JPH114679A - Tool for chemical and micro-organic test - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a tool for tests that can reduce the deviation in the amount of a liquid sample held in a plurality of the sample-holding parts by one operation. SOLUTION: In this tool for testing, the bottom member 21 works in cooperation with the sample vessel to provide the area receiving the liquid-absorbing body 2, the sample vessel is equipped with the peripheral part positioning lower than the sample-holding part 3 and the through-hole 12 bored in the peripheral area and the through-hole 12 liquid-links to the receiving area arranged between the sample vessel and the bottom member. At this time, the bottom member 2 is equipped with a plurality of cylindrical upright-standing walls 23 that have the sample-holding part 3 on its top face and the top faces of the upright- standing walls 23 pass through the opening part 8 for forming the sample-holding part and project upward and the side face of the standing wall 23 is linked to the opening for forming the sample-holding part.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a test device which can be suitably used for a chemical test or a microbiological test.

2. Description of the Related Art A chemical test including a step of reacting a chemical sample with some reagent, or a microbiological test including a step of culturing a microorganism in the sample, particularly for treating a large number of samples, In a test that requires a reaction with the lipase or culture in a large number of systems, a plastic microplate provided with a plurality of wells as a reaction or culture vessel is generally used. For example, in immunological tests such as ELISA, it is common to use a plastic plate provided with many small wells. For the measurement of antimicrobial susceptibility by the microfluidic dilution method, that is, the measurement of the minimum inhibitory concentration (MIC) of the antimicrobial agent, the agar plate dilution method and the one-concentration disk method by the agar diffusion method, which have been widely used, have been used. Furthermore, the standard method of MIC measurement by the microfluidic dilution method set by the Japan Society of Chemotherapy in 1989 as a labor-saving and automated method is a micro method having a U-shaped well similar to that used in immunological tests. Specifies the use of a plate.

In a test using such a microplate, individual reactions, culture of microorganisms, and the like are performed in each well of 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. As a product to be used for such a test, a predetermined amount of an antibacterial agent is dispensed in advance into each well of a microplate, and dried and frozen products are commercially available, but the cost of the microplate itself is low. Due to the high price, inexpensive products cannot be supplied, and there is a strong demand from users for lower prices. In such a microplate, a predetermined amount of a liquid sample or the like must be dispensed into each well at the time of the test, and it takes time and effort to perform the test. It is preferable to have tools that can be used.

In response to such demands, the present inventors have already proposed a chemical test or microbiological test device having a plurality of sample holding portions on a substrate (Japanese Patent Application No. 7-1532).
No. 39). This tool holds a predetermined amount of sample in the sample holding portion by absorbing the liquid sample by the sample holding portion provided on the base and absorbing the excess sample by the liquid absorber. The answer is excellent in practicality. However, there is a need for a more sophisticated chemical or microbiological test device with less variation in the amount of liquid sample held in the sample holding portion.

SUMMARY OF THE INVENTION According to the present invention, it is possible to reduce the fluctuation of the amount of the liquid sample held by a plurality of sample holding portions by one operation, and to easily manufacture the liquid sample. It is intended to provide a chemical or microbiological test device.

[0004]

SUMMARY OF THE INVENTION The present invention relates to Japanese Patent Application No. 7-1.
In the test device disclosed in Japanese Patent No. 53239, a plurality of upwardly extending cylindrical upright walls having a sample holding portion on a top surface are provided on a bottom member, and side surfaces of the upright walls are engaged with the sample holding portion forming openings. And the top surface of the upright wall is made to protrude upward through the sample holding portion forming opening, or the upright wall having a plurality of sample holding portions protruding from the top surface and extending upward is provided. Based on the knowledge that the above problem can be efficiently solved when each of the plurality of projecting sample holding portions provided on the top surface of the upright wall is engaged with the sample holding portion forming opening. It was done.

That is, the present invention provides a sample container for at least temporarily storing a liquid sample, a plurality of sample-holding-portion-forming openings arranged on a hydrophobic bottom surface in the container at intervals. A liquid absorber for absorbing the liquid sample around the sample holding portion when the liquid sample is stored in the sample container; and a bottom portion disposed below the sample container and capable of supporting the liquid absorber. And the bottom member,
In cooperation with the sample container, a storage area for the liquid absorber is formed, and the sample container includes a peripheral portion located below the sample holding portion, and a through hole provided in the peripheral portion. Wherein the through-hole is a chemical and microbiological test tool in fluid communication with the storage area disposed between the sample container and the bottom member, wherein the bottom member comprises: A plurality of upwardly extending cylindrical upright walls having a sample holding portion on a top surface, the top surface of the upright wall projecting upward through the sample holding portion forming opening, and a side surface of the upright wall Is engaged with the sample holding portion forming opening, or
(2) An upright wall having a plurality of sample holding portions protruding from the top surface thereof and extending upward, and each of the plurality of projecting sample holding portions provided on the top surface of the upright wall is the sample holding portion. A chemical and microbiological test device is provided that is engaged with a forming opening.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION The test device of the present invention will be described in detail below. The sample container used to store the liquid sample at least temporarily used in the present invention has a flat surface in the peripheral portion, the inside is depressed to form a hydrophobic bottom surface, and a plurality of sample holding portions are spaced apart from this bottom surface. It is preferable to use one having an opening for formation. This sample container can be easily molded by a method such as integral molding using a material described later. At this time, in the case of molding with a plastic material, if a corona discharge treatment or a surfactant is applied, the wettability of the surface is improved, and bubbles are generated when the test liquid as a liquid sample is put. It is preferable because generation can be prevented and generated air bubbles can be eliminated in a short time. or,
If a spout for a liquid sample is provided, for example, by providing an inclined gradient from the outside to the inside of the sample container at a part of the flat part around the sample container, the liquid (well) is placed in the well (sample holding part) when the liquid sample is introduced. Bouncing can be effectively prevented. It is also preferable to provide an arrow on the inclined pouring port in the direction in which the liquid is poured by rough surface finishing, convex surface treatment, or the like. further,
If a symbol such as A or B is attached to the part corresponding to the row of wells (sample holding parts) on the flat part around the sample container, it is possible to determine which well row corresponds to which test It is preferable because it is easy to do.

The edge of the opening for forming the sample holder is located above the peripheral portion, and the peripheral portion is formed by a funnel-shaped wall extending downward from the edge toward the through hole. This embodiment is preferable because an excess liquid sample among the liquid samples poured into the sample container can be promptly absorbed by the liquid absorber below. It is preferable that a part of the flat part in the peripheral part of the sample container is roughened so that the sample number can be described with a pencil or the like. Also, if a small hole is made in a part of the plane part of the peripheral part of the sample container to make it conductive to the space formed by the bottom member and the sample container, the liquid absorbing the liquid sample is placed on the bottom member. Since the absorber is accommodated, drying when the lid member is put on the sample container can be prevented.

The bottom member used in the present invention is arranged below the sample container, and can form a liquid absorber holding space integrally with the sample container. In the embodiment (1) of the present invention, it is necessary that the bottom member has a plurality of cylindrical upright walls having a sample holding portion on the top surface and extending upward. Wherein the bottom member is formed in the form of a top open container,
On the bottom wall of the bottom member, a plurality of upwardly extending cylindrical upright walls are formed, the top surface of the upright wall forms the sample holding portion, and the upright wall penetrates the liquid absorber. Preferably, it is in the form of a plurality of possible holes. Further, the upright wall is formed in a cylindrical shape, and the outer periphery of the top portion of the upright wall is formed in a size to be engaged with the inner peripheral edge of the opening for forming the sample holding portion of the sample container, or The tip wall surface portion of the cylindrical upright wall 33 is formed to have a size to be engaged with the inner peripheral edge of the sample holding portion forming opening, and to be engaged with the lower end of the inner peripheral edge of the sample holding portion forming opening. A step portion is formed, and the top end of the upright wall projects from the upper end of the opening for forming the sample holding portion preferably by 0.1 to 2 mm, particularly preferably by about 0.3 to 1 mm. Is preferred.

On the other hand, in the embodiment (2) of the present invention, the bottom member is
It has one or more (preferably one) upwardly extending upstanding walls, and has a plurality of projecting sample protruding portions on its top surface. Then, it is necessary that each of the plurality of protruding sample holding portions provided on the top surface of the upright wall is to be engaged with the sample holding portion forming opening. Here, it is preferable that the upper surface of the sample holding unit is located at the same position as the upper surface of the sample holding unit forming opening or at a position lower than the upper surface of the sample holding unit forming opening.
In particular, it is preferable that the upper surface of the sample holder is located between the upper surface and the lower surface of the opening for forming the sample holder. Also in the mode (2), it is preferable that the liquid absorber has a form in which a hole through which the upright wall can penetrate is formed, and the liquid absorber is located at a place other than the upright wall of the bottom member (that is, (Recessed place). In embodiments (1) and (2) of the present invention, it is preferable to provide a recess for holding the sample on the upper surface of the sample holding unit. In addition, if the sample holding portion formed on the top surface of the upright wall is roughened or a vertical groove is formed, bubbles generated when the liquid sample is poured are easily escaped, and bubbles are easily generated. Is preferred because it can be removed. At this time, it is more effective if the sample holding portion is further subjected to ionizing radiation treatment such as electron beam, UV, corona discharge or the like, or a treatment for improving surface wettability such as application of a surfactant.

In the present invention, the "sample holding portion" on the top surface of the plurality of upwardly extending cylindrical upright walls is used in a limited sense such as a portion for holding only a test sample only. Rather, it refers to a part for holding an arbitrary reaction system, culture system, or the like used in a desired chemical test or microbiological test. This sample holding portion is a hydrophilic or water-absorbing portion and may not contain any reagent, but it is preferable that the sample holding portion contains a reagent necessary for the test from the beginning. . For example, in the case of a tool used for measuring antimicrobial susceptibility by a microfluidic dilution method, a predetermined amount of a target antimicrobial agent is contained in a sample holding portion so that an appropriate Antimicrobial susceptibility can be measured simply by adding a bacterial solution in which a specified amount of bacteria is suspended in a medium. However, in such an embodiment,
The sample is not immediately eluted when the liquid sample is poured into the sample container, but the reagent is retained so that the excess sample is removed and a predetermined amount of sample is held in each sample holding part, and then eluted. It is desirable that it is. In the present invention, if an overcoat layer such as PVP (polyvinylpyrrolidone) is provided on the surface of a reagent such as an antibacterial drug, the storage stability of the drug is excellent and the drug is effectively prevented from flowing out.

In the case of a device for measuring antimicrobial susceptibility, for example, a single test device can include a plurality of sample holding portions which can contain the same amount of medium and whose antimicrobial content changes stepwise. If so, a fixed amount of the same bacterial sample is held in each sample holding portion of the device, and the antimicrobial susceptibility can be easily measured by observing the growth state. If the content of the antibacterial agent is changed, for example, by two times for each sample holding portion, it is suitable for the purpose of the above test. The shape of the sample holding portion on the top surface of the cylindrical upright wall is preferably provided as a concave portion, but the shape of the concave portion is not particularly limited, and the shape of the concave portion is not limited to a curved surface provided in an ordinary microplate. The concave portion may be a concave portion having a cylindrical shape or a conical shape. In this aspect, the size of the sample holding portion needs to be such that a certain amount of sample remains in the portion when an extra sample is absorbed and removed by the liquid absorber.
The specific size is not particularly limited as long as the above conditions are satisfied, but the diameter is 3 to 10 mm and the depth is 0.5.
It is preferable to use a sample having a size of about 5 mm and capable of holding a sample of about 0.01 to 0.2 ml.

The liquid absorber held on the bottom member of the test device of the present invention absorbs the liquid relatively quickly when it comes into contact with an aqueous liquid, usually a test liquid. Specifically, polymer fibers such as filter paper, water-absorbing paper, nonwoven fabric, and acetate filaments; and fibrous materials having water absorption and water retention properties, such as cotton, such as Bel Oasis (manufactured by Kanebo Co., Ltd.), (2 ) Sponge-like porous material composed of various polymers, etc., (3) polyvinyl alcohol / polyacrylate gel, crosslinked polyacrylate gel, crosslinked polyvinyl alcohol gel, crosslinked polyethylene oxide gel, crosslinked polydiethylacrylamide gel and crosslinked poly Water-absorbing gels composed of materials such as a cross-linked polyacrylamide-based gel such as isopropylacrylamide gel, a cross-linked polyvinyl pyrrolidone-based gel, and a cross-linked Paogen ™ gel (these water-absorbing gels can be used alone or in combination. ).

(4) A water-absorbing sheet in which the above-mentioned water-absorbing gel is sandwiched between non-woven fabric, paper, etc., such as Aqua Keeper (manufactured by Sumitomo Seika Co., Ltd.) and a drip sheet (Dai Nippon Printing Co., Ltd.) Water-absorbing sheet marketed under the trade name
(5) a water-absorbing sheet in which the above-mentioned water-absorbing gel coexists with the materials of the above (1), (2) and (4), for example, a sheet of paper coated with powder of a water-absorbing gel; Gel is used as a binder, for example, acrylic resin, polyvinyl butyral, polyester resin, polyurethane resin, fluorine resin, silicone resin, styrene resin sold by Mitsubishi Rayon Co., Ltd. under the trademark of Dianal BR-Resin.
Hydrophobic resins (insoluble in water) such as butadiene latex resin, and water-soluble and organic solvent-soluble (miscible) resins such as polyvinylpyrrolidone and hydroxypropylcellulose (these binders can be used alone or Can be used as a mixture)
For example, polyethylene, polypropylene, ethylene-vinyl acetate copolymer, ethylene-vinyl alcohol copolymer, polystyrene, polyacrylate, polyvinyl chloride, polyvinyl alcohol, polyvinyl butyral, polyamide, polyester, polycarbonate, polyurethane, polyimide, triacetyl cellulose, etc. (7) A portion of polyethylene glycol, such as that marketed under the trade name of Aqua Coke (Sumitomo Seika Co., Ltd.) Crosslinkable materials that are soluble in organic solvents and swell in water, polyacrylamide, sodium polyacrylate, polyethylene oxide, polypropylene oxide, polyvinyl alcohol, and polyvinyl alcohol A water-absorbing sheet prepared by coating the above-mentioned base material with various water-soluble polymers such as lolidone, etc., by irradiating with ionizing radiation and crosslinking to make it water-insoluble, (8) above (1) to (1). 7) Materials composed of any combination of the above materials can be mentioned. Among these, a water-absorbing felt (Bell Oasis), a pulp nonwoven fabric and a sponge are particularly preferred.

The liquid absorber composed of these materials is provided in such an amount that the excess amount of the liquid stored in the sample container except for the amount held in the sample holding portion can be sufficiently absorbed. preferable. In the present invention, in addition to the sample container, the bottom member, and the liquid absorber, a transparent or opaque lid that covers the upper portion of the sample container can be provided.
Protrusions are provided on the upper surface of the lid, preferably in parallel with the longitudinal direction of the outer upper surface of the lid, or in parallel with the short direction, and are provided in a belt-like shape. It is preferable to provide a concave portion which can be engaged with the projection, since a plurality of test tools can be vertically aligned and stacked. This structure is particularly effective for automation of measurement.

The base of the test device of the present invention, for example, the sample container, the bottom member and the lid can be formed of any material, but from the viewpoint of strength, workability, cost, etc., for example, polyethylene, polypropylene, The material can be selected from materials such as polystyrene, polyacrylate, polyvinyl chloride, polyamide, polyester, polycarbonate, and polyurethane. The size, thickness, and shape of the base are not particularly limited, and the size of the provided sample holding portion,
Selection should be made according to the test procedure used, such as number, shape, etc., and according to the size of the liquid absorber provided on the bottom member, and so as to obtain appropriate handling of the selected material. Can be. For example, a rectangle having a side of 2 to 25 cm, a length of 5 to 40 cm, and a thickness of about 0.1 to 5 mm is generally used. In addition, the material of the substrate can be appropriately selected in color, transparency, and the like so that a change indicating a test result such as the growth of bacteria and the progress of a color-forming reaction can be appropriately detected. Substrates such as black can be used.

Next, the present invention will be described with reference to the drawings. FIG. 1 is an exploded perspective view of the test tool according to the embodiment (1) of the present invention, and FIG. 2 is a cross-sectional view of the test tool shown in FIG. FIG.
The test tool shown in Fig. 1 includes a base 1 constituting a sample container, a liquid absorber 2 disposed below the base 1, and a bottom member 21 capable of accommodating the liquid absorber 2. A recess is formed in the central region of the base 1 by the continuous wall 9 bent downward,
A plurality of circular sample-holding-portion-forming openings 8 are arranged on the bottom surface of the concave portion at predetermined intervals. The diametrically extending linear portions 5 of the openings 8 are arranged to interconnect the perimeters of the openings 8, each linear portion 5 being at a plane or height where the perimeter of the opening 8 is located. It extends between the openings 8. The bottom surface of the base 1 extends obliquely downward in a region surrounded by the peripheral edge of the opening 8 and the linear portion 5 to form a funnel-shaped lower portion 6. In the center of the lowermost portion 6 located at the lowermost position, a through hole 1 through which the upper region and the lower region of the base 1 can be in fluid communication.
2 are formed.

Further, a spout 7 for a liquid sample is provided in a part of a plane portion around the base 1 constituting the sample container. The number 10 can be described. The liquid absorber 2 has an outer shape that can be stored in the bottom member 21 and has the same number of holes 31 as the circular openings 8 in the base 1 in the central region. Each hole 31 has a circular cross-sectional shape having the same or slightly larger diameter as the circular opening 8 and is formed in the liquid absorber 2 at substantially the same interval as the circular opening 8. . A bottom member 21 formed generally in the form of a top opening container includes a bottom wall 22, an outer peripheral wall 25, and an outer peripheral edge 26. A linear step 29 that can be aligned with the outer periphery of the base 1 is formed at each edge of the outer periphery 26. Also,
At the center of the bottom wall 22, the same number of the cylindrical upright walls 23 as the circular openings 8 or the holes 31 are formed. The outer diameter of the upright wall 23 substantially matches the inner diameter of the hole 31. Also, the standing wall 23
Has an outer diameter substantially matching the inner diameter of the sample holding portion forming opening 8. The top of the upright wall 23 is closed by a concave top wall 24 that is recessed downward, and the upper surface of the top wall 24 forms a concave sample holding portion 3.

In use, the liquid absorber 2 is housed in a bottom member 21 as shown in FIG. 2, and each cylindrical upright wall 23 of the bottom member 21 penetrates each circular hole 31 of the liquid absorber 2. Further, the base 1 is provided with a bottom member 21 such as a lid of the bottom member 21.
And the outer peripheral edge of the base 1 is placed on the outer peripheral edge 2 of the bottom member 21.
6. The circular openings 8 for forming the sample holding portions of the base 1 are aligned with the walls of the upright walls 23 of the bottom member 21. As shown in FIG. 2, the wall surface of the upright wall 23 is engaged with the opening 8 of the base 1, and the outer peripheral edge of the top of the upright wall 23 is the upper surface of the base 1, that is, the upper surface of the opening 8 for forming the sample holder. A concave sample holding portion 3 protruding upward is formed. Bottom member 21
Is provided with a concave portion 11 which can engage with a protrusion on the upper surface of a lid (not shown) covering the base 1 constituting the sample container. The height (thickness) of the liquid absorber 2 is preferably set such that the upper surface of the liquid absorber 2 is close to a hole formed at the lowermost part of the funnel-shaped lower portion 6. In the embodiment shown in FIGS. 1 and 2, a plurality of protrusions 28 are arranged adjacent to each upright wall 23. Each projection 28 is provided with a respective lower section 6 to ensure that the liquid absorber 2 is close to the hole of the lower section 6.
It is preferable to be positioned immediately below the through hole.

In this test device, when the base 1 and the absorber container 21 are formed of a transparent member, the degree of color development in the sample holding portion 3 after the test is automatically determined by transmitting light from the A or B direction. It has the advantage that it can be easily measured. FIG. 3 shows a longitudinal sectional view of a bottom member 34 having a cylindrical upright wall 33 in another form. In this embodiment, the distal end wall portion of the cylindrical upright wall 33 is formed to have a size to be engaged with the inner peripheral edge of the sample holding portion forming opening of the sample container, and the lower end of the inner peripheral edge of the sample holding portion forming opening is formed. A step 35 having a size to be engaged with the section is formed. FIG. 4 is an exploded perspective view of the test device according to the embodiment (2) of the present invention, and FIG. 5 is a cross-sectional view of the test device shown in FIG. The test device shown in FIG. 4 includes a base 51 constituting a sample container, a liquid absorber 52 disposed below the base 51, and a bottom member 7 capable of housing the liquid absorber 52.
1 is provided. A concave portion is formed in the central region of the base body 51 by a continuous wall 59 bent downward, and a plurality of circular sample-holding-portion-forming openings 58 are arranged at predetermined intervals on the bottom surface of the concave portion. I have. A diametrically extending linear portion 55 of the opening 58 is arranged to interconnect the periphery of each opening 58, and each linear portion 55 is
8 at the plane or height where the periphery of
8 between. The bottom surface of the base 51 is formed in a region surrounded by the periphery of the opening 58 and the linear portion 55.
It extends diagonally downward and forms a funnel-shaped lower part 56. At the center of the lowermost portion 56 located at the bottom,
A through-hole 62 that allows fluid communication between the upper region and the lower region of the base 51 is formed.

Further, a spout 57 for a liquid sample is provided in a part of a plane portion around the base 51 constituting the sample container. A number 60 can be described. The liquid absorber 52 has an outer shape that can be stored in the bottom member 71 and is provided with a hole 81 having the same size as or slightly larger than the outer peripheral dimension of the upright wall 73 of the bottom member 51.
Bottom member 51 formed generally in the form of a top opening container
Has a bottom wall 72, an outer peripheral wall 75 and an outer peripheral edge 76. The linear step portion 79 that can be aligned with the outer peripheral edge of the base body 51 has the outer peripheral edge 7.
6 at each edge. An upright wall 73 is formed at the center of the bottom wall 72, and its outer diameter substantially matches the inner diameter of the hole 81. At the top of the upright wall 73, the same number of sample holding portions 5 as the circular sample holding portion forming openings 58 are provided.
3 is provided so as to protrude, and the sample holding portion 53 is provided.
Has an outer diameter substantially matching the inner diameter of the sample holding portion forming opening 58. The top of the sample holding portion 53 is closed by a concave top wall 74 that is recessed downward, and the upper surface of the top wall 74 forms a concave sample holding portion 53.

In use, the liquid absorber 52 is housed in the bottom member 71, and the upright wall 73 of the bottom member 71 passes through the hole 81 of the liquid absorber 52. Further, the base 51 is placed on the bottom member 71 like a lid of the bottom member 71, and the outer peripheral edge of the base 51 is placed on the outer peripheral edge 76 of the bottom member 71. Substrate 5
The circular opening 58 for forming each sample holding part of the
Is aligned with the sample holding portion 53 provided on the upright wall 73. As shown in FIG. 2, the wall surface 74 of the sample holding portion 53
The top outer peripheral edge of the sample holding portion 53 engages with the opening 58 of the base 51, and is concave at the same height as the upper surface of the base 51, that is, the upper surface of the sample holding portion forming opening 58 or lower. Is formed. The bottom of the bottom member 71 is provided with a concave portion 61 that can engage with a projection on the upper surface of a lid (not shown) that covers the base 51 constituting the sample container. The height (thickness) of the liquid absorber 52 is preferably set so that the upper surface of the liquid absorber 52 is close to the hole formed at the bottom of the funnel-shaped lower portion 56. In the embodiment shown in FIGS. 4 and 5, the plurality of protrusions 78 are arranged adjacent to the upright wall 73. Each projection 78 is provided on the liquid absorber 52.
May be positioned directly below the through holes in each lower portion 56 to ensure that they are close to the holes in the lower portions 56.

In this test device, when the base 51 and the absorber container 71 are formed of a transparent member, the degree of color development in the sample holding portion 53 after the test is automatically determined by transmitting light from the A or B direction. It has the advantage that it can be easily measured. When the test device of the present invention is used, a reaction solution, a culture solution, and the like are added to a sample holding portion of the device to perform a reaction, culture, and the like. And the test result is detected by color development, emission, generation of fluorescence, change in turbidity, etc. according to the principle of the test. If a water-soluble silicone compound is added to the culture solution in advance, foaming can be effectively prevented when the culture solution is poured into the sample container. Examples of such a water-soluble silicone compound include the aforementioned organosilicon compounds in which an alkylene oxide such as ethylene oxide or propylene oxide has been introduced into the molecule.

For example, when measuring the minimum inhibitory concentration of an antibacterial drug, a method of visually detecting the growth of bacteria in a medium, a method of detecting the turbidity of a medium by an absorbance of 500 to 700 nm, and a method of detecting fluorescence in a medium There is a method in which a substrate is put in, a fluorescent substance is generated by a bacterial enzyme that accumulates in the medium as the bacteria grow, and the amount is measured by measuring the fluorescence intensity to correlate with the bacterial growth. The addition of a reaction solution, a culture solution, or the like to the sample holding portion can be performed simply by injecting an appropriate amount of the reaction solution or the like into the sample container using a pipette or the like (at least an extent that the entire sample holding portion is immersed in the sample). As a result, a predetermined amount of the sample can be held in the sample holding portion as described above. Test results are most easily detected by the naked eye, but when more accurate results are required, or when the results cannot be detected by the naked eye, especially when the sample holding part is made fine. A desired test result can be obtained by detecting the above-described turbidity, color development, light emission, generation of fluorescence, and the like by a conventionally used detection device. The measurement of the absorbance, the fluorescence intensity, and the like in the measurement of the minimum growth inhibitory concentration of the antibacterial agent can also be performed by a commercially available absorbance meter, fluorometer, or the like. The test device of the present invention can be manufactured to be compatible with these known automated measuring instruments.

The test device of the present invention uses a plastic microplate to sample a biological substance such as an antigen, an antibody or an enzyme, or a non-biological substance such as a coloring compound. Alternatively, it can be used for various chemical tests used as reagents and microbiological tests including cultivation of microorganisms, and a large number of reaction systems can be rapidly formed very easily without dispensing with a micropipette or the like. Moreover, it is possible to effectively prevent the sample from scattering into the surrounding environment at the time of the test. Hereinafter, the present invention will be further described with reference to Examples, but the present invention is not limited thereto. All percentages in the examples are percentages by weight.

[0025]

Example 1 A test device of the present invention shown in FIGS. 1 and 2 was produced as follows. A sample container base 1 of 6 cm long, 8 cm wide and 1 cm high and 1 mm thick, and a bottom member 21 having a thickness of 1 mm, 1 cm high and 12 sample holding portions are respectively injected using polystyrene. It was produced by molding. A test tool was prepared by combining the base 1 and the bottom member 21 of the sample container, and a water-absorbing felt or a pulp nonwoven fabric was accommodated therein as the liquid absorber 2 (test tool A of the present invention). On the other hand, except that the top surface of the upright wall having the sample holding portion is engaged with the lower portion of the inner edge of the sample holding portion forming opening (not protruding above the sample holding portion forming opening). A comparative test device similar to test device A of the present invention (Japanese Patent Application No. 7-1).
No. 53239 (FIG. 16). 10 ml of a culture solution (Mura Hinto Sobros) was added to each of these test devices, and the amount of the culture solution contained in each well was determined. 1 for each of the 4 test devices
The average volume, minimum volume, maximum volume and standard deviation of the two wells were determined. The results are shown in Table 1.

[0026]

Table 1 Table 1 (the present invention) Average (μl) 51.1 50.2 50.8 50.4 49.9 Highest 52.4 51.4 52.4 51.4 50.6 Lowest 49.3 48.6 49.9 49.8 49.1 Standard deviation 0.896 0.851 0.655 0.496 0.487

[0027]

[Table 2] Table 1 (continuation: comparative example) Average (μl) 57.2 50.9 50.3 52.8 57.4 Maximum 62.6 59.0 53.1 59.4 62.2 Minimum 49.4 46.2 45.8 43.9 50.9 Standard deviation 4.44 3.73 2.09 5.47 3.15 Thus, according to the present invention, it is possible to hold a plurality of sample holding portions in one operation. It is possible to reduce the fluctuation width (variation) of the amount of the sample to be performed.

Example 2 Using the same test tool A as used in Example 1, six kinds of antibacterials were applied to the sample holding portion in nine gradations, and the remaining one row was not coated with the antibacterials. Controls were used. The types of antibacterial agents are shown in Table 2, and all of them contain polyvinylpyrrolidone (PVP) L type (Nippon Soda Co., Ltd.)
5% isopropyl alcohol solution as an ink,
Each 0.01 ml was applied and dried using a dispenser.
The antimicrobial content of the solution was the same as the applied amount of PVP and the antimicrobial content was 1600, 800, 400, 200,
It was changed to be 100, 50, 25, 12.5, 6.25, 3.13 and 1.56 ng. Further, a pattern to which a PVP solution containing no antibacterial agent was applied was also prepared as a control.

Staphylococcus aureus ATCC as a test bacterium
Using 25923, the test cells cultured overnight on an agar medium were suspended in sterile physiological saline to about 10 ⁸ CFU / ml, and this was diluted 1000-fold with Mueller Hinton broth (about 10 ⁵ CFU / ml). /
ml) and 10 ml of the obtained broth inoculated with the test cells were poured into the container at one time. The container is once filled with broth, but gradually begins to be absorbed by the liquid absorber, a certain amount of broth remains in each well coated with the drug, and after about 30 seconds, excess broth is completely removed. Was. To prevent evaporation, a lid was placed on the top of the container, and the cells were cultured at 35 ° C for 16 to 18 hours. After the cultivation, the MIC value was defined as the minimum drug concentration in the sample-holding portion where growth of the bacteria was not visually observed. In this experimental example, MI
The C value was determined as shown in Table-2.

[0030]

[Table 3] Table 2 薬 剤 Drug name MIC value (μg / ml) ─────── ──────────────────── Piperacillin (PIPC) 0.5 Oxalysin (MPIPC) 0.25 Cefazolin (CEZ) 0.5 Cefmetazol (CMZ) 1.0 Minocycline (MINO) 0.25 Ofloxacin (OFLX) 0.5 ───────────────────────────

[Brief description of the drawings]

FIG. 1 is an exploded perspective view of a test device of the present invention.

FIG. 2 is a longitudinal sectional view of the test device of FIG.

FIG. 3 is a longitudinal sectional view of another embodiment of a bottom member used in the test device of the present invention.

FIG. 4 is an exploded perspective view of a test device according to another embodiment of the present invention.

5 is a longitudinal sectional view of the test device of FIG. In the figures, 1 and 51 are bases 2 and 52 are liquid absorbers 3 and 53 are sample holders. 8 and 58 are sample holder forming openings 12 and 62 are through holes 21 and 71 are The bottom member 23 is a cylindrical upright wall 73, and the upright wall 73 is an upright wall.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI G01N 33/543 521 G01N 33/543 521 (72) Inventor: Mutsuru Shibuya 2-17-11 Kyobashi, Chuo-ku, Tokyo Showa Pharmaceutical Inside (72) Inventor Masaho Hayashi 1-1-1, Ichigaya-Kagacho, Shinjuku-ku, Tokyo Dai Nippon Printing Co., Ltd. (72) Inventor Yasushi Yasuta 1-1-1, Ichigaya-Kaga-cho, Shinjuku-ku, Tokyo Large Within Nippon Printing Co., Ltd. (72) Inventor Chizuruko Oshina 1-1-1, Ichigaya-Kaga-cho, Shinjuku-ku, Tokyo Within Nippon Printing Co., Ltd. (72) Inventor Kiyoshi Oguchi 1-1-1-1, Ichigaya-ka-cho, Shinjuku-ku, Tokyo No. 1 Dai Nippon Printing Co., Ltd.

Claims (9)

[Claims] 1. A sample container for temporarily storing a liquid sample at least temporarily, a plurality of openings for forming a sample holding portion arranged at intervals on a hydrophobic bottom surface in the container, and a liquid sample in the container. A liquid absorber for absorbing a liquid sample around the sample holding portion when stored is provided, and a bottom member arranged below the sample container and capable of supporting the liquid absorber. The bottom member cooperates with the sample container to form a storage area for the liquid absorber, and the sample container includes a peripheral portion located below the sample holding portion and a peripheral portion located below the sample holding portion. And a chemical and microbiological test tool in fluid communication with the receiving area disposed between the sample container and the bottom member, the through-hole comprising: The material extends upward with (1) a sample holding part on its top surface. A plurality of cylindrical upright walls are provided, a top surface of the upright walls protrudes upward through the sample holding portion forming opening, and a side surface of the upright wall engages with the sample holding portion forming opening. Or (2) an upright wall having a plurality of sample holding portions protruding from the top surface thereof and extending upward, and the plurality of protruding sample holding portions provided on the top surface of the upright wall. A tool for chemical and microbiological tests, wherein each tool is engaged with the opening for forming a sample holder. 2. The method according to claim 1, wherein an edge of the opening for forming the sample holding portion is located above the peripheral portion, and the peripheral portion extends downward from the edge toward the through hole. 2. The tool according to claim 1, wherein the tool is formed by a funnel-shaped wall. 3. The method according to claim 1, wherein the bottom member is formed in the form of a top-opening container, and a plurality of upwardly extending cylindrical rising walls are formed on the bottom wall of the bottom member. The tool according to claim 1, wherein a top surface of the wall forms the sample holding portion, and the liquid absorber has a plurality of holes formed through the upright wall. 4. The method according to (1), wherein the upright wall is formed in a cylindrical shape, and a top outer periphery of the upright wall is engaged with a lower end of an inner peripheral edge of an opening for forming a sample holding portion of the sample container. 2. A step portion is formed, and a top end of the upright wall projects 0.1 to 2 mm upward from an upper end of the opening for forming a sample holding portion.
A tool as described in. 5. The tool according to claim 1, wherein in (2), the upper surface of the sample holder is located at the same position as the upper surface of the opening for forming the sample holder. 6. The tool according to claim 1, wherein in (2), the upper surface of the sample holder is lower than the upper surface of the opening for forming the sample holder. 7. The tool according to claim 1, wherein in (2), the upper surface of the sample holder is located between the upper surface and the lower surface of the opening for forming the sample holder. 8. The tool according to claim 5, wherein in (2), the upper surface of the sample holder has a recess for holding the sample. 9. The tool according to claim 1, further comprising a transparent lid that covers an upper part of the sample container.