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WO2025206266A1 - Plaque de réaction - Google Patents

Plaque de réaction

Info

Publication number
WO2025206266A1
WO2025206266A1 PCT/JP2025/012679 JP2025012679W WO2025206266A1 WO 2025206266 A1 WO2025206266 A1 WO 2025206266A1 JP 2025012679 W JP2025012679 W JP 2025012679W WO 2025206266 A1 WO2025206266 A1 WO 2025206266A1
Authority
WO
WIPO (PCT)
Prior art keywords
reaction
reaction plate
reaction area
area
flow path
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/JP2025/012679
Other languages
English (en)
Japanese (ja)
Inventor
健二 植村
敬太 塚本
誠也 示野
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nippon Chemiphar Co Ltd
Original Assignee
Nippon Chemiphar Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nippon Chemiphar Co Ltd filed Critical Nippon Chemiphar Co Ltd
Publication of WO2025206266A1 publication Critical patent/WO2025206266A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N37/00—Details not covered by any other group of this subclass

Definitions

  • the present invention relates to a reaction plate that is incorporated into a biochemical reaction substrate for measuring the amount of a sample substance that has physiological activity based on a biochemical reaction process such as an immunoassay.
  • Testing for biological substances which checks the presence and amount of substances in the body, is important for understanding health conditions and determining treatment methods. Furthermore, in today's world where living environments are becoming increasingly diverse, there is a need to perform a wide variety of tests for biological substances, and there is a demand for faster and more sensitive testing. For example, when treating an allergic disease, it is important to first understand which allergic disease the person is suffering from. In recent years, there have been a wide variety of causes of allergies, and it is necessary to identify the cause in order to receive appropriate treatment, such as using the appropriate medication.
  • a common method is to quantify IgE antibodies against specific allergens in a blood sample collected from a subject using solid-phase sandwich immunoassay.
  • a solid-phase support such as a glass filter is prepared by adsorbing a ligand capture antibody and blocking protein adsorption sites other than the ligand capture antibody adsorption site with a blocking agent such as casein.
  • a specific allergen such as dust mite or pollen, is then bound to the ligand.
  • This is then mixed with a blood sample to form a complex between the specific allergen bound to the ligand and the IgE antibody against the specific allergen in the blood sample.
  • the mixture containing this complex is then added to the solid-phase support on which the ligand capture antibody has been adsorbed, allowing the ligand portion of the complex to bind to the ligand capture antibody.
  • an anti-IgE antibody labeled with an enzyme or the like is added, allowing the IgE antibody portion of the complex to bind to the labeled anti-IgE antibody.
  • excess labeled anti-IgE antibody that did not bind to the complex is removed, and a color reaction appropriate to the type of label is carried out to detect the labeled anti-IgE antibody that has bound to the IgE antibody.
  • the detection results obtained are compared with a calibration curve previously prepared using labeled IgE antibody, and the amount of IgE antibody against the specific allergen in the blood sample is quantified.
  • a method and device for testing for the presence or absence of allergies is known, using a biological reaction substrate in which a specific allergen is bound to a porous filter of the biological reaction substrate.
  • Another example of a method and device for testing for the presence or absence of allergies using a biological reaction substrate is the biochemical reaction substrate described in Patent Document 2.
  • the substrate includes a reaction plate, an absorbent, a storage container equipped with a reaction plate storage section for storing the reaction plate and an absorbent storage section for storing the absorbent, and an insulated section, and a cover attached to the storage container so as to cover at least a portion of the reaction plate and absorbent stored in the storage container.
  • the reaction plate has a reaction area in which a specific binding substance that reacts specifically with the test substance in the sample is immobilized, and a flow path section connecting the absorbent and the reaction area.
  • the cover has an injection hole for injecting the sample or the like into the reaction plate.
  • the test using the reaction plate described in Patent Document 2 has a reaction area where a specific binding substance (e.g., an antigen (allergen)) that reacts specifically with the test substance in the sample is immobilized, and a flow path section that connects the absorber and the reaction area, and the cover has an injection hole for injecting the sample, etc. into the reaction plate, so the number of test steps can be reduced (e.g., the step of absorbing cleaning liquid from the reaction area in order to drain the cleaning liquid can be eliminated), making the test easier to perform.
  • a specific binding substance e.g., an antigen (allergen)
  • allergen allergen
  • the (4) labeled antibody washing step washing is performed multiple times (for example, about 10 times) to wash the labeled antibody, and the more times this labeled antibody washing step is performed, the longer the washing step takes, making it impossible to shorten the test time.
  • the "(4) labeled antibody washing step” refers to a series of steps that start from a state in which the labeled antibody added in step (3) has been removed (discharged) from the reaction area, and include injecting a washing solution into the reaction area, agitating the washing solution, and removing (discharged) the washing solution
  • the "number of labeled antibody washing steps” refers to the number of times this series of steps is performed.
  • the reaction area of the reaction plate described in Patent Document 2 is coated in advance with an immobilizing agent such as a polymer, and a specific binding substance (e.g., an antigen (allergen)) that reacts specifically with the test substance in the sample is immobilized in the reaction area via the immobilizing agent.
  • an immobilizing agent such as a polymer
  • a specific binding substance e.g., an antigen (allergen)
  • allergen an antigen
  • the immobilizing agent is not coated to a uniform thickness over the entire surface of the reaction area, the amount and density of the specific binding substance immobilized in the reaction area will vary depending on the immobilization site, and it has been found that there is an issue in that even if the same specific binding substance is used, the test results will vary depending on the site.
  • the reaction area in the reaction plate described in Patent Document 2 is surrounded on its outer edge by an outflow prevention wall. Therefore, when the reaction plate is coated with an immobilizing agent, the immobilizing agent is attracted by the surface tension of the inner wall of the outflow prevention wall, resulting in a thick coating film of the immobilizing agent in the reaction area around the outflow prevention wall, which can be thicker than the coating film in the center of the reaction area. This can result in a difference in detection sensitivity between the center of the reaction area and the outer periphery around the outflow prevention wall, resulting in different test results.
  • the present invention has been made to solve these problems; its objectives are: 1) to provide a reaction plate that allows stable testing by ensuring reliable cleaning even when the number of times the reaction area is cleaned is reduced, and/or 2) to provide a reaction plate that allows similar testing to be performed in both the outer and central parts of the reaction area by matching the detection sensitivity between the outer and central parts.
  • the reaction plate of the present invention which solves the above problems, is a reaction plate incorporated into a biochemical reaction substrate, and has a reaction area in which a specific binding substance that reacts specifically with an analyte in a sample is immobilized, and an outflow prevention wall formed to surround the reaction area, with the area where the reaction area and the outflow prevention wall meet forming a corner R, and the corner R being greater than 0.10 mm.
  • the corner R is greater than 0.12 mm.
  • the angle formed between the outflow prevention wall and the reaction area is 91 degrees or greater.
  • the angle between the outflow prevention wall and the reaction area is between 95 degrees and 130 degrees.
  • the reaction plate according to the present invention has a flow path section that is connected to the reaction area, prevents the sample, etc. from flowing out when the sample, etc. in the reaction area is stirred, and is capable of discharging the sample, etc. after stirring.
  • the flow path portion has a slope formed so as to rise from the reaction area.
  • the flow path section is preferably formed from the reaction area to the edge of the reaction plate, and the width of the flow path of the flow path section at the edge is preferably narrower than the width of the reaction area.
  • the flow path section includes a flow path wall that is continuous with the outflow prevention wall, and that the flow path wall is formed in a streamlined shape along the flow path of the flow path section.
  • the outflow prevention wall has an opening that opens toward any edge of the reaction plate, and the flow path portion is formed between the opening and the edge.
  • the surface of the reaction area has a coating film that fixes the specific binding substance to the reaction area.
  • the method for manufacturing a reaction plate according to the present invention is a method for manufacturing a reaction plate to be incorporated into a biochemical reaction substrate, and the reaction plate has a reaction area in which a specific binding substance that reacts specifically with a test substance in a sample is immobilized, and an outflow prevention wall formed to surround the reaction area, the portion where the reaction area and the outflow prevention wall abut forming a corner R, and the corner R is greater than 0.10 mm, and is characterized in that the reaction area is coated with an immobilizing agent for immobilizing the specific binding substance that reacts specifically with the test substance in the sample in the reaction area by rotating the reaction plate.
  • FIG. 1 is a perspective view of an analyzer that performs testing using a reaction plate according to an embodiment of the present invention.
  • 1 is a perspective view of a biochemical reaction substrate in which a reaction plate according to an embodiment of the present invention is incorporated.
  • FIG. 2 is an exploded view of a biochemical reaction substrate into which a reaction plate according to an embodiment of the present invention is incorporated.
  • FIG. 2 is a perspective view of a reaction plate according to an embodiment of the present invention.
  • 5 is a cross-sectional view taken along the line AA in FIG. 4.
  • FIG. 6 is an enlarged view of part B in FIG. 5 .
  • FIG. 10 is a perspective view showing a modified example of the reaction plate according to the embodiment of the present invention.
  • FIG. 10 is a perspective view showing a modified example of the reaction plate according to the embodiment of the present invention.
  • FIG. 10 is a plan view showing a modified example of the reaction plate according to the embodiment of the present invention.
  • FIG. 2 is a top view illustrating the arrangement of antigens (allergens) in the reaction area of the reaction plate.
  • 1A and 1B are diagrams illustrating the detection results of a test using a reaction plate according to an embodiment of the present invention, and
  • FIG. 1B is a diagram illustrating the detection results of a conventional reaction plate.
  • FIG. 1 is a perspective view of an analytical device that performs testing using a reaction plate according to an embodiment of the present invention
  • FIG. 2 is a perspective view of a biochemical reaction substrate into which a reaction plate according to an embodiment of the present invention is incorporated
  • FIG. 3 is an exploded view of a biochemical reaction substrate into which a reaction plate according to an embodiment of the present invention is incorporated
  • FIG. 4 is a perspective view of a reaction plate according to an embodiment of the present invention
  • FIG. 5 is a cross-sectional view taken along line A-A in FIG. 4
  • FIG. 6 is an enlarged view of portion B in FIG. 5
  • FIG. 7 is a perspective view of a modified reaction plate according to an embodiment of the present invention
  • FIG. 1 is a perspective view of an analytical device that performs testing using a reaction plate according to an embodiment of the present invention
  • FIG. 2 is a perspective view of a biochemical reaction substrate into which a reaction plate according to an embodiment of the present invention is incorporated
  • FIG. 3
  • FIG. 8 is a plan view of a modified reaction plate according to an embodiment of the present invention
  • FIG. 9 is a top view illustrating the arrangement of antigens (allergens) in the reaction area of a reaction plate
  • FIG. 10 are diagrams illustrating the detection results of a test, where (a) is a diagram illustrating the detection results of a test using a reaction plate according to an embodiment of the present invention, and (b) is a diagram illustrating the detection results of a conventional reaction plate.
  • the reaction plate 10 As shown in FIG. 1, the reaction plate 10 according to this embodiment is incorporated into a biochemical reaction substrate 5 and is inserted into the input section 3 of the analyzer 1 together with a reagent cartridge 6 containing a specimen and a cleaning solution for testing.
  • the analyzer 1 comprises a housing 4 having an operation panel 2 and the input section 3 in which the reagent cartridge 6 and biochemical reaction substrate 5 are mounted.
  • the input section 3 can be opened and closed freely by an attached door (not shown).
  • the interior of the analytical device 1 is divided into an installation area, barcode reading area, dispensing area, mixing area, detection area, and drainage area. Arranging each area on the same straight line increases area efficiency, allowing the analytical device 1 to be made more compact.
  • a guide means e.g., a moving table
  • the reaction plate 10 of this embodiment can be incorporated into a biochemical reaction substrate 5 for use.
  • the biochemical reaction substrate 5 need only be capable of housing the reaction plate 10 of this embodiment therein, and may further include a cover 30 and/or a storage container 40.
  • the cover 30 has a substantially circular injection hole 31 formed therein, and is configured to be positioned corresponding to the reaction area 11 of the reaction plate 10 when the reaction plate 10 of this embodiment is housed therein.
  • the storage container 40 may also have a concave heat-insulating portion 43 formed at the bottom, which can be configured to allow a heater from an analytical device (described later) to contact and heat the reaction area 11.
  • the cover 30 may also have an uneven anti-slip means 35 formed on the outer peripheral sidewall, which can be configured to allow an examiner to securely grip the biochemical reaction substrate 5.
  • the storage container 40 is formed with a reaction plate storage section 41 for storing the reaction plate 10 according to this embodiment, and may further be provided with an absorber storage section 42 for storing the absorber 20 made of a porous material such as a sponge that can sufficiently absorb liquid. Note that when the reaction plate 10 and absorber 20 according to this embodiment are stored in the storage container 40, the tip of the flow path section 12 formed on the reaction plate 10 according to this embodiment is positioned so as to abut the absorber 20.
  • the reaction plate 10 need only fit smoothly into the reaction plate storage section 41 formed in the storage container 40, and may have, for example, a plate-shaped base section 14 that can fit into the reaction plate storage section 41. Furthermore, the reaction plate 10 is provided with an outflow prevention wall 13, which may be erected from the base section 14. The outflow prevention wall 13 is, for example, a roughly annular wall, and the interior of the outflow prevention wall 13 defines the reaction area 11. Furthermore, the reaction plate 10 is provided with a flow path that communicates with the reaction area 11, and the flow path includes a slope 15 and constitutes at least a part of the flow path section 12.
  • the outflow prevention wall 13 may have any shape that does not cause stagnation during drainage, and may be formed, for example, in a circular or elliptical shape.
  • the reaction area 11 inside the outflow prevention wall 13 may also be formed in a circular or elliptical shape.
  • a slope 15 that forms a flow path is provided at a position facing the absorbent 20 from the reaction area 11, and the flow path is defined by a pair of flow path walls 12X and 12Y extending from the outer periphery of the outflow prevention wall 13. It is preferable that the pair of flow path walls 12X and 12Y are curved in a streamlined shape along the flow path so that the width of the flow path (including the slope 15) (i.e., the distance between the pair of flow path walls 12X and 12Y) changes continuously. For example, as shown in Figures 4 and 5, the distance between the pair of flow path walls 12X and 12Y is gradually narrowed and then gradually widened from the reaction area 11 toward the end side 15' of the slope 15.
  • the pair of flow path walls 12X and 12Y define a flow path (including the slope 15) for draining the sample, etc., dispensed into the reaction area 11, and the drained liquid discharged through the flow path is absorbed by the absorbent 20.
  • the pair of flow path walls 12X and 12Y have slopes 15 formed to rise from the reaction area 11 so that specimens and the like can be discharged efficiently from the reaction area 11 through the flow path toward the absorber 20 by tilting the biochemical reaction substrate 5.
  • the flow path walls 12X and 12Y are preferably formed continuously with the outflow prevention wall 13 and configured in a curved shape from the outflow prevention wall 13 to the flow path walls 12X and 12Y, and preferably configured so as to have no sharp corners. If there is a part that can be seen as a "seam" where the inner wall portion of the flow path walls 12X and 12Y and the inner wall portion of the outflow prevention wall 13 meet, for example, if there is an uneven shape such as a sharp protruding shape, a concave or recessed shape, or a stepped shape, the liquid that is being discharged is likely to remain in the seam, and the liquid, etc., is likely to remain in the seam without being completely discharged.
  • the part that connects the outflow prevention wall 13 to the flow path walls 12X and 12Y be curved in a streamlined shape, and it is preferable that there are no parts that can be seen as seams.
  • the top surfaces of the flow path walls 12X and 12Y and the outflow prevention wall 13 are different (there is a gap) at the contact points between the two walls, specimens and the like are likely to remain in the gap and not be able to be completely discharged. Therefore, it is preferable that the top surfaces are also formed so that there is no gap between them.
  • the inclined surface 15 that constitutes the flow path is preferably formed at an angle of approximately 5° to 70° from the base portion 14 (or reaction area 11), with a more preferred angle of 12° to 50°, an even more preferred angle of 17° to 35°, and a suitable angle of 23°.
  • the flow path section 12 includes a flow path formed from the reaction area 11 to the end side 15', and the length of the end side 15' of the flow path formed by the flow path section 12 in the width direction (the short side direction of the base section 14, i.e., the distance between the flow path walls 12X and 12Y) is formed so that it is narrower than the length of the longest point in the reaction area 11, and the end side 15' of the flow path formed by the flow path section 12 is formed so that the slope 15 abuts at an acute angle against the upper surface of the absorber 20 in side view (the angle formed by the upper surface of the absorber 20 and the extended surface of the slope 15 is an acute angle).
  • the side (inner surface) of the outflow prevention wall 13 facing the reaction area 11 is preferably formed so that its side surface (inner surface) is inclined outward relative to the vertical direction from the base portion 14 on which the reaction area 11 is formed.
  • the angle ⁇ between the inner surface of the outflow prevention wall 13 and the surface of the reaction area 11 is preferably 91° or more when viewed from the reaction area 11 side, and an angle of 91° to 135° can prevent samples, etc., dispensed into the reaction area 11 from leaking out of the reaction area 11 when they are stirred.
  • the angle ⁇ is 95° to 130°, and even more preferably 95° to 125°, 95° to 120°, 97 to 115°, 97 to 113°, etc.
  • the angle ⁇ is 95° to 130°, and even more preferably 95° to 125°, 95° to 120°, 97 to 115°, 97 to 113°, etc.
  • this corner R it is possible to prevent the sample or other liquid dispensed into the reaction area 11 from remaining on the periphery of the reaction area 11 (around the outflow prevention wall 13 in the reaction area) when it is being drained. For example, this prevents labeled antibodies that are being drained from remaining on the periphery of the reaction area 11. As a result, it is possible to improve testing accuracy.
  • the corner R (the radius of the corner R) is preferably greater than 0.10 mm, more preferably greater than 0.12 mm, and even more preferably 0.25 mm or greater.
  • the corner R may be formed to be 0.75 to 0.11 mm, 0.60 to 0.12 mm, 0.50 to 0.12 mm, 0.40 to 0.20 mm, or 0.35 to 0.25 mm.
  • the reaction plate 10 has an outflow prevention wall 13 formed to surround the reaction area 11, and a predetermined corner R is formed where the inner surface of the outflow prevention wall 13 abuts the reaction area 11.
  • a predetermined corner R is formed where the inner surface of the outflow prevention wall 13 abuts the reaction area 11.
  • the surface of the reaction area 11 is preferably coated with an immobilizing agent for immobilizing the specific binding substance.
  • the immobilizing agent can be, for example, a polymer (copolymer), and is not particularly limited as long as it can immobilize the specific binding substance.
  • a conventionally known polymer containing multiple photoreactive groups can be used.
  • a method for coating the surface of the reaction area 11 with the immobilizing agent can be used (sometimes called spin coating), in which a reaction area 11 of a reaction plate 10 comprising an uncoated reaction area 11 and an outflow prevention wall 13 (preferably a flow path section 12) is filled with an amount of immobilizing agent sufficient to coat the reaction area 11 at any location within the area, and then the reaction plate 10 is rotated.
  • spin coating not only the surface of the reaction area 11 but also the inner wall surfaces of the outflow prevention wall and the flow path wall can be coated with the immobilizing agent. This also has the effect of smoothing the flow of liquid when discharging cleaning liquid and the like during the washing process.
  • the thickness of the coating layer of the immobilizing agent can be made more consistent than when there are no rounded corners. If there are no rounded corners, the immobilizing agent being coated during rotation may bounce off the outflow prevention wall 13; as a result, the thickness of the coating layer on the periphery of the reaction area 11 (near the outflow prevention wall 13) may be thicker than the thickness of the coating layer in the center of the reaction area.
  • the rotation speed for coating the fixing agent can be any speed that ensures a uniform thickness of the coating layer after coating, and any speed can be selected as appropriate.
  • the rotation speed may be constant, or may be increased gradually. If the speed is increased gradually, three or more speed stages may be set.
  • the rotation conditions for producing the fixing agent coating layer are preferably divided into two stages, with the first stage being a low speed and the second stage being a high speed.
  • a blocking agent may be applied to the entire surface of the reaction area 11 in advance.
  • the blocking agent can suppress non-specific adsorption in the reaction area 11.
  • Synthetic polymers that are not derived from animals or plants, such as polyethylene glycol, can be used as blocking agents using common techniques, and can be selected appropriately depending on the material of the reaction plate 10, the properties of the target substance (e.g., antigen), the specimen (e.g., blood), and the reagent (e.g., washing solution).
  • the inside of the outflow prevention wall 13 is defined as the reaction area 11
  • coating the inner wall surfaces of the reaction area 11 and the outflow prevention wall 13 with a blocking agent will make the affinity (e.g., hydrophilic, hydrophobic, or water-repellent) for the sample or other liquid the same between the inside of the outflow prevention wall 13 and the reaction area 11, which has the secondary effect of making it easier to discharge the sample.
  • a blocking agent e.g., hydrophilic, hydrophobic, or water-repellent
  • the reaction plate according to this embodiment may include components other than the reaction area 11, flow path section 12, outflow prevention wall 13, base 14, and inclined surface 15.
  • the reaction plate 10' may be formed so that the end side of the flow path section 12 is spaced apart so that the diameter thereof expands further outward.
  • the reaction plate 10' may have a pair of protrusions 16 extending from the base 14 near the position where the flow path walls 12X and 12Y of the flow path section 12 connect to the outflow prevention wall 13. Furthermore, the tip portion of the flow path section 12 may be formed to have an upper surface 17 that is approximately parallel to the upper surface of the protrusions 16.
  • Such protrusions 16 and upper surface 17 facilitate handling of the reaction plate 10'. For example, when coating the reaction area 11 with an immobilizing agent or immobilizing a specific binding substance in the reaction area 11, the reaction plate 10' can be handled by grasping the protrusions 16 with a tool or machine. Furthermore, when storing multiple reaction plates 10', even if they are stacked on top of each other, the protrusions 16 and upper surface 17 abut against the back surface of the base 14, allowing for stable storage.
  • the protruding portion 16 of the reaction plate 10' it is preferable to position the protruding portion 16 of the reaction plate 10' in a position where it does not come into contact with the inner walls of the flow path walls 12X and 12Y and/or the outflow prevention wall 13. If the protruding portion 16 comes into contact with the inner walls of the flow path section 12 and/or the outflow prevention wall 13, the protruding portion 16 will protrude into the inner wall, which may result in liquid remaining in the protruding portion when the sample, etc. is discharged, increasing the number of discharge steps.
  • a specific binding substance e.g., an antigen
  • a specific binding substance is immobilized in the reaction area 11 of the reaction plate 10 according to this embodiment. Only one type of specific binding substance may be immobilized, but preferably multiple types are immobilized. This is because multiple types of substances can be detected in a single test. Furthermore, one type of specific binding substance may be immobilized not just in one location, but in multiple locations. This is because this allows for increased detection accuracy for a single type of substance.
  • the reaction plate 10 of FIG. 9 25 types of antigens 50 are arranged and immobilized in the reaction area 11; and each type of antigen 50 is immobilized in three locations (three spots).
  • the reaction plate of FIG. 9 has multiple sets 51 (e.g., 3 spots/set) of antigens 50 arranged in a predetermined number of sets 51 (e.g., 75 spots/25 sets).
  • n number the number of spots
  • the area within the reaction area 11 can be used effectively (wasted space can be eliminated).
  • the analyzer 1 that performs allergy testing using a biochemical reaction substrate 5 incorporating the reaction plate 10 according to this embodiment.
  • the biochemical reaction substrate 5 and reagent cartridge 6 are set in the input unit 3.
  • the reagent cartridge 6 is a component that contains specimens (including diluted specimens) and reagents.
  • the analyzer 1 is equipped with a dispensing nozzle, and the reagent cartridge 6 is also equipped with a tip.
  • the tip housed in the reagent cartridge 6 is fitted to the tip of the dispensing nozzle, and the specimen, reagent, etc. in the reagent cartridge 6 is aspirated from the reagent cartridge 6.
  • the aspirated specimen, reagent, etc. can then be dispensed into the reaction area 11 via the injection hole 31 of the biochemical reaction substrate 5.
  • This dispensing is performed through a diluted sample addition step in which the sample is dispensed into the reaction area of the reaction plate of the biochemical reaction substrate and allowed to bind to the IgE immobilized in the reaction area; a sample washing step in which washing is performed with a washing solution to remove unreacted sample components in the reaction area; a labeled antibody addition step in which a labeled antibody is added and the IgE antibody binds to the labeled antibody; a labeled antibody washing step in which washing is performed with a washing solution to remove unreacted labeled antibody; and a chemiluminescent substrate addition step in which a chemiluminescent substrate is added.
  • the tip After use during dispensing, the tip is separated from the dispensing nozzle and returned to a designated position in the reagent cartridge 6. By returning the used tip to its original position, there is no need to provide a location for collecting used tips, which allows the size of the reagent cartridge to be reduced.
  • the movable table can be reciprocated in this state to agitate the specimen, reagent, and cleaning solution dispensed into the reaction area 11 of the biochemical reaction substrate 5 as appropriate during the reaction process.
  • the direction of the reciprocating motion of the movable table is along the short side direction of the biochemical reaction substrate 5.
  • This drainage process involves tilting the substrate holder of the moving table, tilting the biochemical reaction substrate 5 held in the substrate holder so that the absorber 20 is on the bottom, and draining the specimens and other liquids dispensed in the reaction area 11 into the absorber 20 via the flow path of the flow path section 12.
  • Any configuration can be used for the mechanism to tilt the substrate holder.
  • a stopper can be installed at the end of the guide member that guides the moving table
  • a tilting cam can be installed on the substrate holder, and the moving table can be moved to a position where the tilting cam abuts the stopper, causing the tilting cam to rise. Since the substrate holder is connected to the moving table via its rotation axis, the substrate holder can be tilted around the rotation axis in response to the operation of the tilting cam.
  • the reaction area 11 of the biochemical reaction substrate 5 is moved to the detection camera.
  • the detection camera is then lowered and the light-shielding member attached to its tip is brought into close contact with the biochemical reaction substrate 5, blocking external light from entering the biochemical reaction substrate 5 and the testing device.
  • the luminescence of the labeled antibody is caused by a luminescent substrate that is luminescent by the labeled anti-IgE antibody, which is dispensed in the dispensing process.
  • This luminescence substrate is allowed to react in the reaction area 11 under specified environmental conditions for a specified time, and the intensity of the luminescence obtained as a result of this reaction is measured by the detection camera.
  • Figure 10(a) is a photograph showing the test results when the reaction plate 10 according to this embodiment was used and the number of washes was reduced to less than half (3 times) of the conventional number.
  • Figure 10(b) is a photograph showing the test results when a conventional reaction plate was used and the number of washes was reduced to less than half (3 times) of the conventional number, as mentioned above.
  • Each wash should be sufficient to ensure that the sample or other object to be washed is sufficiently mixed with the washing solution, and the time and stirring method can be set as appropriate.
  • allergy testing using the reaction plate 10 according to this embodiment not only increases the sensitivity of the test, but also reduces the testing time and labor required, making it possible to perform the test efficiently.
  • the corner R of the example was set to approximately 0.3 mm, the angle ⁇ of Example 1 was 100°, the angle ⁇ of Example 2 was 110°, and the angle ⁇ of Example 3 was 120°; the corner R of the comparative example was 0.1 mm, and the angle ⁇ of Example 3 was 100°.
  • multiple sets 51 e.g., 3 spots/set
  • sets 51 e.g., 75 spots/25 sets.
  • the method for confirming the intensity of the emitted light was to measure the intensity of the emitted light resulting from the reaction using a detection camera with the reaction area shielded from light after the dispensing and reaction steps described above.
  • the average luminescence intensity of the "periphery” was calculated by setting the average luminescence intensity of the "center” when using the reaction plates of each Example and Comparative Example as 100.
  • the "periphery” in the Examples and Comparative Examples was defined as the area located within 1.2 mm from the outflow prevention wall 13, and the "center” was defined as the remaining area.
  • the central and peripheral parts of the reaction area 11 are defined as follows: the peripheral part is the area located within 1.2 mm of the outflow prevention wall 13, and the central part is the remaining area.
  • the "center” of the reaction area 11 refers to the region within any distance (for example, the radius (including the minor and major axes)) from the center point of the reaction area 11 when the reaction area 11 is approximately circular (including circular and elliptical shapes), and the "periphery" of the reaction area 11 refers to the region outside the center.
  • the peripheral region can be the portion where the luminescence intensity is 75 or less, 71 or less, 70 or less, 65 or less, or 60 or less, when the average luminescence intensity of the center is 100.
  • the specific binding substance immobilized on the boundary between the peripheral region and the center is defined as the peripheral region.
  • antigens were used as specific binding substances, and the antigens were arranged in three spots per set.
  • specific binding substances immobilized on the boundary between the outer periphery and the center were considered to be located on the outer periphery, and if even one of the three spots in a set was located on the outer periphery, the three spots per set were evaluated as being located on the outer periphery.
  • the average ratio of the luminescence intensity of the outer periphery to the luminescence intensity of the center was 86% for Example 1, 97% for Example 2, and 92% for Example 3.
  • the ratio for the comparative example was 71%, making it clear that the luminescence intensity of the outer periphery is approximately 30% lower than that of the center.
  • a 30% variation in luminescence intensity affects the detection results when measuring the luminescence intensity with a detection camera, and therefore it is clear that the detection results for the outer periphery cannot be used for effective inspection.
  • the luminescence intensities of a set of three spots spot A, spot B, spot C
  • spot A was closest to the center of the reaction area
  • spot C was furthest from the center of the reaction area. If the luminescence intensity of spot A in the set was taken as 100%, then in the Comparative Example, the luminescence intensities of spots B and C were 84% and 61%, while in Example 2, the luminescence intensities of spots B and C were 100% and 91%.
  • the reaction area 11 can be effectively utilized up to the vicinity of the outflow prevention wall 13, which allows for space-saving in the reaction area 11 and makes it possible to reduce the size of the reaction plate 10. Furthermore, by making the reaction plate 10 the same size as conventional plates, it becomes possible to place more antibodies 50 on them, thereby increasing the number of allergens that can be tested with a single specimen.
  • reaction plate 10 has been described above with the injection hole 31 and reaction area 11 of the biochemical reaction substrate 5 formed in a circular shape, but these shapes are not limited to circular and may be formed in an elliptical shape, for example. It is clear from the claims that such modified or improved forms are also included within the technical scope of the present invention.
  • reaction plate 10 according to the present embodiment described above can be a transparent or colored plate, but using a black plate allows for more sensitive measurements. Furthermore, by applying the blocking agent to the reaction plate 10 to the reaction area 11, including the inside of the outflow prevention wall 13, the slope 15, and the inside of the flow path section 12, non-specific adsorption can be further suppressed. As mentioned above, it is clear from the claims that such modified or improved embodiments are also within the technical scope of the present invention.

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Abstract

L'invention concerne une plaque de réaction incluse dans un substrat de réaction biochimique utilisé pour un test d'allergie ou similaire, 1) un test stable pouvant être effectué en effectuant un nettoyage suffisant même lorsque le nombre de fois où une zone de réaction est nettoyée est réduit, et/ou 2) le même test peut être effectué dans une section périphérique externe et une section centrale de la zone de réaction par mise en correspondance de la sensibilité de détection de la section périphérique externe et de la sensibilité de détection d'une section périphérique interne de la zone de réaction. La plaque de réaction incluse dans un substrat de réaction biochimique comprend : une zone de réaction dans laquelle une substance de liaison spécifique qui réagit spécifiquement avec une substance de test dans un échantillon est immobilisée ; et une paroi de prévention d'écoulement formée de façon à entourer la zone de réaction. Une partie où la zone de réaction et la paroi de prévention d'écoulement viennent en butée est supérieure à un coin R de 0,10 mm.
PCT/JP2025/012679 2024-03-29 2025-03-28 Plaque de réaction Pending WO2025206266A1 (fr)

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JP2024-055314 2024-03-29
JP2024055314 2024-03-29

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WO2025206266A1 true WO2025206266A1 (fr) 2025-10-02

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004505231A (ja) * 1999-08-06 2004-02-19 サーモ　バイオスター　インコーポレイテッド 完全サンプル処理能力を備える、医療現場用検出自動化装置
JP2009140131A (ja) * 2007-12-05 2009-06-25 Fuji Electric Retail Systems Co Ltd 自動販売機
JP2019010630A (ja) * 2017-06-30 2019-01-24 株式会社エッチアールディ 磁力増強具及び水活性化システム
JP2022118238A (ja) * 2018-02-02 2022-08-12 日本ケミファ株式会社 反応用プレート
JP2023051957A (ja) * 2016-12-22 2023-04-11 イラミーナ インコーポレーテッド フローセルパッケージおよびその製造方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004505231A (ja) * 1999-08-06 2004-02-19 サーモ　バイオスター　インコーポレイテッド 完全サンプル処理能力を備える、医療現場用検出自動化装置
JP2009140131A (ja) * 2007-12-05 2009-06-25 Fuji Electric Retail Systems Co Ltd 自動販売機
JP2023051957A (ja) * 2016-12-22 2023-04-11 イラミーナ インコーポレーテッド フローセルパッケージおよびその製造方法
JP2019010630A (ja) * 2017-06-30 2019-01-24 株式会社エッチアールディ 磁力増強具及び水活性化システム
JP2022118238A (ja) * 2018-02-02 2022-08-12 日本ケミファ株式会社 反応用プレート

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