JP2000035428A - Pollen detecting device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To easily collect indoor dust and to easily detect the presence of pollen by putting the collected dust in a detecting device. SOLUTION: This detecting device 1 comprises a base material 101 made of a porous material in which a pollen sample liquid can be developed. The base material 101 has a sample liquid application part 102 in which the pollen sample liquid is applied, a first antibody carrying-on part 103 in which a first antibody capable of specific connection with protein derived from pollen is fixed, a sample liquid development delay part 104, a positive decision part 105 in which a second antibody capable of specific connection with the protein derived from the pollen different from the first antibody is fixed, a negative decision part 106, and water-insoluble parts 107a, 107b in which the sample liquid can not be developed so that the sample liquid converges to the positive decision part 105.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus which can easily detect the presence or absence of pollen, and can be used for detecting pollen floating in the air.

[0002]

2. Description of the Related Art Pollen floating in the air is considered to cause asthma and various allergies. Therefore, knowing the presence or absence of pollen or the type of pollen is considered useful for preventing and treating asthma and various allergies. As a method for detecting pollen, a technician with specialized knowledge collects dust in the air and directly observes it with a microscope.However, devices or instruments that can be easily handled by people without specialized knowledge are In particular, there is a demand for a device or an instrument that can easily detect pollen even at home.

[0003]

The pollen floating in the air causes asthma and various allergies, and it is difficult to know the presence or absence of the indoor pollen or the type of pollen.
Useful for prevention and treatment of asthma and allergies.
However, in order to detect indoor pollen, it is necessary to collect dust on the air on filter paper or a microfilter using a suction device and observe it directly with a microscope or the like. Very difficult for those who do not. Further, the detection device is expensive, and its handling method also requires considerable skill. Therefore, it is easy to collect indoor dust,
A device that can easily detect the presence or absence of pollen by putting the dust into a detection device has been desired.

[0004]

According to the present invention, in order to more easily determine the presence or absence of pollen, the configuration of the detection device is as follows. The first detection device according to the present invention is a pollen detection device that determines the presence or absence of a pollen-derived protein in a sample liquid, wherein the base material of the detection device is capable of developing a pollen sample liquid. Consisting of a porous material,
A sample liquid application site to which the pollen sample liquid is applied, a first antibody-carrying site on which a first antibody capable of specifically binding to a pollen-derived protein, and a second antibody capable of specifically binding to a pollen-derived protein A pollen detection device is characterized by having a positive determination site carrying a second antibody different from one antibody. Further, on the base material of the detection device, a development delay section that delays the development of the sample liquid between the first antibody-carrying site and the positive determination site, and a sample liquid development that cannot expand the sample liquid in a part near the positive determination site. The pollen detection device provided with an impossible part. Further, the base material of the detection device is provided with a third substance-immobilized site on which a third substance capable of specifically binding to the first antibody is immobilized, on the downstream side in the developing direction of the sample liquid from the positive determination site. The device.

[0005] A second detection device according to the present invention is a container having an open one side capable of accommodating pollen sample liquid, a hermetic lid mounted on one side opening of the container, and mounted on the hermetic lid. The sealing lid has a sealing film, the sealing lid has a sealing film, and the detecting body has a cutting portion for breaking the sealing film when the detecting body is mounted on the sealing lid; And a determining unit that is provided continuously to the unit and determines the presence or absence of a protein derived from pollen. Further, the determination unit, a detection plate that allows the pollen sample liquid to penetrate, a first antibody carrying unit that carries a first antibody that can specifically bind to a pollen-derived protein connected to the detection plate, Unlike one antibody, it consists of a second antibody immobilization part on which a second antibody that can specifically bind to pollen-derived protein is immobilized, and an absorber for absorbing the pollen sample liquid that has passed through. Pollen detector.

A third detecting device according to the present invention comprises a container capable of containing pollen sample liquid, one side of which is open, and a detector mounted on one side of the container. Having a communication hole sealed with a conductive film, having a case attached to one side opening of the container, and a determination unit provided in the case and determining the presence or absence of pollen-derived protein And a pollen detecting device characterized by the following. Also,
The determination unit, a detection plate that allows the pollen sample liquid to penetrate, a first antibody carrying unit that carries a first antibody that can specifically bind to a pollen-derived protein coupled to the detection plate, and the first antibody Unlike the second antibody immobilization portion in which the second antibody capable of specifically binding to the pollen-derived protein is immobilized, the solid matter in the pollen sample liquid is provided between the communication hole and the determination portion in the case. A pollen detector comprising: a filter unit for removing the pollen; and an absorber for absorbing the passed pollen sample liquid and the antibody-sensitized particles.

A fourth detector according to the present invention comprises a container capable of accommodating a pollen sample liquid, one side of which is open and the other side of which is sealed, and a hermetically sealed lid attached to the opening of the container. A solution chamber is provided on the opening side and a detection body chamber is provided on the closed side,
A pollen detection device, wherein the solution chamber and the detection body chamber are separated by a partition that is openably and closably provided, and a determination unit that determines the presence or absence of pollen-derived protein in the detection body chamber is provided. did. Further, the determination unit, a detection plate that allows the pollen sample liquid to penetrate, a first antibody carrying unit that carries a first antibody that can specifically bind to a pollen-derived protein connected to the detection plate, Unlike one antibody, between the solution chamber and the detection body chamber, a second antibody-fixed portion in which a second antibody capable of specifically binding to pollen-derived protein is fixed, and impurities in the pollen sample solution are removed. A pollen detection device comprising a filter section to be removed and an absorber for absorbing the passed pollen sample liquid and antibody-sensitized particles.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The pollen detection device according to the present invention is based on an immunological method called a sandwich method using an antibody against a pollen-derived protein. That is, the presence or absence of a pollen-derived protein is determined using a first antibody and a second antibody that can specifically bind to the protein, and thereby the presence or absence of pollen is determined. Therefore, in this specification,
The first antibody and the second antibody are intended to mean, for example, antibodies against different epitopes of a pollen-derived protein. The first antibody and the second antibody may be either monoclonal antibodies or polyclonal antibodies produced by a known method which has been already established, but are preferably monoclonal antibodies in view of their binding specificity. Furthermore, the pollen-derived protein that is the antigen may be appropriately selected depending on the pollen to be detected. It is preferably a protein derived from Akinokirin grass pollen and Camellia pollen.

Further, either the first antibody or the second antibody is labeled, and in the detection device described below, it is preferable that the first antibody be labeled. In the detection device according to the present invention, the signal from the label is preferably one that can be visually confirmed, and the label is preferably present in the first antibody. Specific examples of preferred labels include enzyme labels such as alkaline phosphatase, β-galactosidase and peroxidase; fluorescent labels such as fluorescein rhodamine; dye labels such as erythrocytes and colored latex; metals such as gold, silver and selenium; And inorganic particles.

[0010] In the present invention, dust, dirt or dust that is supposed to contain pollen is collected, a pollen sample solution is prepared therefrom, and the sample solution is applied to a detection device to detect the presence of pollen. Know if there is. That is, dust, dust, dirt, etc. that may contain pollen collected by a vacuum cleaner, or further obtained by passing it through a mesh of appropriate fineness (for example, 16 mesh), an air conditioner filter, A sample obtained by attaching an adhesive tape to a filter of an air purifier or an indoor floor or the like is used as a specimen. Water, saline,
Stir in an aqueous solution such as a phosphate buffer. The solution thus obtained may be applied as it is to the detection device of the present invention, but it is more preferable to use the solution by stirring it with a stick or the like so as to crush dust, dirt or dust.

That is, since the detection device according to the present invention detects pollen-derived proteins as described above, it is desirable that a large amount of the pollen proteins be dissolved in the solution by crushing the individual pollen. It is. Furthermore, fillers (for example, glass fillers, metal fillers, ceramic fillers, etc.) may be simultaneously present and agitated so that the pollen is efficiently crushed, and the pollen may be reduced by using a container provided with irregularities on the inner wall. You may stir so that an individual may be crushed. The pollen sample solution thus prepared is applied to the pollen detection device according to the present invention.

A first detection device according to the present invention will be described. As shown in FIG. 1, the detection apparatus 1 includes a sample liquid application site 10 to which a pollen sample liquid is applied in order from one end of a substrate 101.
2, the first antibody carrying site 103, the sample liquid development delay site 104
A positive determination site 105 and a negative determination site 106 are provided, and non-water-solubilized sites 107a and 107b in which a sample liquid cannot be developed are provided near the positive determination site 105.

The base material 101 of the detection device 1 according to the present invention is made of a porous material on which the pollen sample liquid can be spread. Here, that the pollen sample liquid can be expanded means that the pollen sample liquid can move through the porous material of the base material by capillary action. Preferred examples of the substrate include glass fiber, cellulose, and filter paper made of them, nylon nonwoven fabric,
Examples thereof include a polyester nonwoven fabric, nitrocellulose, polyvinylidene fluoride, regenerated cellulose, cellulose acetate, and a porous material of charge-introducing nylon, and a material having a function of immobilizing a protein is particularly preferable.

The sample solution application site 102 is a site to which the pollen sample solution is applied. After the sample solution is applied, the sample solution spreads in the porous material of the substrate 101 toward the first antibody carrying site 103 by capillary action. Start moving. First antibody carrying site 103
Is a portion where the first antibody is supported on the base material 101,
The first antibody has an ability to specifically bind to a pollen-derived protein. Therefore, when a pollen-derived protein is present in the pollen sample solution that has developed and moved from the sample solution application site 102, the protein and the first antibody are combined at the first antibody carrying site 103 to form a complex.
After the first antibody binds to the substance to be detected (pollen-derived protein), it needs to move with the development of the pollen sample solution, and the first antibody is impregnated without binding to the base material. preferable. Details of the method of carrying the first antibody will be described later.

The pollen sample liquid that has spread and moved from the sample liquid application site 102 passes through a later-described expansion delay site 104, and then reaches a positive determination site 105. The positive determination site 105 is a portion where the second antibody is fixed to the substrate 101. Positive site 10 where the complex of pollen-derived protein and primary antibody is positive
Upon reaching 5, a binding reaction between this complex and the second antibody occurs. The presence or absence of pollen-derived protein can be determined by recognizing the binding substance using a labeling substance described later. For example, when the first antibody is labeled with colloidal gold, the complex of the substance to be detected (pollen-derived protein) and the first antibody binds to the second antibody of the positive determination site 105 and the positive determination site 105 And accumulate. As a result, the color of the gold colloid
Is observed in On the other hand, when the substance to be detected does not exist, the first antibody labeled with colloidal gold does not bind to the second antibody and passes through the positive determination site 105. No color is obtained. As described above, the presence or absence of the pollen-derived protein in the pollen sample liquid, that is, the presence or absence of pollen can be known.

As shown in FIG. 1, the first detection device according to the present invention is provided with a deployment delay portion before the positive determination portion 105.
104 are provided. The expansion delay portion 104 is a portion that delays the expansion of the pollen sample liquid, that is, a part that acts to reduce the expansion speed of the sample liquid. As the developing speed decreases, it takes more time for the pollen sample liquid that has contacted the first antibody at the first antibody carrying site 103 to reach the positive determination site 105. As a result, the time during which the pollen-derived protein and the first antibody are in contact with each other is prolonged. Increases body production.

The development delay portion 104 in the present invention may be, for example, to support a water-soluble polymer on the base material, reduce the porosity of the base material, or to provide a site where the sample liquid cannot be spread on a part of the base material. It is formed by reducing the effective cross-sectional area of the base material on which the sample liquid can be spread, for example, by providing it. For example, when a water-soluble polymer is carried on a substrate, the sample liquid that has spread and moved is absorbed by the water-soluble polymer when it comes into contact with the water-soluble polymer. The sample liquid that has developed and moved continues to be absorbed by the water-soluble polymer until it exceeds the absorption capacity of the water-soluble polymer. During this time, the sample liquid continues to stay in the polymer, and during this stay, The reaction between the sample substance and the first antibody proceeds sufficiently.

As the water-soluble polymer, those which do not affect the analyte, the first antibody and the second antibody, and the binding reaction thereof are preferable. Specific examples thereof include polyacrylic acid, polyvinyl alcohol, and polyvinylpyrrolidone. , Polyacrylamide, polydimethylacrylamide, polyethylene glycol, polypropylene glycol, polyvinyl methyl ether, casein, gelatin,
Hydroxypropyl cellulose and the like.

As a method of forming the development delay portion 104 by reducing the porosity of the base material, for example, the base material 10
There is a method in which particles or fillers smaller than the voids are present in the porous material. When the porosity of the base material is reduced as described above, the resistance when the sample liquid spreads the base material is increased, and the spreading speed is reduced. When the developing speed decreases, the contact time between the substance to be detected and the first antibody becomes longer, and the reaction proceeds sufficiently. Preferred examples of the particles or fillers for decreasing the porosity include fine polymer particles such as polystyrene and acrylic, and inorganic particles such as microsilica and alumina sol.

Further, the undeployable portion 104 may be formed by providing an undeployable portion of the sample liquid in a part of the base material.
A specific example is shown in FIG. FIG. 2 is a schematic cross-sectional view of the development delay portion 104, in which the base 101 is provided with a portion where the pollen sample liquid cannot be expanded, that is, a sample liquid non-deployable portion 104a. Due to the presence of the sample liquid non-deployable portion 104a, the pollen sample liquid is restricted in its development flow path (arrow in the figure), and the development speed is reduced.

Such a site where the sample liquid cannot be spread can be constituted by rendering a part of the base material insoluble in water with a water-insoluble polymer, for example. The water-insoluble polymer is preferably one that is substantially insoluble in water and does not affect the analyte, the first antibody and the second antibody, and the binding reaction thereof. Specific examples thereof include polyacrylate, polymethyl methacrylate, polyvinyl acrylate, polybutyl methacrylate, polystyrene, α
-Methyl polystyrene, chloromethylated polystyrene,
Copolymers composed of vinyl or polymers such as polyvinyl acetate, polybutadiene, polyisoprene, polyacrylonitrile, polyvinyl butyral, polyacrylamide, and polyvinyl carbazole, alone or in combination of two or more monomers, ring-opening polymerization and condensation polymerization. Examples of the obtained polyamide, polyester, polyurethane, and the like, celluloses such as hydroxypropylmethylcellulose phthalate, carboxymethylethylcellulose, and ethylcellulose, shellac, and the like.

Further, a polymer obtained by treating with ionizing radiation after having been used alone or in combination of two or more kinds of ionizing radiation functional monomers or oligomers having one or more functionalities in the molecule, or mixed with the above-mentioned polymer, is also used. As the compound having a functional group in the molecule, a compound which basically reacts with ionizing radiation such as an ultraviolet ray and an electron beam, particularly a compound having a radical polymerizable unsaturated bond is preferable.

As the monofunctional monomer, acrylic acid, acrylic acid dimer, 2-hydroxyethyl acrylate,
Acrylic acid and its esters such as 2-hydroxypropyl acrylate, methyl acrylate, 2-ethylhexyl acrylate, methoxyethyl acrylate, butyl acrylate, methoxybutyl acrylate and phenyl acrylate; methacrylic acid, 2-hydroxyethyl methacrylate, 2-hydroxypropyl methacrylate ,
Glycerol methacrylate, methyl methacrylate,
Methacrylic esters such as ethyl methacrylate, propyl methacrylate, methoxyethyl methacrylate, ethoxyethyl methacrylate, phenyl methacrylate, lauryl methacrylate; acrylamide, propyl acrylamide, propyl methacrylamide, unsaturated carboxylic amides of methacrylamide; 2- (N, N -Dimethylamino) ethyl acrylate, 2- (N, N-dimethylamino) ethyl methacrylate, 2- (N, N-dibenzylamino) ethyl acrylate, (N, N-dimethylamino) methyl methacrylate, 2- (N, Substituted amino alcohol esters of unsaturated acids such as N-diethylamino) propyl acrylate; N-methylcarbamoyloxyethyl acrylate, N-ethylcarbamoyloxyethyl acrylate,
Carbamoyloxyalkyl acrylates such as N-butylcarbamoyloxyethyl acrylate, N-phenylcarbamoyloxyethyl acrylate, 2- (N-methylcarbamoyloxy) ethyl acrylate, 2-carbamoyloxypropyl acrylate, and other vinylbenzyltrimethyl Examples include cationic vinyl monomers such as ammonium chloride, N-methyl-4-vinylpyridinium chloride, 2-methacryloyloxyethyltrimethylammonium chloride, 2-acryloyloxydimethylsulfonium chloride, and acryloylmorpholine.

The polyfunctional monomer has two or more radically polymerizable unsaturated groups, and is particularly preferably a polyfunctional (meth) acrylate having two or more (meth) acryloyl groups. Specific examples include, for example, ethylene glycol di (meth) acrylate, polyethylene glycol di (meth) acrylate, hexanediol di (meth) acrylate, trimethylolpropane tri (meth) acrylate, trimethylolpropane di (meth) acrylate, Pentaerythritol tetra (meth) acrylate, pentaerythritol tri (meth)
Acrylate, dipentaerythritol hexa (meth)
Acrylate, ethylene glycol diglycidyl ether (meth) acrylate, polyethylene glycol diglycidyl ether di (meth) acrylate, propylene glycol diglycidyl ether di (meth) acrylate, polypropylene glycol diglycidyl ether di (meth) acrylate, sorbitol tetraglycidyl ether Tetra (meth) acrylate, melamine (meth)
And polyfunctional (meth) acrylates such as acrylate.

Examples of the oligomer or polymer having a functional group include, for example, urethane acrylate obtained by bonding diisocyanate, a diol having a hydroxyl group at both terminals and an acrylate compound having a hydroxyl group at the terminal and having a radical polymerizable unsaturated group. The urethane acrylate having an arbitrary molecular weight can be obtained by changing the molecular weight of the diol used. In addition, acrylic acid chloride or acrylic acid is added to the hydroxyl group of a copolymer of polyester acrylate, epoxy acrylate, alkyl acrylate and acrylic acid-2-hydroxy ester.
Copolymer in which an acryloyl group is introduced by adding a 1: 1 adduct of 2-hydroxyethyl and diisocyanate, or copolymer in which glycidyl methacrylate is added to a carboxyl group of a copolymer of alkyl acrylate and acrylic acid It is also possible to preferably use a compound obtained by adding a 1: 1 adduct of acrylic acid chloride or 2-hydroxyethyl acrylate and diisocyanate to the remaining hydroxyl groups of polyvinyl butyral. Note that, unlike the first antibody, the second antibody is preferably bound and carried on the substrate 101 with a certain strength. Details of preferred specific examples of the supporting method will be described later.

As shown in FIG. 1, the first detection apparatus of the present invention comprises a sample liquid non-developable part (a part corresponding to 107a and 107b in FIG. 1) on a part of the substrate near the positive judgment part 105. Is provided. The undeployable portion of the sample liquid is formed such that all or a part of the pollen sample solution that moves and converges near the positive determination portion 105. FIG. 1 shows a water-insoluble portion 107a on a part of the base material on both sides of the positive determination portion 105.
And 107b are provided as sites where the sample liquid cannot be developed. Due to the presence of the sample liquid undeployable portion, the effective cross-sectional area of the base material is reduced so that the sample liquid converges in the vicinity of the positive determination portion 105. As a result, more sample liquid passes near the positive determination site 105 as compared to the case where the substrate having no sample liquid undeployable site is used. As a result, when a complex of the substance to be detected and the first antibody is contained in the sample liquid, more complex is advantageously involved in binding to the second antibody.

Another example of the sample liquid non-deployable portion will be described with reference to FIG. FIG. 3 is a cross-sectional view of the vicinity of the positive determination site 105 of the first detection device according to the present invention. As shown in FIG. 3, this is an example in which a sample liquid non-deployable portion 107 is provided by making the back surface of a positive determination portion 105 of a base material 101 water-insoluble with a certain thickness. The sample solution converges on the positive determination site 105 as shown by the arrow in FIG. The formation of the water-insoluble region can be performed by using the water-insoluble polymer used to form the development delay site.

Further, the first detecting device of the present invention has a structure shown in FIG.
As shown in the figure, a negative judgment site 106 is provided. The negative determination site 106 carries a third antibody that can specifically bind to the first antibody. If no pollen-derived protein is present in the pollen sample solution, even if the sample solution passes through the first antibody carrying site 103 due to the development of the pollen sample solution,
Since the first antibody has no protein to bind to, the first antibody develops and moves independently as the sample solution develops. Then, when the first antibody reaches the negative judgment site 106, the first antibody binds to the third antibody. When the first antibody or the third antibody carries a label that generates a signal such as coloration by this binding, the first antibody is covered by the coloration caused by the binding of the first antibody and the third antibody. It can be confirmed that the unfolded and moved independently without reacting with the test substance (pollen-derived protein) and that the unfolding and moving was completed in the detection device. That is, by confirming the label signal at the negative determination site 106, it is possible to reliably determine the end of the development and the absence of the substance to be detected.

In the case where the first antibody is predominantly present in the first antibody-carrying site 103 more than the substance to be detected, the two sites, the positive judgment site 105 and the negative judgment site 106, cause color reaction and the like. Since the label signal can be visually recognized, the end of the development and the presence of the substance to be detected can be reliably determined.

In the above-described first detection apparatus, an example has been described in which both the development delay portion 104 and the sample liquid undeployable portion 107 are provided. The detection device may be provided with any one of them. FIG. 4 shows a specific example of a detection device provided with each of the above two embodiments independently. FIG. 4 (a)
(F) is a top view of the detection device, and the right column is a cross-sectional view of that part. That is, the right side of FIG. 4A shows a cross section taken along line XX on the left side of FIG. 4A. Similarly,
The right side of FIGS. 4B to 4F is a diagram showing a cross section of the left side diagram.

FIG. 4A shows the first antibody carrying site 103 of the substrate 101.
This is an example in which a water-soluble polymer is carried and a deployment delay site 104 is provided between the positive determination site 105 and the positive determination site 105. Figure 4 (b) shows the substrate 101
By making a part of the water insoluble in water,
This is an example in which is provided. Fig. 4 (c) shows the substrate of the positive judgment site 105.
The other side of 101 is made water-insoluble and the sample liquid
This is an example in which the development of the sample liquid is made to converge on the positive determination site 105. Fig. 4 (d) shows that the both sides of the substrate 101 of the positive judgment site 105 are made water-insoluble,
7 is an example in which the development of the sample liquid is caused to converge to the positive determination site 105. Fig. 4 (e) shows Fig. 4 (c) and Fig. 4
This shows an example in which (d) is combined. Figure 4
(f) shows a conventional detection device without the development delay part 104 and the sample liquid non-deployment part 107.

The method for forming the first antibody-carrying site, the second antibody-carrying site, the development delay site, and the sample liquid-impossible site on the base material includes a first antibody, a second antibody, a water-soluble polymer and a water-insoluble polymer. The conductive polymer is dissolved or dispersed in an appropriate solvent, and the solution is formed by dropping the solution at a predetermined position on a substrate by pipetting. Further, by forming these parts by using a printing technique, mass production becomes possible. For example,
The first antibody, the second antibody, the water-soluble polymer or the water-insoluble polymer can be dissolved in a suitable solvent (eg, water, alcohols, esters, polyesters, aldehydes, ketones, penzens, paraffins, etc.). (Single or mixed solvent) is supported on the substrate 101 by a technique such as gravure printing, screen printing, blade coating, or slide coating. When such a printing method is used, as shown in FIG. 5, a first antibody carrying site 103, a development delay site 104, a positive judgment site
105, negative judgment part 106, sample liquid undevelopable part 107
(Water-insoluble portion) and the like, and then, as shown in FIG. 5, by separating the substrate 101 along the dotted line,
The detection device can be efficiently manufactured in large quantities.

Since the first antibody is required to move with the development of the sample solution, the functional group present on the first antibody or the base material when loaded on the base material 101 is replaced with, for example, bovine serum albumin. , Collagen, casein, or the like, and the first antibody is preferably supported so as not to be strongly fixed to the substrate. On the other hand, unlike the first antibody, it is necessary that the second antibody at the positive determination site does not move with the development of the sample solution. Therefore, in order to firmly fix the positive judgment site to the base material, it is preferable to dry or heat it after printing.

Next, a second detector according to the present invention will be described. As shown in FIG. 6, the second detection device 2
A synthetic resin container 211 having an opening on one side;
It comprises a hermetic lid 215 made of synthetic resin, which is mounted on the opening, and a detector 220 mounted on the hermetic lid 215. The sealing lid 215 is mounted on the container 211 by engaging an internal screw 215a provided on the inner peripheral surface with an external screw 211a provided on the outer peripheral surface of the container 211. In addition, sealing lid 215
Has a sealing film 217 at the end opposite to the container 211, and further has an external thread 215 b on the outer periphery of the sealing lid 215. The end of the sealing lid 215 on the container 211 side is
It is in contact with the flange 219 provided in 1 to maintain the sealing performance.

Further, a detector 220 is mounted on the closed lid 215. The detection body 220 includes a detection plate having a cutting portion 223, a filter portion 225, a first antibody carrying portion 229, and a second antibody fixing portion 230 inside a case 221 made of a transparent synthetic resin.
227 and an absorbing pad 232 are provided. Of these, a determination unit is configured by the detection plate 227 having the second antibody fixing unit 230 and the absorption pad 232. Case 221
Is provided with an inner screw 221a. The inner screw 221a is
215. Cutting section 22
As shown in FIG. 7, 3 is supported by a case 221 via a support portion 223a. When the detection body 220 is mounted on the closed lid 215, the seal film 217 is broken.

The filter section 225 is for removing solid matter in the pollen sample liquid flowing in from the container 211 side, and various filters are configured as a single layer to a multilayer structure. Figure
The device described in 6 is a case where it is configured in three stages.
The filter unit 225 is composed of a millipore filter, a little cellulose, a nylon mesh, a Teflon mesh, a glass filter, a filter paper, a PVDF (trade name), and the like. In this case, the filtration efficiency can be improved by making the pore size of the downstream layer smaller than that of the upstream layer. In addition, a filtration efficiency can be improved by interposing a powder such as silica or alumina between the upstream layer and the downstream layer.
Further, between the detection plate 227 and the filter unit 225, a first antibody carrying unit 229 carrying a first antibody capable of specifically binding to a pollen-derived protein is provided.

The detection plate 227 is arranged along the longitudinal direction of the case 221 between the filter unit 225 and the absorption pad 232, and allows the pollen sample liquid to penetrate and the first antibody passing with the pollen sample liquid. Positive second antibody immobilization site 23
It is set to trap at 0. In addition, this detection plate
As shown in FIG. 8, 227 is fixed to the center of the case 221. Examples of the material include nitrocellulose, glass fiber filters, chromatography paper, PVDF (trade name), and nylon or polyester nonwoven fabric. Used. In addition, extra sample solution and antibody sensitized particles
The light passes through the detection plate 227 and is absorbed by the absorption pad 232.

Next, a method for detecting pollen using the second detection device will be described. First, close the lid 21 from the container 211.
5 Remove. Next, a separately prepared pollen sample solution is placed in the container 211. Also, without preparing the pollen sample liquid, put dust or dust that may contain pollen in this container 211, further put water, physiological saline, a buffer solution, etc., and in this container, put the pollen sample liquid in this container. May be prepared. At this time, it is preferable that the above-mentioned filler is further placed in the container, or that the inner wall of the container 211 is provided with irregularities and stirred with a stick or the like so that the pollen is more efficiently crushed.

Next, the container 211 containing the pollen sample liquid is sealed with the sealing lid 215. The pollen may be further crushed by further shaking the container 211 sealed by the sealing lid 215. Thereafter, the case 221 of the detection body 220 is attached to the closed lid 215. That is, the outer screw 21 of the sealing lid 215
The inner screw 221a of the case 221 is engaged with 5b, and the case 221 of the detector 220 is pushed into the outside of the sealing lid 215 while rotating. When the case 221 is completely attached to the closed lid 215, the cut portion 223 breaks the seal film 217, and the pollen sample liquid in the container 211 flows into the detector 220 from the broken portion. From the pollen sample liquid flowing into the detection body 220, solid matters such as dust and pollen in the pollen sample liquid are removed by the filter unit 225.

Next, the pollen sample liquid that has passed through the filter part 225 passes through the first antibody carrying part 229 and permeates inside the detection plate 227 toward the absorption pad 232 side. Here, if a pollen-derived protein is present in the pollen sample solution, the protein and the first antibody bind to each other to form a complex, and the detection plate 227 is deployed and moved. Since the protein and the first antibody also develop and move with the development and movement of the sample solution, the binding reaction between the protein and the first antibody proceeds even while the sample solution is developing and moving on the detection plate 227. Then, the sample liquid reaches the second antibody fixing portion 230, and when a complex of the protein and the first antibody is present in the sample solution, a binding reaction between the complex and the second antibody occurs here. As described above, by recognizing this binding using the labeling substance present in the first antibody, the presence or absence of a protein derived from pollen, that is, the presence or absence of pollen can be known. The excess pollen sample solution is absorbed by the absorption pad 232, so that the pollen sample solution can smoothly penetrate the detection plate 227.

Another example using the second detection device will be described. 9 and 10 are the same as those shown in FIGS. 6 to 8 except that the configurations of the detection plate 227 and the absorption pad 232b provided in the detection body 220 are different. That is, as shown in FIGS. 9 and 10, a filter unit 225 is provided in the case 221.
A deployment spacer 232a is disposed adjacent to the 225. The case 221 is enlarged at an end opposite to the container 211 (FIG. 6), and an annular pad chamber 233 is formed at the enlarged end. An absorption pad 232b is provided in the pad chamber 233, and a detection plate 227 having a second antibody fixing portion 230 is provided between the developing spacer 232a and the absorption pad 232b.
Are arranged at right angles to the longitudinal direction.

The filter part 225 and the developing spacer 23
A first antibody-carrying site 229 is provided between the first and second antibodies 2a. In this case, the solid matter in the pollen sample liquid is
Further, the pollen sample liquid can be sent to the detection plate 227 side more reliably by the developing spacer 232a.
When the complex in which the pollen-derived protein and the first antibody are bound reaches the second antibody fixing site 230, a binding reaction with the second antibody occurs. As described above, this binding reaction is carried out using the labeling substance of the first antibody and at the bottom side of the detector 220 (right side in FIG. 9).
Can be easily confirmed.

The absorption pad of the detection device used in the present invention may be formed using a water-absorbing gel. For example, as shown in FIG. 11A, the absorbent pad 232 may be formed by dispersing a water-absorbing gel 242 inside a nonwoven fabric 241 and attached to the detection plate 227. In this case, Condon REO-65 or Bemliese RE750 (both manufactured by Asahi Kasei Corporation) can be used as the nonwoven fabric.

As the water-absorbing gel, a commercially available gel can be used. For example, Sumika gel (manufactured by Sumitomo Chemical Co., Ltd.)
PVA / polyacrylate copolymer), Diamond Wet (Mitsubishi Yuka Co., Ltd., crosslinked polyacrylate), Aqua Reserve (Nippon Synthetic Chemical Co., Ltd., PVA copolymer), Aqua Coke (Sumitomo) Seika Chemical Co., Ltd., crosslinked polyethylene oxide type), water absorbing paogen (Daiichi Kogyo Seiyaku Co., Ltd., crosslinked polypropylene oxide type), Aquaprene (Meisei Chemical Co., Ltd., nonionic type), Sunwet (Sanyo Chemical Co., Ltd.) Made, starch / acrylic acid graft copolymer), KI gel (made by Kuraray Co., Ltd., isobutylene / maleic anhydride copolymer), Arasorp (made by Arakawa Chemical Co., Ltd., polyacrylate), Aqualic (Nippon Shokubai) Chemical Co., Ltd., cross-linked polyacrylate), Alonzap (Toa Gosei Chemical Co., Ltd.), Wonder Gel (Kao Corporation) And cross-linked polyacrylate).

Further, as shown in FIG.
232 may be configured by storing a water-absorbing gel 242 inside a non-woven bag 243, which may be attached to the detection plate 227. In this case, the aforementioned nonwoven fabric and water-absorbing gel are used. Further, as shown in FIG. 11 (c), first, a composition containing the water-absorbing gel 242 was prepared by mixing the water-absorbing gel 242, a binder, and a solvent, and this composition was applied to the detection plate 227.
It is good also as an absorption pad 232 by apply | coating on it and drying.

As shown in FIG. 11D, first, a water-absorbing gel solution is prepared by mixing the water-absorbing gel 242 and a solvent (eg, alcohol + water).
This water-absorbing gel solution is applied on a film (hereinafter referred to as PET) 245) and dried. Next, the sheet may be irradiated with an electron beam to crosslink the water-absorbing gel to form a sheet-like absorbing pad 232. The sheet-shaped absorption pad 232 is attached to the detection plate 227 with the water-absorbing gel 242 side abutting on the detection plate 227 side.

Next, a third detector according to the present invention will be described. The third detection device 3, as shown in FIG.
Cylindrical synthetic resin container 311 with one side open and detector 32
Consists of zero. The container 311 has an internal screw 31
1a is provided, and an external screw 311b is provided on the outer surface on the opening side. Also, an inner screw 313a is provided on the inner surface on the opening side of the container 311.
The inner cover 313 having the inner screw 313a is connected to the outer screw 31 of the container 311.
It is mounted by engaging with 1b. Further, the sealing lid 313 is removed from the container 311 at the time of use, and the detection body 320 is attached to the container 311 instead.

Next, the detector 320 will be described in detail. The detection body 320 is composed of a cylindrical case 321 provided with an external screw 321a. By engaging the external screw 321a with the internal screw 311a of the container 311, the case 321 is mounted on the container 311. I have. The case 321 is made of a transparent synthetic resin, and a water-soluble film is provided on the case 311 side of the case 321.
A communication hole 322 sealed by 323 is formed. Also,
In the case 321, the filter section is arranged in order from the communication hole 322 side.
325 and a deployment spacer 332a are provided. Further, the case 321 is enlarged at an end opposite to the communication hole 322, and an annular pad chamber 333 is formed at the enlarged end. Absorbing pad 332b is arranged in pad room 333,
A detector 327 to which a second antibody fixing part 330 is attached is provided between a case 321 and a deployment sensor 332a and an absorption pad 332b.
Are arranged orthogonally to the axial direction (longitudinal direction). Among them, the detection plate 327 to which the second antibody fixing part 330 is attached,
A determination unit is configured by the absorption pad 332b. Further, a first antibody holding portion 329 carrying the first antibody is provided between the filter portion 325 and the developing spacer 332a.

Next, each component will be described. The filter unit 325 is for removing solid matter in the pollen sample liquid flowing from the container 311 side, and is configured in three stages in FIG. 12, but the filter unit 325 is not limited to three stages and may be smaller. Or more. Detection plate 327
As shown in FIG. 13, the pollen sample liquid is disposed between the developing spacer 332a and the absorption pad 332b at right angles to the longitudinal direction of the case 321 to allow the pollen sample liquid flowing from the container 311 to penetrate, and The complex between the pollen-derived protein and the first antibody that passes along with the sample liquid is fixed to the second antibody
It is designed to trap. The detection plate 32
7 is made of the same material as that used in the second detection device. The surplus pollen sample liquid passes through the detection plate 327 and is absorbed by the absorption pad 332b.

Next, a method for detecting pollen using the third detection device 3 will be described. First, close the lid from container 310
Remove 313. Next, the pollen sample liquid is put into this container 310. As described above, the pollen sample liquid may be prepared in the container 310. Next, the case 321 is attached to the container 310 containing the pollen sample liquid. Then, after a while, the water-soluble film 323 sealing the communication hole 322 of the case 321 is dissolved by the pollen sample liquid, the pollen sample liquid in the container 310 passes through the communication hole 322, and the solid matter is removed by the filter unit 325. Then, it flows into the case 321 side.

Next, the pollen sample liquid having passed through the filter section 325 passes through the inside of the developing spacer 332a together with the first antibody carried on the first antibody carrying portion 329, passes through the inside of the detection plate 327 toward the absorption pad 332b. Infiltrate. Here, if a pollen-derived protein is present in the pollen sample solution, the protein and the first antibody bind to form a complex, and this reaction proceeds even while the sample solution expands and moves the spacer 332a. . Then, the sample liquid reaches the second antibody fixing portion 330, and when the complex is present in the sample solution, a binding reaction between the complex and the second antibody occurs here. As described above, by recognizing this binding using the labeling substance present in the first antibody, the presence or absence of a pollen-derived protein, that is, the presence or absence of pollen can be known. The extra pollen sample liquid that has permeated into the detection plate 327 is absorbed by the absorption pad 332b, so that the pollen sample liquid can be smoothly permeated into the detection plate 327.

Next, another example using the third detecting device will be described. This detection device differs from the above-described detection device only in the configuration of the detection body 320, as shown in FIGS. 14 and 15, and is otherwise substantially the same as the examples shown in FIGS. FIG. 15 is a sectional view taken along line IV-IV in FIG. As shown in FIGS. 14 and 15, the detection body 320 has a cylindrical case 321. The case 321 has a communication hole 322 sealed with a water-soluble film 323. In addition, the filter portion is provided in the case 321 in order from the communication hole 322 side.
325, a detection plate 327 to which the second antibody fixing part 330 is attached, and an absorption pad 332 are provided. Furthermore, the filter section
Between the 325 and the detection plate 327, a first antibody holding portion 329 is provided, and the detection plate 327 is arranged along the axial direction of the case 321.

When pollen is detected in the same manner as described above using the present detection apparatus, if a pollen-derived protein is present in the pollen sample solution, a complex consisting of this protein and the first antibody is immobilized on the second antibody. The color is trapped in the section 330 to show the color. In this case, the detection plate to which the second antibody fixing portion 330 is attached
Since the 327 is arranged along the axial direction of the case 321, the color change of the second antibody fixing portion 330 can be easily and clearly recognized from the side of the case 321. Note that the absorption pad 332 of the third detection device is formed using a water-absorbing gel, as described above.

Next, a fourth detector according to the present invention will be described. The fourth detection device 4 is, as shown in FIGS. 16 and 17,
Transparent container 41 made of synthetic resin or glass that can be divided into two
1a and 411b. The containers 411a and 411b are rotatable relative to each other, and each of the containers 411a and 411b has partition walls 413a and 413b that slide relative to each other. That is, FIG. 16, FIG.
The partition 413 in 17 includes a partition 413a of the container 411a and a partition 413b of the container 411b. Further, one side of the container 411a is open, and a sealing lid 421 is attached to the opening. Also, as shown in FIG. 19, the partition wall 41 of the container 411a
3a and the partition wall 413b of the container 411b have communication holes 431a and 431b, respectively.
When the container 411a is rotated to a predetermined position with respect to the container 411b, the position of the communication hole 431a and the position of the communication hole 431b coincide with each other, and the inside of the container 411a and the inside of the container 411b communicate with each other. It has become. The partition 413 in FIG.
It is composed of a partition wall 413a and a partition wall 413b, and shows a case where the communication hole 431a and the communication hole 431b do not match.

When the containers 411a and 411b are combined, as shown in FIG. 16, combined containers 411a and 411b having one side open and the other side sealed are formed.
411b is a partition wall 413 composed of partition walls 413a and 413b sliding on each other.
The solution chamber 414 in the container 411a and the detection body chamber in the container 411b
It is divided into 415. The pollen sample liquid can be stored in the solution chamber 414 in the container 411a. Further, in the container 411b, in order from the partition 413 side, the filter unit is provided.
418, a detection plate 417 (determination unit) having a second antibody fixing unit 430, and an absorption pad 420.

The filter section 418 is made up of a millipore filter, a little cellulose, a nylon mesh, a Teflon mesh, a glass filter, a filter paper, a PVDF (trade name), and the like. Be composed. In this case, the filtration efficiency can be improved by making the pore size of the downstream layer smaller than that of the upstream layer. In addition, a filtration efficiency can be improved by interposing a powder such as silica or alumina between the upstream layer and the downstream layer.

Between the filter 418 and the detection plate 417, there is provided a first antibody carrier 419 carrying a first antibody capable of specifically binding to pollen-derived protein. In addition, the first antibody carrying section 419 is
It can be provided inside or on the detection plate 417. Detection plate
417 is a container between the filter section 418 and the absorption pad 420.
It is arranged along the axial direction of 411b so that the pollen sample liquid can penetrate. The detection plate 417 is made of a hydrophobic porous film material such as nitrocellulose, glass fiber filter, chromatography paper, PVDF (trade name), nylon or polyester nonwoven fabric, and the like. Excess pollen sample liquid passes through the detection plate 417 and is absorbed by the absorption pad 420. As the absorbent pad 420, cellulose, rayon, polyethylene, polypropylene microporous plastic, glass microfiber, Sumikagel, or the like can be used.

Next, a case where pollen is detected using the fourth detection device will be described. First, the solution chamber 414 and the detection body chamber 415 are constituted by the partition 413 by combining the container 411a and the container 411b. At this time, the partition 413a of the container 411a and the partition 413b of the container 411b are set not to communicate with each other. In the solution chamber 414, put the pollen sample liquid 425,
Attach 1. The pollen sample liquid 425 may be prepared in this solution chamber 414 as in the case described above.

Next, the container 411a is rotated with respect to the container 411b so that the position of the communication hole 431a formed in the partition 413a of the container 411a matches the position of the communication hole 431b formed in the partition 413b of the container 411b. Then, the solution chamber 414 in the container 411a and the detection body chamber 415 in the container 411b communicate with each other. As a result, the pollen sample liquid 425 flows from the communication hole 431a of the container 411a into the detection body chamber 415 through the communication hole 431b of the container 411b. Detection body chamber 41
5 The pollen sample liquid 425 flowing into the
8, during which the solids in the pollen sample liquid are removed by the filter unit 418.

Next, the pollen sample liquid that has passed through the filter section 418 passes through the first antibody holding section 419 and permeates the detection plate 417 toward the absorption pad 420 side. Here, if the protein of the pollen origin is present in the pollen sample solution, the protein and the first antibody are combined in the first antibody holding section 419 to form a complex, and the sample plate 417 is developed and moved. Then, the sample liquid reaches the second antibody fixing part 430, and if the complex is present in the sample liquid, a binding reaction between the complex and the second antibody occurs here. By recognizing this binding using the labeling substance present in the first antibody as described above, it is possible to know the presence or absence of a protein that is not pollen-specific, that is, the presence or absence of pollen.

Since the extra pollen sample liquid is absorbed by the absorption pad 420, the pollen sample liquid can be smoothly permeated into the detection plate 417. In the above example, after the solution chamber 414 and the detection body chamber 415 are communicated, the containers 411a, 41
Although an example in which 1b is left standing in the vertical direction has been described, as shown in FIG. 18, the opening of the container 411a may be closed with the sealing lid 421, and the containers 411a and 411b may be left standing in the horizontal direction. Further, the containers 411a and 411b are rotated with respect to each other to separate the partitions 413a and 413b.
However, the present invention is not limited to this, and the partition 413 may be formed of a film or the like, a stick may be attached to the sealing lid 421, and the partition 413 may be broken by this.

Next, another example of the fourth detector will be described. The detection device shown in FIGS. 20 and 21 has a detection plate 417 provided with a second antibody fixing portion 430 and a first antibody carrying portion.
The only difference is the configuration of the detection device 419, and the rest is substantially the same as the detection device shown in FIGS. 20 and 21, the same parts as those of the detection device shown in FIGS. 16 to 18 are denoted by the same reference numerals, and detailed description will be omitted. As shown in FIG. 20, a first antibody holding portion 419 is provided in a filter portion 418 in the detection body chamber 415, so that the pollen sample liquid 425 passes through the first antibody holding portion 419. Further, a detection plate 417 is provided below the first antibody carrying portion 419 at a predetermined interval. The detection plate 417 is arranged in a direction orthogonal to the axial direction of the container 411b, and a second antibody fixing portion 430 is attached to a substantially central portion of the detection plate 417.

Next, a method for detecting pollen using this detection device will be described. First, the pollen sample liquid 425 is placed in the solution chamber 414 of the container 411a, and a closed lid 421 is attached to the opening. Next, the partition 413a and the partition 413b are aligned to make the solution chamber 414 communicate with the detection body chamber 415, and the containers 411a and 411b are allowed to stand vertically with the container 411a facing upward. Then
The pollen sample liquid 425 in the solution chamber 414 flows into the detection body chamber 415, passes through the filter section 418, and passes through the first antibody holding section 419. After that, the pollen sample liquid 425 is filled with the first antibody carrying part 419
The liquid drops downward from the central part of the sensor and drops on the detection plate 417, and as shown in FIG. 21, penetrates into the inside of the second antibody fixing part 430 attached to the detection plate 417.

When the pollen-free protein is present in the pollen sample solution 425, a complex of the protein and the first antibody is formed in the first antibody carrying portion 419 as described above, and the second antibody The presence of the complex can be confirmed by the binding reaction between the complex and the second antibody in the fixing portion 430.
Incidentally, even in this fourth detection device, the absorption pad 420
May be configured using a water-absorbing gel, as in the second detection device.

When producing the detection device of the present invention, it is preferable to use a biodegradable material as a material constituting any of the above detection devices. Such biodegradable materials include, for example, bionol (manufactured by Showa Polymer Co., Ltd., aliphatic polyester), biopol (manufactured by ICI, microbial production polyester), matabbie (manufactured by Novamont, polyvinyl alcohol / starch based). ).

[0066]

EXAMPLES Example 1 An absorber made of cellulose pulp was provided on the short side of a rectangular (10 × 80 mm) glass fiber paper (GF / DVA, manufactured by Whatman Co., Ltd.), and 20 mm from the absorber. Using a dispenser, 1 μl of an anti-protein polyclonal antibody solution against a protein derived from cedar pollen was dropped in a circular pattern. afterwards,
After drying at 50 ° C. for 2 minutes, a second antibody-immobilized site was formed to obtain a test device. 10 μg of protein antigen derived from cedar pollen,
Anti-protein monoclonal solution labeled with colloidal gold
A solution was prepared by dissolving 10 μl in 5 ml of physiological saline.
In 5 ml of this solution, the short side on the opposite side of the test device provided with the absorber was immersed. As a result, after 5 minutes, the round pattern portion (the second antibody-immobilized site) into which the anti-protein polyclonal antibody solution was dropped was colored pink.

(Example 2) As a detecting device as shown in FIG. 6, a transparent container 211 made of polystyrene and a sealing lid 215 were used.
The case 221 was manufactured. The container is 14mm inside diameter and 80m long
m, the sealing lid had an inner diameter of 14 mm, an outer screw forming portion of 15 mm, an inner screw forming portion of 15 mm, and the case had an inner screw forming portion of 15 mm and a detector body of 30 mm. In the case, a cut portion 223 for cutting the sealing film of the sealing lid was provided in a shape as shown in FIG. The detection plate 232 is made of the same material as that of the first embodiment and has a width of 10 mm and a length of 25 mm.
A second antibody fixing portion 230 was formed at a position 15 mm from the top of 2 in the same manner as in Example 1. Furthermore, 5m at the lower end of the detection plate 232
The m-part was sprayed with the same anti-protein monoclonal solution labeled with colloidal gold as in Example 1 and dried at room temperature to form the first antibody-carrying site 229. Then, FIG.
As shown in (1), a filter 225, a first antibody carrying portion 229, a detection plate 232, and an absorption pad 232 were mounted on the case 221 to form a detection body 220.

5 g of garbage collected by a vacuum cleaner from an indoor carpet into which cedar pollen enters into the container 211 of the detection device, and 10 ml of physiological saline are added thereto.
After that, the protein in the pollen was extracted into the solution using an ultrasonic vibrator. Note that a polyethylene film having a thickness of 30 μ was used as the seal film 217 of the sealing lid 215. Next, the hermetic lid 215 of the container 211 containing the pollen sample liquid is placed.
The detection body 220 was attached to the portion, and the seal filler 217 of the hermetic lid 215 was broken by the cutting portion 223 so that the cedar pollen detection liquid in the container 211 permeated toward the detection body 220. As a result, after 5 minutes, the second antibody fixing portion 230 was colored pink, and it was confirmed that cedar pollen was present in the garbage collected from the carpet.

[0069]

According to the present invention, pollen causing asthma and various allergies can be detected at home with a simple device without having specialized knowledge.
Useful for prevention and treatment of asthma and allergies.
In particular, pollen floating in a room, pollen adhering to tatami mats, carpets, and the like are collected using a vacuum cleaner or the like, and the collected dust is easily applied to the device of the present invention, so that the presence of pollen can be easily determined. It is very useful for preventing asthma and allergy in the home because it is possible to know the presence or absence of pollen and the type of pollen.
Further, the pollen detection device of the present invention is a pollen determination unit,
That is, since a base material having a first antibody carrying site, a sample solution delaying site, a second antibody immobilizing site, a sample solution undeployable site (water-insoluble portion) and the like can be mass-produced using a printing technique, The product can be produced at low cost with little variation.

Since the first antibody and the second antibody used in the present invention are monoclonal or polyclonal antibodies produced by a known method which has been already established, a very small amount of pollen can be detected. can do. In addition, pollen-derived proteins that are antigens are derived from cedar pollen, ragweed pollen, kanagura pollen, hygema pollen, chrysanthemum pollen, akinokirin grass pollen, and duckweed pollen, which are particularly present in many homes as antigens. Therefore, the type of pollen can be determined. Further, either the first antibody or the second antibody is labeled, and the binding reaction between the complex of the first antibody and the protein derived from pollen and the binding reaction of the second antibody can be determined by coloration. The presence or absence of
No special equipment is required and can be checked with the naked eye.

[Brief description of the drawings]

FIG. 1 is a schematic diagram illustrating an example of a first detection device of a pollen detection device of the present invention.

FIG. 2 is a diagram showing that, in the first detection device according to the present invention, the development speed of the pollen sample liquid is reduced by providing a water-insoluble portion in a part of the base material.

FIG. 3 is a diagram showing that the development of a pollen sample solution converges on a positive determination site by providing a water-insoluble portion in a part of the base material in the first detection device according to the present invention.

FIG. 4 is an explanatory diagram when a water-insoluble portion is provided in a part of a substrate in the first detection device according to the present invention.

FIG. 5 is an explanatory diagram when a first detection device according to the present invention is manufactured.

FIG. 6 is a schematic sectional view showing a second detection device according to the present invention.

FIG. 7 is a sectional view taken along line II-II in FIG.

FIG. 8 is a sectional view taken along line III-III in FIG. 6;

FIG. 9 is a schematic sectional view showing another embodiment of the second detection device according to the present invention.

FIG. 10 is a sectional view taken along line VV in FIG. 9;

FIG. 11 is a schematic cross-sectional view of various types of absorbent pads constituted by using an absorbent gel.

FIG. 12 is a schematic sectional view of a third detection device according to the present invention.

FIG. 13 is a sectional view taken along line II-II of FIG.

FIG. 14 is a schematic sectional view showing another embodiment of the third detection device according to the present invention.

FIG. 15 is a sectional view taken along the line IV-IV in FIG. 14;

FIG. 16 is a schematic sectional view of a fourth detection device according to the present invention.

FIG. 17 is a perspective view of the detection device of FIG.

FIG. 18 is a schematic sectional view in which the detection device of FIG. 16 is left standing in the horizontal direction.

FIG. 19 is a plan view showing a partition 413 of the detection device of FIG.

FIG. 20 is a schematic sectional view showing another embodiment of the fourth detection device according to the present invention.

21 is a perspective view of a lower portion of the detection device of FIG.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 1st detection apparatus 2 2nd detection apparatus 3 3rd detection apparatus 4 4th detection apparatus 101 base material 102 sample liquid application part 103 1st antibody carrying part 104 expansion delay part 104a undeployment part 105 positive determination part 106 Negative judgment part 107 Undeployable part (near the positive judgment part) 107a, 107b Non-water-soluble part 211 Container 211a External screw (of container) 215 Sealing lid 215a Internal screw (of sealing lid) 215b External screw (of sealing lid) 217 Seal film 219 Flange 220 Detector 221 Case 221a Inner screw (of case) 223 Cutting part 225 Filter part 227 Detection plate 229 First antibody carrying part 230 Second antibody fixing part 232 Absorption pad 232b Absorption pad (annular) 232a Deployment spacer 233 Pad room 241 Non-woven fabric 24 Water-absorbing gel 243 Bag (made of non-woven fabric) 245 PET film 311 Container 311a Internal screw (of container) 311b External screw (of container) 313 Sealing lid 313a Internal screw (of sealing lid) 320 Detector 320a External screw (of detector) 321 Case 323 Water-soluble film 322 Communication hole 325 Filter section 327 Detection plate 329 First antibody holding section 330 Second antibody fixing section 411a Container (upper) 411b Container (lower) 413 Partition wall 413a Partition wall (container 411a) 413b Partition wall (of container 411b) 414 Solution chamber 415 Detection body chamber 417 Detection plate 418 Filter section 419 First antibody support section 420 Absorption pad 421 Sealing lid 425 Pollen specimen liquid 430 Second antibody fixing section 431a Communication hole (of partition wall 413a) 431b Communication hole (of partition 413b)

Claims (9)

[Claims] 1. A device for detecting pollen by determining the presence or absence of a pollen-derived protein, wherein a base material of the detection device is made of a porous material in which a pollen sample liquid can be developed, The substrate, a sample liquid application site to which a pollen sample liquid is applied, a first antibody-carrying site on which a first antibody capable of specifically binding to pollen-derived protein is supported, and a pollen-derived protein specific A pollen detection device comprising a positive determination site carrying a second antibody different from the first antibody capable of binding to a pollen. 2. A development delay section for delaying the development of the sample liquid between the first antibody-carrying part and the positive determination part, and a sample liquid non-deployable part in which the sample liquid cannot be developed is provided in a part near the positive determination part. The pollen detection device according to claim 1, wherein: 3. A third substance-immobilized part, on which a third substance capable of specifically binding to the first antibody is immobilized, is provided downstream of the positive determination part in the developing direction of the sample liquid. The pollen detection device according to claim 1 or 2. 4. An apparatus for detecting pollen by determining the presence or absence of a pollen-derived protein, the detection apparatus comprising: a container having one side open and capable of accommodating a pollen sample liquid; A sealing lid attached to the one-side opening, and a sealed detection body attached to the sealing lid, wherein the sealing lid has a sealing film, and the detection body seals the detection body with the sealing body. It has a cutting portion for breaking the seal film when attached to the lid, and a determining portion provided continuously to the cutting portion and for determining the presence or absence of pollen-derived protein. Pollen detector. 5. The detection unit, comprising: a detection plate that allows a pollen sample liquid to penetrate; and a first antibody holding unit that holds a first antibody capable of specifically binding to a pollen-derived protein connected to the detection plate. Unlike the first antibody, the second antibody is fixed to a second antibody capable of specifically binding to a pollen-derived protein, and an absorber for absorbing the pollen sample liquid that has passed therethrough. The pollen detection device according to claim 4, wherein: 6. An apparatus for detecting pollen by determining the presence or absence of a pollen-derived protein, the detection apparatus comprising: a container having one side open and capable of accommodating a pollen sample liquid; A detection body attached to one side opening of the container, the detection body has a communication hole sealed with a water-soluble film, a case mounted on one side opening of the container, and a case inside the case. A pollen detection device provided with a determination unit for determining the presence or absence of a protein derived from pollen. 7. The detection unit, comprising: a detection plate that allows a pollen sample liquid to penetrate; and a first antibody holding unit that holds a first antibody that can specifically bind to a pollen-derived protein connected to the detection plate. Unlike the first antibody, a pollen sample is provided between a second antibody-fixed portion in which a second antibody capable of specifically binding to a pollen-derived protein is fixed, and a communication hole and a determination portion in the case. The pollen detecting device according to claim 6, comprising a filter unit for removing solid matter in the liquid, and an absorber for absorbing the pollen sample liquid and the antibody-sensitized particles that have passed therethrough. 8. A device for detecting pollen by determining the presence or absence of pollen-derived protein, wherein the detection device is capable of accommodating a pollen sample liquid, one side being open and the other side being sealed. And a closed lid attached to the opening of the container, a solution chamber is provided on the opening side of the container and a detection body chamber is provided on the closed side, and the solution chamber and the detection body chamber are openable and closable. A pollen detection device, wherein the pollen detection device is divided by a partition wall, and provided with a determination unit for determining the presence or absence of pollen-derived protein in the detection body chamber. 9. The detection unit, comprising: a detection plate that allows a pollen sample liquid to penetrate; and a first antibody holding unit that holds a first antibody that can specifically bind to a pollen-derived protein connected to the detection plate. Unlike the first antibody, a second antibody-fixed portion in which a second antibody capable of specifically binding to pollen-derived protein is fixed, and between the solution chamber and the detection body chamber, a pollen sample liquid is contained. 9. The pollen detecting device according to claim 8, comprising: a filter section for removing the solid matter, and an absorber for absorbing the passed pollen sample liquid and the antibody-sensitized particles.