JP2006136274A - Adhesive sheet for examining microorganism, method for producing the same, and method and kit for examining the microorganism by using the adhesive sheet - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a kit for examining microorganisms by collecting the microorganisms, growing the microorganisms on an adhesive sheet, and detecting the proliferated microorganisms. <P>SOLUTION: The adhesive sheet and kit for examining the microorganisms present in a specimen by pressing an adhesive layer of the adhesive sheet obtained by laminating the water-soluble adhesive layer on a base material to the surface of the specimen, separating the adhesive layer to collect the microorganisms or cells on the surface of the adhesive layer, culturing the microorganisms on the adhesive sheet, and detecting the collected and cultured microorganisms are provided. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a pressure-sensitive adhesive sheet for microbiological examination capable of culturing and growing microorganisms collected on a pressure-sensitive adhesive layer and a method for producing the same. The present invention also relates to a microorganism testing method and a testing kit, characterized in that microorganisms are detected using the adhesive sheet.

  Conventionally, in order to estimate the number of bacteria in a test material, for example, an agar stamp method using a commercially available food stamp or the like, it is applied that one bacterium forms one colony, There is a method in which a solid medium is pressed against a test material to cultivate bacteria, and a colony expressed on an agar plate is measured with the naked eye. In addition, as in the membrane filter method using a commercially available membrane filter for microorganisms / microparticle inspection, etc., the test material is washed out with physiological saline, collected by the membrane filter, and absorbed on the liquid medium. There is also a method of forming a colony. Furthermore, Non-Patent Document 1 below reports an example in which a film is coated on a film and a film-coated medium capable of collecting a sample is prepared by simply culturing the film on a test surface.

  However, with the agar stamp method, the size (area) of the agar medium is limited, and since it can usually be used only once per test surface, the collection efficiency varies depending on the moisture content of the agar medium, and reproducibility. In many cases, it is inconvenient in the collection efficiency of bacteria. Furthermore, there is also a problem that the test surface is contaminated by moisture in the agar medium.

  In addition, the membrane filter method requires two steps, a washing-out step and a filtration step with a filter, and thus there are inconveniences such as complicated operations and concern about secondary contamination. That is, in this method, bacteria cannot be collected directly from the test substance, and sterilization work before use, sampling with a cotton swab, adjustment of the washing solution, filter filtration, etc. require a lot of labor, and an increase in cost is inevitable. .

  Further, the film-coated medium disclosed in Non-Patent Document 1 below has a problem that the test surface is contaminated by moisture, as in the agar stamp method. Moreover, since the bending strength of the agar culture medium used is small, utilization as a sheet | seat is restrict | limited and there exist various faults, such as insufficient adhesive force of a culture medium and a film.

On the other hand, the present applicant contains one or more coloring materials capable of staining microorganisms after pressure-bonding and peeling the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet and collecting (accumulating) the microorganisms on the pressure-sensitive adhesive layer. The test method which can detect the microorganisms which exist in a test subject by making the aqueous solution to contact the adhesive layer surface and observing and counting (image analysis) the dyed microbial cell was proposed (patent document 1 below). In this inspection method, the microorganisms present on the surface of the solid subject (solid surface) can be easily detected by using the adhesive layer of the adhesive sheet as a microorganism collecting means and test surface. Is. However, for example, when the surface of the subject is relatively clean, there is a problem that sufficiently high detection accuracy cannot be obtained because the amount of collected microorganisms is small. Moreover, in the method of the following patent document 1, there also existed a problem that it was difficult to distinguish between the collected microorganisms and the minute garbage.
JP 2002-142797 A Medical Examination, 1992, Vol. 41, No. 3, p352

  The present invention has been regarded as a matter of the above circumstances, and its purpose is to test microorganisms that can culture and proliferate microorganisms (specimens) collected on an adhesive layer by pressure bonding to and peeling from a subject. An adhesive sheet, a method for producing the same, and a microorganism testing method and a testing kit characterized by detecting microorganisms using such an adhesive sheet.

  As a result of diligent research to solve the above-mentioned problems, the present inventors have found that the pressure-sensitive adhesive layer obtained by mixing a microbial nutrient with a water-soluble pressure-sensitive adhesive can be a medium-like medium capable of growing microorganisms. As a result of further research based on the finding and such findings, the present invention has been completed.

That is, the present invention
(1) A pressure-sensitive adhesive sheet for microbiological examination, comprising an adhesive layer made of a water-soluble pressure-sensitive adhesive provided on at least one surface of a support, containing nutrients for microorganisms,
(2) The adhesive sheet for microbiological examination according to (1), wherein the water-soluble adhesive is an acrylic copolymer,
(3) The pressure-sensitive adhesive sheet for microbiological examination according to (2), wherein the acrylic copolymer is an acrylic copolymer mainly comprising alkoxy acrylate and N-vinyl lactam.
(4) The pressure-sensitive adhesive sheet for microbial testing according to any one of (1) to (3), wherein the pressure-sensitive adhesive layer further contains a selection agent capable of selecting a microorganism to be tested.
(5) The pressure-sensitive adhesive sheet for microbial examination according to any one of (1) to (4), further comprising a microbial colorant in the pressure-sensitive adhesive layer,
(6) The adhesive sheet for microbial examination according to (5), wherein the microbial colorant is a fluorescent dye,
(7) The adhesive sheet for microbiological examination according to any one of (1) to (6), wherein the support is porous,
(8) A pressure-sensitive adhesive for microorganism testing, characterized in that a pressure-sensitive adhesive composition prepared by adding and mixing at least microbial nutrients to a water-soluble pressure-sensitive adhesive is coated on a support and dried to form a pressure-sensitive adhesive layer. Manufacturing method of the adhesive sheet,
(9) The adhesive composition for microbiological examination according to (8), wherein the adhesive composition is prepared by adding and mixing a water-soluble adhesive with a microbial nutrient and a selective agent capable of selecting a microorganism to be examined. Sheet manufacturing method,
(10) Microorganisms collected on the pressure-sensitive adhesive layer by collecting the microorganisms by pressure-bonding the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet for microorganism testing according to any one of (1) to (7) to a subject. A method for examining microorganisms present in a subject, the method comprising culturing and proliferating, and detecting the proliferated microorganisms,
(11) A solution containing microorganisms to be tested is dropped on the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet for microorganism testing according to any one of (1) to (7), and the microorganisms on the pressure-sensitive adhesive layer are cultured. A method of examining microorganisms present in a subject, the method comprising growing and detecting the grown microorganisms;
(12) The method according to (10) or (11), wherein the grown microorganisms are detected by visual observation, a sensitizer or an automatic detector.
(13) A pressure-sensitive adhesive sheet for microorganism testing according to any one of (1) to (7), a colorant capable of coloring the microorganism to be examined and / or a selection agent capable of selecting the microorganism to be examined. Microbiological test kit, characterized by
About.

  In the present specification, “microorganism inspection” means “microorganism detection inspection”.

  According to the pressure-sensitive adhesive sheet for microbiological examination of the present invention, since the pressure-sensitive adhesive layer contains microbial nutrients, the microorganisms collected on the pressure-sensitive adhesive layer can be cultured and propagated as they are. Therefore, even when the amount of microorganisms (specimen) collected on the adhesive layer is small, adhesiveness can be obtained without performing troublesome operations such as transferring and culturing the microorganisms collected on the adhesive layer to another medium. It is possible to increase the concentration of microorganisms (specimen) on the sheet and perform a highly accurate detection operation using the pressure-sensitive adhesive layer surface as a test surface. In addition, when a subject with relatively high cleanliness is to be examined, the examination range can be expanded on the same adhesive surface, so that microorganisms can be detected economically and labor-saving. In addition, since the collection of microorganisms depends on the adhesive strength of the adhesive, a relatively high collection effect can be stably obtained, and the problem of contamination of the test surface due to moisture, which has been a problem with conventional culture media, Can also be resolved.

  In addition, since the adhesive sheet for microbiological examination of the present invention is excellent in operability, handling, stability and storage as a sheet, other reagents, materials and instruments necessary for the growth and / or detection of microorganisms In combination with the above, a kit for microbiological examination can be constituted, and the kit allows the inspection operator to carry out microbiological examination more easily.

Hereinafter, the present invention will be described in more detail.
FIG. 1 is a cross-sectional view showing, in a simplified manner, an example of an adhesive sheet for microbiological examination of the present invention (hereinafter sometimes simply referred to as “adhesive sheet”). The pressure-sensitive adhesive sheet for microbiological examination of the present invention has a pressure-sensitive adhesive layer 1 formed on at least one surface of a support 2 as shown in the microbe-testing pressure-sensitive adhesive sheet 10 of the example, and the pressure-sensitive adhesive layer 1 has at least a microorganism. It contains nutrients necessary for growth.

  In this example, the pressure-sensitive adhesive sheet 10 is provided with the pressure-sensitive adhesive layer 1 on one surface of the support 2. In the pressure-sensitive adhesive sheet of the present invention, the pressure-sensitive adhesive layer 1 is formed on both surfaces of the support 2 (one and the other). It may be provided on the other side.

  In the pressure-sensitive adhesive sheet of the present invention, the pressure-sensitive adhesive used for the pressure-sensitive adhesive layer 1 is not particularly limited as long as it is water-soluble, and a known water-soluble pressure-sensitive adhesive can be used without limitation.

  Examples of the water-soluble pressure-sensitive adhesive (hereinafter also referred to as “water-soluble polymer”) constituting the pressure-sensitive adhesive layer include agar, carrageenan, gum arabic, polyvinyl pyrrolidone, polyvinyl alcohol (PVA), gelatin, carboxymethyl cellulose, and alginic acid. , Methylcellulose, polyacrylic acid and / or polyacrylate polymer compounds, acrylic copolymers, hydrophilic polyether urethane polymers, stilbazolated polyvinyl alcohol, xanthan gum and other polysaccharides, polyethylene oxide, poly (N Acrylamides such as -isopropylacrylamide) are preferred.

  More preferable water-soluble polymers include acrylic copolymers mainly composed of alkoxy acrylate and N-vinyl lactam from the viewpoint of the speed of colony growth and separation performance. Here, “acrylic copolymer having alkoxy acrylate and N-vinyl lactam as main structural units” means a binary copolymer of alkoxy acrylate and N-vinyl lactam or other monomers in the binary copolymer. Further, it means a copolymerized copolymer.

  As the alkoxyalkyl acrylate which is a monomer component of the acrylic copolymer, an acrylate having an alkoxy group having 1 to 4 carbon atoms and an alkyl group having 2 to 4 carbon atoms or an alkylene glycol group is preferable. Specifically, for example, 2-methoxyethyl acrylate, 2-ethoxyethyl acrylate, 2-butoxyethyl acrylate, 3-methoxypropyl acrylate, 3-methoxybutyl acrylate, 3-ethoxypropyl acrylate, 3-ethoxybutyl acrylate, Examples include alkoxyalkylene glycol acrylates such as butoxytriethylene glycol, 2- (2-ethoxyethyl) ethyl acrylate, methoxytriethylene acrylate, and methoxydipropylene glycol acrylate, which have a low glass transition point, high hydrophilicity, and reactivity. From the point of being high, 2-methoxyethyl acrylate and 2-ethoxyethyl acrylate are preferable.

  As N-vinyl lactam, 5 to 7-membered N-vinyl lactam is used. For example, N-vinyl-2-pyrrolidone, N-vinyl-2-piperidone, N-vinyl-2-caprolactam and the like can be mentioned, and N-vinyl-2-pyrrolidone is preferable from the viewpoint of safety and versatility.

  The compounding ratio of the alkoxyalkyl acrylate is preferably 60 to 80% by weight, more preferably 65 to 75% by weight, based on the entire acrylic copolymer. When the blending ratio of the alkoxyalkyl acrylate is less than 60% by weight, the obtained pressure-sensitive adhesive layer may have low flexibility. On the other hand, if it exceeds 80% by weight, the pressure-sensitive adhesive layer may have increased adhesiveness and lower strength.

  Further, the blending ratio of N-vinyl lactam is preferably 20 to 40% by weight, more preferably 25 to 35% by weight, based on the entire acrylic copolymer. When the blending ratio of N-vinyl lactam is less than 20% by weight, the strength and water absorption capacity of the obtained pressure-sensitive adhesive layer may be lowered. On the other hand, if it exceeds 40% by weight, the resulting adhesive layer has low flexibility, and the acrylic copolymer may dissolve in moisture and flow may occur.

  The acrylic copolymer may be copolymerized with vinyl carboxylic acid in order to improve water absorption ability. Examples of the vinyl carboxylic acid include acrylic acid, methacrylic acid, crotonic acid, itaconic acid, maleic acid, fumaric acid, maleic acid half ester, and fumaric acid half ester. Among these, acrylic acid is preferable from the viewpoint of reactivity and versatility.

  The blending ratio of the vinyl carboxylic acid is preferably 10% by weight or less, more preferably 5% by weight or less, based on the entire acrylic copolymer. When the blending ratio of the vinyl carboxylic acid exceeds 10% by weight, the pressure-sensitive adhesive layer tends to have increased water absorption capacity and lower flexibility.

  An acrylic copolymer obtained by blending 0.01 to 1% by weight of polyfunctional acrylate or methacrylate and copolymerizing is also useful.

  Examples of the polyfunctional acrylate or methacrylate used in the present invention include 1,4-butanediol diacrylate, 1,4-butanediol dimethacrylate, neopentyl glycol diacrylate, 1,6-hexanediol diacrylate, 1,6. -Alkyl diacrylate or dimethacrylate such as hexanediol dimethacrylate, diethylene glycol diacrylate, diethylene glycol dimethacrylate, triethylene glycol diacrylate, triethylene glycol dimethacrylate, polyethylene glycol 200 diacrylate, polyethylene glycol 400 diacrylate, polyethylene glycol 600 Diacrylate, polyethylene glycol 200 dimethacrylate, polyethylene glycol 400 dimethacrylate, polyethylene glycol 600 dimethacrylate, polyethylene glycol 1000 dimethacrylate, polypropylene glycol 400 diacrylate, polypropylene glycol 400 dimethacrylate and other alkylene glycol diacrylates or dimethacrylates, hydroxypivalate neopentyl glycol diacrylate, etc. Trimethylolpropane type tri and tetraacrylates or trimethacrylates such as ester type diacrylate or dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, ethoxylated trimethylolpropane triacrylate, ditrimethylolpropane tetraacrylate, pentaerythritol Pentaerythritol type tri and tetra acrylates such as acrylate and pentaerythritol tetramethacrylate or tri and tetra methacrylate, isocyanurate type triacrylate or trimethacrylate such as tris (acryloxyethyl) isocyanurate, tris (methacryloxyethyl) isocyanurate, ethoxy Bisphenol A diacrylate or dimethacrylate such as bisphenol A diacrylate and ethoxylated bisphenol A dimethacrylate, and the like. From the viewpoint of hydrophilicity and safety, alkylene glycol diacrylate or dimethacrylate is preferable, and crosslinking is moderate. Polyethylene glycol 600 dimethacrylate, polyethylene glycol 400 dimethacrylate More preferred.

  The blending ratio of the polyfunctional acrylate or methacrylate is preferably 0.01 to 1% by weight, more preferably 0.05 to 0.5% by weight, based on the whole acrylic copolymer. When the amount of this component is less than 0.01% by weight, the crosslinking is insufficient, the strength is lowered, and it tends to be water-soluble. On the other hand, when it exceeds 1% by weight, flexibility and adhesiveness are lowered, hydrophilicity is remarkably lowered, and whitening tends to occur after water absorption.

  The water-soluble pressure-sensitive adhesive of the present invention can be produced by a method known per se.

  The acrylic copolymer may be produced by any copolymerization method such as solution polymerization, emulsion polymerization, suspension polymerization, bulk polymerization, and photopolymerization. Solution polymerization is carried out by stirring the monomer mixture in the presence of a medium and a polymerization initiator. Examples of the solvent include water, methanol, ethanol, isopropanol, acetone, methyl ethyl ketone, methyl acetate, ethyl acetate, toluene, xylene, ethylene glycol monoalkyl ether, and the like. These may be used alone or in combination of two or more. Can be used. Examples of the polymerization initiator include benzoyl peroxide, cumene hydroperoxide, diisopropyl peroxydicarbonate, azobisisobutyronitrile and the like, and these can be used alone or in combination of two or more. . Polymerization preferably proceeds at 0.01 to 5 parts by weight with respect to 100 parts by weight of the monomer, and more preferably 0.1 to 0.5 parts by weight.

  The acrylic copolymer can also be polymerized by photopolymerization performed by irradiating the monomer mixture with UV light in the presence of a photoinitiator. In photopolymerization, a mixture of monomers and photoinitiators excluding polyfunctional acrylate or methacrylate is sufficiently purged with nitrogen, and then irradiated with a certain dose of UV light, and the resulting partially polymerized syrup is mixed with polyfunctional acrylate or A mixture of methacrylate and a plasticizer described later is applied or cast into a frame, and then irradiated with sufficient UV light to obtain a copolymer. Alternatively, after thoroughly replacing the monomer, photoinitiator, and if necessary, a mixture of plasticizers with nitrogen, irradiate a certain dose of UV light, and apply or cast the resulting partially polymerized syrup into a frame A method of obtaining a copolymer by irradiating with sufficient UV light is preferable.

In this case, the total irradiation dose of the partial polymerization is preferably 20 mJ / cm ² or more 300 mJ / cm ² or less, completely polymerized total irradiation dose at the time of preferably 1000 mJ / cm ² or more 10000 mJ / cm ² or less, 2000 mJ / cm ² or more 5000 mJ / cm ² or less is more preferable.

  Photoinitiators include, for example, 4-phenoxydichloroacetophenone, 4-t-butyl-dichloroacetophenone, 4-t-butyl-trichloroacetophenone, diethoxyacetophenone, 2-hydroxy-2-methyl-1-phenylpropane-1- ON, 1- (4-isopropylphenyl) -2-hydroxy-2-methylpropan-1-one, 1- (4-dodecylphenyl) -2-hydroxy-2-methylpropan-1-one, 4- (2 Acetophenones such as -hydroxyethoxy) -phenyl- (2-hydroxy-2-propyl) ketone, 1-hydroxycyclohexyl phenylketone, 2-methyl-1- [4- (methylthio) phenyl] -2-morpholinopropane-1 Photoinitiator, benzoin, benzoin methyl ether, benzoin ethyl Benzoin photoinitiators such as ether, benzoin isopropyl ether, benzoin isobutyl ether, benzyl dimethyl ketal, benzophenone, benzoylbenzoic acid, methyl benzoylbenzoate, 4-phenylbenzophenone, hydroxybenzophenone, 4-benzoyl-4'-methyldiphenyl sulfide Benzophenone photoinitiators such as 3,3′-dimethyl-4-methoxybenzophenone, thioxanthone, 2-chlorothioxanthone, 2-methylthioxanthone, 2,4-dimethylthioxanthone, isopropylthioxanthone, 2 , 4-dichlorothioxanthone, 2,4-diethylthioxanthone, 2,4-diisopropylthioxanthone, and other thioxanthone photoinitiators. Although the photoinitiator varies depending on the extinction coefficient and the like, the polymerization preferably proceeds at 0.005 parts by weight or more and 0.5 parts by weight or less based on 100 parts by weight of the monomer, and 0.01 parts by weight or more and 0.2 parts by weight or less Is more preferable.

  In this case, a photoinitiator aid may be added as necessary. Photoinitiator is, for example, triethanolamine, methyldiethanolamine, triisopropanolamine, 4,4′-dimethylaminobenzophenone, 4,4′-diethylaminobenzophenone, ethyl 2-dimethylaminobenzoate, 4-dimethylaminobenzoic acid Examples include ethyl, ethyl 4-dimethylaminobenzoate (n-butoxy), isoamyl 4-dimethylaminobenzoate, 2-ethylexyl 4-dimethylaminobenzoate, and the like. These may be used alone or in combination of two or more. Can be used. Polymerization proceeds preferably at 0.01 to 1 part by weight, and more preferably 0.02 to 0.5 part by weight, with respect to 100 parts by weight of the monomer.

  A plasticizer can be added to the water-soluble polymer as necessary. By adding a plasticizer, flexibility and adhesiveness can be further improved. As the plasticizer, it is desirable to use a hydrophilic compound having good compatibility with the water-soluble polymer to be used.

  For example, when a plasticizer is added to an acrylic copolymer, the plasticizer is preferably 5 to 40 parts by weight, more preferably 10 to 30 parts by weight, based on 100 parts by weight of the acrylic copolymer. preferable. Further, when a plasticizer is added to another water-soluble polymer, the plasticizer is preferably 5 parts by weight or more and 500 parts by weight or less, more preferably 10 parts by weight or more and 300 parts by weight or less with respect to 100 parts by weight of the water-soluble polymer. preferable.

  If the plasticizer exceeds 40 parts by weight with respect to 100 parts by weight of the acrylic copolymer, the acrylic copolymer properties such as a decrease in cohesive force may be lost, or radiation or chemical crosslinking such as an electron beam described later. Insolubilization by an agent may be difficult, or insolubilization by polyfunctional acrylate or methacrylate crosslinking may be difficult. From these viewpoints, the plasticizer is more preferably 40 parts by weight or less with respect to 100 parts by weight of the acrylic copolymer. In order to further improve flexibility and tackiness, the plasticizer is preferably 5 parts by weight or more with respect to 100 parts by weight of the acrylic copolymer.

  Examples of the plasticizer used in the present invention include ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, glycerin, diglycerin, 2,3-butanediol, 1,2-butanediol, and 3-methyl. Low molecular weight polyhydric alcohols such as 1,3-butanediol, 3-methyl-1,3,5-pentanetriol, 2-ethyl-1,3-hexanediol, polyethylene glycol having an average molecular weight of 1000 or less, Polypropylene glycol, polyoxyethylene butyl ether, polyoxyethylene, polyoxypropylene, polyoxypropylene glyceryl ether, polyoxypropylene glycol, polyoxypropylene sorbitol, polyoxyethylene polyoxypro Renpentaerythritol ether and the like can be mentioned, but polyoxypropylene glyceryl ether having an average molecular weight of 200 or more and 800 or less, and an average molecular weight of 200 or more and 800 or less from the viewpoint of obtaining excellent flexibility, adhesiveness and hydrophilicity. A certain polyoxypropylene glycol, polyoxypropylene sorbitol having an average molecular weight of 200 to 800 is preferable.

  Moreover, in order to maintain an appropriate moisture content, a hydrophilic low molecular weight substance can also be added. Examples of hydrophilic low-molecular substances include high-boiling liquid substances and / or deliquescent inorganic salts. As the high-boiling liquid substance, a compound having a boiling point of 100 to 400 ° C., preferably 200 to 350 ° C. is preferable. Specific examples thereof include polyhydric alcohol, sugar alcohol and the like. Examples of the polyhydric alcohol include ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol, propylene glycol, polypropylene glycol and glycerin. Examples of the sugar alcohol include sorbitol and the like. Is mentioned. Specific examples of deliquescent inorganic salts include lithium nitrate and lithium chloride.

  The amount of the hydrophilic low-molecular substance used is preferably 20 to 300 parts by weight with respect to 100 parts by weight of the water-soluble polymer, and this ratio is particularly preferable in the polyacrylic acid polymer compound. If the amount of the hydrophilic low-molecular substance used is less than 20 parts by weight, a remarkable moisture absorption effect or moisturizing effect cannot be obtained, and if it exceeds 300 parts by weight, the viscosity of the water-soluble composition is too high and workability is poor. There is nothing.

  Examples of the sterilization method for the water-soluble adhesive of the present invention include sterilization by irradiation with an electron beam or γ-ray. In this case, the radiation irradiation dose is preferably 25 kGy or more and 50 kGy or less, although it varies depending on the water solubility of an acrylic copolymer, for example. When the dose is less than 25 kGy, sterilization is difficult to ensure, and when the irradiation dose is 25 kGy or more, sterilization defined by medical regulations can be used. When it exceeds 50 kGy, the modification such as coloring is remarkable and the use is limited.

  The above water-soluble polymer, plasticizer and / or hydrophilic low-molecular substance are added with water and a crosslinking agent as necessary, and mixed and stirred to form a gelled product. Alternatively, it may be used as a frozen or thawed gel like PVA. Moreover, the gelation by the radiation irradiation bridge | crosslinking of an electron beam or a gamma ray is also mentioned. In particular, in the case of an acrylic copolymer, UV photopolymerization is effective.

  Preferable crosslinking agents include compounds having at least two epoxy groups in the molecule, such as polyethylene glycol diglycidyl ether, ethylene glycol diglycidyl ether, glycerin triglycidyl ether, triglycidyl isocyanurate, and the like. These cross-linking agents are particularly preferable for polyacrylic acid polymer compounds.

  Examples of the cross-linking treatment method include insolubilization by irradiation cross-linking such as electron beam or γ-ray. In this case, the radiation dose varies depending on the degree of water solubility of, for example, an acrylic copolymer, but is preferably 1 kGy or more and 50 kGy or less.

  Moreover, also when water solubility increases by mix | blending a plasticizer with an acrylic type copolymer, the insolubilization by a crosslinking process may be needed.

  Furthermore, when the plasticizer is added to the above acrylic copolymer, the blending ratio of the acrylic copolymer is 60% by weight or more in the total amount of the acrylic copolymer and the plasticizer. In some cases, it is necessary to realize the shape retention by realizing insolubilization of the acrylic copolymer by crosslinking treatment, including the case of consisting of the copolymer alone, that is, 100% by weight of the acrylic copolymer.

  The water content in the aqueous composition for the pressure-sensitive adhesive layer thus obtained is preferably 5 to 95% by weight, and the water content of the gel layer after solidification / curing is usually preferably maintained in the range of 5 to 80% by weight. . This moisture content is a preferred moisture content especially in a polyacrylic acid polymer compound. The pH of the gelled product can be adjusted as necessary, but is usually preferably in the range of 6.5 to 7.5.

  In the pressure-sensitive adhesive sheet of the present invention, the nutrients of the microorganisms contained in the pressure-sensitive adhesive layer 1 are not particularly limited, and various nutrients can be selected according to the microorganisms to be detected. For example, standard agar medium, LB medium, SCDA medium (For general bacteria), R2A medium (for water quality test), mannitol salt medium (for S. aureus), and the like. Although the compounding quantity of this nutrient is not specifically limited, About 0.5 to 20 weight% is preferable with respect to an adhesive (solid content) from the point of the proliferation property of microorganisms, and about 1 to 10 weight% is more preferable.

  In addition to microbial nutrients, a functional substance (additive) that suitably acts on detection of microorganisms can be added to the adhesive layer 1. Examples of the functional substance (additive) include a selective agent for detecting a specific microorganism, a colorant capable of coloring the microorganism, and the like.

Specific examples of the selective agent include a selective medium, an antibacterial agent, an antibiotic and the like. Specific microorganisms include Escherichia coli, enterohemorrhagic Escherichia coli, Staphylococcus aureus, Salmonella, Vibrio parahaemolyticus, Listeria, and drug-resistant MRSA, natural water, which is attracting attention in hospital infection problems. Microorganisms present therein, microorganisms present in pharmaceutical water, yeasts, fungi and the like. Specific examples of antibacterial agents added for detection of resistant bacteria include, for example, alcohols, glutaraldehyde, iodophor, popidone iodine, benzalkonium chloride, benzethonium chloride, alkylpolyaminoethylglycine, cyclohexidine gluconate, benzisothiazoline, Organic nitrogen-containing cyclic compounds, organic nitrogen sulfur compounds, organic nitrogen sulfur halogen compounds, pyridine oxide compounds, etc. Specific examples of antibiotics include, for example, cephem, second generation cephem, third generation Cephem antibiotics (cephalosin, cephaloridine, etc.), penicillin antibiotics (methicillin, ampicillin, penicillin G, etc.), tetracycline antibiotics (tetracycline, etc.), peptide antibiotics (vancomycin, etc.), aminoglycoside anticancer agents Substance (arbekacin, spray script mycin, kanamycin, etc.). The blending amount of these antibacterial agents and / or antibiotics is not particularly limited, but is preferably about 1 × 10 ⁻⁶ to 1 × 10 ⁻¹ wt% with respect to the adhesive (solid content), and 2 × 10 ⁻⁵ to 4 ×. ^10-3 wt% is more preferred.

  As the microorganism colorant, a known material used for coloring microorganisms in the microorganism detection test (test) can be used. For example, fluorescent dyes widely used for coloring microorganisms, enzyme substrates specific for growing microorganisms, and colorants such as pH indicators can be used. The fluorescent dye is not particularly limited as long as it is a substance that emits fluorescence by acting on the microorganism or cell to be examined (fluorescent substance), but it does not emit light itself, but is converted by the action of the microorganism to emit light. It is preferable to use a substance such as SYBR Green I, SYBR Green II, SYBR Gold, SYTOX Green, DAPI, 6-Carboxyfluorescein Diacetate (CFDA), TOTO-1 Iodide, YOYO-1 Iodide, SYTO 62, Examples thereof include SYTO 63 and SYTO 64 (both manufactured by Molecular Probes). Among them, SYBR Gold is more preferable because it has good staining ability of microorganisms and hardly inhibits growth of microorganisms. The concentration of the fluorescent dye to be added is preferably 0.01 to 100 ppm, more preferably 0.1 to 10 ppm in the pressure-sensitive adhesive layer. That is, if the concentration is less than 0.01 ppm, the microorganisms are weakly colored and detection is difficult, and if the concentration exceeds 100 ppm, background coloration occurs and detection is difficult, which is not preferable. Moreover, when using an enzyme substrate as a coloring agent, the concentration of the enzyme substrate to be added is preferably 10 μg / ml to 10 mg / ml, more preferably 50 μg / ml to 2 mg / ml in the pressure-sensitive adhesive layer. That is, when the concentration is less than 10 μg / ml, the microorganisms are weakly colored and detection is difficult, and when the concentration exceeds 10 mg / ml, background coloring occurs and detection is difficult, which is not preferable. Further, when a pH indicator is used as the colorant, the concentration of the pH indicator to be added is preferably 1 μg / g to 1 mg / g, more preferably 5 μg / g to 200 μg / g in the pressure-sensitive adhesive layer. That is, when the concentration is less than 1 μg / g, the microorganisms are weakly colored and detection is difficult, and when the concentration exceeds 1 mg / g, background coloration occurs and detection is difficult, which is not preferable.

  The pressure-sensitive adhesive sheet of the present invention is produced by a method known per se.

  For example, an aqueous solution (aqueous composition) of a water-soluble polymer substance containing a plasticizer, a hydrophilic low-molecular substance and / or a crosslinking agent is applied to a support and dried at 10 ° C. to 200 ° C. In the case of non-permeability, it is produced by making a hole in the support. The pressure-sensitive adhesive layer formed into a thin film by drying may be post-treated at a temperature of 10 ° C. to 100 ° C. as necessary. The formed pressure-sensitive adhesive layer is imparted with appropriate tackiness and moisture content.

  In particular, an acrylic copolymer has excellent thermoplasticity and can be processed into a sheet by extrusion. For this extrusion molding, any generally known method such as inflation molding, T-die molding, lamination molding or the like can be used. However, since high temperature extrusion and rapid cooling are possible, a film or sheet having excellent optical properties can be obtained, and cooling efficiency can be obtained. From the standpoints that the production speed can be increased and the sheet thickness can be easily adjusted, T-die molding is preferred. As the extruder, either a single screw extruder or a twin screw extruder may be used. The thickness of the film or sheet can be adjusted by appropriately controlling the molding conditions such as the molding temperature, die lip width, extrusion speed, and take-up speed. In this case, the molding temperature is preferably 140 ° C. or higher and 180 ° C. or lower, more preferably 150 ° C. or higher and 170 ° C. or lower. When the molding temperature is less than 140 ° C., non-melted material is often mixed and the quality may vary. On the other hand, when the molding temperature exceeds 180 ° C., film breakage and bubble entrapment may occur.

  In addition, as a method for forming the film and sheet, methods such as a calendar method and a casting method can be used. However, when the polymerization method is photopolymerization, a monomer mixture containing a photoinitiator, or a partially polymerized syrup thereof, or a monomer mixture containing a photoinitiator and a plasticizer, or a partially polymerized syrup thereof on a peelable film made of a polyester film or the like. After the film is cast and applied in a constant thickness and covered with a transparent peelable film, the film is polymerized and film-formed simultaneously by sufficiently irradiating UV light in an inert atmosphere. The film and sheet thus obtained can be cut into an arbitrary shape and used as an adhesive sheet, and can be used by being bonded to an arbitrary support.

  Furthermore, an acrylic copolymer can be obtained by radical polymerization of a mixture of a monomer, a solvent and a polymerization initiator constituting the acrylic copolymer in the present invention by ordinary operations. This acrylic copolymer is obtained. A solution of the polymer can be cast to form a film. This film is adjusted to have a thickness of 10 to 75 μm, preferably 20 to 50 μm. If the thickness exceeds 75 μm, impurities and residual monomers cannot be sufficiently removed, and the film is remarkably removed during drying described later. On the other hand, if the thickness is less than 10 μm, the water absorption is small and the required strength may not be obtained. The film thus obtained can be dried under certain conditions to obtain a film-like pressure-sensitive adhesive from which the solvent and the like (solvent and / or prepolymer, hereinafter simply referred to as impurities) and residual monomers have been removed.

  In this case, the drying conditions differ depending on the thickness of the film and further drying conditions such as reduced pressure drying or atmospheric pressure drying. In the atmospheric pressure drying, the film thickness is 10 to 75 μm, and the film thickness is 30 to 110 to 150 ° C. Second to 20 minutes, particularly 1 to 15 minutes at 120 to 140 ° C. is desirable.

  As described above, the impurities and residual monomers are removed by drying the film formed of the present polymer. As a result, the quality is stabilized, and adverse effects on microorganisms based on the impurities and residual monomers can be eliminated.

  Except for the method of casting the acrylic copolymer with a solvent, the pressure-sensitive adhesive layer 1 in the pressure-sensitive adhesive sheet for microbiological examination of the present invention has a thickness of 10 μm or more, preferably 20 μm or more and 2000 μm or less, more preferably 30 μm or more. It is preferable to set it to 1000 μm or less. When the thickness is less than 10 μm, the film breakage phenomenon often occurs, which is not preferable. When the thickness exceeds 2000 μm, the water permeability is poor and there is no point in increasing the thickness.

  In the present invention, the water-soluble polymer can be formed into this film or sheet and then subjected to crosslinking by irradiation with radiation such as electron beam or γ-ray. In particular, this radiation crosslinking is effective in acrylic copolymers. The crosslinking treatment can be performed under the same conditions as described above.

  In addition, the adhesive sheet of the present invention can be in a sterilized state, for example, by being sealed in a bacteria-blocking packaging material in a sterilized state. The sterilization method is preferably an EOG (ethylene oxide gas) method, an electron beam method, or a γ ray method. That is, after the adhesive sheet is sealed in a bacteria-blocking wrapping material, the sterilized state can be maintained, and then subjected to crosslinking and sterilization treatment by irradiation with electron beams or γ rays. The polymer and the pressure-sensitive adhesive sheet using the polymer can be distributed.

  In this case, the irradiation dose of radiation such as an electron beam or γ-ray varies depending on the hydrophilicity of the polymer, but the crosslinking treatment can be performed under the same conditions as described above.

  The pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet thus obtained preferably has a uniform water absorbability. That is, the pressure-sensitive adhesive layer has a peel adhesive force in the direction of 180 ° (according to JISK6854) at a speed of 300 mm / min with a SUSBA430 plate of 5 to 500 g / 20 mm, preferably 100 to 400 g / 20 mm.

  In the polymer of the present invention, it is particularly desirable that the 24-hour water absorption is 20% by weight or more and 5000% by weight or less.

  Here, the water absorption rate means that a polymer is formed into a film or sheet, cut into 1.5 cm × 1.5 cm, measured for the weight before water absorption, and then each piece is treated with phosphate buffered saline. It was immersed in 135 ml (pH 7.4, 32 ° C.) for 24 hours. After that, each cut piece is taken out, the surface moisture is wiped off, the weight after water absorption is measured, and the value calculated from the increased weight.

  With this increase in water absorption, flexibility and stress relaxation properties increase. However, if the water absorption exceeds 5000% by weight, the gel strength decreases and becomes brittle or tends to dissolve, making it virtually impossible to use. However, when the water absorption is 5000% by weight or less, a gel state having substantially usable strength can be obtained, and particularly when the water absorption rate is 3000% by weight or less, a gel state having effectively usable strength can be obtained. . On the other hand, when the water absorption is less than 20% by weight, the water permeability is deteriorated and the absorption of moisture and nutrients from the support is deteriorated.

  Although the well-known sheet | seat (film) currently used as a support body for adhesive tapes can be applied to the support body 2 in the adhesive sheet | seat for microorganisms tests of this invention, it does not form a large unevenness | corrugation on the adhesive layer surface. Further, those made of a flexible material that can be freely crimped to a curved surface or a narrow surface are suitable. For example, polyester, polyethylene, polycarbonate, polyurethane, polyvinyl chloride, ethylene-vinyl acetate copolymer, urethane- Examples thereof include solid or porous plastic sheets made of acrylic hybrid resin, cloth, nonwoven fabric, paper, polyethylene laminated paper, and the like. Among these, a polyester film and a polycarbonate film are preferable from the viewpoint of flexibility and mechanical strength, and these porous films are particularly preferable.

  Further, the thickness of the support 2 is not particularly limited as long as sufficient strength can be obtained as the support of the pressure-sensitive adhesive sheet, but is generally about 5 to 5000 μm, preferably 5 to 5 μm. It is about 200 μm.

  The pressure-sensitive adhesive sheet for microbiological examination of the present invention is colored on the surface opposite to the surface of the support 2 that is in contact with the pressure-sensitive adhesive layer by increasing the strength of the base material or by using optical scattering prevention or a coloring material. In order to form a background color that improves the visibility of microorganisms, a backing layer may be further provided.

  The pressure-sensitive adhesive sheet for microbiological examination of the present invention is prepared by adding and mixing microbial nutrients and further various functional substances (additives) according to the purpose to a water-soluble pressure-sensitive adhesive, and preparing a pressure-sensitive adhesive composition. Such a pressure-sensitive adhesive composition can be produced by coating on a support using a known coating means such as an applicator and then drying at about 90 to 130 ° C. The pressure-sensitive adhesive sheet thus obtained can be cut into an arbitrary shape and used.

  The pressure-sensitive adhesive sheet thus prepared can be in a sterilized state, for example, by being sealed in a bacteria-blocking packaging material in a sterilized state. The sterilization method is preferably γ sterilization. That is, the adhesive sheet of the present invention can be distributed by applying a γ-ray sterilization treatment after the adhesive sheet is sealed in a bacteria-blocking packaging material, etc. so that it can be maintained in a sterile state. . In addition, when a fluorescent pigment | dye is contained in the adhesive layer as a coloring agent of microorganisms, in order to prevent decline of a fluorescent pigment | dye, it is preferable to store the produced adhesive sheet light-shielding.

  Hereinafter, a method for detecting microorganisms will be described using the adhesive sheet for microorganism testing of the present invention. First, the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet is pressed against the subject, and microorganisms attached to the subject are collected on the pressure-sensitive adhesive layer. Alternatively, an aqueous test substance (that is, an aqueous solution containing microorganisms to be tested, such as drinking water, food suspension, or a microorganism-containing liquid collected by a wiping method) on the adhesive layer of the adhesive sheet Inoculate directly. Next, the microorganisms on the adhesive layer are cultured and grown. Since the pressure-sensitive adhesive layer contains nutrients necessary for the growth of microorganisms, the microorganisms on the pressure-sensitive adhesive layer can be grown on the pressure-sensitive adhesive layer without having to be transferred (contacted with the medium) to the medium.

  For the purpose of preventing secondary contamination of human bodies and equipment, the pressure-sensitive adhesive sheet of the present invention is preferably kept in a state cut off from these pollution source articles and the environment, for example, a pressure-sensitive adhesive sheet that collects microorganisms. It is preferable to take measures such as immediately putting the in a petri dish or covering the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet with a film through which microorganisms cannot permeate. FIG. 2 shows an example of an embodiment in which microorganisms are cultured by putting an adhesive sheet collecting microorganisms in a petri dish. Moreover, FIG. 3 has shown an example of the embodiment covered with the film which cannot permeate | transmit microorganisms instead of putting in a petri dish. Here, the “film through which microorganisms cannot permeate” refers to a film that does not have pores of a size that allows microorganisms to move, such as plastic films such as polyester, polyurethane, polyethylene, Examples include a membrane filter of 0.2 μm or less.

In the present invention, microorganisms are cultured in an environment where microorganisms can be cultured as they are on an adhesive sheet in which microorganisms are collected (attached) on the adhesive layer, that is, in a temperature and humidity environment where microorganisms can grow. Can be done by putting on. The temperature / humidity environment in which microorganisms can grow on the adhesive sheet is, for example, an environment obtained by a constant temperature incubator or a wet box. Moreover, you may use incubators, such as an incubator. Nutrients contained in the pressure-sensitive adhesive layer and various functional substances (additives) are taken up by microorganisms, and the growth of microorganisms proceeds. In addition, it is preferable to culture microorganisms in the presence of appropriate moisture. For example, a method of moistening the inside of an apparatus (such as an incubator) used for incubation, sterilized water or sterilized physiological saline on the adhesive layer surface to which the microorganisms have adhered The method of dripping etc. is mentioned. When sterilized water, sterilized physiological saline, or the like is dropped, the sterilized water or the like may contain a selective agent for detecting a specific microorganism. In this case, the amount of the selective agent is 1 × 10 ⁻⁶ to 1 ×. About 10 ⁻¹ wt% is preferable, and 2 × 10 ⁻⁵ to 4 × 10 ⁻³ wt% is more preferable.

  The microorganisms detected by the adhesive sheet of the present invention are not particularly limited. As described above, it is possible to detect only specific bacteria by adding a selective medium or antibiotics.

  The microorganisms to be detected in the present invention include prokaryotes such as bacteria and actinomycetes, eukaryotes such as yeast and mold, lower algae and the like. Microorganisms include colonies and microcolonies.

  A means for detecting an aggregate (colony) of microorganisms grown on the adhesive sheet of the present invention is not particularly limited. You may observe visually or you may observe with an optical microscope, a phase contrast microscope, a fluorescence microscope, an electron microscope, a laser microscope etc. In addition, a commercially available colony counter or an automatic detection device for various images (for example, a combination of a simple microscope and an autoscan device, and by simply placing the adhesive sheet on the stage, it automatically captures images and detects microorganisms. May also be used.

  The size of the colony of microorganisms grown on the adhesive sheet of the present invention is not particularly limited. You may make it grow to the magnitude | size which can be observed visually. Moreover, if it is an aggregate of at least 4 or more, it can be determined as a grown colony.

  When detecting microorganisms grown on the adhesive sheet of the present invention, the microorganisms may be observed directly, or the microorganisms may be colored before observation. In addition, as described above, in the pressure-sensitive adhesive sheet of the present invention, the pressure-sensitive adhesive layer can contain a microbial colorant in advance, but the pressure-sensitive adhesive layer contains a microbial colorant to grow the microorganism. Thereafter, the microorganisms may be colored by attaching an adhesive layer to the colorant before observing the microorganisms. As the colorant used for coloring the microorganisms before the observation of such microorganisms, the same colorants as those contained in the pressure-sensitive adhesive layer can be used, and specific examples thereof are the same as those described above. It is done. In addition, when a grown microorganism can be sufficiently colored by adding a colorant to the adhesive in advance, the labor of coloring the microorganism before observation can be saved.

  There are no particular limitations on the subject to which the microorganism test is performed by applying the adhesive sheet of the present invention. For example, floors and walls, food and medicine manufacturing devices, meat, fish and vegetable surfaces, homogenized food, cooking utensils, medical utensils, water, and the like.

  As an example of application of the pressure-sensitive adhesive sheet of the present invention, if the pressure-sensitive adhesive sheet is used as a roller, colonies can be expressed on the pressure-sensitive adhesive surface of the sheet that has trapped bacteria, and the cleanliness of the test substance can be easily measured. It can be used for in-hospital environment surveys. Moreover, it can also be used for the detection use of resistant bacteria, such as MRSA, by containing a specific antibacterial agent such as methicillin or an antibiotic in the adhesive medium.

  The contents of the kit for testing microorganisms using the adhesive sheet of the present invention are: a colorant that can color microorganisms, an additive that can select microorganisms, a petri dish for culturing after collecting microorganisms, and a culture after collecting microorganisms In this case, a film, a small incubator, or the like that does not allow microorganisms covering the adhesive sheet to permeate can be considered, and a set of environmental survey instruments that anyone can do anywhere can be provided.

  The present invention will be described more specifically with reference to the following examples. However, this is merely an example and does not limit the scope of the present invention.

[Example 1]
(1) Preparation of pressure-sensitive adhesive sheet 70 parts by weight of 2-methoxyethyl acrylate, 30 parts by weight of N-vinyl-2pyrrolidone, 0.175 parts by weight of azobisisobutyronitrile as a polymerization initiator and distillation in a closed reactor equipped with a stirrer First, 250 parts by weight of a water-methanol-isopropanol mixed solvent (weight ratio = 16: 23: 1) was charged and purged with nitrogen at a flow rate of 10 ml / min for 2 hours, and then the reactor was maintained at 60 to 62 ° C. Stir for 5 hours. Subsequently, the reaction was completed by stirring at 75 ° C. for 2 hours, and then cooled to room temperature to obtain an acrylic copolymer solution.
To this, 20 parts by weight of polyoxypropylene glyceryl ether (average molecular weight 400) as a plasticizer was added and mixed to obtain an adhesive composition.

  1 g of SCD Broth (manufactured by Nissui) was added to and mixed with 81.3 g (solid content 41%) of the pressure-sensitive adhesive composition obtained as described above. After applying this mixture on a release paper to a thickness of 20 μm using an applicator, the solvent and unreacted monomer are removed by drying at 110 ° C. for 3 minutes, and the adhesive is composed mainly of a water-soluble polymer. An agent was obtained. Furthermore, a polycarbonate type membrane filter (manufactured by Whatman, product name Nucleopore, pore size 0.2 μm) was attached as a support.

  This was sealed in a bacterial-resistant packaging material and then sterilized by irradiation with gamma rays (irradiation dose 25 kGy) using cobalt 60 as a radiation source. Finally, it was cut into a size suitable for adhesive collection (about 3 × 4 cm) to obtain an adhesive sheet of the present invention.

(2) Collection and staining of microorganisms Using the kit of the present invention, microorganisms were detected from an aqueous sample.
10 μl of Staphylococcus ATCC strain No. 155 diluted to about 6 × 10 ⁷ cells / ml with sterile physiological saline was dropped onto the adhesive surface of the medium-added adhesive sheet prepared in (1). The adhesive sheet collected by the above method was put in a petri dish, covered, and cultured at 32 ° C. for 16 hours. After culturing, the filter paper soaked with 300 μl of 6-CFDA solution (Molecular Probes) prepared to 1 ppm with phosphate buffer is brought into contact with the support (the support is water-permeable, so the staining solution is applied to the adhesive surface). After being stained at room temperature for 3 minutes and staining the microcolony grown on the pressure-sensitive adhesive surface, the support surface was washed with a phosphate buffer.

(3) Detection (measurement)
When the surface of the pressure-sensitive adhesive layer (microbe collection culture surface) was observed with an epi-fluorescence microscope (Olympus, BX51) (1000 times, blue excitation: excitation wavelength of 460 nm to 490 nm), The staphylococcus grew and the formed microcolonies could be observed.

It is a cross-sectional view which shows one embodiment of the adhesive sheet of this invention. It is a cross-sectional view which shows one embodiment example at the time of culture | cultivating the adhesive sheet of this invention. It is a cross-sectional view which shows another example of an embodiment at the time of culture | cultivating the adhesive sheet of this invention.

Explanation of symbols

1 Adhesive 2 Support 3 Petri dish 4 Film

Claims (13)

  A pressure-sensitive adhesive sheet for microbiological examination, wherein a pressure-sensitive adhesive layer made of a water-soluble pressure-sensitive adhesive provided on at least one surface of a support contains microbial nutrients.   The pressure-sensitive adhesive sheet for microbiological examination according to claim 1, wherein the water-soluble pressure-sensitive adhesive is an acrylic copolymer.   The pressure-sensitive adhesive sheet for microbiological examination according to claim 2, wherein the acrylic copolymer is an acrylic copolymer mainly comprising alkoxy acrylate and N-vinyl lactam.   The pressure-sensitive adhesive sheet for microbiological examination according to any one of claims 1 to 3, wherein the pressure-sensitive adhesive layer further contains a selection agent capable of selecting a microorganism to be examined.   The pressure-sensitive adhesive sheet for microbiological examination according to any one of claims 1 to 4, further comprising a microbial colorant in the pressure-sensitive adhesive layer.   The pressure-sensitive adhesive sheet for microbial examination according to claim 5, wherein the microbial colorant is a fluorescent dye.   The pressure-sensitive adhesive sheet for microorganism testing according to any one of claims 1 to 6, wherein the support is porous.   A pressure-sensitive adhesive sheet for microbiological examination characterized in that a pressure-sensitive adhesive composition prepared by adding and mixing at least microbial nutrients to a water-soluble pressure-sensitive adhesive is coated on a support and dried to form a pressure-sensitive adhesive layer. Production method.   The pressure-sensitive adhesive composition according to claim 8, wherein the pressure-sensitive adhesive composition is prepared by adding and mixing a water-soluble pressure-sensitive adhesive with a microbial nutrient and a selection agent capable of selecting a microorganism to be tested. Method.   A pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet for microorganism testing according to any one of claims 1 to 7 is pressure-bonded to a subject to collect microorganisms, and the microorganisms collected on the pressure-sensitive adhesive layer are cultured. A method for examining microorganisms present in a subject, the method comprising growing and detecting the grown microorganisms.   Dropping a solution containing microorganisms to be inspected on the pressure-sensitive adhesive layer of the pressure-sensitive adhesive sheet for microorganism testing according to any one of claims 1 to 7, and culturing and growing microorganisms on the pressure-sensitive adhesive layer; And a method for examining microorganisms present in a subject, comprising detecting the grown microorganisms.   12. The method according to claim 10, wherein the grown microorganism is detected by visual observation, a sensitizer or an automatic detector.   The pressure-sensitive adhesive sheet for microorganism testing according to any one of claims 1 to 7, and a colorant capable of coloring the microorganism to be examined and / or a selection agent capable of selecting the microorganism to be examined. Microorganism testing kit.

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