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WO2018199168A1 - Support de membrane, son procédé de production et kit de test d'échantillon liquide - Google Patents

Support de membrane, son procédé de production et kit de test d'échantillon liquide Download PDF

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Publication number
WO2018199168A1
WO2018199168A1 PCT/JP2018/016815 JP2018016815W WO2018199168A1 WO 2018199168 A1 WO2018199168 A1 WO 2018199168A1 JP 2018016815 W JP2018016815 W JP 2018016815W WO 2018199168 A1 WO2018199168 A1 WO 2018199168A1
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WIPO (PCT)
Prior art keywords
substance
charge
detection
flow path
membrane carrier
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PCT/JP2018/016815
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English (en)
Japanese (ja)
Inventor
佳那恵 原
門田　健次
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デンカ株式会社
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Application filed by デンカ株式会社 filed Critical デンカ株式会社
Priority to JP2019514578A priority Critical patent/JP7025413B2/ja
Publication of WO2018199168A1 publication Critical patent/WO2018199168A1/fr

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    • 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

Definitions

  • the present invention relates to a membrane carrier, a manufacturing method thereof, and a liquid sample inspection kit.
  • POCT Point of Care Test
  • reagents have been attracting attention, which measure antigen morbidity, pregnancy, blood glucose levels, etc. by using antigen-antibody reactions and the like.
  • POCT is a test performed beside the subject in the POCT guidelines emitted by the Japan Society for Clinical Laboratory Automation, or a test performed by the subject himself, shortening the test time and testing on the spot (test visible to the subject) Is defined as an inspection that has the advantage of being possible.
  • POCT reagents are characterized by rapid testing, simple usage, and inexpensive reagents. Because of these characteristics, it is often used for medical examinations and regular medical examinations at a mild stage, and it is an important diagnostic tool in home medical care that is expected to increase in the future.
  • determination is performed by introducing a liquid sample such as blood into a test kit and detecting a specific substance to be detected contained therein.
  • An immunochromatography method is often used as a method for detecting a specific substance to be detected from a liquid sample.
  • the immunochromatography method is that the liquid dropped on the membrane carrier of the test kit moves on the membrane carrier, and the substance to be detected and the subject to be detected in the liquid sample suspended or dissolved in the liquid sample.
  • Labeled particles bound with antibodies or antigen-binding fragments thereof that specifically react with the substance bind to each other, and these bind specifically with the substance immobilized in the test kit (hereinafter referred to as detection substance). This is a method of detecting a change in color or mass that occurs as a result.
  • the visual sensitivity in the immunochromatography method may decrease.
  • the detection substance for example, solid phase protein
  • An antibody is composed of a base Fc site and a Fab site that reacts with an antigen.
  • the outermost surface of the flow path of the POCT reagent is a random surface composed of a Fab site and an Fc site.
  • the outermost surface of the POCT reagent channel is the Fab site, and the adhesive surface between the channel and the antibody is the Fc site. Therefore, in the POCT reagent in which the orientation of the antibody is controlled, the probability of reaction between the Fab site and the antigen on the outermost surface of the flow path is higher than that in the current non-controlled POCT reagent, so that the visual sensitivity in the immunochromatography method is increased. I think that. That is, it is expected that the visual sensitivity of the POCT reagent is increased by controlling the orientation of the detection substance.
  • Patent Document 1 when immobilizing a micro object such as a protein by light irradiation on a solid phase containing a photoresponsive component, the micro object can be fixed with a fixed orientation with respect to the solid phase. By providing an operable component in the solid phase, the orientation of the micro object is controlled and immobilized on the solid phase.
  • a POCT reagent is produced using the light immobilization method of Patent Document 1, the light stability of the POCT reagent is lowered, and thus practical application is difficult.
  • Patent Document 2 and Patent Document 3 show that protein crystal growth is promoted by placing a concentrated protein aqueous solution in an electric field. However, Patent Document 2 and Patent Document 3 do not describe a liquid sample inspection kit.
  • Patent Document 4 succeeds in adhering microparticles to the surface of ceramic particles by utilizing the charge of a cationic polymer and an anionic polymer as an electrical effect.
  • a specific method is to apply an aqueous solution of a cationic polymer and an anionic polymer to the surface of the ceramic particles, thereby imparting a charge to the surface of the ceramic particles and generating electrostatic charges between the charged fine particles. It is strongly bonded by mechanical interaction.
  • Patent Document 4 does not describe a liquid sample inspection kit.
  • Patent Document 5 is a membrane carrier for a test kit that detects a substance to be detected in a liquid sample, and is provided with at least one flow path capable of transporting the liquid sample, and the liquid sample is provided on the bottom surface of the flow path.
  • a membrane carrier for a liquid sample test kit is described, which is provided with a microstructure that causes a capillary action for transporting the sample.
  • Patent Document 5 does not describe the orientation control of the detection substance.
  • an object of the present invention is to provide a membrane carrier capable of highly sensitive determination.
  • the present invention is as follows.
  • a flow path, a charge imparting substance, and a detection substance are provided, the detection substance is held in the flow path via the charge imparting substance, and the mass ratio of the charge imparting substance to the detection substance (charge imparting substance / detection substance) is A membrane carrier that is 0.01-100.
  • the charge-imparting substance is at least one selected from the group consisting of a cationic polymer and an anionic polymer.
  • the liquid sample inspection kit (hereinafter also simply referred to as “inspection kit”) detects a substance to be detected in the liquid sample.
  • FIG. 1 is a schematic top view of an inspection kit.
  • the test kit 10 includes a membrane carrier 4 and a housing 10 a that houses the membrane carrier 4.
  • the membrane carrier 4 has, on its surface, a dropping zone 4x where a liquid sample is dropped and a detection zone 4y for detecting a substance to be detected in the liquid sample.
  • the dripping zone 4x is exposed at the first opening 10b of the housing 10a.
  • the detection zone 4y is exposed at the second opening 10c of the housing 10a.
  • the dripping zone 4x and the detection zone 4y are located on the upstream side and the downstream side in the transport direction of the liquid sample, respectively.
  • the membrane carrier is a membrane carrier for a liquid sample inspection kit that detects a substance to be detected in the liquid sample.
  • the substance to be detected is not limited at all, and may be any substance capable of antigen-antibody reaction with an antibody such as various pathogens and various clinical markers.
  • viral antigens such as influenza virus, norovirus, adenovirus, RS virus, HAV, HBs, HIV, bacterial antigens such as MRSA, group A streptococci, group B streptococci, legionella, toxins produced by bacteria, etc.
  • hormones such as Mycoplasma, Chlamydia trachomatis, human chorionic gonadotropin, C-reactive protein, myoglobin, cardiac troponin, various tumor markers, agricultural chemicals, and environmental hormones.
  • the substance to be detected may be an antigen capable of inducing an immune reaction alone, or a hapten capable of binding an antibody by an antigen-antibody reaction although it cannot induce an immune reaction alone.
  • the substance to be detected may be in a suspended or dissolved state in the liquid sample.
  • the liquid sample may be, for example, a sample in which the substance to be detected is suspended or dissolved in a buffer solution.
  • FIG. 2 is a schematic top view of the membrane carrier.
  • the membrane carrier 4 includes a flow path 2, a charge imparting substance 1, and a detection substance 3.
  • the detection substance 3 is held in the flow path 2 via the charge imparting substance 1.
  • a region where the detection substance 3 exists constitutes a detection zone.
  • the liquid sample is transported from the dropping zone to the detection zone via the flow path 2.
  • FIG. 3 is a schematic cross-sectional view (cross-sectional view taken along the line II-II in FIG. 2) of the membrane carrier of one embodiment.
  • a membrane carrier 4A shown in FIG. 3 is composed of a flow path 2, a charge-providing substance 1 present on at least a part of the main surface of the flow path 2, and a detection substance 3 present on the charge-provided substance 1. ing.
  • the membrane carrier 4A is obtained, for example, by applying the charge-providing substance 1 to at least a part of the surface of the flow path 2 and further applying the detection substance 3 on the charge-providing substance 1.
  • FIG. 4 is a schematic cross-sectional view of a membrane carrier according to another embodiment.
  • a membrane carrier 4B shown in FIG. 4 includes a flow path 2, a charge-providing substance 1 present in at least a part of the flow path 2, and a detection substance 3 present on the charge-providing substance 1 and on the surface of the flow path 2. It consists of and.
  • the membrane carrier 4B is formed on the surface of the flow path 2 after applying the charge imparting substance 1 to at least a part of the surface of the flow path 2 and infiltrating the charge imparting substance 1 into the flow path 2. It is obtained by further applying the detection substance 3 to the area where the charge imparting substance 1 is applied.
  • FIG. 5 is a schematic cross-sectional view of a membrane carrier of another embodiment.
  • a membrane carrier 4C shown in FIG. 5 includes a flow path 2, a charge-providing substance 1 present in at least a part of the flow path 2, and a detection substance present on the charge-providing substance 1 and in and on the surface of the flow path 2. 3. That is, the detection substance 3 exists inside the flow path 2 and is also exposed on the surface of the flow path 2.
  • the membrane carrier 4C is formed on the surface of the flow channel 2 after applying the charge imparting material 1 to at least a part of the surface of the flow channel 2 and infiltrating the charge imparting material 1 into the flow channel 2. It is obtained by further applying the detection substance 3 to the area where the charge imparting substance 1 is applied and infiltrating the detection substance 3 into the flow path 2.
  • the channel 2 functions as a channel for transporting the liquid sample in the membrane carrier 4.
  • the surface of the flow path 2 may be flat or curved.
  • a fine structure (uneven structure) may be formed on the surface of the flow path 2.
  • the fine structure is provided on the surface of the flow path 2
  • the liquid sample is transported through the fine structure by the capillary action of the fine structure.
  • the flow path 2 may be comprised with the porous base material.
  • the liquid sample is composed of a porous substrate, the liquid sample is transported by moving through the pores of the porous substrate by capillary action.
  • the overall shape of the flow path is not particularly limited, but may be, for example, a polygon such as a quadrangle, a circle, or an ellipse.
  • the vertical width (length in the short direction) of the flow path may be, for example, 2 mm or more and 100 mm or less
  • the horizontal width (length in the longitudinal direction) of the flow path is, for example, 2 mm. It may be 100 mm or less.
  • the thickness of the flow path 2 is preferably 100 ⁇ m to 5 mm, more preferably 500 ⁇ m to 2 mm. When the uneven structure is provided on the flow path 2, the thickness of the flow path 2 does not include the height of the convex portion.
  • the material forming the flow path 2 is preferably a resin, and more preferably a thermoplastic resin.
  • the thermoplastic resin include nitrocellulose, polyester resin, polyolefin resin, polystyrene resin, polycarbonate resin, fluorine resin, and acrylic resin.
  • PET polyethylene terephthalate
  • COP cycloolefin polymer
  • PP polypropylene
  • PS polystyrene
  • MS methyl methacrylate-styrene copolymer
  • MBS methyl methacrylate-butadiene-styrene copolymer
  • MABS Methyl methacrylate-acrylonitrile-butadiene-styrene
  • SBC polycarbonate
  • PC polyvinylidene fluoride
  • PVDF polymethyl methacrylate
  • the material of the channel 2 is preferably nitrocellulose.
  • a nitrocellulose membrane can be used as the flow path 2.
  • the material of the flow path 2 is preferably a thermoplastic resin.
  • the fine structure has a concavo-convex structure and has a plurality of convex portions.
  • the flow path 2 has a concavo-convex structure (fine structure) on at least a part of the surface from the viewpoint of facilitating highly sensitive inspection. That is, it is preferable that the membrane carrier 4 includes the flow path 2 provided with an uneven structure on at least a part of the surface.
  • the concavo-convex structure may be located at least between the dropping zone and the detection zone. An uneven structure may be provided over the entire surface of the flow path 2.
  • the fine structure can be formed by imprinting.
  • thermal imprinting is a method for producing a substrate having a fine structure by pressing a mold (mold) having a fine structure against the substrate and transferring the fine structure to a substrate softened by heating. This thermal imprint enables nano-order microfabrication.
  • the mold (mold) having a fine structure has an uneven structure.
  • a structure in which the recesses 5 are regularly arranged in a triangular arrangement on the surface of the mold 7 is preferable.
  • a structure in which the convex portions 6 are regularly arranged in each direction as protrusions on the surface of the flow path 2 is preferable.
  • the structure in which the convex portions 6 are regularly arranged as protrusions is preferably a structure in which the protrusions 6 are arranged in a planar lattice pattern.
  • the planar lattice is preferably one or more of an orthorhombic lattice, a hexagonal lattice, a tetragonal lattice, a rectangular lattice, and a parallel lattice, and more preferably a hexagonal lattice. Of the hexagonal lattice, an equilateral triangular lattice is preferable.
  • FIG. 8 shows a structure in which the convex portions 6 are arranged in an equilateral triangular lattice shape.
  • the convex part 6 is preferably made of the same material as the flow path 2.
  • the cross section of the convex portion 6 is preferably a triangle, and more preferably a regular triangle.
  • a cone shape or a pyramid shape is preferable, and a cone shape is more preferable.
  • the pitch 11 of the convex portions 6 is preferably 1 to 300 ⁇ m, more preferably 5 to 100 ⁇ m, and most preferably 10 to 50 ⁇ m.
  • the pitch 11 of the convex portions 6 is 1 ⁇ m or more, the light reflectivity is more excellent.
  • the pitch 11 of the convex portions 6 is 300 ⁇ m or less, the transparency is more excellent.
  • the pitch 11 of the convex portions 6 refers to the dimension between the convex portions 6 and 6 adjacent to each other (the distance between the centers of the adjacent convex portions 6).
  • the pitch 11 means the dimension between the vertex of a cone and the vertex of a cone as shown in FIG.
  • the height of the convex portion 6 is preferably 1 to 1000 ⁇ m, and more preferably 10 to 50 ⁇ m. If it is 1 ⁇ m or more, the flow path volume becomes large, and it does not take a long time to develop the liquid sample. When the thickness is 1000 ⁇ m or less, it does not take much time and cost to produce the fine structure, and the production of the fine structure becomes easy.
  • the height of the convex portion 6 is defined as the maximum length of the convex portion 6 in the direction orthogonal to the bottom surface of the convex portion 6. That is, the height of the convex portion 6 is a dimension from the bottom surface of the convex portion 6 to the apex of the convex portion 6.
  • the diameter of the bottom surface of the convex portion 6 is preferably 1 to 1000 ⁇ m, and more preferably 10 to 50 ⁇ m.
  • the diameter of the bottom surface of the convex portion 6 is 1 ⁇ m or more, the microfabrication cost of the mold is reduced, and it is easy to uniformly produce an infinite number of fine structures on the surface of the flow path 2 of the membrane carrier 4 having a large area. Capillary force necessary to move is increased.
  • the diameter of the bottom surface of the convex portion 6 is 1000 ⁇ m or less, the volume of the metal scraped from the metal member during the production of the mold is reduced, the production cost of the mold and the membrane carrier 4 is reduced, and the area of the flow path in the membrane carrier 4 , The membrane carrier 4 is downsized, and the membrane carrier 4 itself can be easily transported.
  • the diameter of the bottom surface of the convex portion 6 is defined as the representative length at the bottom surface of the convex portion 6.
  • the representative length at the bottom is the diameter when the shape of the bottom is a circle, the length of the shortest side when it is a triangle or a quadrangle, the length of the longest diagonal line when it is a pentagon or more polygon, and other shapes In this case, the maximum length at the bottom is used.
  • the convex part 6 is a cone (refer FIG. 8) or a cylinder
  • the diameter of the bottom face of the convex part 6 is a diameter of the bottom face (circle) of the convex part 6.
  • a polymer is preferable.
  • the polymer is preferably at least one selected from the group consisting of cationic polymers and anionic polymers.
  • the cationic polymer is a polymer compound having a positive charge in an aqueous solution
  • the anionic polymer is a polymer compound having a negative charge in an aqueous solution.
  • the weight average molecular weight of the polymer is preferably 10,000 to 500,000, more preferably 50,000 to 200,000.
  • the weight average molecular weight is a value in terms of standard polyethylene oxide measured by a gel permeation chromatography (GPC) method.
  • GPC gel permeation chromatography
  • the weight average molecular weight can be obtained by preparing a calibration curve with commercially available standard polyethylene oxide using GPC system (SC-8010 manufactured by Tosoh Corporation) using water as a solvent.
  • cationic polymer for example, poly (diallyldimethylammonium chloride) (PDDA) can be preferably used.
  • PDDA poly (diallyldimethylammonium chloride)
  • Other cationic polymers that can be used in the present embodiment include polyethyleneimine (PEI), polyvinylamine (PVAm), poly (vinylpyrrolidone / N, N-dimethylaminoethylacrylic acid) copolymer, and the like. It is done. However, it is an example as a cationic polymer, and is not limited thereto.
  • anionic polymer for example, polystyrene sulfonic acid (PSS) can be preferably used.
  • Other anionic polymers that can be used in the present embodiment include polyvinyl sulfate (PVS), polyacrylic acid (PAA), polymethacrylic acid (PMA), and the like.
  • PVS polyvinyl sulfate
  • PAA polyacrylic acid
  • PMA polymethacrylic acid
  • this is an example of an anionic polymer and is not limited thereto.
  • the charge imparting substance may exist in at least a part of the detection zone, may exist throughout the detection zone, or may exist outside the detection zone.
  • the detection substance may be, for example, a solid phase protein.
  • the solid phase protein include antibodies.
  • the antibody is an antibody that reacts with a substance to be detected by antigen-antibody reaction, and may be a polyclonal antibody or a monoclonal antibody.
  • the detection substance only needs to be present on the charge imparting substance, may be present in a part of the region where the charge imparting substance is present, or may be present throughout.
  • the mass ratio of the charge imparting substance to the detection substance is preferably 0.01 to 100, more preferably 0.1 to 10.
  • the charge-providing substance / detecting substance may be 0.01 or more, 0.1 or more, 0.3 or more, 0.5 or 0.8 or more, and is 100 or less, 10 or less, 8 or less, 5 or less, or 3 or less. It may be.
  • the detection substance 3b can be strongly fixed.
  • the charge imparting substance / detection substance is 100 or less, non-specific interaction with the substance to be detected does not occur.
  • the mass ratio of the charge imparting substance to the detection substance is defined as the mass ratio in the region where both the charge imparting substance and the detection substance exist.
  • the mechanism of the orientation control of the detection substance will be described with reference to FIG. 6, taking a solid phase protein as an example.
  • the solid phase protein constituting the detection substance 3 has a solid phase protein base portion 3a and a solid phase protein end portion 3b.
  • the charge-imparting substance 1 exhibits an electrostatic interaction with the surface charge localized in the solid phase protein base 3a.
  • the solid phase protein base 3a faces the direction of the charge imparting substance 1 (channel 2) and the solid phase protein end 3b is located on the outermost surface, the orientation of the detection substance (solid phase protein) 3 can be controlled. It becomes.
  • the detection substance may be oriented by an electric field generated by electrostatic interaction between the detection substance and the charge imparting substance. In one embodiment, it can also be said that the detection substance is oriented by the potential of the surface of the substrate (flow path) holding the detection substance.
  • the detection zone on the flow path shows a color change when a substance to be detected is detected.
  • the color change in the detection zone occurs when the detection target substance is held in the detection zone by the detection substance (reacts with the detection substance).
  • the color change may be a color change that can be confirmed by an optical method.
  • the optical method there are mainly two methods: a visual determination and a method of measuring fluorescence intensity.
  • a visual determination the color difference between two color stimuli (described in JIS Z8781-4: 2013) when the color before detection and after detection is measured in the color system of CIE1976L * a * b * color space.
  • ⁇ E a color change occurs such that ⁇ E) is 0.5 or more.
  • this color difference is 0.5 or more, it becomes easy to visually confirm the color difference.
  • the ratio (Fl1 / Fl2) 10 of the fluorescence intensity (Fl1) in the detection zone and the fluorescence intensity (Fl2) in the upstream region and downstream region adjacent to the detection zone. It is preferable that a color change that is 1 or more occurs. When this ratio is 10/1 or more, separation of signal and noise becomes easy.
  • the film carrier has a step of applying a charge-imparting substance on at least a part of the surface of the flow path (first application step), and a step of applying a detection substance on the applied charge-imparting material (second application step). It can manufacture by the method provided with these.
  • the charge imparting substance is applied to at least a part of the surface of the flow path.
  • the charge imparting substance may be applied to at least a part of the detection zone, the charge imparting substance may be applied over the entire detection zone, or may be applied outside the detection zone.
  • the charge imparting substance may be implemented by applying an aqueous solution containing the charge imparting substance on the charge imparting substance.
  • a step (first drying step) of volatilizing the solvent in the aqueous solution containing the charge imparting substance is provided between the first coating step and the second coating step.
  • the first drying step can be performed using a vacuum dryer or the like.
  • the concentration of the charge-providing substance in the aqueous solution containing the charge-providing substance is preferably 0.01 to 20% by mass, and more preferably 0.1 to 5% by mass.
  • the detection substance is applied on the applied charge imparting substance.
  • the detection substance may be applied to a part of the charge-providing substance, or may be applied over the entire region where the charge-providing substance exists, or may be applied so as to protrude from the area where the charge-providing substance exists.
  • the detection substance can be held on the charge imparting substance by applying a solution containing the detection substance (detection substance solution) on the charge imparting substance.
  • a step of volatilizing the solvent in the detection substance solution is provided after the second application step.
  • the detection substance solution is preferably an aqueous solution in which the detection substance is dissolved in purified water.
  • other compounds such as sodium chloride and trishydroxymethylaminomethane may be added to the detection substance solution.
  • the concentration of the detection substance in the detection substance solution is preferably 0.00001 to 1 w / v%, more preferably 0.0001 to 0.1 w / v%.
  • the appropriate charge imparting substance 1 can vary depending on the pH of the detection substance solution.
  • the charge imparting substance 1 to be used is preferably a cationic polymer in terms of enhancing electrostatic interaction.
  • the charge imparting substance 1 to be used is preferably an anionic polymer in terms of enhancing electrostatic interaction.
  • the pH of the detection substance solution is 7.0, either a cationic polymer or an anionic polymer can be used as the charge imparting substance 1.
  • a step of forming an uneven structure in the flow path by imprinting may be further provided.
  • the ratio of the application amount of the charge-imparting substance to the application amount of the detection substance is preferably 0.01 to 100, more preferably 0.1 to 10.
  • the charge-providing substance / detecting substance may be 0.01 or more, 0.1 or more, 0.3 or more, 0.5 or 0.8 or more, and is 100 or less, 10 or less, 8 or less, 5 or less, or 3 or less. It may be.
  • the charge application amount / detection material application amount is a value calculated based on the mass (solid content) of each substance.
  • the membrane carrier 4 of the present embodiment has a limit magnification that can be visually determined compared to a membrane carrier in which no charge-providing substance is present and a membrane carrier in which the mass ratio of the charge-providing substance to the detection substance is outside the range of 0.01 to 100. Since it is large, it can be evaluated as a membrane carrier capable of highly sensitive determination.
  • a charge imparting substance such as a cationic polymer or an anionic polymer is applied to the surface of the base material (flow path 2), so that the base material surface is charged and charged. Due to the electrostatic interaction that occurs between the detection substance (for example, solid phase protein), the charge-providing substance strongly adheres the detection substance.
  • the orientation of the detection substance can be controlled by using the localized charge existing in the detection substance. In the present embodiment, the orientation of the detection substance can be controlled using the electric field generated by the charge imparting substance.
  • PDDA diallyldimethylammonium chloride
  • Anti-influenza A virus NP antibody suspension in which anti-influenza A virus NP antibody is suspended in water as a solid phase protein in the area where PDDA is applied on the nitrocellulose membrane (the concentration of anti-influenza A virus NP antibody is 0 .0033 w / v%) was applied so as to have the same area as that of PDDA, and dried well under warm air.
  • the solid phase protein coating amount is a mass (solid content) obtained by subtracting the mass of water from the mass of the anti-influenza A virus NP antibody suspension.
  • Example 1 the mass ratio of the charge-imparting substance to the solid phase protein (charge-giving substance coating amount / solid-phase protein coating amount) (solid content conversion ratio) was 1.
  • a purified anti-influenza A virus NP antibody (an antibody different from the above) was used.
  • the purified anti-influenza A virus NP antibody is labeled with 0.2 ⁇ m red latex particles (SC-042-R manufactured by JSR Life Sciences) with a covalent bond, and added to a Tris buffer containing sugar, surfactant and protein.
  • An anti-A-type labeling body was prepared by suspending it so that the concentration of latex particles was 0.025 w / v%, and performing sonication to sufficiently disperse and float.
  • a specimen suspension attached to Quick Navi-Flu manufactured by Denka Seken Co., Ltd. was used as a diluted solution.
  • the dilution factor of influenza A virus A / Beijing / 32/92 (H3N2) is increased from 2 ⁇ 10 4
  • the dilution factor (visual determination) that makes it impossible to visually check the presence of a colored line 10 minutes after the start of the test Possible limit magnification).
  • Table 1 shows the results when the flow path is a membrane.
  • Example 2 The experiment was performed under the same conditions as in Example 1 except that the mass ratio of the charge-imparting substance to the solid-phase protein in Example 1 (charge-imparting substance application amount / solid-phase protein application amount) was 100.
  • Example 3 The experiment was performed under the same conditions as in Example 1 except that the mass ratio of the charge-imparting substance to the solid-phase protein in Example 1 (charged substance application amount / solid-phase protein application amount) was 0.01.
  • PSS polystyrene sulfonic acid
  • Example 5 The same conditions as in Example 1 except that the charge-providing substance in Example 1 is PSS and the mass ratio of the charge-providing substance to the solid-phase protein is 100 (charge-providing substance coating amount / solid-phase protein coating amount). The experiment was conducted.
  • Example 6 The charge imparting substance in Example 1 is PSS, and the mass ratio of the charge imparting substance to the solid phase protein (charge imparting substance coating amount / solid phase protein coating amount) is 0.01. The experiment was conducted under conditions.
  • Example 1 The same as Example 1 except that the mass ratio of the charge-imparting substance to the solid phase protein (charge-giving substance application amount / solid-phase protein application amount) was set to 0 without applying the charge-giving substance in Example 1. The experiment was conducted under the following conditions.
  • Example 2 The experiment was performed under the same conditions as in Example 1 except that the charge-providing substance in Example 1 was set to a mass ratio of the charge-providing substance to the solid phase protein (charged substance coating amount / solid phase protein coating amount) of 1000. It was.
  • Example 3 The charge imparting substance in Example 1 is PSS, and the mass ratio of the charge imparting substance with respect to the solid phase protein (charge imparting substance coating amount / solid phase protein coating amount) is 0.001. The experiment was conducted under conditions.
  • Example 7 ⁇ Mold preparation> The mold was produced by laser processing and mechanical cutting. This mold is made of aluminum alloy A5052. A conical recess having a diameter of 25 ⁇ m, a pitch of 30 ⁇ m, and a depth of 30 ⁇ m is machined in the center of the mold in a triangular array form as shown in FIG. 7 in a range of 3 cm ⁇ 3 cm. A release treatment was performed on the uneven surface of the mold in order to easily and reliably peel off the mold and the thermoplastic resin when transferred. The mold release treatment was performed by immersing in OPTOOL HD-2100TH manufactured by Daikin Industries, Ltd. for about 1 minute, drying, and allowing to stand overnight.
  • OPTOOL HD-2100TH manufactured by Daikin Industries, Ltd.
  • thermoplastic resin polystyrene (PS, Denka styrene sheet manufactured by Denka Co., Ltd., film thickness: 300 ⁇ m) was used.
  • Thermal imprinting was used as a processing method, and an X-300 manufactured by SCIVAX was used as the apparatus.
  • the molding temperature was 120 ° C.
  • the applied pressure was 5.5 MPa
  • transfer was performed for 10 minutes.
  • the thermoplastic plastic and the mold were cooled to 80 ° C. while applying pressure, and then the pressure was removed to produce a flow path having a conical convex portion on the surface.
  • conical convex portions 6 having a pitch of 30 ⁇ m, a diameter of 25 ⁇ m, and a height of 30 ⁇ m are processed into a 3 cm ⁇ 3 cm range in a triangular arrangement form as shown in FIG.
  • Example 2 The subsequent operation was conducted under the same conditions as in Example 1 except that the flow path in Example 1 was made of polystyrene.
  • Table 2 shows the results when the flow path is made of polystyrene.
  • Example 8 The experiment was performed under the same conditions as in Example 7, except that the mass ratio of the charge-imparting substance to the solid-phase protein in Example 7 (charge-imparting substance coating amount / solid-phase protein coating amount) was 100.
  • Example 9 The experiment was performed under the same conditions as in Example 7 except that the mass ratio of the charge-imparting substance to the solid-phase protein in Example 7 (charged substance application amount / solid-phase protein application amount) was 0.01.
  • Example 10 The experiment was performed under the same conditions as in Example 7 except that PSS was used as the charge-providing substance in Example 7.
  • Example 11 The same conditions as in Example 7 except that the charge-providing substance in Example 7 was PSS and the mass ratio of the charge-providing substance to the solid phase protein (charge-providing substance coating amount / solid-phase protein coating amount) was 100. The experiment was conducted.
  • Example 7 is the same as Example 7 except that the charge-providing substance in Example 7 is PSS, and the mass ratio of the charge-providing substance to the solid-phase protein (charge-providing substance coating amount / solid-phase protein coating amount) is 0.01. The experiment was conducted under conditions.
  • Example 7 is the same as Example 7 except that the mass ratio of the charge-providing substance to the solid phase protein (charge-applying substance application amount / solid-phase protein application quantity) was set to 0 without applying the charge-providing substance in Example 7. The experiment was conducted under the following conditions.
  • Example 5 The experiment was performed under the same conditions as in Example 7, except that the mass ratio of the charge-imparting substance to the solid-phase protein in Example 7 (charge-imparting substance coating amount / solid-phase protein coating amount) was 0.0001.
  • Example 6 Under the same conditions as in Example 7, except that the charge-providing substance in Example 7 was PSS and the mass ratio of the charge-providing substance to the solid phase protein (charged substance coating amount / solid phase protein coating amount) was 10,000. The experiment was conducted.
  • a highly sensitive liquid sample inspection kit in which the orientation of the detection substance is controlled can be obtained.
  • the liquid sample test kit of this embodiment has a test score 1.5 times higher than that of a liquid sample test kit that does not use this embodiment, and can achieve high sensitivity.
  • SYMBOLS 1 Charge provision substance 2 Flow path 3 Detection substance 3a Solid phase protein base 3b Solid phase protein edge part 4,4A, 4B, 4C Membrane carrier 4x Dropping zone 4y Detection zone 5 Recess 6 Protrusion 7 Mold (mold) 10 Liquid Sample Inspection Kit 10a Case 10b First Opening 10c Second Opening 11 Pitch

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Abstract

La présente invention concerne un support de membrane ayant un trajet d'écoulement, une substance conférant une charge et une substance de détection. La substance de détection est retenue dans le trajet d'écoulement par l'intermédiaire de la substance conférant une charge. Le rapport en masse de la substance conférant une charge à la substance de détection (substance conférant une charge/substance de détection) est de 0,01 à 100.
PCT/JP2018/016815 2017-04-25 2018-04-25 Support de membrane, son procédé de production et kit de test d'échantillon liquide WO2018199168A1 (fr)

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KR20210142702A (ko) * 2019-04-24 2021-11-25 덴카 주식회사 막 담체 및 검사 키트
CN115485547A (zh) * 2020-04-28 2022-12-16 电化株式会社 检测装置和检测方法

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JPH06160394A (ja) * 1992-11-25 1994-06-07 Iatron Lab Inc 不溶性担体及びその担体を用いる免疫学的検出方法
JP2003528321A (ja) * 2000-03-20 2003-09-24 マサチューセッツ インスティテュート オブ テクノロジー 無機粒子結合体
JP2004264052A (ja) * 2003-02-10 2004-09-24 Japan Science & Technology Agency ターゲット認識素子及びターゲット認識素子を利用したバイオセンサ
JP2004346209A (ja) * 2003-05-23 2004-12-09 National Institute Of Advanced Industrial & Technology イオン性ポリマーおよびポリマー含有基板
JP2007051998A (ja) * 2005-07-22 2007-03-01 Toyota Central Res & Dev Lab Inc 光固定化方法及び構造体
JP2007171054A (ja) * 2005-12-22 2007-07-05 National Institute Of Advanced Industrial & Technology 免疫センサ用デバイス
JP2012505418A (ja) * 2008-10-17 2012-03-01 紅電醫學科技股▲分▼有限公司 液体試験ストリップ及び方法
WO2016098740A1 (fr) * 2014-12-15 2016-06-23 デンカ株式会社 Support de membrane pour kit de test d'échantillon liquide, kit de test d'échantillon liquide et procédé permettant de produire un kit de test d'échantillon liquide

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Publication number Priority date Publication date Assignee Title
JPH06160394A (ja) * 1992-11-25 1994-06-07 Iatron Lab Inc 不溶性担体及びその担体を用いる免疫学的検出方法
JP2003528321A (ja) * 2000-03-20 2003-09-24 マサチューセッツ インスティテュート オブ テクノロジー 無機粒子結合体
JP2004264052A (ja) * 2003-02-10 2004-09-24 Japan Science & Technology Agency ターゲット認識素子及びターゲット認識素子を利用したバイオセンサ
JP2004346209A (ja) * 2003-05-23 2004-12-09 National Institute Of Advanced Industrial & Technology イオン性ポリマーおよびポリマー含有基板
JP2007051998A (ja) * 2005-07-22 2007-03-01 Toyota Central Res & Dev Lab Inc 光固定化方法及び構造体
JP2007171054A (ja) * 2005-12-22 2007-07-05 National Institute Of Advanced Industrial & Technology 免疫センサ用デバイス
JP2012505418A (ja) * 2008-10-17 2012-03-01 紅電醫學科技股▲分▼有限公司 液体試験ストリップ及び方法
WO2016098740A1 (fr) * 2014-12-15 2016-06-23 デンカ株式会社 Support de membrane pour kit de test d'échantillon liquide, kit de test d'échantillon liquide et procédé permettant de produire un kit de test d'échantillon liquide

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210142702A (ko) * 2019-04-24 2021-11-25 덴카 주식회사 막 담체 및 검사 키트
EP3961220A4 (fr) * 2019-04-24 2022-06-15 Denka Company Limited Support de film et kit de test
KR102553911B1 (ko) 2019-04-24 2023-07-10 덴카 주식회사 막 담체 및 검사 키트
CN115485547A (zh) * 2020-04-28 2022-12-16 电化株式会社 检测装置和检测方法

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