JPH10136972A - Tool for test - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a test tool enabling to simply and rapidly carry out chemical sand biological tests by subjecting a water-absorbing sheet excluding its water-absorbing parts to a hydrophobic treatment in order to dispose plural mutually separated water-absorbing parts for reacting test liquids. SOLUTION: This sheet-like test tool is produced by subjecting a water- absorbing sheet having a thickness of 300-3000μm and comprising a water- absorbing fabric piece such as an acrylic nonwoven fabric to a hydrophobic treatment in order to form plural mutually separated water-absorbing parts 1 for reacting test liquids, and subsequently laminating a water-diffusing sheet to the partially treated sheet to form diffusion parts 2 in a water-absorbing ratio (water-absorbing part/water-diffusing part) of >=2 between the water- absorbing parts 1 and the water-diffusing parts 2. The hydrophobic treatment is carried out by a method comprising impregnating parts excluding the water- absorbing parts 1 and test liquid-supplying parts 3 in the water-absorbing sheet with an ink consisting mainly of a hydrophobic resin by a screw printing method, and subsequently drying the printed sheet. Thus, the objective sheet-like test tool enabling to simply and rapidly carry out chemical and biological tests requiring many items of reactions can profitably be obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention makes it possible to carry out chemical and microbiological tests requiring a large variety of reactivity more easily and quickly than conventional microplates.
In addition, the present invention relates to a test tool that can be industrially advantageously mass-produced.

[0002]

2. Description of the Related Art In a chemical test including a step of reacting a chemical sample with some kind of reagent and a microbiological test including a step of culturing a microorganism in the sample, particularly, treatment of a large number of samples, In a test requiring a reaction with a reagent or culture in a large number of systems, a plastic microplate provided with a plurality of wells is usually used as a reaction or culture vessel.

In a test using such a microplate, individual reactions, culture of microorganisms, and the like are performed in each well on the plate. For example, M by a microfluidic dilution method using a microplate
In IC measurement, various concentrations of antimicrobial agents are added to a plurality of wells, and the growth of the target bacterium in each well is observed to determine the minimum growth inhibitory concentration.

[0004] As a test tool used for such a test, for example, a product in which a predetermined amount of an antibacterial agent is previously dispensed into each well of a microplate, which has been dried, and which has been frozen and stored as it is, has been commercialized. However, such a test device has a problem in that the cost of the microplate itself is high, so that it becomes an expensive product, and a reduction in price has been desired. In addition, even when a large number of tests are performed with one type of sample, a predetermined amount of the sample must be dropped in each well, so that there is a problem that the operation is complicated.

[0005]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to mass-produce at low cost, and to carry out a chemical test and a microbiological test for testing various items of reaction performance in a simple and rapid manner. It is to provide a test tool that can be used.

[0006]

Under such circumstances, the present inventors have conducted intensive studies and as a result, have found that using a water-absorbent sheet, a portion other than a portion (water-absorbing portion) corresponding to each well portion of a microplate is made of a hydrophobic resin. Etc.
It has been found that there is no leakage of the test liquid from the water absorbing portion, the amount of water absorbed in the water absorbing portion can be made constant, and mass production can be performed at low cost due to hydrophobic treatment such as printing. In addition, if the water absorption part thus created is used in combination with a diffusion part for supplying a test liquid, a multi-item test can be performed by dropping the test liquid to one diffusion part. It was also found that the present invention was completed.

That is, according to the present invention, a portion of the water-absorbent sheet other than the water-absorbing portion is subjected to a hydrophobic treatment so that a plurality of mutually isolated water-absorbing portions for reacting the test liquid are formed. It is intended to provide a sheet-shaped test device characterized by the following (hereinafter, referred to as the sheet-shaped test device of the present invention).

Further, the present invention is characterized in that it has the above-mentioned sheet-like test device and a diffusion portion for contacting at least a part of each of the water absorbing portions and supplying a test liquid to the water absorbing portion. A test tool (hereinafter, referred to as a test tool of the present invention) is provided.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION In the sheet-like test device of the present invention, as a material of the water-absorbent sheet, for example, rayon,
Polyester, acrylic, vinylon, cotton, glass fiber,
Other examples include fibers made of other chemical and natural materials. Among them, acrylic nonwoven fabrics are preferred because of their high water absorption. The water-absorbing sheet preferably has a water absorption of 0.020 g / cm ² or more, preferably 0.08 to 0.2 g / cm ² .

The amount of water absorption used in the present invention is
Crow et al. (INDA-TEC, 207 (199)
The value obtained by partially modifying 1)) according to the following method. 1) A sponge (polyurethane) having a size of 8.5 cm × 14 cm and a thickness of 3 cm is sufficiently hydrated in advance and immersed in a dish containing purified water. 2) A 1 cm × 1 cm test piece is placed on a water-containing sponge, and the same type of sponge is stacked. 3) Put the whole dish in a wet box and place at room temperature (20-25 ° C) for 5 minutes.
After standing for a period of time, the sample weight is measured. 4) The value obtained by subtracting the sample weight before water absorption from the sample weight after water absorption is defined as the water absorption.

The thickness of the water-absorbent sheet is 300 to 3
000 μm is preferable, and it is particularly preferable to use one having a thickness of 500 to 2000 μm. If the thickness is less than 300 μm, the amount of the test liquid held in the water absorbing portion is not sufficient, and the reaction is not sufficient. As a result, accurate measurement may not be performed. On the other hand, when the thickness exceeds 3000 μm, it becomes difficult to perform the hydrophobic treatment described below.

In the sheet-like test device of the present invention, for example, only a portion other than the water absorbing portion is subjected to hydrophobic treatment so that a plurality of mutually isolated water absorbing portions are formed on the water absorbing sheet. Is obtained by

The hydrophobizing treatment is preferably performed by a printing method, that is, by infiltrating a liquid containing a hydrophobic resin as a main component into a predetermined portion (a portion other than the water absorbing portion) of the water-absorbing sheet and then drying. . Examples of the hydrophobic resin used here include a polyurethane resin, a polyacrylic resin, a polyester resin, a polyamide resin, a polyvinyl chloride resin, a polystyrene resin, a fluorine resin, and other rubber modified products. As long as the resin is not particularly a water-soluble resin or a resin having a high hygroscopicity, any known resin conventionally used as a binder of an ink can be used. In addition, the solvent for dissolving or dispersing these hydrophobic resins differs depending on the hydrophobic resin used, but is selected from various organic solvents and used, for example,
Examples include toluene and cyclohexanone. In addition, a liquid containing a hydrophobic resin as a main component (hydrophobic ink) may include a coloring agent, an inorganic or organic filler, a water repellent such as a fluororesin or silicone, a surfactant, and a viscosity adjusting agent, if necessary. Additives such as agents can be added.

In order to make the hydrophobic ink permeate a predetermined portion of the water-absorbing sheet, it is preferable to use a usual silk screen printing method. That is, it is preferable that the hydrophobic ink is applied to a predetermined portion of the water-absorbent sheet, penetrated, and then dried. The application of the hydrophobic ink may be performed once, but is preferably performed a plurality of times to ensure the hydrophobic treatment.

In order for the hydrophobic ink thus applied to sufficiently penetrate into the back surface of the water-absorbent sheet, (1) a method of applying pressure to the coated surface, and (2) a method of sucking from the surface opposite to the coated surface. , (3) a method of blowing air from the application surface side, and (4) a method of appropriately combining two or more of these (1), (2) and (3).

(1) The method of applying pressure to the coating surface is not particularly limited, but a preferable method is, for example, a roll press method.

In this method, for example, at least two mirror-finished metal rolls and rubber rolls are arranged in a vertical combination, and a water-absorbing sheet coated with a hydrophobic ink is placed between the rolls with the coated surface facing the metal roll. It is a method of inserting and rotating both rolls while continuously pressing between them. At this time, the mirror-finished metal roll can be heated, but when the hydrophobic ink is not dried, the hydrophobic ink can be permeated to the back surface of the hydrophobic sheet only by pressing without heating. Can be. According to this method, it is not necessary to prepare a mold or the like having a predetermined shape for pressing, and pressurization can be easily performed with high productivity only by adjusting the rotation speed of the rolls and the gap or nip pressure between the rolls. . Further, when pressurizing, it is preferable that a film that does not absorb the hydrophobic ink is overlaid on the application surface on the water-absorbent sheet, and an absorbent material such as paper is overlaid on the surface on the opposite side of the application surface. By adopting such a method, excess ink that has permeated to the back surface by pressure can be absorbed by the absorbent material, and the adhesion of the ink to the roll and the spread of the water-absorbent sheet in the lateral direction can be reduced. Can be prevented.

As another method of applying pressure to the ink-coated surface of the water-absorbent sheet, a flat press method using a flat plate at room temperature or heated or a flat plate patterned into a shape corresponding to the application pattern of the hydrophobic ink is mentioned. be able to. When the flat press method is employed, when the ink applied to the water-absorbent sheet is not dried, the pressure plate may be at room temperature as in the case of the roll press method, but the pressure is applied after drying the ink. In this case, it is necessary to heat the pressure plate in order to soften or melt the hydrophobic ink.

When pressure is applied using a patterned pressure plate, the pressure plate and the application pattern of the water-absorbent sheet are, of course, positioned and pressed.

Also in this method, when the hydrophobic ink on the water-absorbent sheet is not dried at the time of pressurizing, a film which does not absorb the hydrophobic ink is overlaid on the coated surface, and the surface opposite to the coated surface is also applied. In this case, it is preferable that an ink-absorbing material such as paper is overlaid and pressurized. Also, if the applied ink is dried and then pressed with a heated flat plate, heat-resistance is applied to both to prevent the softened or melted hydrophobic ink from sticking to the pressed flat plate and the receiving surface plate. It is preferable to perform a release treatment.

In the method (2) of sucking from the surface opposite to the application surface, when the application means is a silk screen printing method, a vacuum suction device for fixing the sheet to be printed to the stage on the printing machine itself. This is used as it is, and if necessary, by adjusting the hole diameter of suction holes and their intervals or their arrangement pattern, suction can be performed simultaneously with printing and ink can permeate.
Also, if the printing machine does not have a vacuum suction device,
Separately, a similar suction device can be prepared so that the ink is sucked and permeated before printing before the ink dries.

In the case of the method (3), in which the air is blown from the coating surface side to pressurize and permeate, for example, a hooded blower and a wire net-like pedestal are prepared, and the coated water-absorbent sheet is prepared. The ink can be permeated by placing it on a receiving table and applying wind pressure from above the application surface with a blower. In addition to the above, there is an air pressure / vacuum forming machine for a plastic sheet as a ready-made device, but a chamber for this forming can also be used. In this case, a wire net-shaped pedestal is prepared separately, this is replaced with a female mold for molding, installed in the chamber, a coated water-absorbing sheet is placed thereon, and after closing the chamber, no vacuum is used. The ink can be penetrated by sending only air for pressurized air from above the application surface and pressurizing only by exhausting.

In the case of a combination of (2) and (3), that is, a method of performing both blowing from the coating surface and suction from the surface opposite to the coating surface, the method of (3) is used. The molding chamber of the pressurized / vacuum molding machine described can be used as it is. Also in this case, instead of a female mold for molding, a wire net-shaped pedestal is mounted in the chamber, a coated water-absorbent sheet is placed on it, and after closing the chamber, pressure is applied from above the application surface by compressed air. And ink is allowed to penetrate by performing suction from the lower side by vacuum.

Usually, in the pressurizing / vacuum forming machine, a heating device and a forming device for the plastic sheet are lined up. However, in the above-mentioned methods (2) and (3), heating is not required. It is also possible to separately prepare a device, connect the device after the silk screen printing machine, and if necessary, further connect a drying device thereafter, so that the application, the permeation, and the drying can be continuously performed.

The shape and size of the water-absorbing portion left by the above-mentioned hydrophobic treatment are not particularly limited, and can be selected according to the amount of the test liquid, the amount of water absorption, and the like. Usually, a circular or square shape is preferable, and for example, a shape having a diameter or a side length of about 0.2 to 1 cm is preferably used. In this case, the amount of the test liquid to be absorbed by one water absorbing portion is generally about 0.025 to 0.2 ml.

The number of such water absorbing portions is not particularly limited as long as it is plural, and may be arbitrarily determined from the amount of the test liquid, the number of test items, and the like.

In order to adjust the amount of water absorbed by the water absorbing section,
A material having the water-absorbing polymer fixed thereon can be used as the water-absorbing portion. As the water-absorbing polymer used here,
For example, soluble starch, mannan, agar, sodium alginate, plant mucilage (such as gum arabic), microbial mucilage (dextran, xanthan gum, gellan gum, etc.), cellulosic substances (methyl cellulose, carboxymethyl cellulose, other cellulose), polyvinyl alcohol, Examples thereof include sodium polyacrylate and a graft polymer of these polymers. As a method of fixing these water-absorbing polymers to the water-absorbing member, for example, a water-absorbing polymer is dissolved or emulsified in water, an organic solvent, or the like, a printed water-absorbing sheet is immersed, and then dried and fixed. And a method in which the water-absorbing polymer liquid is dropped onto the water-absorbing section and then dried and fixed.

The test device of the present invention has the above-mentioned sheet-like test device and a diffusion portion for contacting at least a part of the water absorbing portion and supplying a test liquid to the water absorbing portion. The ratio of the water absorption amount of the diffusion part to the water absorption part (water absorption part / diffusion part) is preferably 2 or more, particularly preferably 5 to 15. If this ratio is less than 2, it may be difficult to diffuse the test liquid into the diffusion section and cause the water absorption section to absorb water.

Examples of the material of the diffusion portion include the same materials as those of the water-absorbent sheet, and a cotton-based nonwoven fabric having high diffusivity and low water absorption is preferable.

[0030] Of these materials, the diffusion portion serves as an absorbing member.
The amount of water is 0.005 to 0.05 g / cm ^Two, Especially 0.008
~ 0.02g / cm^TwoIs preferable, and 1cm × 10
One end of the short side of the rectangular sheet of cm was immersed in purified water
At this time, moisture spreads over the entire surface within 15 minutes, especially 30 seconds to 3 minutes.
Those having a dispersing rate of water diffusion are preferred.

The diffusing section used in the present invention is for supplying a test liquid and diffusing it to the water absorbing section. The shape, size, and thickness of the diffusion portion are not particularly limited, and can be selected according to the amount of water to be used in the test solution and the number of the water absorption portions. Usually, a sheet-like thing is preferable, and a thing about 0.05-0.6 mm in thickness is used preferably.

In the test device of the present invention, the water-absorbing portion of the sheet-shaped test device is arranged so as to be at least partially in contact with the diffusion portion.

The method of fixing the water-absorbing portion of the sheet-shaped test device to the diffusion portion is not particularly limited. For example, a part of the water-absorbing portion and the diffusion portion may be bonded with an adhesive substance (hydrophilic adhesive, water-soluble polymer, etc.). A method of fixing with water; a method of placing a water-absorbing part on the diffusion part and bonding a part by (heat) crimping; a method of sewing the diffusion part and the water-absorption part; There is a method of fitting.

In a more preferred form of the test device of the present invention, the sheet-shaped test device has a test liquid supply hole in the center of a plurality of water absorbing portions, and the diffusion portion is a water-diffusible sheet. A sheet-shaped test tool is placed on the upper side of the section so as to connect the test liquid supply hole and the plurality of water absorbing sections (see FIG. 1).

The test device of the present invention is preferably distributed in a waterproof case. As a form stored in a particularly preferable waterproof case, a waterproof bottom portion,
The sheet-like test device has a plurality of water-absorbing portions and a test-solution supply hole, and a diffusion portion. Is a water-diffusible sheet, and a test tool on which a sheet-shaped test tool is mounted so as to connect the test liquid supply hole and the plurality of water-absorbing sections on the upper side of the diffusion section is fixed and stored. Are particularly preferred (see FIG. 2). For distribution, it is preferable to further provide an upper lid on this lid.

As a waterproof material used for such a bottom and a lid, for example, a polyethylene, polypropylene, polystyrene or the like plastic plate, glass or the like is used.

In order to conduct a test using the sheet-shaped test device of the present invention, a predetermined amount of a test liquid may be dropped on each water absorbing portion. The amount of the test liquid is determined according to the size of the water absorption part, the amount of water absorption, and the like.

In order to conduct a test using the test device of the present invention, a predetermined amount of a test liquid may be poured into any one of the diffusion sections, whereby the test liquid diffuses in the diffusion section. ,
Water is absorbed by each of the plurality of water absorbing portions. In the case of the preferred embodiment described above, the solution may be dropped into the test solution supply hole provided in the water absorbing portion. The test liquid amount is determined according to the size of the diffusion part and the water absorption part, the water absorption amount, and the like.

In the present invention, the test liquid absorbed in each water absorbing section reacts with various reaction reagents and the like here. The reaction reagent or the like may be contained in the test device in advance, or may be added after water absorption. If there is no problem in stability or the like, it is preferable to include the reaction reagent in advance for performing a rapid test. preferable. The reaction reagents used here are not particularly limited, and are, for example, reagents for measuring enzyme activities such as various chromogenic substrates and fluorescent substrates; reagents for identifying microorganisms of sugars, amino acids and fatty acid sugars; and measuring drug sensitivity to microorganisms such as various antibiotics. Reagents;
Antimicrobial test reagents for various antibacterial substances and the like can be mentioned. These reaction reagents can be fixed, for example, by adding a reagent solution to a previously sterilized water-absorbing part so as to have a water absorption per unit area or less, and then drying under reduced pressure.
At this time, the above-described water-absorbing polymer may be added together with the reaction reagent.

[0040] Reactive reagents of different types and concentrations can be immobilized on the plurality of water-absorbing parts of the test device of the present invention by such a method, so that a multi-item reaction performance can be tested at a time. The reaction reagent at this time is, for example, a test solution (a liquid such as a bacterial suspension, a body fluid, a cerebrospinal fluid, or a food emulsion).
L-alanine used for enzyme activity measurement
p-nitroanilide and L-proline
For chromogenic substrates such as -β-naphthylamide,
The amount of the test solution absorbed in the water-absorbing piece is 0.01 to 0.1%.
5% by weight; 4-methylumbelliferyl
-Β-D-galactopyranoside, L
-Alanine-7-amido-4-methyll
In the case of a fluorescent substrate such as coumarin, it is preferable to use the fluorescent substrate in the range of 0.001 to 0.02% by weight based on the amount of the test liquid absorbed in the water absorbing piece. In the case of sugars, amino acids, fatty acids or their intermediate metabolites used for the microorganism identification test, the amount of the test liquid absorbed by the water absorption part is set at 0.
In the case of an antibiotic used for measuring drug sensitivity to microorganisms or an antibacterial substance used for an antibacterial test, the drug concentration can be arbitrarily set depending on the target drug and the target microorganism species. it can.

The water-absorbing part may contain a gelling agent so as to increase the viscosity of the test liquid having absorbed water and to obtain sufficient gel strength in the water-absorbing part. Examples of such gelling agents include natural polymers such as xanthan gum, carrageenan, gellan gum, gum arabic, locust bean gum, guar gum, tragacanth gum, and crystalline cellulose;
Examples include synthetic polymers such as polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyacrylamide, polyacrylic acid, hydroxyethyl cellulose, and hydroxypropyl cellulose. These gelling agents can be fixed to the water-absorbing part in the same manner as for fixing the above-mentioned reagent. The gelling agent at this time is 0.1 to 1% by weight for a natural polymer and 0.05 to 0. 0% for a synthetic polymer, based on the amount of the test liquid absorbed by the water absorbing portion. Preferably, it is used in the range of 5% by weight.

Further, when the above-mentioned reaction reagent or gelling agent is fixed to the water absorbing portion, various nutrients such as peptone and the like, and the gel strength are increased for culturing the bacteria in the test solution, thereby improving the reaction sensitivity. For this purpose, a cation such as an alkali metal salt or an alkaline earth metal salt is blended in the reagent solution, so that these can be contained in the water absorbing portion.

Further, various nutrients, surfactants, cations such as alkali metal salts and the like can be mixed in the test solution.

[0044]

EXAMPLES The present invention will be further described with reference to examples, but the present invention is not limited to these examples.

Example 1 The entire surface of a water-absorbing piece (A-200; manufactured by Kuraray Trading Co., Ltd.) of acrylic nonwoven fabric having a thickness of 1000 μm is shown in Table 1 below.
Screen printing was carried out using the printing method and ink type shown in Table 1 to obtain a test device having a non-printed portion having a diameter of 7.5 mm. Test of sealing effect by printing using hydrophobic ink; outer edge 12 mm × 12 mm of this non-printed portion was cut to obtain a sample for measuring water absorption. As a control sample, a non-printed water-absorbing piece was cut into a piece of 10 mm x 10 mm to obtain a sample for measuring the water absorption. The measurement was performed according to the method of Crow et al. However, in the control group, the measured water absorption “surface area of 7.5 mm in diameter” /
It is shown as a value multiplied by the “10 mm × 10 mm area” ratio. Table 1 shows the results.

[0046]

[Table 1]

Ink type Acrylic resin: Teikoku Ink Manufacturing Sericol 13
Medium Solvent: Teikoku Ink Manufacturing Sericol G-002 Solvent Fluorine: Soken Chemical Comb Polymer LFB-40
15 Silicon: Soken Chemical Comb Polymer LSI-6
0

[0048]

Printing method Plate: Number of lines: 70 mesh Resist thickness: 90 μm Printing (by a known screen printing method) F2B1 Printing twice continuously from the front side, printing once from the back side after drying F2 to F4 Continuous 2 to 4 from the front side One-time printing F1B1 One-time printing from the front side, one-time printing from the back side after drying

Roll Press Method After printing, an OPP film was placed on the upper side with the print side up, and a wiper was placed on the lower side, and roll press was performed to infiltrate the ink.

From the above results, a good printing method can be selected by measuring the amount of water absorption. 2, 3, 4, 5, 6, and 8 could be selected as good printing methods.

Example 2 Absorbent piece in section; acrylic nonwoven fabric (A-200 sheet)
0.114 g / cm of water ^TwoNo. Uniform with 11 printing methods
Production of a whole printed piece with 12 non-printed areas with a diameter of 7.5 mm
Then, the printed piece is punched into a rectangular piece of 106 mm x 38 mm
And then punch out the central part into a 13mm x 18mm ellipse
(FIG. 1).

Developing layer: Cotton-based nonwoven fabric (PS-14)
A PET film (thickness: 75 μm) was adhered to one side having a water absorption of 0.008 g / cm ² (adhesive: Cobonyl 5406D, manufactured by Nippon Synthetic Chemical Industry Co., Ltd.).
Punch 5mm x 110mm. Water-impermeable substrate and lid: The dimensions are such that the above-mentioned water-absorbing pieces and the spreading layer can be stored. The lid is open at the same position as the non-printing part and the center punching part. Manufacturing method: Place the spreading layer, compartment water-absorbing pieces, lid on the substrate in order,
The water absorption piece and the spreading layer are fixed in place with the upper and lower substrates and lids. Further, the substrate and the lid are bonded to ensure the fixation. In the test device obtained in this manner, since the developing layer is exposed at a part of the section water-absorbing piece punched out, if the test liquid is dropped here, it reaches the water absorbing portion through the developing layer ( (Fig. 2).

Test Example 2 Using the test device obtained in Example 2, a test was performed according to the following test method. As a control, a test by the microplate method was also performed. The results are shown in Tables 2 and 3.

Test method: Test strain: E. Coli ATCC11775, K. pneumoniae ATCC138
83

Reaction items and preparation method: ALA: L-alanine-p-nitroanilide (SIGMA, 6m
M pH 7.0), 2. PNP: p-nitrophenyl-β-D-galactopyranoside (S
2. IGMA, 6 mM pH 7.0) 3. PYR: Pyrroglutamyl-β-naphthylamide (manufactured by SIGMA, 3 mM pH 7.0); 4. GLR: p-nitrophenyl-β-D-glucronide (manufactured by SIGMA, 6 mM pH 7.0); PHE: p-nitro-L-phenylalanine (manufactured by SIGMA, 4 mM)
pH 7.3), 6. PRO: Proline-β-naphthylamide (SIGMA, 3m
M pH 7.0), 7. IND: Tryptophan (manufactured by Wako Pure Chemical Industries, 0.1% pH
7.4), 8. BGL: p-nitrophenyl-β-D-glucopyranoside (SIG
8. MA, pH 6.3), 9. NGL: p-nitrophenyl-N-acetyl-β-D-glucosamin
ide (manufactured by SIGMA, 2 mM pH 7.0); NIT: KN0 ₃ (manufactured by Wako Pure Chemical Industries, Ltd., 0.5% pH7.0), 11. BIP: 5-Bromo-3-indolyl phosphate (manufactured by Wako Pure Chemical Industries, Ltd., 4 mM pH 8.0), 12. XYL: p-nitrophenyl-β-xyloside (SIGMA, 6
(mM pH7.0)

Regarding the above items, the final concentration is the substrate concentration shown in parentheses, beef meat peptone 0.5% (manufactured by DIFCO), casein peptone (manufactured by Markova) 1%, and xanthan gum (manufactured by Saneigen FFI) ) Adjust to 0.1%. This solution is sterilized, and the diameter of the test device is 7.5.
50 μl of the substrate solution was directly dispensed to one portion of the non-printing portion of the mm circle and dried under reduced pressure. As a control, 100 μl per well was dropped on the microplate and fixed similarly.

Culture test method and determination method: Sheep blood agar medium
N “Nissui” (manufactured by Nissui Pharmaceutical Co., Ltd.)
The cultured fresh bacteria are suspended in a 0.1% gellan gum (MERCK) solution so that the turbidity becomes equivalent to that of the standard turbid liquid McFarland # 0.5. 1 ml of this bacterial solution is injected into the test device from an injection port opened at the center. After the injection, cover the outer lid to maintain the airtightness.
At this time, the germ solution spreads quickly, and furthermore, 6
Water was absorbed within minutes. The amount of test solution absorbed is 50 μl ±
10 μl. To the microplate serving as a control, 100 μl is dropped into each well using a pipette. After culturing at 37 ° C. for 4 hours, positive (+) and negative (−) were determined from the color development of the substrate with the naked eye. 3,
6, 7, 10 are detection reagents (3, 6: cinnamaldehyde reagent (manufactured by BioMerieux Biotech), 7: Kovac reagent (manufactured by Nissui Pharmaceutical Co., Ltd.), 10: nitrite detection reagent solution I and nitrite detection Test solution II (manufactured by Nissui Pharmaceutical Co., Ltd.) was added dropwise in an amount of 25 to 50 μl, and judgment was made after 5 to 10 minutes.

[0059]

[Table 2]

[0060]

[Table 3]

As is clear from the above table, the method using the test device of the present invention showed the same results as the microplate method. Therefore, by using the test device of the present invention, it is possible to easily and quickly test the reactivity of many items by pouring a fixed amount of the test solution into the injection port. In addition, since the test liquid is trapped in the water absorption section even when the test equipment is overturned, the test liquid is not scattered, ensuring safety in biohazard compared to microplate-type equipment. it can.

[0062]

The test device of the present invention can be manufactured by hydrophobic treatment such as printing using a hydrophobic ink, so that it can be industrially advantageously mass-produced. Further, if a diffusion unit is further provided, there is no need to inject the test liquid into each well as in a conventional microplate. The water can be absorbed in the part, and the test can be performed very simply and quickly. Further, the necessary reagents and the like can be contained in the water absorbing portion in advance, so that the test can be performed more quickly.

[Brief description of the drawings]

FIG. 1 is a view showing a test device of the present invention manufactured in Example 2.

FIG. 2 is a view showing a test device (including a container) of the present invention.

[Explanation of symbols]

 1. Water absorption part 2. Diffusion unit 3. Sample supply hole

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Masaho Hayashi 1-1-1, Ichigaya-Kagacho, Shinjuku-ku, Tokyo Inside Dai Nippon Printing Co., Ltd. No. 1 Dai Nippon Printing Co., Ltd. (72) Inventor Kiyoshi Oguchi 1-1-1, Ichigaya Kagacho, Shinjuku-ku, Tokyo Dai Nippon Printing Co., Ltd.

Claims (9)

[Claims] 1. A water-absorbing sheet is characterized in that a portion other than the water-absorbing portion is subjected to a hydrophobic treatment so that a plurality of mutually isolated water-absorbing portions for reacting a test liquid are formed. Sheet-like test tool. 2. The water-absorbent sheet has a thickness of 300 to 300.
The sheet-like test device according to claim 1, which has a thickness of 0 µm. 3. The sheet-like test according to claim 1, wherein the hydrophobizing treatment is carried out by permeating a liquid containing a hydrophobic resin as a main component into a predetermined portion of the water-absorbent sheet and drying it. Tools. 4. A test comprising: the sheet-like test device according to claim 1; and a diffusion portion for contacting at least a part of each of the water absorbing portions and supplying a test liquid to the water absorbing portion. Tools. 5. The water-absorbent sheet has a thickness of 300 to 300.
The test device according to claim 4, which has a thickness of 0 µm. 6. The test device according to claim 4, wherein the hydrophobizing treatment is performed by allowing a liquid containing a hydrophobic resin as a main component to permeate a predetermined portion of the water-absorbent sheet and drying. 7. The test device according to claim 4, wherein the water absorption ratio between the water absorbing portion and the diffusing portion (water absorbing portion / diffusing portion) is 2 or more. 8. A sheet-like test device having a plurality of water absorbing portions and a test liquid supply hole, a diffusion portion being a water diffusible sheet, and a test liquid supply hole and a plurality of test liquid supply holes provided above the diffusion portion. The test device according to any one of claims 4 to 7, wherein a sheet-like test device is placed so as to communicate with the water absorbing portion. 9. A test in which the test device according to claim 8 is fixed and accommodated by a waterproof bottom portion, and a lid having holes for the test liquid supply hole and the plurality of water absorbing portions of the sheet-shaped test device. Tools.