TW201627669A - Membrane support for liquid sample test kit, liquid sample test kit, and method for producing liquid sample test kit - Google Patents

Abstract

A membrane support (3) can be used in a test kit (18) for detecting a substance of interest in a liquid sample. The membrane support (3) has, provided thereon, at least one flow path that can transport a liquid sample, and microstructures (14) that can cause a capillary action for transporting the liquid sample are formed on the bottom of the flow path. The membrane support (3) can be produced by a thermal imprinting technique at low cost, and can move a liquid sample with the aid of a capillary force. In a detection zone (3y) in the test kit (18), the change in color of the substance of interest during the detection can be confirmed with naked eyes.

Description

Membrane carrier for liquid sample inspection kit, liquid sample inspection kit and liquid sample inspection kit

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

In recent years, a point-of-care (POCT) reagent for detecting an infection or a pregnancy, or measuring a blood sugar level or the like by using an antigen-antibody reaction or the like has been attracting attention. In the examination and measurement using the POCT reagent, the determination of the result can be performed in a short time. Moreover, the method of using the POCT reagent is relatively simple, and the POCT reagent is relatively inexpensive. Since the POCT reagent has such characteristics, it is used more frequently for examinations at a less severe stage or for regular examinations. In addition, POCT reagents have become an important diagnostic tool for predicting home care that will increase in the future.

In the inspection or diagnosis using an inspection kit which is one of the POCT reagents, a liquid sample such as blood is introduced into the inspection kit to detect a specific test substance contained in the liquid sample. As a method of detecting a specific substance to be detected from a liquid sample, immunochromatography is often used. In the immunochromatography method, a liquid sample is dropped on the membrane carrier provided in the inspection kit, and the liquid to be detected is combined with the labeling substance during the movement of the liquid sample on the membrane carrier. Further, the substance to be detected is specifically and selectively bound to a substance (hereinafter referred to as a detection substance) immobilized in the test kit. As a result, changes in color or weight generated by the inspection kit are detected. The test substance can also be referred to as a reagent (reagant).

A nitrocellulose membrane is often used as a membrane carrier for moving a liquid sample (see Patent Document 1 below). The nitrocellulose membrane has a plurality of fine pores having a diameter of about several μm, and the liquid sample moves in the pore by capillary force.

However, the nitrocellulose membrane is derived from a natural product, and the pore diameter or pores of the membrane are not connected to each other, and the flow rate of the liquid sample in the membrane varies depending on the membrane. If the flow rate is different, the time taken for the detection of the detected substance also changes. As a result, there is a possibility that an erroneous determination that the detected substance is not detected is performed before the substance to be detected is combined with the labeling substance or the reagent. In order to solve the above problems, the industry has considered a method of manually producing a fine flow path of a liquid sample (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 of erroneous determination that the detected substance is not detected before the substance to be detected is combined with the labeling substance or reagent.

[Previous Technical Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2014-062820

[Patent Document 2] Japanese Patent No. 4957664

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2012-524894

As a method of artificially producing a flow path, hot stamping is mentioned. The hot stamping is a method in which a mold having a fine structure is pressed against a substrate (thermoplastic plastic before processing), and the fine structure is transferred onto the surface of the substrate softened by heating. A film carrier having a fine structure is produced. This fine structure functions as a flow path of the liquid sample. When hot stamping is used, a fine structure can be produced in a wide range of several tens of nm to several hundreds of μm. Furthermore, for hot stamping, there is no need for a vacuum device or the like. Large-scale equipment makes it possible to mass-produce a membrane carrier having a uniform structure inexpensively and simply. Further, the fine structure of the mold is formed in numerous recesses on the surface of the mold pressed against the substrate. The fine structure of the film carrier is a number of convex portions (protrusions) protruding from the surface of the film carrier, and has a shape corresponding to the concave portion formed on the surface of the mold. That is, the portion of the substrate filled in the concave portion of the mold serves as a fine structure (convex portion) of the film carrier.

However, the above-described hot stamping is difficult to produce a structure in which the aspect ratio of the fine structure 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 of the size Lh in the horizontal direction and the size Lv in the vertical direction is higher than 2/1 by hot stamping. The higher the aspect ratio of the fine structure, the more easily one part of the film carrier remains on the mold side when the film carrier is peeled off from the mold, or the fine structure formed on the surface of the film carrier is more likely to collapse or deform. Therefore, the higher the aspect ratio of the fine structure, the lower the productivity of the film carrier.

Further, in the case where a uniform fine structure is produced by hot stamping, it is necessary to perform fine processing of the mold with high precision and uniformity. Examples of the method of performing such microfabrication include etching, photolithography, mechanical cutting, and laser processing. However, either method requires a considerable processing fee. Further, in the case of any of the methods, the larger the volume of the metal which is cut from the flat surface of the metal member when the mold is made of the metal member, the more the processing cost increases. Therefore, the mold can be produced at low cost by minimizing the volume removed from the metal member. As a result, a film carrier having a fine structure can be produced inexpensively by hot stamping. Furthermore, the volume of metal removed from the flat surface of the metal member may also be the volume formed in the recess of the finished mold surface. Further, the volume of the concave portion formed on the surface of the mold may be, in other words, the volume of each convex portion (fine structure) formed on the surface of the film carrier by thermal embossing.

Furthermore, the more 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, the use of a liquid sample as the flow rate of the liquid sample becomes advantageous for the detection of the substance to be detected. In order to increase the flow rate of the liquid sample in the membrane carrier, the voids in the membrane carrier for the liquid sample to flow must be large. Therefore, the demand void ratio is large The fine structure. When the total volume of the voids in the fine structure of the membrane carrier is Vv, and the total volume of the fine structure itself (the convex portion itself) of the membrane carrier is expressed as Vf, the porosity of the fine structure is Rv. Expressed as 100‧Vv/(Vv+Vf). Vv can also be said to be the total of the volume of the space between the convex portions.

In summary, when the membrane carrier for the liquid sample inspection kit for the POCT reagent is produced, as a flow path for generating a flow of the liquid sample by capillary action, it is required to form an aspect ratio of less than 1: 2 (that is, 2/1), and a fine structure having a large void ratio. In the production of a mold for forming a fine structure, it is required to minimize the volume of metal scraped from the metal member.

As a method of detecting a substance to be detected, a method of detecting a color change of a detection area by an optical measuring device such as an absorbance measuring device due to coloring latex particles, fluorescent particles, or A substance to be detected in which a labeling substance such as a metal colloidal particle is combined is produced in combination with a reagent immobilized in the detection area.

However, in the above method, since the optical measuring device for performing the determination must be prepared, the method of using the POCT reagent is complicated. Further, the use of an optical measuring device is a factor for increasing the manufacturing cost of the POCT reagent.

Therefore, in order to exhibit the characteristics of the POCT reagent (inspection kit) which is capable of determining the result in a short period of time and which is simple and inexpensive to use, it is necessary to increase the color change when detecting the substance to be detected by visual inspection. degree.

In summary, when preparing a liquid sample inspection kit for the POCT reagent, the color change during detection must be increased to the extent that it can be confirmed by visual inspection. Further, as a flow path for generating a flow of the liquid sample by capillary action, it is required to form a fine structure having an aspect ratio of less than 1:2 (that is, 2/1) and a larger void ratio. In the production of a mold for forming a fine structure, it is required to minimize the volume of metal scraped from the metal member.

The present invention has been made in view of the above circumstances, and an object thereof is to provide a It is produced by hot stamping and is inexpensive, and has a flow path through which the liquid sample can be moved by capillary force, and a liquid sample inspection cover which can visually confirm the color change of the detected substance in the liquid sample at the time of detection. a membrane carrier for use; a liquid sample inspection kit using the same; and a method for preparing a liquid sample inspection kit.

That is, the present invention is as follows.

(1) A film carrier for a liquid sample inspection kit, which is a film carrier for a test kit for detecting a substance to be detected in a liquid sample, and is provided with at least one flow path for transporting a liquid sample, The bottom surface of the flow path is provided with a fine structure for generating a capillary action for transporting a liquid sample.

The fine structure is a plurality of convex portions (protrusions) that generate capillary action, or a total of a plurality of convex portions. Therefore, the above invention can also be referred to as follows.

A film carrier for a liquid sample inspection kit according to an aspect of the present invention includes a flat portion corresponding to a bottom surface of a flow path and a plurality of convex portions (protrusions) protruding from the flat portion. That is, the surface of the film carrier includes a flat portion and a plurality of convex portions (protrusions) protruding from the flat portion. The pores in the fine structure (the space between the plurality of convex portions) function as a flow path for transporting the liquid sample along the surface of the membrane carrier by the capillary action caused by the fine structure.

(2) The film carrier for a liquid sample inspection kit according to (1), which comprises a thermoplastic plastic having a glass transition temperature (Tg) of 80 to 180 °C.

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

(4) The film carrier for a liquid sample inspection kit according to (2) or (3), wherein the storage modulus of the thermoplastic plastic is 1.0 at a temperature 20 ° C higher than a glass transition temperature or a melting point. ×10 ⁷ Pa or less.

The so-called "temperature of glass transition temperature or melting point of 20 ° C" can also be said to be The glass transition temperature is 20 ° C higher, or 20 ° C higher than the melting point.

(5) The film carrier for a liquid sample inspection kit according to any one of (1) to (4), wherein the fine structure has a shape of a cone.

That is, the fine structure (convex portion) may be a cone.

(6) The film carrier for a liquid sample inspection kit according to any one of (1) to (5), wherein a diameter of a bottom surface of the fine structure is 10 to 1000 μm.

For example, in the case where the fine structure is a cone, the diameter of the bottom surface of the fine structure may be, in other words, 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, in other words, the height of the convex portion (protrusion) 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 fine structure has an aspect ratio of 10:1 to 1:2.

When the length (thickness) of the horizontal direction (short side direction) of the fine structure is expressed as Lh, and the length of the vertical direction (longitudinal direction) of the fine structure is expressed as Lv, the aspect ratio is expressed as Lv/Lh. In other words, the aspect ratio Lv/Lh of the fine structure may be 1/10 or more and 2/1 or less. For example, in the case where the microstructure is a cone, Lh may also be the diameter of the bottom surface of the cone, and Lv may also be the height of the cone from the flat portion of the membrane carrier.

(9) The film carrier for a liquid sample inspection kit according to any one of (1) to (8), wherein a ratio of a diameter of a bottom surface of the fine structure to a distance between a center of the fine structure and a center of the fine structure is larger than 1 and 5 or less.

The term "the distance between the fine structures and the closest center" may be, in other words, the distance between the centers of adjacent ones of the fine structures (convex portions). In other words, the term "the distance between the fine structures and the center closest to each other" may be referred to as the distance between the fine structures. The diameter of the bottom surface of the fine structure may be, for example, the diameter of the bottom surface of the convex portion. For example, in a fine structure as a cone In the case of a body, the so-called "the distance between the fine structures and the closest center" may be the distance between the vertices of adjacent cones.

(10) A liquid sample inspection kit using the film carrier for a liquid sample inspection kit according to any one of (1) to (9), and having a liquid test for detecting the liquid in the film carrier In the detection area of the detected substance in the sample, when the detected substance is detected in the detection area, it is possible to confirm the detected color change by visual inspection.

In other words, the liquid sample inspection kit of one aspect of the present invention comprises the above-mentioned membrane carrier of the present invention, and has a dropping region for dropping a liquid sample on the surface of the membrane carrier for detecting a substance to be detected in the liquid sample. The detection area and the above-described fine structure at least between the drop area and the detection area. The liquid sample is transported from the dropping area to the detection area by capillary action of the fine structure. If the detected substance in the liquid sample is detected in the detection area, the color of the detection area changes.

(11) The liquid sample inspection kit according to (10), wherein the detection substance is fixed in the detection area, and the detection substance is capable of detecting a substance to be detected in the detection area The color change detected has been confirmed by visual inspection.

The detection substance is, in other words, a reagent that selectively binds to the substance to be detected, or a labeling substance (color-developing 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 is displayed by a distance meter between the RGB (Red Green Blue) coordinates before and after detection. A change of more than 30.

That is, the coordinate of the color of the detection area before detecting the detected substance in the RGB color space is represented as C1, and the color of the detection area after detecting the detected substance is represented by coordinates in the RGB color space. When it is C2, the distance between the coordinate C1 and the coordinate C2 may be 30 or more.

(13) A method of producing a liquid sample inspection kit according to any one of (1) to (9), wherein the film carrier for a liquid sample inspection kit according to any one of (1) to (9) is produced by hot stamping.

That is, the method of manufacturing the liquid sample inspection kit of the present embodiment may further include the following steps (hot stamping step): by forming a surface of a mold in which a plurality of recesses are formed and a film containing thermoplastic plastic. The substrate is brought into contact with each other, and the substrate is heated to form a film carrier having a fine structure (a plurality of convex portions) corresponding to the shape of the concave portion and a flat portion.

(14) The method for producing a liquid sample inspection kit according to (13), wherein the detection substance is fixed in the detection area of the liquid sample inspection kit according to any one of (10) to (12) The detection substance is capable of confirming the detected color change by visual inspection when the detected substance is detected in the detection area.

That is, the manufacturing method of the liquid sample inspection kit of one aspect of the present invention may further comprise the step of: fixing the reagent selectively bonded to the substance to be detected in the liquid sample to the surface of the membrane carrier having the fine structure. .

The method for manufacturing a liquid sample inspection kit according to an aspect of the present invention may further comprise the step of: fixing at least one of a reagent and a labeling substance selectively bound to the substance to be detected in the liquid sample to have fineness The surface of the membrane carrier constructed.

According to the film carrier, the liquid sample inspection kit and the liquid sample inspection kit of the present invention, since the determination of the detected substance in the liquid sample can be performed by visual inspection, the inspection kit is It is easier to use. Further, since the fine structure of the film carrier can be produced by hot stamping, the film carrier is relatively inexpensive, and the inspection kit including the film carrier is useful for a POCT reagent (inspection kit) that can be used at one time.

1‧‧‧Mold with fine structure

2‧‧‧Film-like substrate containing thermoplastic plastic

3‧‧‧Film carrier with fine structure

3a‧‧‧membrane carrier

3b‧‧‧membrane carrier

3x‧‧‧Drip area

3y‧‧‧Detection area

4‧‧‧The bottom of the fine structure

5‧‧‧The closest distance between the fine structures and the center

6‧‧‧The height of the fine structure

7‧‧‧Width of the groove of the line and gap structure

8‧‧‧Ditch depth of the line and gap structure

9‧‧‧The distance between the line and the gap structure

10‧‧‧diameter of the cylindrical recesses regularly arranged

11‧‧‧The distance between the cylindrical recesses regularly arranged

12.‧‧Dr. the depth of the cylindrical recesses that are regularly arranged

13, 13a, 13b‧‧‧ flat

14‧‧‧Cone (fine structure)

14a‧‧‧tetragonal cone (fine structure)

14b‧‧‧hexeral cone (fine structure)

15‧‧‧Slots of line and gap structure

16‧‧‧ exists in the flat part between the cylindrical recesses

17‧‧‧Regularly arranged cylindrical recesses

18‧‧‧Check kits for liquid samples

18a‧‧‧Shell

18b‧‧‧First opening

18c‧‧‧second opening

(a) is a bird's-eye view (top view) of the fine structure provided in the film carrier of one embodiment of the present invention, and (b) of FIG. 1 is a fine structure described in (a) of FIG. Stereogram.

(a) is a bird's-eye view (top view) of the fine structure provided in the film carrier of one embodiment of the present invention, and (b) of FIG. 2 is a fine structure described in (a) of FIG. Stereogram.

(a) is a bird's-eye view (top view) of the fine structure provided in the film carrier of one embodiment of the present invention, and (b) of FIG. 3 is a fine structure described in (a) of FIG. Stereogram.

Fig. 4 is a cross-sectional view showing the film carrier shown in (a) and (b) of Fig. 1, which is a cross-sectional view perpendicular to the surface (flat portion) of the film carrier.

Fig. 5 (a), (b) and (c) are schematic views showing the steps of producing a film carrier according to an embodiment of the present invention by thermal imprinting.

(a) of FIG. 6 is a bird's-eye view (top view) of the structure of the mold used in the calculation of Reference Example 6, and (b) of FIG. 6 is the AA line of the structure described in (a) of FIG. Cutaway view.

(a) of FIG. 7 is a bird's-eye view (top view) of the structure of the mold used in the calculation of Reference Example 6, and (b) of FIG. 7 is the BB line of the structure described in (a) of FIG. Cutaway view.

Figure 8 is a schematic plan view of an inspection kit according to an embodiment of the present invention.

Embodiments of the present invention will be described below. The inspection kit of the present embodiment detects the substance to be detected in the liquid sample. For example, as shown in FIG. 8, the inspection kit 18 is provided with a film carrier 3 and a casing 18a accommodating the film carrier 3. On the surface of the membrane carrier 3, there is a dropping region 3x for dropping a liquid sample, and a detecting region 3y for detecting a substance to be detected in the liquid sample. The dropping region 3x is exposed in the first opening portion 18b of the casing 18a. The detection area 3y is exposed in the second opening portion 18c of the casing 18a. At least one flow path for transporting the liquid sample is provided in the membrane carrier 3, and a fine structure is provided on the bottom surface of the flow path. The fine structure is located at least between the dropping area 3x and the detecting area 3y. It may have a fine structure throughout the entire surface of the film carrier 3. The entire surface of the membrane carrier 3 may also be a flow path for a liquid sample. The fine structure produces a capillary action. The liquid sample is self-contained via a fine structure by capillary action of a fine structure The dropping area 3x is conveyed to the detection area 3y. If the detected substance in the liquid sample is detected in the detection area 3y, the color of the detection area 3y changes. As shown in Fig. 1, Fig. 2, or Fig. 3, the microstructure is a projection (14, 14a or 14b) or a plurality of projections (14, 14a or 14b). That is, the film carrier (3, 3a or 3b) is provided with 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) acts as a flow path for transporting the liquid sample along the surface of the film carrier 3 by capillary action. In other words, the voids in the fine structure (14, 14a or 14c) function as a flow path for transporting the liquid sample along the surface of the membrane carrier 3 by capillary action. The plurality of convex portions (14, 14a or 14c) may also be arranged regularly or symmetrically symmetrically on the surface of the film carrier 3.

The inspection method of the liquid sample according to one aspect of the present invention is an inspection method using the inspection kit 18, for example, the inspection kit 18 is provided with a film carrier 3 and a casing 18a accommodating the film carrier 3, and the film carrier 3 The surface has a dropping region 3x for dropping a liquid sample, a detecting region 3y for detecting a substance to be detected in the liquid sample, and a fine structure (at least a plurality of structures between the dropping region 3x and the detecting region 3y) The convex portion) is exposed in the first opening portion 18b of the casing 18a, and the detection region 3y is exposed in the second opening portion 18c of the casing 18a, and the film carrier including the fine structure (plurality of convex portions) The whole of 3 includes a thermoplastic plastic. For example, the above inspection method may include the steps of: dropping a liquid sample onto the dropping portion 3x in the surface of the film carrier 3; and forming a fine structure on the surface of the film carrier 3 The capillary action exerted by 14 (plurality of convex portions), the step of transporting the liquid sample from the dropping region 3x to the detecting region 3y via the fine structure 14, and the detection of the liquid sample during the conveying process The substance is combined with the labeling substance to Detecting binding substance immobilized on the detection region in the reagent 3y visually determining the presence or absence of the step of detecting color change in the region 3y (labeled substances color) of.

The film carrier can comprise a thermoplastic plastic. In other words, the film-form substrate comprising the thermoplastic plastic is processed by hot stamping, whereby a film carrier having a fine structure can be produced. The thermoplastic material constituting the film carrier may be, for example, at least one selected from the group consisting of a polyester resin, a polyolefin resin, a polystyrene resin, a polycarbonate resin, a fluorine resin, and an acrylic resin. The specific thermoplastic plastic may be, for example, polyethylene terephthalate (PET), cycloolefin polymer (COP), polypropylene (PP), polystyrene (PS), polycarbonate (PC), and polyposition. At least one of difluoroethylene (PVDF) and polymethyl methacrylate (PMMA).

The glass transition temperature Tg or the melting point Tm of the above thermoplastic plastic may be 80 to 180 °C. The storage modulus of the thermoplastic plastic at a temperature 20 ° C higher than the glass transition temperature Tg may be 1.0 Pa or more and 1.0 × 10 ⁷ Pa or less. The storage 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 transfer or melting of the thermoplastic plastic occurs at a temperature less than 80 ° C, and the storage modulus of the thermoplastic plastic at a temperature 20 ° C higher than the glass transition temperature or melting point is 1.0 × 10 ⁷ Pa or less, It is practically difficult to use a thermoplastic plastic in the form of a solid at room temperature, so that it is difficult to form a film carrier by hot stamping. When the glass transfer or melting of the thermoplastic plastic occurs at a temperature higher than 180 ° C, the molding temperature at the time of hot stamping is increased, and the productivity of the film carrier is lowered. That is, when the temperature required to soften the thermoplastic plastic at the time of hot stamping is higher than 180 ° C, the productivity of the film carrier is lowered. When the storage modulus of the thermoplastic plastic at a temperature 20 ° C higher than the glass transition temperature or the melting point is 1.0 × 10 ⁷ Pa or less, the molding pressure required for the production of the fine structure can be suppressed to be small, and can be relatively The fine structure is produced under mild conditions, so that the production efficiency of the membrane carrier is improved.

The fine structure provided in the film carrier may be, for example, a structure in which the cones are regularly arranged. That is, the fine structure may also be a cone. The regularly arranged cones can be formed by hot stamping using a mold. When the mold is used to form a cone, compared with the case where a groove-shaped flow path (line and gap structure) is formed using a mold, the mold is fabricated from the metal structure. The volume of metal scraped off the surface of the piece is greatly reduced, and the processing cost of the mold is reduced. In contrast, in the fabrication of a mold for forming a line and gap configuration, a large amount of metal must be cut from the metal member.

Also, the upper portion of the cone is thinner than the bottom surface of the cone. Therefore, in the case of forming a cone using a mold, the volume of metal scraped from the surface of the metal member is greatly reduced in the production of the mold as compared with the case of forming a cylinder having the same bottom surface as the cone by the mold. The processing cost of the mold is reduced.

Further, the void ratio of the fine structure in which the cones are regularly arranged is larger than the void ratio of the line and the gap structure. Further, the porosity of the fine structure in which the pyramids are regularly arranged is larger than the porosity of the structure in which a plurality of columns having the same bottom surface as the pyramid 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, and it becomes advantageous to detect the detected substance.

The shape of the bottom surface of the cone (fine structure) can be freely selected. For example, as shown in (a) and (b) of FIG. 1, the fine structure may be a cone 14. For example, as shown in (a) and (b) of FIG. 2, the fine structure may be a quadrangular pyramid 14a. For example, as shown in (a) and (b) of FIG. 3, the fine structure may be a hexagonal cone 14b. In order to facilitate the processing of the mold and to suppress the processing cost, the bottom surface of the cone is preferably circular or polygonal (for example, square, diamond, rectangular, triangular, or hexagonal, etc.).

The diameter of the bottom surface of the above fine structure may be 10 to 1000 μm. When the diameter of the bottom surface of the fine structure is less than 10 μm, the micro-machining cost of the mold becomes high, and it is difficult to uniformly produce a myriad of fine structures on the surface of the film carrier having a large area. Therefore, the microstructure which is too small is not suitable for practical use. Further, when the diameter of the bottom surface of the fine structure is less than 10 μm, the capillary force required 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 off from the metal member at the time of production of the mold increases, and the production cost of the mold and the film carrier becomes high. Further, when the diameter of the bottom surface of the fine structure is larger than 1000 μm, the area of the flow path in the film carrier must also be increased, and the liquid The sample inspection kit is enlarged, which becomes unfavorable for the delivery of the liquid sample inspection kit itself. As shown in FIG. 1(a) or FIG. 4, in the case where the fine structure is the cone 14, the diameter of the bottom surface of the fine structure may be the diameter 4 of the bottom surface (circle) of the cone 14.

The height of the above fine structure may be 10 to 500 μm. When the height of the fine structure is less than 10 μm, there is a tendency that the capillary force required to move the liquid sample is weakened. When the height of the fine structure is more than 500 μm, it is difficult to completely fill the thermoplastic into the concave portion of the mold (the recess corresponding to the shape of the fine structure) at the time of hot stamping. As shown in FIG. 1(a) or FIG. 4, in the case where the fine structure is the cone 14, the height of the fine structure may be the height 6 of the cone 14 from the flat portion 13.

The shape of the entire film carrier 3, 3a or 3b is not particularly limited, and may be, for example, a polygonal shape such as a square shape, a circular shape, or an elliptical shape. In the case where the film carrier 3, 3a or 3b is quadrangular, the longitudinal width of the film carrier 3, 3a or 3b may be, for example, 2 to 100 mm, and the lateral width of the film carrier 3, 3a or 3b may be, for example, 2 to 100 mm. The thickness of the film carrier 3, 3a or 3b from which the height of the fine structure is removed may be, for example, 0.1 to 10 mm.

The aspect ratio of the above fine structure 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 less than 10:1 (i.e., 1/10), the contact area between the liquid sample and the flow path is small, and the capillary force is reduced, so that the liquid sample is less likely to move. When the aspect ratio is greater than 1:2 (i.e., 2/1), the productivity of the film carrier using hot stamping is lowered. As shown in FIG. 1(a) or FIG. 4, when the fine structure is a cone, the length Lh of the fine structure in the horizontal direction may be the diameter 4 of the bottom surface of the cone 14. Further, the length Lv of the fine structure in the vertical direction may be the height 6 of the cone 14 from the flat portion 13 of the film carrier 3.

The ratio D2/D1 of the diameter (D1) of the bottom surface of the fine structure to the closest center-to-center distance (D2) of the fine structures may be greater than 1 and 5 or less. It is impossible to be 1 or less than D2/D1. When the ratio D2/D1 is larger than 5, the contact area between the liquid sample and the flow path is reduced, the capillary force is reduced, and there is a tendency that the liquid sample is not easily moved. As shown in Figure 1 (a) or Figure 4, In the case where 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 distance D2 closest to the center may also be the distance between the vertices of the adjacent pair of cones 5 . The diameter D1 of the bottom surface of the fine structure may coincide with the length Lh of the fine structure in the horizontal direction. Therefore, the aspect ratio Lv/Lh can also be expressed as Lv/D1.

As shown in (a), (b) and (c) of FIG. 5, the method for manufacturing a liquid sample inspection kit of the present embodiment may include the following steps (hot stamping step): forming a plurality of recesses The surface of the mold (mold 1) is in contact with the substrate 2 including the thermoplastic plastic film, and the substrate 2 is heated to produce a fine structure (plurality of convex portions (14)) corresponding to the shape of the concave portion. The film carrier 3 with the flat portion 13. The method of manufacturing a liquid sample inspection kit may further comprise the step of immobilizing the reagent or labeling substance in a detection area in the surface of the membrane carrier having a fine structure. If the substance to be detected in the liquid sample is combined with the labeling substance, and the substance to be detected is selectively combined with the reagent of the detection area, the color of the detection area changes. Therefore, by visually detecting the color change of the detection area, it is confirmed that the detected substance in the liquid sample has been detected.

The microfabrication method of the mold used in the hot stamping step may be, for example, etching, photolithography, mechanical cutting, or laser processing. Microfabrication methods suitable for processing dimensions or processing ranges can be selected.

It is preferable to perform mold release treatment of the mold before performing hot stamping. Regarding the mold release treatment, for example, a single molecule film is formed on the surface of the mold to reduce the surface energy. As a result, after the thermal imprinting, the film carrier 3 containing the thermoplastic plastic is easily peeled off from the surface of the mold 1.

The hot stamping method may be either a flat press type or a roll type. The hot stamping method shown in Fig. 5 is a flat plate press type. With respect to the flat plate press type, the mold 1 is overlapped with the substrate 2 containing the thermoplastic plastic between the platforms which are opposed to each other in parallel, and the sheets are sandwiched between the platforms. Then, the mold 1 and the substrate 2 are heated via a platform and pressurized. Such a flat plate press type is excellent in terms of precision in molding. Roller type is heated The roll mold is formed by the pressing pressure of the rolls. The roll type is excellent in productivity.

The conditions such as the molding temperature, the molding pressure, and the transfer time at the time of hot embossing 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 temperature Tg or a temperature 20 to 50 ° C higher than the melting point Tm. The molding pressure can be 1 to 10 MPa. The transfer time (the time for which the mold 1 and the substrate 2 are held while being pressurized) may be 3 to 10 minutes. The fine structure of the mold 1 is easily and accurately transferred to the surface of the substrate 2 by the hot embossing under the above conditions.

Depending on the type of the thermoplastic material constituting the film carrier 3 and the type of the reagent (detection substance), it may be difficult to fix the reagent (detection substance) which can be determined by visual inspection to the detection area of the film carrier 3. in. In this case, the reagent (detection substance) is easily fixed in the detection area of the membrane carrier 3 by performing appropriate surface treatment only on the detection area in advance.

The surface treatment method of the detection area is not limited, and may be, for example, various plasma treatment, UV (Ultra Violet) treatment, UV/ozone treatment, or using 3-aminopropyltriethoxydecane (3- Various methods such as surface modification of Aminopropyltriethoxysilane or Glutaraldehyde.

The reagent (detection substance) immobilized in the detection area may be, for example, an antibody. The anti-system reacts with the antigen-antibody of the substance to be detected. The antibody may be a plurality of antibodies or a monoclonal antibody. The substance to be detected is not limited, and may be any substance capable of reacting with an antigen-antibody of an antibody, such as various pathogens and various clinical markers. Specific examples of the test substance may be influenza virus, Norovirus, adenovirus, RS (Respiratory Syncytial Virus), HAV (Hepatitis A Virus), and HBs (Hepatitis B). Surface antigen, hepatitis B surface antigen), HIV (Human Immunodeficiency Virus) original. The substance to be detected may also be produced by MRSA (Methicillin Resistant Staphylococcus Aureus, methicillin-resistant Staphylococcus aureus), Group A Streptococcus, Group B Streptococcus, Legionella vulgaris, and other bacterial antigens, bacteria, and the like. toxin. The substance to be tested may also be a hormone such as mycoplasma, Chlamydia trachomatis, human chorionic gonadotropin. The substance to be detected may also be a C-reactive protein, a myoglobin, a cardiac troponin, various tumor markers, a pesticide, and an environmental hormone. In particular, when the detection of a substance to be detected such as influenza virus, norovirus, C-reactive protein, myoglobin, and cardiac troponin is urgently required, and the treatment measures for diseases caused by the same, the present embodiment is The usefulness of the inspection kit is particularly large. Furthermore, the substance to be detected may be an antigen which can induce an immune response alone. The substance to be detected may be a hapten that can induce an immune response alone, but can bind to the antibody by reacting the antibody with the antigen-antibody.

The coordinate of the color of the detection area before detecting the detected substance in the RGB color space (RGB coordinate space) is represented as C1, and the color of the detection area after detecting the detected substance is in the RGB color space. When the coordinate is expressed as C2, the distance between the coordinate C1 and the coordinate C2 in the RGB color space is preferably 30 or more. The coordinate C1 is obtained by measuring the R value (Red value), the G value (Green value), and the B value (Blue value) of the detection area before detection. The coordinate C2 is obtained by measuring the R value, the G value, and the B value of the detected detection area. The distance between the coordinate C1 and the coordinate C2 in the RGB color space is calculated by calculation. The distance between the RGB coordinates may also be the chromaticity difference of the detection area before and after the detection of the detected substance. When the chromaticity difference is less than 30, it is difficult to visually confirm the difference in color and the tendency to change.

[Examples]

Hereinafter, the present invention will be specifically described by way of examples and comparative examples of the invention, but the invention is not limited to the examples.

[Example 1]

<Preparation of the mold>

A mold is produced by laser processing of a metal member. The metal member is made of aluminum alloy A5052. In the laser processing, a plurality of inverted conical recesses (i.e., fine structures) are formed at the center portion of the flat surface of the mold (the range of a square of 3 cm × 3 cm). Any of the recesses has the same shape. The diameter of the recess is 10 μm. The diameter of the recess is equal to the diameter (D1 or Lh) of the bottom surface of the fine structure (cone) of the film carrier finally obtained. The distance between the centers of one of the adjacent pairs of recesses was 15 μm. The distance between the centers of the recesses is equal to the distance (D2) between the apexes of the fine structures (cones) adjacent to the surface of the film carrier. The depth of the recess is 10 μm. The depth of the recess is equal to the height (hv) of the fine structure (cone) of the film carrier. The plurality of recesses are regularly arranged in the center portion of the mold in the same manner as the arrangement of the cones 14 (triangular arrangement) shown in FIG.

The surface (the uneven surface) of the mold in which the concave portion is formed is subjected to a mold release treatment. In the mold release treatment, the surface (concave surface) of the mold was immersed in the treatment liquid for about 1 minute, and then the surface of the mold was dried. The dried mold was allowed to stand overnight. The treatment liquid used in the above-mentioned mold release treatment was OPTOOL HD-2100TH manufactured by Daikin Industries.

<Hot embossing step (transfer of fine structure)>

The fine structure of the surface of the mold is transferred to the surface of the substrate containing the thermoplastic plastic film by the following hot stamping step. In the hot stamping step, the X-300 manufactured by SCIVAX Corporation was used. In the hot stamping step, the surface of the mold on which the fine structure (plurality of recesses) is formed is brought into contact with the substrate containing the thermoplastic plastic film, and the mold and the substrate are heated while being heated. The molding temperature was 120 °C. The applied pressure was 5.5 MPa. The transfer time is 5 minutes. After the transfer of the fine structure, the mold and the substrate were cooled to 80 ° C while applying pressure to the mold and the substrate. The pressure is released after cooling. The film carrier of Example 1 was obtained by the above thermal imprinting step. The film carrier has a surface comprising a plurality of cones (fine structures) and flat portions. The shape and size of the convex portion (cone) present on the surface of the film carrier coincide with the shape and size of the concave portion (inverted cone) formed in the mold.

A film-like substrate comprising a thermoplastic plastic is a film comprising polystyrene (PS) (electrification) Denka styrene sheet manufactured by Industrial Industries). The thickness of the substrate was 188 μm. The substrate (membrane carrier) has a square shape. The substrate (film carrier) had a longitudinal width of 50 mm and the substrate (membrane carrier) had a lateral width of 50 mm.

The glass transition temperature Tg of the polystyrene constituting the substrate (film carrier) was measured using DSC3100 manufactured by Bruker AXS. In the measurement of the glass transition temperature Tg, the substrate was heated at a temperature elevation rate of 10 ° C/min under a nitrogen atmosphere. The glass transition temperature Tg of polystyrene was 106 °C.

The storage modulus of the polystyrene constituting the substrate (membrane carrier) in the tensile mode was measured. RSAIII manufactured by TA Instruments was used for the measurement of the storage modulus. The frequency at which the storage modulus was measured was 1 Hz. The storage modulus of polystyrene at 126 ° C was 1.8 × 10 ⁶ Pa.

[Embodiment 2]

The diameter of the concave portion formed on the surface of the mold of Example 2 was 10 μm. The distance between the centers of the adjacent ones of the recesses on the surface of the mold of Example 2 was 50 μm. The depth of the concave portion formed on the surface of the mold of Example 2 was 10 μm. The film carrier of Example 2 was produced by the same method as in Example 1 except that the mold was different.

[Example 3]

The diameter of the concave portion formed on the surface of the mold of Example 3 was 10 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Example 3 was 15 μm. The depth of the concave portion formed on the surface of the mold of Example 3 was 20 μm. The film carrier of Example 3 was produced by the same method as in Example 1 except that the mold was different.

[Example 4]

The diameter of the concave portion formed on the surface of the mold of Example 4 was 10 μm. The distance between the centers of adjacent ones of the recesses on the surface of the mold of Example 4 was 50 μm. The depth of the concave portion formed on the surface of the mold of Example 4 was 20 μm. The film carrier of Example 4 was produced by the same method as in Example 1 except that the mold was different.

[Example 5]

The diameter of the concave portion formed on the surface of the mold of Example 5 was 100 μm. The distance between the centers of adjacent ones of the recesses on the surface of the mold of Example 5 was 110 μm. The depth of the concave portion formed on the surface of the mold of Example 5 was 10 μm. The film carrier of Example 5 was produced in the same manner as in Example 1 except that the mold was different.

[Embodiment 6]

The diameter of the concave portion formed on the surface of the mold of Example 6 was 100 μm. The distance between the centers of the adjacent ones of the recesses on the surface of the mold of Example 6 was 500 μm. The depth of the concave portion formed on the surface of the mold of Example 6 was 10 μm. The film carrier of Example 6 was produced by the same method as in Example 1 except that the mold was different.

[Embodiment 7]

The diameter of the concave portion formed on the surface of the mold of Example 7 was 1000 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Example 7 was 1010 μm. The depth of the concave portion formed on the surface of the mold of Example 7 was 100 μm. The film carrier of Example 7 was produced in the same manner as in Example 1 except that the mold was different.

[Embodiment 8]

The diameter of the concave portion formed on the surface of the mold of Example 8 was 1000 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Example 8 was 5000 μm. The depth of the concave portion formed on the surface of the mold of Example 8 was 100 μm. The film carrier of Example 8 was produced by the same method as in Example 1 except that the mold was different.

[Embodiment 9]

The diameter of the concave portion formed on the surface of the mold of Example 9 was 250 μm. The distance between the centers of adjacent ones of the recesses on the surface of the mold of Example 9 was 260 μm. The depth of the concave portion formed on the surface of the mold of Example 9 was 500 μm. The film carrier of Example 9 was produced by the same method as in Example 1 except that the mold was different.

[Embodiment 10]

The diameter of the concave portion formed on the surface of the mold of Example 10 was 250 μm. In the embodiment The distance between the centers of one of the adjacent recesses on the surface of the mold of 10 is 1250 μm. The depth of the concave portion formed on the surface of the mold of Example 10 was 500 μm. The film carrier of Example 10 was produced by the same method as in Example 1 except that the mold was different.

[Example 11]

The diameter of the concave portion formed on the surface of the mold of Example 11 was 1000 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Example 11 was 1010 μm. The depth of the concave portion formed on the surface of the mold of Example 11 was 500 μm. The film carrier of Example 11 was produced by the same method as in Example 1 except that the mold was different.

[Embodiment 12]

The diameter of the concave portion formed on the surface of the mold of Example 12 was 1000 μm. The distance between the centers of adjacent ones of the recesses on the surface of the mold of Example 12 was 5000 μm. The depth of the concave portion formed on the surface of the mold of Example 12 was 500 μm. The film carrier of Example 12 was produced by the same method as in Example 1 except that the mold was different.

[Examples 13 to 24]

In Examples 13 to 24, a film containing a polycarbonate (PC) was used as a film comprising a thermoplastic plastic film instead of a film containing polystyrene. As a film containing polycarbonate, Panlite manufactured by Teijin Co., Ltd. was used. The polycarbonate substrate was included to have a thickness of 200 μm. The substrate (membrane carrier) has a square shape. The substrate (film carrier) had a longitudinal width of 50 mm and the substrate (membrane carrier) had a lateral width of 50 mm.

The glass transition temperature Tg of the polycarbonate constituting the substrate (film carrier) was measured using DSC3100 manufactured by Bruker AXS. In the measurement of the glass transition temperature Tg, the substrate was heated at a temperature elevation rate of 10 ° C/min under a nitrogen atmosphere. The glass transition temperature Tg of the polycarbonate was 160 °C.

The storage modulus of the polycarbonate constituting the substrate (film carrier) in the tensile mode was measured. RSAIII manufactured by TA Instruments was used for the measurement of the storage modulus. The frequency at which the storage modulus was measured was 1 Hz. The storage modulus of the polycarbonate at 180 ° C was 4.5 × 10 ⁶ Pa.

The film carrier of Example 13 was produced in the same manner as in Example 1 except that the film-form substrate was different. The film carrier of Example 14 was produced in the same manner as in Example 2 except that the film-form substrate was different. The film carrier of Example 15 was produced by the same method as in Example 3 except that the film-form substrate was different. The film carrier of Example 16 was produced in the same manner as in Example 4 except that the film-like substrate was different. The film carrier of Example 17 was produced in the same manner as in Example 5 except that the film-form substrate was different. The film carrier of Example 18 was produced in the same manner as in Example 6 except that the film-form substrate was different. The film carrier of Example 19 was produced in the same manner as in Example 7 except that the film-form substrate was different. The film carrier of Example 20 was produced in the same manner as in Example 8 except that the film-form substrate was different. The film carrier of Example 21 was produced in the same manner as in Example 9 except that the film-form substrate was different. The film carrier of Example 22 was produced in the same manner as in Example 10 except that the film-like substrate was different. The film carrier of Example 23 was produced in the same manner as in Example 11 except that the film-form substrate was different. The film carrier of Example 24 was produced by the same method as in Example 12 except that the film-form substrate was different.

<Fixation of reagent (antibody) to detection area>

In the surface of the film carrier (the surface having the fine structure) of each of the examples, only the detection area A and the detection area B described below were exposed by the following method, and the other portions were covered with a mask. Then, the detection area A and the detection area B are subjected to UV processing.

Detection area A: a linear portion having a distance of 0.6 cm from the lower end of the film carrier and a width of about 1 mm.

Detection area B: a line-shaped portion having a distance of 1.0 cm from the lower end of the film carrier and a width of about 1 mm.

Suspension A of anti-influenza A NP (Nuclear Protein) antibody (Reagent A) was applied to the detection area A. Then, the detection area A is sufficiently dried under warm air, and the anti-influenza A NP antibody is immobilized in the detection area A. The coating amount of the suspension A was 18 μL. Investigate The length of the portion of the measurement area A coated with the suspension A was 3 cm.

Suspension B of anti-influenza B NP antibody (Reagent B) was applied to detection zone B. Then, the detection area B is sufficiently dried under warm air, and the anti-influenza B NP antibody is immobilized in the detection area B. The coating amount of the suspension B was 18 μL. The portion of the detection area B coated with the suspension B was 3 cm in length.

<Preparation of labeled substance>

A purified anti-influenza A virus NP antibody (refined antibody A) different from the above-described anti-influenza A NP antibody (Reagent A) was prepared. Further, a purified anti-influenza B virus NP antibody (purified antibody B) different from the above-described anti-influenza B NP antibody (reagent B) was prepared.

The purified antibody A was labeled by the covalent bond between the purified antibody A and the blue latex particles (manufactured by CM/BL CERADYNE). The particle size of the blue latex particles was 0.394 μm. Prepare a trishydroxymethylaminomethane buffer containing sugar, surfactant and protein. The labeled purified antibody A was added to trishydroxymethylaminomethane buffer in such a manner that the concentration of the latex particles in the trimethylolamine methane buffer was 0.025 w/v% to make trimethylol. The amino methane buffer was suspended. Then, sonication of trishydroxymethylaminomethane buffer was carried out to prepare an anti-type A marker which was sufficiently dispersed and suspended in trishydroxymethylaminomethane buffer.

An anti-B-type marker sufficiently dispersed in a buffer of trishydroxymethylaminomethane was prepared by the same method as in the case of the anti-type A label, except that the purified antibody B was used instead of the purified antibody A.

A mixture of an anti-type A marker and an anti-type B marker is prepared. The mixture was applied to glass fibers. The size of the glass fiber is 3 cm x 1 cm. The coating amount per 1 cm 2 of the glass fiber mixture was 50 μL. As the glass fiber, 33GLASS No. 10539766 manufactured by Schleicher & Schuell was used. The glass fiber coated with the mixed solution was sufficiently dried under a warm air to prepare a marker mat. The label material pad was overlapped at the end of each of the film carriers of Examples 1 to 24 on the side close to the detection region. The width of the end of the film carrier where the marking substance pads overlap is 2mm. The film carrier in which the marker substance pads were overlapped by a cutter was cut into short strips having a width of 5 mm, and a liquid sample inspection kit including the integrated film carrier and the marker substance pad was prepared.

<Evaluation of detection performance>

As a diluted solution, a sample suspension attached to QuickNavi-Flu manufactured by DENKA SEIKEN was prepared. Influenza A virus A/Beijing/32/92 (H3N2) was used as a test substance. Liquid sample A was prepared by diluting the influenza A virus to 4 × 10 ⁴ times with a sample suspension. Influenza B virus B/Shangdong/7/97 was used as another test substance. Liquid sample B was prepared by diluting the influenza B virus to 4 × 10 ³ times with a sample suspension.

100 μL of each of the liquid samples A and B were individually dropped at the ends of the liquid sample inspection kits of Examples 1 to 24, respectively. The end of the inspection kit to which the liquid sample is dropped is adjacent to the end of the detection area A and the detection area B side. The digital camera is used to record the movement of the dropped liquid sample on the inspection set from the top. Based on the animation, the flow rate of the liquid sample moving on the inspection set is calculated. The evaluation results of the flow rates of the liquid samples of the test kits of Examples 1 to 24 are shown in Tables 1 and 2.

Embodiments in which the double circle symbols are marked in each table are superior to the embodiments marked with circle symbols in terms of the effect of the invention. Embodiments marked with a circle symbol in each table are superior to the embodiment marked with a triangular symbol in terms of the effect of the invention. The embodiment in which the triangular symbols are attached in each table is superior to the reference example marked with the X symbol in terms of the effect of the invention.

<Determination of detection>

5 minutes after the liquid samples A and B were added to each of the inspection kits of Examples 1 to 24, the presence or absence of the color of each of the detection area A and the detection area B in each inspection set was visually observed. . From this observation, it was determined whether or not each of the influenza A virus and the influenza B virus was detected.

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

[Example 25]

The four liquid sample inspection kits of Example 25 were produced in exactly the same manner as in the case of Example 1. Then, the dilution ratios of the influenza A virus were prepared to be four liquid samples of 1 × 10 ⁴ , 2 × 10 ⁴ , 4 × 10 ^{4 ,} and 8 × 10 ⁴ . The preparation method of the liquid sample is the same as the preparation method of the above liquid sample A except for the dilution ratio.

By the same method as in the case of Examples 1 to 24, only one of the four liquid samples was dropped on the inspection set of one of the examples 25, and was detected in the detection area A of each inspection set. To the influenza A virus. As shown in Table 3, the detection area A of the inspection set in which only the liquid sample having the dilution ratio of 8 × 10 ⁴ was dropped was not shown to have a color change confirmed by visual inspection. In any of the detection areas A of the inspection set to which the other three liquid samples were dropped, the color change was confirmed by visual inspection.

Before the detection of the influenza A virus, the R value, the G value and the B value of the detection area A of the four inspection kits were measured by a laser microscope. As a laser microscope, OPLETICS HYBRID manufactured by Lasertec Corporation was used. Further, after the detection of the influenza A virus, the R value, the G value, and the B value of the detection regions A of the four test kits were measured by a laser microscope. Based on the measurement results, the distance between the RGB coordinates before and after the detection of each of the detection areas A of the four inspection sets is calculated. The calculation results are shown in Table 3.

[Example 26]

The four liquid sample inspection kits of Example 26 were produced in exactly the same manner as in the case of Example 1. Then, the dilution ratios of the preparation of the influenza B virus were four liquid samples of 1 × 10 ³ , 2 × 10 ³ , 4 × 10 ³ and 8 × 10 ³ . The preparation method of the liquid sample is the same as the preparation method of the above liquid sample B except for the dilution ratio.

By the same method as in the case of Examples 1 to 24, only one of the four liquid samples was dropped on one of the inspection kits of Example 26, and was detected in the detection area B of each inspection set. To the influenza B virus. As shown in Table 3, the detection area B of the inspection set in which only the liquid sample having the dilution ratio of 8 × 10 ³ was dropped was not shown to have a color change confirmed by visual inspection. In any of the detection areas B of the inspection set to which the other three kinds of liquid samples were dropped, the color change was confirmed by visual inspection.

Before the detection of the influenza B virus, the R value, the G value, and the B value of each of the detection regions B of the four inspection kits were measured using the above-described laser microscope. Further, after the detection of the influenza B virus, the R value, the G value, and the B value of the detection regions B of the four test kits were measured by a laser microscope. Based on the measurement results, the distance between the RGB coordinates before and after the detection of each of the detection areas B of the four inspection sets is calculated. The calculation results are shown in Table 3.

[Reference Example 1]

In Reference Example 1, a film containing polytetrafluoroethylene (PTFE) was used as a substrate containing a film of a thermoplastic plastic. As a film containing polytetrafluoroethylene, Polyflon F-104 manufactured by Daikin Industries, Inc. was used. The thickness of the polytetrafluoroethylene material was 200 μm. The substrate (membrane carrier) has a square shape. The substrate (film carrier) had a longitudinal width of 50 mm and the substrate (membrane carrier) had a lateral width of 50 mm. The melting point Tm of the polytetrafluoroethylene constituting the substrate (film carrier) was measured using DSC3100 manufactured by Bruker AXS. With respect to the measurement of the melting point Tm, the substrate was heated at a temperature elevation rate of 10 ° C/min under a nitrogen atmosphere. The melting point Tm of polytetrafluoroethylene was 327 °C. The molding temperature in the hot imprinting step of Reference Example 1 was 250 °C as the limit value of the apparatus. A film carrier of Reference Example 1 was produced in the same manner as in Example 1 except the above. The characteristics of the membrane carrier of Reference Example 1 are shown in Table 4 below.

[Reference Example 2]

The diameter of the concave portion formed on the surface of the mold of Reference Example 2 was 10 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Reference Example 2 was 15 μm. The depth of the concave portion formed on the surface of the mold of Reference Example 2 was 30 μm. The film carrier of Reference Example 2 was produced by the same method as in Example 1 except that the mold was different. Each of the membrane carriers of Reference Example 2 The characteristics are shown in Table 4 below.

[Reference Example 3]

The diameter of the concave portion formed on the surface of the mold of Reference Example 3 was 1000 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Reference Example 3 was 1010 μm. The depth of the concave portion formed on the surface of the mold of Reference Example 3 was 2000 μm. The film carrier of Reference Example 3 was produced by the same method as in Example 1 except that the mold was different. The characteristics of the membrane carrier of Reference Example 3 are shown in Table 4 below.

The film carriers of Reference Examples 1 to 3 were observed. In the film carrier of Reference Example 1, the desired fine structure was not formed. The reason for this is that the melting point of PTFE is higher than 180 ° C, and the elastic modulus of PTFE is still high even if the molding temperature in the hot stamping step is set to the upper limit of the apparatus. In the film carrier of Reference Example 2, the breakage and bending of the fine structure were observed. The reason for this is that the aspect ratio of the fine structure is more than 2, and the film carrier cannot be smoothly peeled off from the mold. In the film carrier of Reference Example 3, the height of the fine structure did not reach 2000 μm. The reason for this is that the depth of the concave portion formed on the surface of the mold of Reference Example 3 is more than 500 μm, and PS is not sufficiently filled into the concave portion of the mold.

[Reference Example 4]

The diameter of the concave portion formed on the surface of the mold of Reference Example 4 was 100 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Reference Example 4 was 1000 μm. Formed on The depth of the concave portion of the surface of the mold of Reference Example 4 was 200 μm. The film carrier of Reference Example 4 was produced by the same method as in Example 1 except that the mold was different. The characteristics of the membrane carrier of Reference Example 4 are shown in Table 5 below.

[Reference Example 5]

The diameter of the concave portion formed on the surface of the mold of Reference Example 5 was 1000 μm. The distance between the centers of the adjacent pairs of the recesses on the surface of the mold of Reference Example 5 was 1010 μm. The depth of the concave portion formed on the surface of the mold of Reference Example 5 was 50 μm. The film carrier of Reference Example 5 was produced by the same method as in Example 1 except that the mold was different. The 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 to the fine structure of each of the film carriers of Reference Examples 4 and 5 was observed from the upper side by a digital camera. In any of the film carriers of Reference Examples 4 and 5, none of the liquid samples moved. The reason why the liquid sample has not moved is as follows. In Reference Example 4, the ratio D2/D1 of the diameter D1 of the bottom surface of the fine structure to the closest center-to-center distance D2 between the fine structures is more than 5, and no capillary force sufficient to move the liquid sample is generated. In Reference Example 5, the aspect ratio of the fine structure was less than 0.1, and no capillary force sufficient to move the liquid sample was generated.

[Reference Example 6]

Calculated by forming a line and gap structure on the surface of the film carrier by a metal member The volume of metal that is removed from the surface of the metal member during the mold. The line and gap structure has a structure in which a plurality of groove-shaped flow paths are arranged in parallel. The mold itself for forming the line and gap structure also has a line and gap configuration as shown in (a) and (b) of Fig. 6. The width 7 of the groove 15 used for the calculation is 10 μm. The depth 8 of the groove 15 used for the calculation is 10 μm. The distance between the grooves 15 for calculation is 9 μm. In the calculation, the range of the surface of the machined mold is assumed to be a square of 3 cm x 3 cm. Further, the void ratio of the line and gap structure formed using the mold of Fig. 6 was also calculated. The calculation results are shown in Table 6.

Further, the volume of the metal scraped off from the surface of the metal member when the mold is formed of a metal member to form a plurality of cylinders regularly arranged on the surface of the film carrier is calculated. The mold has a flat portion 16 and a plurality of cylindrical recesses 17 as shown in (a) and (b) of FIG. The diameter 10 of the recess 17 for calculation is 10 μm. The depth 12 of the recess 17 used for the calculation is 10 μm. The distance 11 between the recesses 17 for calculation was 15 μm. In the calculation, the range of the surface of the machined mold is assumed to be a square of 3 cm x 3 cm. The calculation results are shown in Table 6. Further, the void ratio of the structure in which a plurality of columns formed by using the mold of Fig. 7 are regularly arranged is also calculated. The calculation results are shown in Table 6.

The volume of the metal removed from the metal member at the time of producing the mold of Example 1 is shown in Table 6. The void ratio of the fine structure of the membrane carrier of Example 1 is shown in Table 6.

As shown by the results of Tables 1 to 5, the liquid sample inspection kit of the present invention can be produced by hot stamping, and the dropped liquid sample can be moved by capillary force. Moreover, according to the present invention, the color change in the detection area is also large enough to be confirmed by visual inspection. Enter From the results of Table 6, it is shown that by using the fine structure as a cone, the volume of the metal scraped off during the mold production is reduced, and the void ratio of the fine structure of the film carrier is increased.

[Industrial availability]

According to the film carrier and the liquid sample inspection kit of the present invention, since the determination of the detected substance in the liquid sample can be performed by visual inspection, the method of using the inspection kit is relatively easy. Further, since the fine structure of the film carrier can be produced by hot stamping, the film carrier is relatively inexpensive, and the inspection kit including the film carrier is useful for a POCT reagent (inspection kit) that can be used at one time.

3‧‧‧Film carrier with fine structure

4‧‧‧The bottom of the fine structure

5‧‧‧The closest distance between the fine structures and the center

6‧‧‧The height of the fine structure

13‧‧‧ Flat Department

14‧‧‧Cone (fine structure)

Claims (14)

A membrane carrier for a liquid sample inspection kit, which is a membrane carrier for a test kit for detecting a substance to be detected in a liquid sample, and is provided with at least one flow path for transporting the liquid sample, in the flow path The bottom surface is provided with a fine structure for generating a capillary action for transporting the liquid sample. The film carrier for a liquid sample inspection kit of claim 1, which comprises a thermoplastic plastic having a glass transition temperature of 80 to 180 °C. A film carrier for a liquid sample inspection kit according to claim 1, which comprises a thermoplastic plastic having a melting point of 80 to 180 °C. The film carrier for the liquid sample inspection kit of claim 2 or 3, wherein the storage modulus of the thermoplastic plastic is 1.0 × 10 ⁷ Pa or less at a temperature 20 ° C higher than the glass transition temperature or melting point. The film carrier for a liquid sample inspection kit according to any one of claims 1 to 3, wherein the shape of the fine structure is a cone. The film carrier for a liquid sample inspection kit according to any one of claims 1 to 3, wherein a diameter of a bottom surface of the fine structure is 10 to 1000 μm. The film carrier for a liquid sample inspection kit according to any one of claims 1 to 3, wherein the height of the fine structure is 10 to 500 μm. The film carrier for a liquid sample inspection kit according to any one of claims 1 to 3, wherein the fine structure has an aspect ratio of 10:1 to 1:2. The film carrier for a liquid sample inspection kit according to any one of claims 1 to 3, wherein a ratio of a diameter of a bottom surface of the fine structure to a distance between a center of the fine structures and a center of the fine structure is greater than 1 and 5 or less. A liquid sample inspection kit using the liquid according to any one of claims 1 to 9 The sample inspection kit uses a membrane carrier, and the membrane carrier has a detection region for detecting a substance to be detected in the liquid sample, and when the detected substance is detected in the detection region, the generation can be Visually confirm the detected color change. The liquid sample inspection kit of claim 10, wherein the detection substance is fixed in the detection area, and the detection substance is capable of confirming by visual inspection when the detection substance is detected in the detection area The color change detected. The liquid sample inspection kit of claim 10 or 11, wherein the color change shows a change of more than 30 by the distance between the detected and the detected RGB coordinates. A method for producing a liquid sample inspection kit, which is characterized in that the film carrier for a liquid sample inspection kit according to any one of claims 1 to 9 is produced by hot stamping. The method of manufacturing the liquid sample inspection kit of claim 13, wherein the detection substance is fixed in the detection area of the liquid sample inspection kit of any one of claims 10 to 12, wherein the detection substance is When the detected substance is detected in the detection area, it is possible to confirm that the color change has been detected by visual inspection.