of cleaning steps during inspection. Therefore, the portion where the outflow prevention wall 13 is continuous with the flow path walls 12X and 12Y is preferably curved in a streamlined shape, and preferably does not have the portion deemed a seam.

[0050] Furthermore, at the locations where the flow path walls 12X and 12Y meet the outflow prevention wall 13, if the heights of the top surfaces of the two walls are different (a gap exists), the sample or the like can easily remain in the gap, making it easy for the sample to not be completely discharged. Therefore, it is preferable to form them continuously without gaps so that the top surfaces also do not have gaps.

[0051] Furthermore, the inclined surface 15 constituting the flow path is preferably formed at an angle of about 5° to 70° from the base portion 14 (or the reaction zone 11). A more preferred angle is 12° to 50°, a further preferred angle is 17° to 35°, and a more preferred angle is 23°. The larger the angle of inclination of the inclined surface 15, the better it can prevent leakage of the solution dispensed into the reaction zone 11, but on the other hand, it is easy to cause a situation where it is difficult to drain.

[0052] Thus, the flow path 12 includes a flow path formed from the reaction zone 11 to the end side 15′. The length of the end side 15′ of the flow path formed by the flow path 12 in the width direction (the short side direction of the base portion 14, i.e., the interval between the flow path walls 12X and 12Y) is formed to be narrower than the length of the longest part in the reaction zone 11. The end side 15′ of the flow path formed by the flow path 12 is formed such that, when viewed from the side, the inclined surface 15 abuts against the upper surface of the absorber 20 at an acute angle (the angle formed between the upper surface of the absorber 20 and the extended surface of the inclined surface 15 is an acute angle).

[0053] like Figure 5 and Figure 6 As shown, the outflow prevention wall 13 preferably has its inner side (side surface) on the reaction zone 11 side inclined outward relative to the vertical direction from the base portion 14 where the reaction zone 11 is formed. The angle θ formed by the inner side surface of the outflow prevention wall 13 and the surface of the reaction zone 11 is preferably 91° or more when viewed from the reaction zone 11 side. If it is set to 91° to 135°, the sample or the like dispensed into the reaction zone 11 can be prevented from flowing out of the reaction zone 11 during stirring. In addition, it is more preferable that the angle θ is 95° to 130°, and even more preferably 95° to 125°, 95° to 120°, 97° to 115°, 97° to 113°, etc. By making the angle θ greater than 90°, the binding of the sample or the like in the reaction zone 11 during stirring and the cleaning of unreacted samples or the like with the cleaning solution can be carried out without liquid residue. On the other hand, if the angle θ is too large, there is a possibility that the sample or other substances may overflow from the anti-stirring wall 13 to the outside of the reaction zone 11 due to stirring. Therefore, by setting an optimal included angle θ, the objective of the present invention can be achieved.

[0054] Furthermore, the portion of the outflow prevention wall 13 that abuts against the reaction zone 11, i.e., the vertical cross-section of the base portion 14 at the continuous portion of the outflow prevention wall 13 erected from the base portion 14 and the base portion 14 on the reaction zone 11 side, is rounded with a radius R.

[0055] By forming this rounded corner R, it is possible to prevent the fluid, such as the sample dispensed into the reaction zone 11, that is intended to be expelled from the reaction zone 11 from remaining at the periphery of the reaction zone 11 (the periphery of the outflow prevention wall 13 in the reaction zone). For example, it can inhibit the retention of labeled antibodies that are intended to be expelled at the periphery of the reaction zone 11. As a result, the accuracy of the examination can be improved.

[0056] The radius of the fillet R is preferably greater than 0.10 mm, more preferably greater than 0.12 mm, and even more preferably greater than 0.25 mm. Furthermore, the radius of the fillet R can be 0.75~0.11 mm, 0.60~0.12 mm, 0.50~0.12 mm, 0.40~0.20 mm, or 0.35~0.25 mm. By making the radius of the fillet R greater than 0.10 mm, the binding of samples, etc., in the reaction zone 11 during stirring can be carried out without liquid residue, and the cleaning of unreacted samples, etc., based on the cleaning solution, can be performed. On the other hand, if the radius of the fillet R is too large, there is a possibility that samples, etc., may overflow through the anti-overflow wall 13 and flow out of the reaction zone 11 due to stirring.

[0057] Thus, in this embodiment, the reaction plate 10 is formed with an outflow prevention wall 13 surrounding the reaction zone 11, and a predetermined rounded corner R is formed at the contact point between the inner surface of the outflow prevention wall 13 and the reaction zone 11. By forming the rounded corner R in this way, the sample can be prevented from flowing out of the reaction zone 11 when the biochemical reaction matrix 5 is stirred in the analytical apparatus 1, and after the sample is discharged after stirring with fewer washing cycles, excess liquid residue in the reaction zone 11 can be reduced.

[0058] Furthermore, it is preferable to coat the surface of the reaction zone 11 with an immobilizing agent for immobilizing the specific binding substance. For the immobilizing agent, a polymer (copolymer) can be used, and there are no particular limitations as long as it can immobilize the specific binding substance; for example, conventionally known polymers containing multiple photoreactive groups can be used.

[0059] As a method for applying the immobilizing agent to the surface of the reaction zone 11, the following method can be used: In the reaction zone 11 of a reaction plate 10 having an uncoated reaction zone 11 and an outflow prevention wall 13 (preferably a flow path 12), an amount of immobilizing agent sufficient to coat the reaction zone 11 is placed at any position within the zone, and then the reaction plate 10 is rotated (sometimes referred to as spin coating). When coating is performed in a rotating manner, not only the surface of the reaction zone 11, but also the inner wall surfaces of the outflow prevention wall and the flow path wall can be coated with the immobilizing agent. As a result, the liquid flow during the discharge of cleaning fluid, etc., in the cleaning process becomes smoother.

[0060] Furthermore, in the reaction plate 10 of the present invention, a fillet R is applied to the portion where the reaction zone 11 abuts against the outflow prevention wall 13. Therefore, compared with the case without fillet R, the thickness of the coating of the immobilizing agent can be kept constant. Without fillet R, it is assumed that the immobilizing agent applied during rotation will bounce back after hitting the outflow prevention wall 13. As a result, the coating thickness at the outer periphery of the reaction zone 11 (near the outflow prevention wall 13) may be thicker than the coating thickness at the center of the reaction zone.

[0061] The rotation speed used for applying the fixing agent only needs to be a speed that ensures a uniform coating thickness; any speed can be appropriately selected. Furthermore, the rotation speed can be constant or gradually increased. Additionally, when gradually increasing the speed, three or more speed stages can be set. As for the rotation conditions when manufacturing the fixing agent coating, it is preferable to divide the speed into two stages, setting the first stage to a low speed and the second stage to a high speed.

[0062] Alternatively, a sealing agent can be pre-coated onto the entire surface of the reaction zone 11 as needed. The sealing agent can inhibit non-specific adsorption in the reaction zone 11. As a sealing agent, synthetic polymers of non-animal and plant origin, such as polyethylene glycol, can be used in general techniques, and the appropriate agent can be selected based on the properties of the material of the reaction plate 10, the target substance such as antigen, the sample such as blood, and the reagents such as washing solution.

[0063] Furthermore, when the interior of the outflow prevention wall 13 is divided into a reaction zone 11, if a sealing agent is applied to the reaction zone 11 and the inner wall surface of the outflow prevention wall 13, the affinity (e.g., hydrophilicity, hydrophobicity, or water-repellentness) with liquids such as the sample can be made the same on the inner side of the outflow prevention wall 13 and the reaction zone 11, thus also achieving the secondary effect of making it easier to expel the sample.

[0064] Furthermore, the reaction plate of this embodiment may also have structures other than the reaction zone 11, the flow path 12, the outflow prevention wall 13, the base 14, and the inclined surface 15. For example, such as Figure 7 and 8 As shown, the reaction plate 10' can also be formed to separate the flow path section 12 by further expanding its diameter outward.

[0065] Furthermore, the reaction plate 10' may also have a pair of protrusions 16 erected from the base 14 near the location where the flow path walls 12X and 12Y of the flow path section 12 connect with the outflow prevention wall 13. Additionally, the front end portion of the flow path section 12 may be formed with an upper surface 17 that is substantially parallel to the upper surface of the protrusions 16. Such protrusions 16 and upper surface 17 facilitate the handling of the reaction plate 10'. For example, when applying a fixing agent to the reaction zone 11, or when immobilizing a specific binding substance in the reaction zone 11, the reaction plate 10' can be handled by holding the protrusions 16 with tools or machinery. Furthermore, when storing multiple reaction plates 10', even when stacked, they can be stably stored by the protrusions 16 and upper surface 17 abutting against the back of the base 14.

[0066] Furthermore, the protrusion 16 of the reaction plate 10' can be positioned so as not to contact the inner walls of the flow path walls 12X and 12Y and / or the outflow prevention wall 13. This is because if the protrusion 16 contacts the inner wall of the flow path section 12 and / or the outflow prevention wall 13, the protrusion 16 protrudes into the inner wall, and therefore, liquid residue may be generated in the protruding part during the discharge of the sample, etc., which may increase the number of discharge steps.

[0067] Furthermore, in this embodiment, a specific binding substance (e.g., an antigen) is fixed in the reaction zone 11 of the reaction plate 10. The fixed specific binding substance may be only one type, but multiple types are preferred. This is because multiple substances can be detected in a single test. Additionally, not only can one specific binding substance be fixed in one place, but it can also be fixed in multiple places. This is because the detection accuracy of a single substance can be improved.

[0068] For example, in Figure 9 In the reaction plate 10, 25 antigens 50 are arranged and fixed in the reaction zone 11; and one antigen 50 is fixed at 3 locations (3 points). That is, in Figure 9 In the reaction plate, multiple groups 51 (e.g., 3 points / group) of antigen 50 are arranged in a predetermined number of groups 51 (e.g., 75 points / 25 groups). This arrangement increases the number of antigens 50 placed in the reaction zone 11 (the so-called n number) by the number of points (e.g., n=3), and effectively utilizes the area within the reaction zone 11 (eliminating unused space). Furthermore, by arranging them in groups 51 as described above, differentiation from other antigens is easily achieved, thus preventing contamination of reaction results with other antigens during testing.

[0069] Next, the operation of the analytical apparatus 1 for allergy testing using the biochemical reaction substrate 5 assembled with the reaction plate 10 of this embodiment will be described. Figure 1 As shown, the biochemical reaction matrix 5 and the reagent kit 6 are placed in the input section 3. The reagent kit 6 is a component that contains the sample (including diluted sample) and reagents.

[0070] Subsequently, within the analytical apparatus 1, the samples and reagents of the reagent kit 6 are dispensed into the reaction zone 11 of the biochemical reaction substrate 5. For example, the analytical apparatus 1 is equipped with a dispensing nozzle, and the reagent kit 6 is also equipped with a pipette tip. The tip of the dispensing nozzle is engaged with the pipette tip housed in the reagent kit 6 to draw the samples and reagents from the reagent kit 6. Furthermore, the drawn samples and reagents can be dispensed into the reaction zone 11 through the injection port 31 of the biochemical reaction substrate 5.

[0071] The aliquoting process involves the following steps: a sample dilution and addition step, in which the sample is aliquoted into the reaction zone of a reaction plate for a biochemical reaction matrix and binds to IgE immobilized in the reaction zone; a sample washing step, in which a washing solution is used to remove unreacted sample components from the reaction zone; a labeled antibody addition step, in which labeled antibodies are added to allow IgE antibodies to bind to the labeled antibodies; a labeled antibody washing step, in which a washing solution is used to remove unreacted labeled antibodies; and a chemiluminescent matrix addition step, in which a chemiluminescent matrix is ​​added.

[0072] After use, the pipette tip detaches from the dispensing nozzle and returns to the designated position on kit 6. At this point, the used pipette tip returns to its original position, eliminating the need for a designated location for recycling used pipette tips, thus enabling kit miniaturization.

[0073] Furthermore, after these dispensing processes, the moving stage can be reciprocated in this state to timely agitate and dispense the samples, reagents, and cleaning solutions dispensed into the reaction zone 11 of the biochemical reaction substrate 5 during the reaction process. The reciprocating motion of the moving stage is along the short side of the biochemical reaction substrate 5. By reciprocating in this direction, agitation is achieved, eliminating the possibility that samples, etc., would originally pass through the flow path 12, and preventing them from being discharged to the absorber 20 side.

[0074] After stirring, the moving stage is moved to the drainage section for draining the samples, etc., dispensed into the reaction zone 11. In this drainage process, by tilting the substrate holding section of the moving stage, the biochemical reaction substrate 5 held in the substrate holding section is tilted so that the absorber 20 is below it, and the samples, etc., dispensed into the reaction zone 11 are discharged into the absorber 20 through the flow path of the flow path section 12. The mechanism for tilting the substrate holding section can have any structure. For example, a stop can be provided at the end of the guide member guiding the moving stage, and a tilting cam can be provided in the substrate holding section. The moving stage is moved to a position where the tilting cam abuts against the stop, thereby raising the tilting cam. The substrate holding section is then connected to the moving stage via its rotation axis. Therefore, it can also be configured such that the substrate holding section tilts about the rotation axis as a base point according to the action of the tilting cam.

[0075] After the sample is drained, the reaction zone 11 of the biochemical reaction substrate 5 is moved to the detection camera. The detection camera is then lowered so that the light-shielding component at its front end is in close contact with the biochemical reaction substrate 5, preventing external light from entering the biochemical reaction substrate 5 and the external examination device. In this state, the presence or absence of an allergic reaction can be detected by detecting whether the labeled antibody luminesces. Regarding the luminescence of the labeled antibody, since the luminescent matrix labeled with anti-IgE antibody is dispensed during the dispensing process, the luminescent matrix is ​​reacted in the reaction zone 11 under specified environmental conditions for a specified time, and the intensity of the luminescence obtained by measuring the reaction result using the detection camera is measured.

[0076] Figure 10 (a) is a photograph showing the inspection results when the number of cleaning cycles (3 times) is set to less than half of the conventional number using the reaction plate 10 of this embodiment. Additionally, Figure 10 (b) is a photograph showing the inspection results when the number of washes was set to less than half (3 times) of the previous number of washes, as described above, using the conventional reaction plate. Furthermore, each wash (each wash) should only be performed to the extent that the sample or other object being washed is thoroughly mixed with the washing solution; the time and stirring method can be set appropriately.

[0077] from Figure 10 (b) It is evident that when the number of washes is less than half of that previously used with conventional reaction plates, undesirable luminescence is observed at the outer edge of reaction zone 11 (near the outflow prevention wall 13). Therefore, luminescence based on labeled anti-IgE antibody cannot be clearly observed at the outer edge of reaction zone 11. This is because residual liquid containing the labeled antibody solution that should be washed out indicates insufficient washing. Furthermore, although the results are not illustrated, when using conventional reaction plates for thorough washing, more than 7 washes are required.

[0078] In contrast, by Figure 10 (a) It can be seen that no unwanted luminescence was observed at the outer edge of the reaction zone 11 of the reaction plate 10 in this embodiment, and there was no liquid residue. Therefore, a distinct luminescence state based on the labeled anti-IgE antibody can be confirmed. It should be noted that the above-mentioned "thorough cleaning" refers to the degree to which there is no noise caused by unreacted residual labeled anti-IgE antibody or which will not cause problems during detection when the luminescence of the labeled antibody is detected by the detection camera.

[0079] As described above, in the allergy test using the reaction plate 10 of this embodiment, high sensitivity of the test can be achieved, and the test can be performed efficiently by reducing the test time and test work hours.

[0080] Example

[0081] Next, with reference to the embodiments, the effects of the "rounded corner R of the part where the reaction zone 11 and the outflow prevention wall 13 abut" and the "angle θ formed by the reaction zone 11 and the outflow prevention wall 13" of the reaction plate 10 of this embodiment will be explained.

[0082] For the reaction plates 10 of each embodiment and comparative example, the fillet radius R of the embodiment is set to about 0.3 mm, the included angle θ of embodiment 1 is 100°, the included angle θ of embodiment 2 is 110°, and the included angle θ of embodiment 3 is 120°; the fillet radius R of the comparative example is 0.1 mm, and the included angle θ is 100°.

[0083] In the reaction zone 11 of the reaction plate 10 in each embodiment and comparative example, as described above, multiple groups 51 (e.g., 3 points / group) of antigen 50 are arranged in a predetermined number of groups 51 (e.g., 75 points / 25 groups). At this time, when the antigen 50 contained in each group 51 binds to the sample and generates luminescence of the labeled antibody, the extent to which the intensity of this luminescence deviates within the reaction zone 11 is confirmed. Regarding the method for confirming the luminescence intensity, after the dispensing and reaction steps described above, with the reaction zone shielded from light, a detection camera is used to measure the intensity of the luminescence obtained as a result of the reaction.

[0084] The average luminous intensity of the "outer periphery" was calculated when the average luminous intensity of the "center portion" when using the reaction plates of each embodiment and comparative example was set to 100. In the embodiments and comparative examples, the "outer periphery" is the portion located within a range of 1.2 mm from the outflow prevention wall 13; the "center portion" refers to the remaining portions.

[0085] [Table 1]

[0086]

[0087] In addition, the center and the outer periphery of the reaction zone 11 are defined as follows: the outer periphery is located within a range of 1.2 mm from the outflow prevention wall 13, and the center is defined as the remaining portion.

[0088] Furthermore, when the reaction zone 11 is approximately circular (including circular and elliptical shapes), the "center" refers to the region inside the reaction zone 11 at any distance (e.g., radius, including minor and major axes) from the center point of the reaction zone 11, and the "peripheral" refers to the region outside the center. The peripheral region can be a portion where, in the case of a fillet radius R of 0.1 mm or less and an included angle of 100°, the luminous intensity decreases to 75 or less, 71 or less, 70 or less, 65 or less, or 60 or less when the average luminous intensity of the center is set to 100. Furthermore, the specific binding material immobilized on the boundary line between the peripheral and center regions is defined as the peripheral region.

[0089] In the reaction plates of the Examples and Comparative Examples, an antigen was used as a specific binding substance. The antigen was a substance configured in a 3-point / 1-group manner. However, the specific binding substance immobilized on the boundary line between the peripheral and central portions was considered to be located in the peripheral portion. Furthermore, if any one of the 3 points in a group was located in the peripheral portion, the 3-point / 1-group was considered to be located in the peripheral portion for evaluation.

[0090] As shown in Table 1 above, the average ratio of the luminous intensity of the outer periphery to that of the center was 86% in Example 1, 97% in Example 2, and 92% in Example 3. In contrast, the ratio of the comparative example was 71%, indicating that the luminous intensity of the outer periphery was approximately 30% lower than that of the center. When the deviation in luminous intensity is 30%, it affects the detection results when measuring the luminous intensity using a detection camera. Therefore, it can be concluded that the detection results of the outer periphery cannot be used for effective inspection.

[0091] Furthermore, the luminescence intensity of three points (point A, point B, and point C) arranged at the same position were compared on the outer periphery of the reaction zone of the reaction plate in both the embodiment and the comparative example. Point A is closest to the center of the reaction zone, and point C is farthest from the center of the reaction zone. In the above group, when the luminescence intensity at point A is set to 100%, the luminescence intensities of points B and C in the comparative example are 84% and 61%, respectively. In contrast, in embodiment 2, the luminescence intensities of points B and C are 100% and 91%, respectively.

[0092] Thus, in Examples 1-3, it can be confirmed that the luminescence intensity of the peripheral portion is approximately the same as that of the central portion, and that the luminescence intensity at each point disposed on the peripheral portion is also constant. Therefore, the vicinity of the outflow prevention wall 13 of the reaction zone 11 can be effectively utilized as a reaction zone, achieving space-saving of the reaction zone 11 and miniaturization of the reaction plate 10. Furthermore, by setting the reaction plate 10 to the same size as before, more antibodies 50 can be disposed, and the number of allergens that can be tested with a single sample can be increased.

[0093] It should be noted that, regarding the reaction plate 10 of this embodiment described above, the injection hole 31 and reaction zone 11 of the biochemical reaction substrate 5 are formed in a circular shape, but their shape is not limited to a circle; for example, they may also be formed in an elliptical shape. As can be seen from the claims, such modifications or improvements are also included within the scope of the present invention.

[0094] Furthermore, while the reaction plate 10 of this embodiment can be transparent or colored, using black allows for more sensitive measurements. Additionally, the sealing agent applied to the reaction plate 10 extends beyond the reaction zone 11 and the inclined surface 15 inside the outflow prevention wall 13, and can also suppress non-specific adsorption by being positioned inside the flow path 12. Similarly, as described above, any modifications or improvements made in this way are also within the scope of the present invention.

[0095] Explanation of reference numerals in the attached figures

[0096] 1: Analytical apparatus

[0097] 2: Operations Department

[0098] 3: Investment Department

[0099] 4: Casing

[0100] 5: Matrix for biochemical reactions

[0101] 6: Reagent kit

[0102] 10: Reaction plate

[0103] 11: Reaction Zone

[0104] 12: Flow path part

[0105] 12X and 12Y: Flow path wall

[0106] 13: Outflow prevention wall

[0107] 14: Base section

[0108] 15: Bevel

[0109] 15': End side

[0110] 16: Protrusion

[0111] 17: Top surface

[0112] 20: Absorber

[0113] 30: Cover

[0114] 31: Injection Hole

[0115] 35: Anti-slip unit

[0116] 40: Storage Containers

[0117] 41: Reaction plate storage section

[0118] 42: Absorbent housing section

[0119] 43: Insulated section

[0120] 46: Opening

[0121] 50: Antigen

[0122] 51: Group

[0123] θ: included angle.

Claims

1. A reaction plate assembled on a biochemical reaction substrate, characterized in that, The reaction plate has: a reaction zone for immobilizing a specific binding substance that reacts specifically with the analyte in the sample; and an outflow prevention wall formed to surround the reaction zone. The area where the reaction zone meets the outflow prevention wall forms a rounded corner R. The fillet radius R is greater than 0.10 mm.

2. The reaction plate according to claim 1, characterized in that, The fillet radius R is greater than 0.12 mm.

3. The reaction plate according to claim 1 or 2, characterized in that, The angle between the outflow prevention wall and the reaction zone is 91 degrees or more.

4. The reaction plate according to any one of claims 1 to 3, characterized in that, The angle between the outflow prevention wall and the reaction zone is 95 degrees to 125 degrees.

5. The reaction plate according to any one of claims 1 to 4, characterized in that, The reaction plate has a flow path that is connected to the reaction zone and prevents the samples from flowing out when the samples are stirred in the reaction zone, and discharges the samples after stirring.

6. The reaction plate according to claim 5, characterized in that, The flow path has an inclined surface formed in a manner that rises from the reaction zone.

7. The reaction plate according to claim 5 or 6, characterized in that, The flow path extends from the reaction zone to the end of the reaction plate, and the length of the end of the flow path in the width direction is narrower than the length of the reaction zone in the width direction.

8. The reaction plate according to any one of claims 5 to 7, characterized in that, The flow path section has a flow path wall that is continuous with the outflow prevention wall. The flow path wall is formed into a streamline shape along the flow path of the flow path section.

9. The reaction plate according to any one of claims 5 to 8, characterized in that, The outflow prevention wall has an opening that opens toward any end of the reaction plate. The flow path is formed between the opening and the end.

10. The reaction plate according to any one of claims 5 to 9, characterized in that, The flow path wall of the flow path portion is formed such that the width of the flow path narrows from the reaction zone toward the end, and the width of the flow path widens at the end side.

11. The reaction plate according to any one of claims 1 to 10, characterized in that, The surface of the reaction zone has a coating that fixes the specific binding substance to the reaction zone.

12. A method for manufacturing a reaction plate, wherein the reaction plate is assembled onto a substrate for biochemical reactions, characterized in that, The reaction plate has: a reaction zone for immobilizing a specific binding substance that reacts specifically with the analyte in the sample; and an outflow prevention wall formed to surround the reaction zone. The reaction zone and the outflow prevention wall form a rounded corner R, which is greater than 0.10 mm. In this reaction plate, an immobilizing agent is applied to the reaction zone by rotating the reaction plate. This immobilizing agent is used to immobilize a specific binding substance that specifically reacts with the analyte in the sample in the reaction zone.

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