JPWO2016098740A1 - Membrane carrier for liquid sample inspection kit, liquid sample inspection kit, and method for producing liquid sample inspection kit - Google Patents

Abstract

The membrane carrier (3) is used in a test kit (18) for detecting a substance to be detected in a liquid sample. The membrane carrier (3) is provided with at least one flow channel capable of transporting a liquid sample, and a fine structure (14) for causing a capillary action for transporting the liquid sample is provided on the bottom surface of the flow channel. . The membrane carrier (3) can be produced at low cost by thermal imprinting, and the liquid sample can be moved by capillary force. In the detection zone (3y) of the inspection kit (18), the color change at the time of detection of the substance to be detected can be confirmed with the naked eye.

Description

  The present invention relates to a membrane carrier for an inspection kit for detecting a substance to be detected in a liquid sample, a liquid sample inspection kit using the same, and a method for producing the liquid sample inspection kit.

  In recent years, the Point of Care Test (POCT) reagent, which uses an antigen-antibody reaction or the like to test for morbidity or pregnancy of an infectious disease or to measure a blood glucose level or the like, has attracted attention. In the inspection / measurement using the POCT reagent, it is possible to discriminate the result in a short time. The method of using the POCT reagent is simple and the POCT reagent is inexpensive. Since the POCT reagent has these characteristics, it is often used for examinations at a stage where symptoms are mild and periodic examinations. The POCT reagent is also an important diagnostic tool in home medical care, which is expected to increase in the future.

  In a test or diagnosis using a test kit that is a kind of POCT reagent, a liquid sample such as blood is introduced into the test kit, and a specific target substance contained in the liquid sample is detected. As a method for detecting a specific substance to be detected from a liquid sample, an immunochromatography method is often used. In the immunochromatography method, a liquid sample is dropped on a membrane carrier provided in a test kit, and a substance to be detected in the liquid sample is bound to a labeling substance in the process of moving the liquid sample on the membrane carrier. Furthermore, the substance to be detected binds specifically and selectively to a substance (hereinafter referred to as a detection substance) immobilized in the test kit. As a result, changes in color and weight generated in the inspection kit are detected. The detection substance may be rephrased as a reagent.

  As a membrane carrier for moving a liquid sample, a nitrocellulose membrane is often used (see Patent Document 1 below). The nitrocellulose membrane has many fine holes with a diameter of about several μm, and the liquid sample moves through the holes by capillary force.

  However, since the nitrocellulose membrane is derived from a natural product and the pore diameter in the membrane and how the pores are connected are not uniform, the flow rate of the liquid sample in the membrane varies depending on the membrane. When a difference occurs in the flow rate, the time required for detecting the detection target substance also changes. As a result, an erroneous determination may be made that the detected substance is not detected before the detected substance is combined with the labeling substance or reagent.

  In order to solve the above problems, a technique for artificially producing a fine flow path of a liquid sample has been devised (see Patent Documents 2 and 3 below). By using this method, a film carrier having a uniform structure can be produced. As a result, it is possible to reduce the possibility that an erroneous determination is made that the detection target substance is not detected before the detection target substance binds to the labeling substance or the reagent.

JP 2014-0662820 A Japanese Patent No. 4597664 Special table 2012-524894 gazette

  Thermal imprinting is an example of a technique for artificially creating a flow path. Thermal imprinting is a technique in which a microstructure (mold) is pressed against a substrate (thermoplastic before processing), and the microstructure is transferred to the surface of the substrate softened by heating. This is a method of producing a film carrier having This fine structure functions as a flow path for the liquid sample. If thermal imprinting is used, it is possible to produce a fine structure in a wide order range of several tens nm to several hundreds μm. Furthermore, since thermal imprinting does not require a large-scale device such as a vacuum apparatus, a membrane carrier having a uniform structure can be mass-produced inexpensively and easily. In addition, the fine structure which a metal mold | die has is a countless recessed part formed in the surface of the metal mold | die pressed against a base material. The fine structure of the film carrier is an infinite number of protrusions (protrusions) protruding on the surface of the film carrier, and has a shape corresponding to the recess formed on the surface of the mold. That is, the portion of the base material filled in the concave portion of the mold becomes the fine structure (convex portion) of the film carrier.

  However, with the thermal imprint described above, it is difficult to produce a structure in which the aspect ratio of the microstructure is higher than 1: 2. That is, it is difficult to produce a film carrier having a fine structure in which the ratio Lv / Lh between the horizontal size Lh and the vertical size Lv is higher than 2/1 by thermal imprinting. The higher the aspect ratio of the microstructure, the easier it is for a part of the membrane carrier to remain on the mold side when the membrane carrier is peeled from the mold, or the microstructure formed on the surface of the membrane carrier collapses or deforms more easily. To do. Therefore, the higher the aspect ratio of the microstructure, the lower the productivity of the membrane carrier.

  Moreover, when producing a uniform fine structure by thermal imprinting, it is necessary that the fine processing of the mold is performed uniformly with high accuracy. Examples of the technique for performing such fine processing include etching, photolithography, mechanical cutting, and laser processing. However, both methods require considerable processing costs. In any of the methods, the processing cost increases as the volume of metal scraped from the flat surface of the metal member when a mold is manufactured from the metal member is increased. Therefore, the mold can be manufactured at low cost by reducing the volume cut out from the metal member as much as possible. As a result, a film carrier having a fine structure can be manufactured at low cost by thermal imprinting. In addition, the volume of the metal cut out from the flat surface of the metal member is paraphrased as the volume of the recess formed on the surface of the completed mold. In addition, the volume of the recess formed on the surface of the mold may be rephrased as the volume of each protrusion (microstructure) formed on the surface of the film carrier by thermal imprinting.

  By the way, the larger the flow rate of the liquid sample in the membrane carrier, the easier it is to detect the substance to be detected in the liquid sample. Therefore, it is advantageous for the detection of the substance to be detected to use a membrane carrier that increases the flow rate of the liquid sample. In order to increase the flow rate of the liquid sample in the membrane carrier, it is necessary that the gap through which the liquid sample can flow is large in the membrane carrier. Therefore, a fine structure having a large porosity is required. When the total volume of voids in the fine structure of the film carrier is expressed as Vv, and the total volume of the fine structure itself (convex part) of the film carrier is expressed as Vf, the void ratio Rv of the fine structure is 100 -It is expressed as Vv / (Vv + Vf). Vv may be rephrased as the total volume of the space located between the convex portions.

  In summary, when a membrane carrier for a liquid sample test kit for use in a POCT reagent is prepared, the aspect ratio is 1: 2 (that is, 2) as a flow path that causes the flow of the liquid sample by capillary action. / 1) It is required to form a fine structure that is smaller than that and has a higher porosity. In the production of a mold used for forming a fine structure, it is required to reduce the volume of metal cut out from a metal member as much as possible.

  As a method for detecting a detected substance, a color change of the detection zone caused by binding of a detected substance bound to a labeling substance such as colored latex particles, fluorescent particles, or metal colloid particles with a reagent fixed in the detection zone Is well known to detect this with an optical measuring instrument such as an absorbance meter.

  However, in the above method, since it is necessary to prepare an optical measuring instrument for performing the determination, the method of using the POCT reagent is complicated. Furthermore, using an optical measuring instrument is a factor that increases the manufacturing cost of the POCT reagent.

  Therefore, in order to take advantage of the characteristics of the POCT reagent (test kit) that the results can be distinguished in a short time, the method of use is simple, and the cost is low, the color change when detecting the substance to be detected is visually observed. It must be large enough to be confirmed by

  In summary, when producing a liquid sample test kit for use in a POCT reagent, it is necessary to increase the color change at the time of detection so that it can be visually confirmed. Furthermore, it is required to form a fine structure having an aspect ratio smaller than 1: 2 (that is, 2/1) and a larger porosity as a flow path for generating a flow of a liquid sample by capillary action. In the production of a mold used for forming a fine structure, it is required to reduce the volume of metal cut out from a metal member as much as possible.

  The present invention has been made in view of the above circumstances, and can be manufactured at low cost by thermal imprinting, has a flow path capable of moving a liquid sample by capillary force, and a substance to be detected in the liquid sample It is an object of the present invention to provide a film carrier for a liquid sample inspection kit capable of visually confirming a color change at the time of detection, a liquid sample inspection kit using the same, and a method for producing the liquid sample inspection kit.

That is, the present invention is as follows.
(1) A membrane carrier for a test kit for detecting a substance to be detected in a liquid sample,
At least one flow path capable of transporting the liquid sample is provided;
A membrane carrier for a liquid sample test kit, wherein a fine structure that causes a capillary action for transporting a liquid sample is provided on a bottom surface of the flow path.
The fine structure is a plurality of convex portions (projections) that cause a capillary action, or a total of the plurality of convex portions. Therefore, the present invention may be rephrased as follows.
The membrane carrier for a liquid sample test kit according to one aspect of the present invention includes a flat portion corresponding to the bottom surface of the flow path and a plurality of convex portions (projections) protruding from the flat portion. That is, the surface of the film carrier includes a flat portion and a plurality of convex portions (projections) protruding from the flat portion. Due to the capillary action caused by the fine structure, the voids (spaces between the plurality of convex portions) in the fine structure function as channels for transporting the liquid sample along the surface of the membrane carrier.

(2) The membrane carrier for a liquid sample inspection kit according to (1), comprising a thermoplastic having a glass transition point (Tg) of 80 to 180 ° C.

(3) The membrane carrier for a liquid sample inspection kit according to (1) or (2), which is made of a thermoplastic having a melting point (Tm) of 80 to 180 ° C.

(4) The storage elastic modulus of the thermoplastic plastic is 1.0 × 10 ⁷ Pa or less at a temperature 20 ° C. higher than the glass transition point or the melting point, described in (2) or (3) Membrane carrier for liquid sample inspection kit.
The term “temperature 20 ° C. higher than the glass transition point or melting point” may be rephrased as a temperature 20 ° C. higher than the glass transition point or a temperature 20 ° C. higher than the melting point.

(5) The membrane carrier for a liquid sample inspection kit according to any one of (1) to (4), wherein the shape of the microstructure is a cone.
That is, the fine structure (convex portion) may be a cone.

(6) The membrane carrier for a liquid sample inspection kit according to any one of (1) to (5), wherein the diameter of the bottom surface of the microstructure is 10 to 1000 μm.
For example, when the fine structure is a cone, the diameter of the bottom surface of the fine structure may be rephrased as the diameter of the bottom surface of the cone.

(7) The film carrier for a liquid sample inspection kit according to any one of (1) to (6), wherein the fine structure has a height of 10 to 500 μm.
The height of the fine structure may be rephrased as the height of the convex portion (projection) from the flat portion of the film carrier.

(8) The film carrier for a liquid sample inspection kit according to any one of (1) to (7), wherein the aspect ratio of the fine structure is 10: 1 to 1: 2.
When the length (thickness) in the horizontal direction (short direction) of the fine structure is represented by Lh, and the length in the vertical direction (longitudinal direction) of the fine structure is represented by Lv, the aspect ratio is Lv / Lh. expressed. That is, the aspect ratio Lv / Lh of the fine structure may be 1/10 or more and 2/1 or less. For example, if the microstructure is a cone, Lh may be the diameter of the bottom surface of the cone and Lv may be the height of the cone from the flat part of the membrane carrier.

(9) The ratio between the diameter of the bottom surface of the microstructure and the distance between the closest centers of the microstructures is greater than 1 and 5 or less, according to any one of (1) to (8). Membrane carrier for liquid sample inspection kit.
The “distance between nearest centers of fine structures” may be rephrased as the distance between the centers of a pair of adjacent fine structures (convex portions). In other words, “the distance between the centers of the nearest neighbors of the microstructures” may be rephrased as the pitch of the microstructures. The diameter of the bottom surface of the fine structure may be paraphrased as, for example, the diameter of the bottom surface of the convex portion. For example, when the fine structure is a cone, the “distance between nearest centers of fine structures” may be rephrased as the distance between the apexes of adjacent cones.

(10) Using the membrane carrier for a liquid sample test kit described in any one of (1) to (9),
A detection zone for detecting a substance to be detected in a liquid sample in the membrane carrier;
A liquid sample inspection kit in which a color change that can be visually confirmed is detected when a substance to be detected is detected in the detection zone.
In other words, a liquid sample inspection kit according to one aspect of the present invention includes the membrane carrier according to the present invention, and a dropping zone where the liquid sample is dropped on the surface of the membrane carrier, and a substance to be detected in the liquid sample There is a detection zone for detecting the above and the fine structure located at least between the dropping zone and the detection zone. Due to the capillary action of the microstructure, the liquid sample is transported from the dropping zone to the detection zone. When the substance to be detected in the liquid sample is detected in the detection zone, the color of the detection zone changes.

(11) The detection substance that causes a color change that can be visually confirmed when the detection target substance is detected in the detection zone is fixed in the detection zone. Liquid sample inspection kit.
The detection substance may be rephrased as a reagent that selectively binds to the substance to be detected, or a labeling substance (coloring substance) that selectively binds to the substance to be detected.

(12) The liquid sample inspection kit according to (10) or (11), wherein the color change indicates a change of 30 or more in the distance between RGB coordinates before and after detection.
That is, the coordinates in the RGB color space of the color of the detection zone before the detection target substance is detected are represented as C1, and the coordinates in the RGB color space of the color of the detection zone after the detection target substance is detected are , C2, the distance between the coordinates C1 and C2 may be 30 or more.

(13) A method for producing a liquid sample inspection kit, wherein the liquid sample inspection kit film carrier according to any one of (1) to (9) is prepared by thermal imprinting.
That is, in the method for manufacturing a liquid sample inspection kit according to the present embodiment, the surface of a mold (mold) in which a plurality of recesses is formed is applied to a film-like base material made of thermoplastic plastic, and the base material is used. You may provide the process (thermal imprint process) of producing the film | membrane carrier which has the fine structure (several convex part) and flat part corresponding to the shape of a recessed part by heating.

(14) When a substance to be detected is detected in the detection zone of the liquid sample inspection kit described in any one of (10) to (12), it can be visually confirmed that the substance is detected. The method for producing a liquid sample inspection kit according to (13), wherein a detection substance that causes a color change is fixed.
That is, the method for manufacturing a liquid sample test kit according to one aspect of the present invention further includes a step of fixing a reagent that selectively binds to a detected substance in a liquid sample on the surface of a membrane carrier having a fine structure. Good.
In the method for producing a liquid sample inspection kit according to one aspect of the present invention, at least one of a reagent and a labeling substance that selectively binds to a detected substance in a liquid sample is fixed to the surface of a membrane carrier having a fine structure. The process of carrying out may be further provided.

  According to the membrane carrier, the liquid sample inspection kit, and the method for producing the liquid sample inspection kit according to the present invention, since the determination at the time of detection of the substance to be detected in the liquid sample can be made visually, the method of using the inspection kit is easy. It is. Further, since the fine structure of the membrane carrier can be produced by thermal imprinting, the membrane carrier is inexpensive, and a test kit including this membrane carrier is useful as a disposable POCT reagent (test kit).

(A) in FIG. 1 is an overhead view (top view) of the fine structure provided in the membrane carrier according to one embodiment of the present invention, and (b) in FIG. 1 is shown in (a) in FIG. It is a perspective view of the fine structure of description. (A) in FIG. 2 is an overhead view (top view) of the fine structure provided in the membrane carrier according to one embodiment of the present invention, and (b) in FIG. 2 corresponds to (a) in FIG. It is a perspective view of the fine structure of description. (A) in FIG. 3 is an overhead view (top view) of the fine structure provided in the membrane carrier according to one embodiment of the present invention, and (b) in FIG. 3 corresponds to (a) in FIG. It is a perspective view of the fine structure of description. 4 is a cross-sectional view of the film carrier described in (a) and (b) of FIG. 1, and is a cross-sectional view perpendicular to the surface (flat portion) of the film carrier. (A), (b), and (c) in FIG. 5 are schematic views showing a process for producing a membrane carrier according to an embodiment of the present invention by thermal imprinting. (A) in FIG. 6 is an overhead view (top view) of the structure of the mold used in the calculation of Reference Example 6, and (b) in FIG. 6 is the structure described in (a) in FIG. It is sectional drawing in the AA of FIG. (A) in FIG. 7 is an overhead view (top view) of the structure of the mold used in the calculation of Reference Example 6, and (b) in FIG. 7 is the structure described in (a) in FIG. It is sectional drawing in the BB line | wire. FIG. 8 is a schematic top view of a test kit according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described. The inspection kit according to the present embodiment detects a substance to be detected in a liquid sample. For example, as shown in FIG. 8, the test kit 18 includes a membrane carrier 3 and a casing 18 a that houses the membrane carrier 3. On the surface of the membrane carrier 3, there are a dropping zone 3x where a liquid sample is dropped and a detection zone 3y for detecting a substance to be detected in the liquid sample. The dripping zone 3x is exposed at the first opening 18b of the housing 18a. The detection zone 3y is exposed at the second opening 18c of the housing 18a. The membrane carrier 3 is provided with at least one channel for transporting a liquid sample, and a fine structure is provided on the bottom surface of the channel. The fine structure is located at least between the dropping zone 3x and the detection zone 3y. There may be a fine structure over the entire surface of the membrane carrier 3. The entire surface of the membrane carrier 3 may be a flow path for the liquid sample. The microstructure causes capillary action. Due to the capillary action of the fine structure, the liquid sample is transported from the dropping zone 3x to the detection zone 3y via the fine structure. When the substance to be detected in the liquid sample is detected in the detection zone 3y, the color of the detection zone 3y changes. As shown in FIG. 1, FIG. 2, or FIG. 3, the microstructure is a convex portion (14, 14 a or 14 b), or a total of a plurality of convex portions (14, 14 a or 14 b). That is, the membrane carrier (3, 3a or 3b) includes a flat portion (13, 13a or 13b) corresponding to the bottom surface of the flow path of the liquid sample and a plurality of convex portions protruding from the flat portion (13, 13a or 13b). (14, 14a or 14c). The space between the plurality of convex portions (14, 14a, or 14c) functions as a flow path for transporting the liquid sample along the surface of the membrane carrier 3 by capillary action. In other words, the gap in the fine structure (14, 14a or 14c) functions as a flow path for transporting the liquid sample along the surface of the membrane carrier 3 due to capillary action. The plurality of convex portions (14, 14a or 14c) may be arranged on the surface of the membrane carrier 3 regularly or translationally symmetrically.

The liquid sample inspection method according to one aspect of the present invention is an inspection method using the inspection kit 18.
For example, the test kit 18 includes a membrane carrier 3 and a casing 18a that accommodates the membrane carrier 3, and a dropping zone 3x in which a liquid sample is dropped on the surface of the membrane carrier 3, and a liquid sample in the liquid sample. There are a detection zone 3y for detecting a substance to be detected, and a fine structure (a plurality of convex portions) positioned at least between the dropping zone 3x and the detection zone 3y. The dropping zone 3x is the first of the casing 18a. The opening 18b is exposed, the detection zone 3y is exposed at the second opening 18c of the housing 18a, and the entire film carrier 3 including the microstructure (plural protrusions) is made of thermoplastic. Consists of
For example, the inspection method includes a step of dropping a liquid sample onto the dropping zone 3x of the surface of the membrane carrier 3, and a capillary action exerted by the fine structure 14 (plural protrusions) formed on the surface of the membrane carrier 3. In the step of transporting the liquid sample from the dropping zone 3x to the detection zone 3y via the fine structure 14, and in the transportation process, the detected substance in the liquid sample is combined with the labeling substance, and the detected substance And a step of visually determining whether or not there is a color change (coloration of the labeling substance) in the detection zone 3y by combining with a reagent fixed in the detection zone 3y.

  The membrane carrier may consist of a thermoplastic. In other words, a film carrier having a fine structure can be produced by processing a film-like base material made of thermoplastic plastic by thermal imprinting. The thermoplastic plastic constituting the film carrier may be at least one selected from the group consisting of polyester resins, polyolefin resins, polystyrene resins, polycarbonate resins, fluorine resins, and acrylic resins, for example. Specific thermoplastics include, for example, polyethylene terephthalate (PET), cycloolefin polymer (COP), polypropylene (PP), polystyrene (PS), polycarbonate (PC), polyvinylidene fluoride (PVDF), and polymethyl methacrylate (PMMA) may be used.

The thermoplastic resin may have a glass transition point Tg or a melting point Tm of 80 to 180 ° C. The storage elastic modulus of the thermoplastic plastic at a temperature 20 ° C. higher than the glass transition point Tg may be 1.0 Pa or more and 1.0 × 10 ⁷ Pa or less. The storage elastic modulus of the thermoplastic plastic at a temperature 20 ° C. higher than the melting point Tm may be 1.0 Pa or more and 1.0 × 10 ⁷ Pa or less. When the glass transition or melting of the thermoplastic occurs at a temperature below 80 ° C., and the storage elastic modulus of the thermoplastic at a temperature 20 ° C. higher than the glass transition point or the melting point is 1.0 × 10 ⁷ Pa or less It is practically difficult to use a thermoplastic plastic as a solid at room temperature, and it becomes difficult to produce a film carrier by thermal imprinting. When the glass transition or melting of the thermoplastic occurs at a temperature higher than 180 ° C., the molding temperature at the time of thermal imprinting becomes high, and the productivity of the film carrier decreases. That is, when the temperature required for softening the thermoplastic during hot imprinting is higher than 180 ° C., the productivity of the film carrier is lowered. When the storage elastic modulus of a thermoplastic plastic at a temperature 20 ° C. higher than the glass transition point or the melting point is 1.0 × 10 ⁷ Pa or less, the molding pressure required for producing a fine structure can be kept small. Since the microstructure can be produced under relatively mild conditions, the production efficiency of the membrane carrier is improved.

  The microstructure provided on the film carrier may be, for example, a structure in which cones are regularly arranged. That is, the fine structure may be a cone. Regularly aligned cones can be formed by thermal imprinting using a mold. When a cone is formed using a mold, the volume of the metal scraped from the surface of the metal member during the production of the mold is significantly larger than when a groove-like channel (line and space structure) is formed using a mold. The mold processing cost is reduced. In contrast, in making a mold to form a line and space structure, a large amount of metal must be scraped from the metal member.

  The top of the cone is thinner than the bottom of the cone. Therefore, when a cone is formed using a mold, the volume of metal scraped from the surface of the metal member during the production of the mold is greatly reduced compared to the case where a pillar having the same bottom surface as the cone is formed by the mold. As a result, the processing cost of the mold is reduced.

  Further, the porosity of the fine structure in which the cones are regularly arranged is larger than the porosity of the line and space structure. Further, the porosity of the fine structure in which the cones are regularly arranged has a larger porosity than the structure in which a plurality of pillars having the same bottom surface as the cone are regularly arranged. Therefore, according to the fine structure in which the cones are regularly arranged, the flow rate of the liquid sample can be increased, which is advantageous for detecting the substance to be detected.

  The shape of the bottom surface of the cone (fine structure) can be freely selected. For example, as shown in FIGS. 1A and 1B, the microstructure may be a cone 14. For example, as shown in FIGS. 2A and 2B, the fine structure may be a quadrangular pyramid 14a. For example, as shown in FIGS. 3A and 3B, the fine structure may be a hexagonal pyramid 14b. In order to easily process the mold and reduce the processing cost, the bottom surface of the cone is preferably circular or polygonal (for example, a square, a rhombus, a rectangle, a triangle, or a hexagon).

  The diameter of the bottom surface of the microstructure may be 10 to 1000 μm. When the diameter of the bottom surface of the fine structure is smaller than 10 μm, the fine processing cost of the mold becomes high, and it is difficult to uniformly produce innumerable fine structures on the surface of the film carrier having a large area. Therefore, a microstructure that is too small is not suitable for practical use. When the diameter of the bottom surface of the microstructure is smaller than 10 μm, the capillary force necessary to move the liquid sample tends to be weakened. When the diameter of the bottom surface of the fine structure is larger than 1000 μm, the volume of the metal scraped from the metal member at the time of producing the mold becomes large, and the production cost of the mold and the film carrier becomes high. When the diameter of the bottom surface of the fine structure is larger than 1000 μm, the area of the flow path in the membrane carrier must be increased, which makes the liquid sample inspection kit huge and disadvantageous for transporting the liquid sample inspection kit itself. As shown in FIG. 1A or 4, when the microstructure is a cone 14, the diameter of the bottom surface of the microstructure may be the diameter 4 of the bottom surface (circle) of the cone 14.

The fine structure may have a height of 10 to 500 μm. When the height of the fine structure is smaller than 10 μm, the capillary force necessary to move the liquid sample tends to be weakened. When the height of the fine structure is larger than 500 μm, it is difficult to completely fill the thermoplastic resin into the concave portion of the mold (the depression corresponding to the shape of the fine structure) at the time of thermal imprinting. As shown in FIG. 1A or 4, when the microstructure is a cone 14, the height of the microstructure may be the height 6 of the cone 14 from the flat portion 13.
The overall shape of the membrane carrier 3, 3a or 3b is not particularly limited, but may be, for example, a polygon such as a quadrangle, a circle, or an ellipse. When the membrane carrier 3, 3a or 3b is square, the vertical width of the membrane carrier 3, 3a or 3b may be, for example, 2 to 100 mm, and the lateral width of the membrane carrier 3, 3a or 3b is, for example, 2 to 2 mm. It may be 100 mm. The thickness of the membrane carrier 3, 3a or 3b excluding the height of the fine structure may be, for example, 0.1 to 10 mm.

  The aspect ratio of the microstructure may be 10: 1 to 1: 2. That is, the aspect ratio Lv / Lh of the fine structure may be 1/10 or more and 2/1 or less. When the aspect ratio is smaller than 10: 1 (that is, 1/10), the contact area between the liquid sample and the channel is small, and the capillary force is reduced, so that the liquid sample tends to be difficult to move. When the aspect ratio is larger than 1: 2 (that is, 2/1), the productivity of the film carrier by thermal imprinting is lowered. As shown in FIG. 1A or 4, when the microstructure is a cone, the horizontal length Lh of the microstructure may be the diameter 4 of the bottom surface of the cone 14. Further, the length Lv in the vertical direction of the fine structure may be the height 6 of the cone 14 from the flat portion 13 of the film carrier 3.

  The ratio D2 / D1 between the diameter (D1) of the bottom surface of the microstructure and the distance (D2) between the nearest centers of the microstructures may be greater than 1 and 5 or less. The ratio D2 / D1 cannot be less than 1. When the ratio D2 / D1 is greater than 5, the contact area between the liquid sample and the flow path decreases, the capillary force decreases, and the liquid sample tends to be difficult to move. As shown in FIG. 1A or 4, when the microstructure is a cone 14, the diameter D1 of the bottom surface of the microstructure may be the diameter 4 of the bottom surface of the cone 14, and the closest center-to-center distance. D2 may be a distance 5 between the apexes of a pair of adjacent cones 14. The diameter D1 of the bottom surface of the microstructure may coincide with the length Lh in the horizontal direction of the microstructure described above. Therefore, the aspect ratio Lv / Lh may be expressed as Lv / D1.

As shown in (a), (b), and (c) of FIG. 5, the manufacturing method of the liquid sample inspection kit according to the present embodiment applies the surface of the mold (mold 1) in which a plurality of recesses are formed. The fine structure (plurality of convex portions (14)) corresponding to the shape of the concave portion and the flat portion 13 are obtained by heating the base material 2 against the film-like base material 2 made of thermoplastic plastic. A step of preparing the membrane carrier 3 (thermal imprinting step) may be provided. The method for producing a liquid sample inspection kit may further include a step of fixing the reagent or the labeling substance to the detection zone in the surface of the membrane carrier having a fine structure. When the substance to be detected in the liquid sample binds to the labeling substance and the substance to be detected selectively binds to the reagent in the detection zone, the color of the detection zone changes. Therefore, by visually observing the change in the color of the detection zone, it can be confirmed that the substance to be detected in the liquid sample has been detected.
The mold fine processing method used in the thermal imprint process may be, for example, etching, photolithography, mechanical cutting, or laser processing. A fine processing method suitable for the processing size and processing range can be selected.

  It is desirable to perform a mold release treatment before performing the thermal imprint. In the release treatment, for example, a monomolecular film may be produced on the mold surface to reduce the surface energy. As a result, it becomes easy to peel the film carrier 3 made of thermoplastic plastic from the surface of the mold 1 after the thermal imprinting.

  The thermal imprinting method may be either a flat plate press type or a roll type. The thermal imprint system shown in FIG. 5 is a flat plate press system. In the flat plate press type, the mold 1 is overlapped with the base material 2 made of thermoplastic plastic between upper and lower stages facing in parallel, and these are sandwiched between the stages. Then, the mold 1 and the substrate 2 are heated and pressurized through the stage. Such a flat plate press type is excellent in terms of good molding accuracy. The roll method is a method in which a heated roll mold is used and molding is performed with a pinching pressure between rolls. The roll type is excellent in productivity.

  Conditions such as molding temperature, molding pressure, and transfer time when performing thermal imprinting may be selected according to the size of the fine processing, the shape of the fine structure, the size of the processing range, and the like. For example, in the case of a flat plate press type, the molding temperature may be a temperature 20 to 50 ° C. higher than the glass transition point Tg, or a temperature 20 to 50 ° C. higher than the melting point Tm. The molding pressure may be 1-10 MPa. The transfer time (the time for holding the mold 1 and the substrate 2 while being pressed) may be 3 to 10 minutes. By heat impressing under the above various conditions, it becomes easy to accurately transfer the microstructure of the mold 1 to the surface of the substrate 2.

  Depending on the type of thermoplastic plastic constituting the membrane carrier 3 and the type of reagent (detection substance), it is difficult to fix a reagent (detection substance) to the detection zone of the membrane carrier 3 so that detection can be judged visually. In this case, it is easy to fix the reagent (detection substance) to the detection zone of the membrane carrier 3 by applying an appropriate surface treatment only to the detection zone in advance.

  The surface treatment method of the detection zone is not limited at all, and may be various methods such as various plasma treatments, UV treatments, UV / ozone treatments, or surface modification with 3-Aminopropyltrioxysilane or Glutaraldehyde.

  The reagent (detection substance) fixed to the detection zone may be, for example, an antibody. An antibody is a substance that causes an antigen-antibody reaction with a substance to be detected. The antibody may be a polyclonal antibody or a monoclonal antibody. The substance to be detected is not limited in any way, and may be any substance capable of causing an antigen-antibody reaction with an antibody, such as various pathogens and various clinical markers. The specific substance to be detected may be, for example, a viral antigen such as influenza virus, norovirus, adenovirus, RS virus, HAV, HBs, and HIV. The substance to be detected may be MRSA, group A streptococci, group B streptococci, bacterial antigens such as Legionella spp., And toxins produced by bacteria. The substance to be detected may be a hormone such as mycoplasma, Chlamydia trachomatis, human chorionic gonadotropin. The substance to be detected may be C-reactive protein, myoglobin, cardiac troponin, various tumor markers, agricultural chemicals, environmental hormones, and the like. In particular, when it is urgent to detect a substance to be detected such as influenza virus, norovirus, C-reactive protein, myoglobin, and cardiac troponin and to treat a disease caused by these, the test kit according to this embodiment Especially useful. The substance to be detected may be an antigen capable of inducing an immune reaction alone. The substance to be detected may be a hapten capable of binding an antibody by an antigen-antibody reaction, although it cannot induce an immune reaction by itself.

  The coordinates in the RGB color space (RGB coordinate space) of the color of the detection zone before the detection target substance is detected is expressed as C1, and the RGB color space of the color of the detection zone after the detection target substance is detected. Is expressed as C2, the distance between the coordinates C1 and C2 in the RGB color space is preferably 30 or more. The coordinate C1 is obtained by measuring the R value (Red value), G value (Green value), and B value (Blue value) of the detection zone before detection. The coordinate C2 is obtained by measuring the R value, G value, and B value of the detection zone after detection. The distance between the coordinates C1 and C2 in the RGB color space is obtained by calculation. This distance between the RGB coordinates may be rephrased as a chromaticity difference in the detection zone before and after the detection of the detection target substance. When this chromaticity difference is smaller than 30, there is a tendency that it is difficult to visually confirm the difference or change in color.

  Hereinafter, the present invention will be specifically described with reference to examples and comparative examples, but the present invention is not limited to these examples.

[Example 1]
<Mold preparation>
A mold was produced by laser processing of a metal member. The metal member was made of aluminum alloy A5052. In the laser processing, a plurality of inverted conical concave portions (that is, fine structures) were formed in the center portion (3 cm × 3 cm square range) of the flat surface of the mold. All the recesses had the same shape. The diameter of the recess was 10 μm. The diameter of the recess is equal to the diameter (D1 or Lh) of the bottom surface of the microstructure (cone) of the finally obtained membrane carrier. The distance between the centers of a pair of adjacent recesses was 15 μm. The distance between the centers of the recesses is equal to the distance (D2) between the apexes of adjacent fine structures (cones) on the surface of the membrane carrier. The depth of the recess was 10 μm. The depth of the recess is equal to the height (hv) of the microstructure (cone) of the membrane carrier. The plurality of concave portions were regularly arranged in the center portion of the mold in the same manner as the arrangement of the cones 14 (triangular arrangement format) shown in FIG.
A mold release treatment was performed on the surface (uneven surface) of the mold in which the recesses were formed. In the mold release treatment, the mold surface (uneven surface) was immersed in the treatment liquid for about 1 minute, and then the mold surface was dried. The dried mold was left overnight. Optool HD-2100TH manufactured by Daikin Industries, Ltd. was used as the treatment liquid used for the above mold release treatment.

<Thermal imprint process (transfer of fine structure)>
The microstructure on the mold surface was transferred to the surface of a film-like substrate made of thermoplastic plastic by the following thermal imprint process. In the thermal imprint process, X-300 manufactured by SCIVAX was used. In the thermal imprinting process, the surface of the mold on which the fine structure (a plurality of concave portions) was formed was applied to a film-like base material made of thermoplastic plastic, and the mold and the base material were pressed while heating. The molding temperature was 120 ° C. The applied pressure was 5.5 MPa. The transfer time was 5 minutes. After the transfer of the fine structure, the mold and the substrate were cooled to 80 ° C. while pressure was applied to the mold and the substrate. The pressure was removed after cooling. The membrane carrier of Example 1 was obtained by the above thermal imprint process. This membrane carrier had a surface containing a plurality of cones (fine structure) and flat portions. The shape and size of the convex portion (cone) on the surface of the membrane carrier was consistent with the shape and size of the concave portion (inverted cone) formed in the mold.
The film substrate made of thermoplastic was a film made of polystyrene (PS) (Denka styrene sheet manufactured by Denki Kagaku Kogyo Co., Ltd.). The thickness of the base material was 188 μm. The substrate (membrane carrier) was square. The vertical width of the base material (membrane carrier) was 50 mm, and the horizontal width of the base material (membrane carrier) was 50 mm.
The glass transition point Tg of polystyrene constituting the substrate (membrane carrier) was measured using DSC3100 manufactured by Bruker AXS. In measuring the glass transition point Tg, the substrate was heated at a rate of temperature increase of 10 ° C./min in a nitrogen atmosphere. The glass transition point Tg of polystyrene was 106 ° C.
The storage elastic modulus in the tensile mode of polystyrene constituting the substrate (membrane carrier) was measured. RSAIII manufactured by TA Instruments was used for the measurement of storage elastic modulus. The frequency at the time of measuring the storage elastic modulus was 1 Hz. The storage elastic modulus of polystyrene at 126 ° C. was 1.8 × 10 ⁶ Pa.

[Example 2]
The diameter of the recess formed on the surface of the mold of Example 2 was 10 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 2 was 50 μm. The depth of the recess formed on the surface of the mold of Example 2 was 10 μm. A membrane carrier of Example 2 was produced in the same manner as in Example 1 except that the mold was different.

[Example 3]
The diameter of the recess formed on the surface of the mold of Example 3 was 10 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 3 was 15 μm. The depth of the recess formed on the surface of the mold of Example 3 was 20 μm. A membrane carrier of Example 3 was produced in the same manner as in Example 1 except that the mold was different.

[Example 4]
The diameter of the recess formed on the surface of the mold of Example 4 was 10 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 4 was 50 μm. The depth of the recess formed on the surface of the mold of Example 4 was 20 μm. A membrane carrier of Example 4 was produced in the same manner as in Example 1 except that the mold was different.

[Example 5]
The diameter of the recess formed on the surface of the mold of Example 5 was 100 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 5 was 110 μm. The depth of the recess formed on the surface of the mold of Example 5 was 10 μm. A membrane carrier of Example 5 was produced in the same manner as in Example 1 except that the mold was different.

[Example 6]
The diameter of the recess formed on the surface of the mold of Example 6 was 100 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 6 was 500 μm. The depth of the recess formed on the surface of the mold of Example 6 was 10 μm. A membrane carrier of Example 6 was produced in the same manner as in Example 1 except that the mold was different.

[Example 7]
The diameter of the recess formed on the surface of the mold of Example 7 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 7 was 1010 μm. The depth of the recess formed on the surface of the mold of Example 7 was 100 μm. A membrane carrier of Example 7 was produced in the same manner as in Example 1 except that the mold was different.

[Example 8]
The diameter of the recess formed on the surface of the mold of Example 8 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 8 was 5000 μm. The depth of the recess formed on the surface of the mold of Example 8 was 100 μm. A membrane carrier of Example 8 was produced in the same manner as in Example 1 except that the mold was different.

[Example 9]
The diameter of the recess formed on the surface of the mold of Example 9 was 250 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 9 was 260 μm. The depth of the recess formed on the surface of the mold of Example 9 was 500 μm. A membrane carrier of Example 9 was produced in the same manner as in Example 1 except that the mold was different.

[Example 10]
The diameter of the recess formed on the surface of the mold of Example 10 was 250 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 10 was 1250 μm. The depth of the recess formed on the surface of the mold of Example 10 was 500 μm. A membrane carrier of Example 10 was produced in the same manner as in Example 1 except that the mold was different.

[Example 11]
The diameter of the recess formed on the surface of the mold of Example 11 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 11 was 1010 μm. The depth of the recess formed on the surface of the mold of Example 11 was 500 μm. A film carrier of Example 11 was produced in the same manner as in Example 1 except that the mold was different.

[Example 12]
The diameter of the recess formed on the surface of the mold of Example 12 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Example 12 was 5000 μm. The depth of the recess formed on the surface of the mold of Example 12 was 500 μm. A membrane carrier of Example 12 was produced in the same manner as in Example 1 except that the mold was different.

[Examples 13 to 24]
In Examples 13 to 24, a film made of polycarbonate (PC) was used instead of a film made of polystyrene as a film-like base material made of thermoplastic. Panlite manufactured by Teijin Limited was used as the polycarbonate film. The thickness of the base material made of polycarbonate was 200 μm. The substrate (membrane carrier) was square. The vertical width of the base material (membrane carrier) was 50 mm, and the horizontal width of the base material (membrane carrier) was 50 mm.
Using a DSC3100 manufactured by Bruker AXS, the glass transition point Tg of the polycarbonate constituting the substrate (membrane carrier) was measured. In measuring the glass transition point Tg, the substrate was heated at a rate of temperature increase of 10 ° C./min in a nitrogen atmosphere. The glass transition point Tg of the polycarbonate was 160 ° C.
The storage elastic modulus in the tensile mode of the polycarbonate constituting the substrate (membrane carrier) was measured. RSAIII manufactured by TA Instruments was used for the measurement of storage elastic modulus. The frequency at the time of measuring the storage elastic modulus was 1 Hz. The storage elastic modulus of the polycarbonate at 180 ° C. was 4.5 × 10 ⁶ Pa.
A membrane carrier of Example 13 was produced in the same manner as in Example 1 except that the film-like substrate was different. A membrane carrier of Example 14 was produced in the same manner as in Example 2 except that the membrane-like substrate was different. A membrane carrier of Example 15 was produced in the same manner as in Example 3 except that the membrane-like substrate was different. A membrane carrier of Example 16 was produced in the same manner as in Example 4 except that the membrane-like substrate was different. A membrane carrier of Example 17 was produced in the same manner as in Example 5 except that the membrane-like substrate was different. A membrane carrier of Example 18 was produced in the same manner as in Example 6 except that the membrane-like substrate was different. A membrane carrier of Example 19 was produced in the same manner as in Example 7 except that the film-like substrate was different. A membrane carrier of Example 20 was produced in the same manner as in Example 8, except that the film-like substrate was different. A membrane carrier of Example 21 was produced in the same manner as in Example 9 except that the membrane-like substrate was different. A membrane carrier of Example 22 was produced in the same manner as in Example 10 except that the membrane-like substrate was different. A membrane carrier of Example 23 was produced in the same manner as in Example 11 except that the membrane-like substrate was different. A membrane carrier of Example 24 was produced in the same manner as in Example 12 except that the film-like substrate was different.

<Immobilization of reagent (antibody) to detection zone>
By the following method, only the following detection zone A and detection zone B of the surface of the film carrier (surface having a fine structure) of each example were exposed, and the other portions were covered with a mask. Subsequently, the detection zone A and detection zone B were subjected to UV treatment.
Detection zone A: A line-shaped portion having a distance of 0.6 cm from the lower end of the membrane carrier and a width of about 1 mm.
Detection zone B: A line-shaped portion having a distance of 1.0 cm from the lower end of the membrane carrier and a width of about 1 mm.
A suspension A of anti-influenza A NP antibody (reagent A) was applied to detection zone A. Subsequently, the detection zone A was thoroughly dried under hot air, and the anti-influenza A influenza NP antibody was fixed to the detection zone A. The application amount of the suspension A was 18 μL. In the detection zone A, the length of the portion where the suspended liquid A was applied was 3 cm.
A suspension B of anti-influenza B NP antibody (reagent B) was applied to detection zone B. Subsequently, the detection zone B was thoroughly dried under warm air, and the anti-B influenza NP antibody was fixed to the detection zone B. The application amount of the suspension B was 18 μL. The length of the portion where the suspension B was applied in the detection zone B was 3 cm.

<Preparation of labeling substance>
A purified anti-influenza A virus NP antibody (purified antibody A) different from the above-described anti-influenza A NP antibody (reagent A) was prepared. In addition, a purified anti-B influenza virus NP antibody (purified antibody B) different from the above-described anti-B influenza NP antibody (reagent B) was prepared.
Purified antibody A was labeled by covalent bond between purified antibody A and blue latex particles (CM / BL Ceradyne). The particle size of the blue latex particles was 0.394 μm. A Tris buffer containing sugar, surfactant and protein was prepared. The labeled purified antibody A was added to the Tris buffer so that the concentration of latex particles in the Tris buffer was 0.025 w / v%, and the Tris buffer was suspended. Subsequently, sonication of the Tris buffer was performed to prepare an anti-A-type label that was sufficiently dispersed and suspended in the Tris buffer.
An anti-B-type labeled body that was sufficiently dispersed and suspended in Tris buffer was prepared in the same manner as in the case of the anti-A-type labeled body except that purified antibody B was used instead of purified antibody A.

  A mixed solution of an anti-A type label and an anti-B type label was prepared. This mixed solution was applied to glass fibers. The size of the glass fiber was 3 cm × 1 cm. The coating amount of the mixed solution per square centimeter of glass fiber was 50 μL. Examples of the glass fiber include 33GLASS NO. Manufactured by Schleicher & Schuell. 10539766 was used. The glass fiber coated with the mixed solution was thoroughly dried under warm air to prepare a marker pad. A labeling substance pad was placed on the end of the membrane carrier of each of Examples 1 to 24 closer to the detection zone. The width of the end portion of the membrane carrier on which the labeling substance pad overlaps was 2 mm. The membrane carrier on which the labeling substance pad overlaps was cut into a strip having a width of 5 mm with a cutter to prepare a liquid sample inspection kit comprising the integrated membrane carrier and the labeling substance pad.

<Evaluation of detection performance>
As a diluting solution, a sample suspension attached to Quick Navi-Flu manufactured by Denka Seken was prepared. As a detection substance, influenza A virus A / Beijing / 32/92 (H3N2) was used. This influenza A virus was diluted 4 × 10 ⁴ times with the sample suspension to prepare liquid sample A. As another detection substance, influenza B virus B / Shangdong / 7/97 was used. This influenza B virus was diluted 4 × 10 ³ times with the sample suspension to prepare liquid sample B.
100 μL of each of the liquid samples A and B was dropped individually on the ends of the liquid sample inspection kits of Examples 1 to 24. The end of the inspection kit to which the liquid sample was dropped was the end closer to the detection zone A and the detection zone B. The state in which the liquid sample after dropping was moved on the inspection kit was recorded with a digital camera from directly above. From this moving image, the flow rate of the liquid sample moving on the test kit was calculated. Tables 1 and 2 show the evaluation results of the flow rate of the liquid sample in each test kit of Examples 1 to 24.
The examples with double circles in each table are superior to the examples with circles in the effects of the invention. The examples with circles in each table are superior to the examples with triangles in the effects of the invention. The examples marked with a triangle in each table are superior to the reference examples marked with an X in the effects of the invention.

<Detection of detection>
Five minutes after the liquid samples A and B were dropped onto the respective inspection kits of Examples 1 to 24, the presence or absence of coloring of the detection zone A and the detection zone B in each inspection kit was visually observed. By this observation, the presence or absence of detection of each of influenza A virus and influenza B virus was determined.

  In any of the examples, the change in color of only the detection zone A to which the anti-influenza A influenza NP antibody was immobilized was confirmed by the dripping of the liquid sample A containing influenza A virus. In any of the examples, the color change of only the detection zone B to which the anti-influenza B influenza NP antibody was immobilized was confirmed by the dripping of the liquid sample B containing influenza B virus.

[Example 25]
Four liquid sample test kits of Example 25 were produced in exactly the same manner as in Example 1. Subsequently, four types of liquid samples with influenza A virus dilution ratios of 1 × 10 ⁴ , 2 × 10 ⁴ , 4 × 10 ⁴ and 8 × 10 ⁴ were prepared. The method for preparing these liquid samples is the same as the method for preparing the liquid sample A described above, except for the dilution factor.
In the same manner as in Examples 1 to 24, only one of the four types of liquid samples was dropped for each test kit of Example 25, and influenza A was detected in detection zone A of each test kit. A virus was detected. As shown in Table 3, only the detection zone A of the test kit in which the liquid sample having a dilution rate of 8 × 10 ⁴ was dropped showed no color change that could be visually confirmed. In any of the detection zones A of the test kit in which the other three types of liquid samples were dropped, a color change was visually confirmed.
Prior to detection of influenza A virus, the R value, G value and B value of each of the detection zones A of the four test kits were measured with a laser microscope. As a laser microscope, OPLETICS HYBRID manufactured by Lasertec Corporation was used. Further, after detection of influenza A virus, the R value, G value, and B value of each of detection zones A of the four test kits were measured with a laser microscope. Based on these measurement results, the distance between the RGB coordinates before and after detection in each of the detection zones A of the four test kits was calculated. Table 3 shows the calculation results.

[Example 26]
Four liquid sample test kits of Example 26 were produced in exactly the same manner as in Example 1. Next, four types of liquid samples with influenza B virus dilution ratios of 1 × 10 ³ , 2 × 10 ³ , 4 × 10 ³ and 8 × 10 ³ were prepared. The method for preparing these liquid samples is the same as the method for preparing the liquid sample B described above, except for the dilution factor.
In the same manner as in Examples 1 to 24, only one of the four types of liquid samples is dropped for each test kit of Example 26, and influenza B is detected in detection zone B of each test kit. A virus was detected. As shown in Table 3, only the detection zone B of the test kit to which the liquid sample having a dilution rate of 8 × 10 ³ was dropped showed no color change that could be visually confirmed. In any of detection zones B of the test kit in which the other three types of liquid samples were dropped, a color change was visually confirmed.
Prior to detection of influenza B virus, the R value, G value, and B value of each of detection zones B of the four test kits were measured with the laser microscope described above. Further, after detection of influenza B virus, the R value, G value, and B value of each of detection zones B of the four test kits were measured with a laser microscope. Based on these measurement results, the distance between the RGB coordinates before and after detection of each of the detection zones B of the four test kits was calculated. Table 3 shows the calculation results.

[Reference Example 1]
In Reference Example 1, a film made of polytetrafluoroethylene (PTFE) was used as a film-like base material made of thermoplastic. As a film made of polytetrafluoroethylene, Daikin Industries, Ltd. polyflon F-104 was used. The thickness of the base material made of polytetrafluoroethylene was 200 μm. The substrate (membrane carrier) was square. The vertical width of the base material (membrane carrier) was 50 mm, and the horizontal width of the base material (membrane carrier) was 50 mm. The melting point Tm of polytetrafluoroethylene constituting the substrate (membrane carrier) was measured using DSC3100 manufactured by Bruker AXS. In measuring the melting point Tm, the substrate was heated at a rate of temperature increase of 10 ° C./min in a nitrogen atmosphere. The melting point Tm of polytetrafluoroethylene was 327 ° C. The molding temperature in the thermal imprint process of Reference Example 1 was 250 ° C., which is the device limit value. A membrane carrier of Reference Example 1 was produced in the same manner as in Example 1 except for the above items. Various characteristics of the membrane carrier of Reference Example 1 are shown in Table 4 below.

[Reference Example 2]
The diameter of the recess formed on the surface of the mold of Reference Example 2 was 10 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Reference Example 2 was 15 μm. The depth of the recess formed on the surface of the mold of Reference Example 2 was 30 μm. A membrane carrier of Reference Example 2 was produced in the same manner as in Example 1 except that the mold was different. Various characteristics of the membrane carrier of Reference Example 2 are shown in Table 4 below.

[Reference Example 3]
The diameter of the recess formed on the surface of the mold of Reference Example 3 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Reference Example 3 was 1010 μm. The depth of the recess formed on the surface of the mold of Reference Example 3 was 2000 μm. A membrane carrier of Reference Example 3 was produced in the same manner as in Example 1 except that the mold was different. Various characteristics of the membrane carrier of Reference Example 3 are shown in Table 4 below.

  The membrane carriers of Reference Examples 1 to 3 were observed. In the film carrier of Reference Example 1, a desired microstructure was not formed. This is because PTFE has a melting point higher than 180 ° C., and the elastic modulus of PTFE remained high even when the molding temperature in the hot impressing process was set to the upper limit of the apparatus. In the film carrier of Reference Example 2, a fine structure fold and inflection were observed. This is because the aspect ratio of the fine structure is larger than 2, and the film carrier cannot be smoothly peeled off from the mold. In the membrane carrier of Reference Example 3, the height of the fine structure did not reach 2000 μm. This is because the depth of the recess formed on the surface of the mold of Reference Example 3 is larger than 500 μm, and PS is not sufficiently filled in the recess of the mold.

[Reference Example 4]
The diameter of the recess formed on the surface of the mold of Reference Example 4 was 100 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Reference Example 4 was 1000 μm. The depth of the recess formed on the surface of the mold of Reference Example 4 was 200 μm. A membrane carrier of Reference Example 4 was produced in the same manner as in Example 1 except that the mold was different. Various characteristics of the membrane carrier of Reference Example 4 are shown in Table 5 below.

[Reference Example 5]
The diameter of the recess formed on the surface of the mold of Reference Example 5 was 1000 μm. The distance between the centers of a pair of adjacent recesses on the surface of the mold of Reference Example 5 was 1010 μm. The depth of the recess formed on the surface of the mold of Reference Example 5 was 50 μm. A membrane carrier of Reference Example 5 was produced in the same manner as in Example 1 except that the mold was different. Various characteristics of the membrane carrier of Reference Example 5 are shown in Table 5 below.

  In the same manner as in Examples 1 to 24, the state of the liquid sample dropped on the fine structure of each of the reference examples 4 and 5 was observed with a digital camera from directly above. In any of the membrane carriers of Reference Examples 4 and 5, the liquid sample did not move. The reason why the liquid sample did not move was as follows. In Reference Example 4, the ratio D2 / D1 between the diameter D1 of the bottom surface of the fine structure and the distance D2 between the closest centers of the fine structures was larger than 5, and sufficient capillary force for moving the liquid sample was not generated. In Reference Example 5, the aspect ratio of the microstructure was less than 0.1, and sufficient capillary force for moving the liquid sample was not generated.

[Reference Example 6]
The volume of metal scraped from the surface of the metal member when a mold for forming a line and space structure on the surface of the film carrier was produced from the metal member was calculated. The line and space structure is a structure having a plurality of groove-shaped flow paths arranged in parallel. The mold itself for forming the line and space structure also has a line and space structure as shown in FIGS. 6A and 6B. The width 7 of the groove 15 used for the calculation was 10 μm. The depth 8 of the groove 15 used for the calculation was 10 μm. The pitch 9 of the grooves 15 used for the calculation was 15 μm. In the calculation, the range of the processed mold surface was assumed to be a 3 cm × 3 cm square. Moreover, the porosity of the line and space structure formed using the mold of FIG. 6 was also calculated. Table 6 shows the calculation results.
In addition, the volume of metal scraped from the surface of the metal member when a mold for forming a structure in which a plurality of cylinders are regularly arranged on the surface of the film carrier is manufactured from the metal member was calculated. As shown in FIGS. 7A and 7B, this mold has a flat portion 16 and a plurality of cylindrical concave portions 17. The diameter 10 of the recess 17 used for the calculation was 10 μm. The depth 12 of the recess 17 used for the calculation was 10 μm. The pitch 11 of the recesses 17 used for the calculation was 15 μm. In the calculation, the range of the processed mold surface was assumed to be a 3 cm × 3 cm square. Table 6 shows the calculation results. Further, the porosity of a structure in which a plurality of cylinders are regularly arranged formed using the mold of FIG. 7 was also calculated. Table 6 shows the calculation results.
Table 6 shows the volume of the metal cut out from the metal member when the mold of Example 1 was manufactured. Table 6 shows the porosity of the fine structure of the membrane carrier of Example 1.

  From the results of Tables 1 to 5, it was shown that the liquid sample inspection kit according to the present invention can be produced by thermal imprinting, and the dropped liquid sample can be moved by capillary force. In addition, according to the present invention, it has been shown that the color change in the detection zone is large enough to be visually confirmed. Furthermore, from the results in Table 6, it was shown that the volume of the metal to be machined out during the mold production was reduced and the porosity of the microstructure of the film carrier was increased by making the microstructure a cone.

  According to the membrane carrier and the liquid sample inspection kit according to the present invention, since the determination at the time of detection of the substance to be detected in the liquid sample can be made visually, the method of using the inspection kit is easy. Further, since the fine structure of the membrane carrier can be produced by thermal imprinting, the membrane carrier is inexpensive, and a test kit including this membrane carrier is useful as a disposable POCT reagent (test kit).

DESCRIPTION OF SYMBOLS 1 Mold which has microstructure 2 Film-like base material which consists of thermoplastics 3 Film carrier provided with microstructure 3x Drip zone 3y Detection zone 4 Diameter of bottom surface of microstructure 5 Distance between nearest centers of microstructures 6 Microstructure height 7 Line and space structure groove width 8 Line and space structure groove depth 9 Line and space structure groove pitch 10 Regularly arranged cylindrical recess diameters 11 Regular array Pitch 12 of cylindrical recesses Depths of regularly arranged cylindrical recesses 13, 13a, 13b Flat portion 14 Cone (fine structure)
14a square pyramid (fine structure)
14b Hexagonal pyramid (fine structure)
15 Line and space structure groove 16 Flat portion 17 between cylindrical recesses Cylindrical recesses 18 regularly arranged 18 Liquid sample test kit 18a Case 18b First opening 18c Second opening

Claims (14)

A membrane carrier for a test kit for detecting a substance to be detected in a liquid sample,
At least one flow path capable of transporting the liquid sample is provided;
A membrane carrier for a liquid sample inspection kit, wherein a fine structure that causes a capillary action for transporting the liquid sample is provided on a bottom surface of the channel.   The membrane carrier for a liquid sample inspection kit according to claim 1, comprising a thermoplastic having a glass transition point of 80 to 180 ° C.   The film carrier for a liquid sample inspection kit according to claim 1 or 2, comprising a thermoplastic having a melting point of 80 to 180 ° C. 4. The liquid sample inspection kit according to claim 2, wherein a storage elastic modulus of the thermoplastic plastic is 1.0 × 10 ⁷ Pa or less at a temperature 20 ° C. higher than a glass transition point or a melting point. 5. Membrane carrier.   The film carrier for a liquid sample inspection kit according to any one of claims 1 to 4, wherein the shape of the fine structure is a cone.   The membrane carrier for a liquid sample inspection kit according to any one of claims 1 to 5, wherein a diameter of a bottom surface of the fine structure is 10 to 1000 µm.   The membrane carrier for a liquid sample inspection kit according to any one of claims 1 to 6, wherein the fine structure has a height of 10 to 500 µm.   The membrane carrier for a liquid sample inspection kit according to any one of claims 1 to 7, wherein an aspect ratio of the fine structure is 10: 1 to 1: 2.   The liquid sample inspection kit according to any one of claims 1 to 8, wherein a ratio between a diameter of a bottom surface of the fine structure and a distance between closest centers of the fine structures is greater than 1 and 5 or less. Membrane carrier. Using the membrane carrier for a liquid sample inspection kit according to any one of claims 1 to 9,
A detection zone for detecting a substance to be detected in a liquid sample in the membrane carrier;
A liquid sample inspection kit in which when a substance to be detected is detected in the detection zone, a color change that can be visually confirmed is detected.   The liquid sample according to claim 10, wherein when the detection target substance is detected in the detection zone, a detection substance that causes a color change that can be visually confirmed to be detected is fixed to the detection zone. Inspection kit.   The liquid sample inspection kit according to claim 10 or 11, wherein the color change indicates a change of 30 or more in a distance between RGB coordinates before detection and after detection.   A method for producing a liquid sample inspection kit, wherein the liquid sample inspection kit film carrier according to any one of claims 1 to 9 is produced by thermal imprinting.   The color that can be visually confirmed to be detected when the detection target substance is detected in the detection zone in the detection zone of the liquid sample inspection kit according to any one of claims 10 to 12. The method for producing a liquid sample inspection kit according to claim 13, wherein a detection substance that causes a change is fixed.

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