JP2007024734A - Evaporation prevention means of biochemical substance - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an evaporation prevention means of a biochemical substance, preventing evaporation of a biochemical substance specimen, such as collected saliva, and freely selecting the time from collection, until the analysis of the specimen. <P>SOLUTION: In a measuring method of the concentration and/or the activity of the biochemical substance, the biochemical substance is collected into a specimen collection part of a sheet-shaped carrier, having at least the specimen collection part, and at least the specimen collection part is covered with a covering object formed from a moisture-impermeable member, in order to prevent transpiration of the moisture in the specimen, and then the cover is opened, to thereby generate a contact between the specimen collection part and a reagent, and to generate a specimen detection reaction. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to means for preventing evaporation of biochemical substances such as collected saliva.

Yamaguchi of the present inventor pays attention to the fact that the activity of α-amylase (enzyme, hereinafter referred to as salivary amylase) in saliva varies with changes in human sensitivity. (Patent Documents 1 to 4) shown in FIG.
The salivary amylase activity analysis method for which a part of the present inventors has already applied for a patent consists of a saliva collection / analysis disposable saliva collection / analysis instrument (hereinafter abbreviated as a test strip) and an analyzer main body. Has been.
Japanese Patent Application No. 2000-364241 (Japanese Patent Laid-Open No. 2002-168860): Stress judgment method and judgment device Japanese Patent Application No. 2003-051150 (Japanese Patent Application Laid-Open No. 2004-24236): Amylase Measuring Reagent and Measuring Method Japanese Patent Application No. 2003-051151 (Japanese Patent Application Laid-Open No. 2004-121214): Reagent, Measurement Method and Apparatus for Measuring Amylase Activity Japanese Patent Application No. 2003-362805 (Japanese Patent Laid-Open No. 2004-267202): Kit for measuring amylase activity

  The salivary amylase activity analysis method needs to analyze the concentration and / or activity of salivary amylase promptly after collecting saliva in order to avoid changes in characteristics due to evaporation of water after collecting saliva. The technology did not consider a test strip saliva sample evaporation prevention method or instrument. Therefore, at present, there is a problem that only one sample can be handled by one analytical instrument. In order to solve this problem, the present inventors have studied various means that can prevent evaporation of water and prevent loss of activity, and have also studied methods for solving the problem at low cost.

The present inventor has studied a means for covering the sample collection part with a water-impermeable coating in order to easily prevent evaporation of moisture from the sample collection part of the sheet-like carrier having at least the sample collection part. In general, the present invention has been found to be effective when measuring biochemical substances as specimens.
That is, this invention consists of the following.
1. In order to collect biochemical substances at least in the sample collection part of the sheet-like carrier having the sample collection part, and then prevent the evaporation of the water component in the sample, at least the sample collection part is made of a water component-impermeable member. A method for measuring the concentration and / or activity of a biochemical substance, in which the coating is opened and then the coating is released to bring the specimen collection part into contact with the reagent, thereby leading to a specimen detection reaction.
2. The method for measuring the concentration and / or activity of the biochemical substance according to item 1, wherein the coating means is selected from the following.
1) Cover the specimen collection part with a suction cup type flat sheet.
2) Cover the specimen collection part with an adhesive-type flat sheet.
3) Cover the specimen collection part with a cap.
4) Place the entire sheet-like carrier having the specimen collection part in an airtight bag or container.
3. The method according to item 2 above, wherein a sheet-like carrier is sandwiched between at least two upper and lower planar sheets.
4). 4. The method according to item 2 or 3 above, wherein a transparent part capable of visually observing at least the specimen collection part is provided on at least one surface of the planar sheet.
5. 5. The method according to any one of items 2 to 4, wherein the sheet portion corresponding to the specimen collecting portion of the planar sheet has a gap or a recess.
6). It is a planar sheet formed by stacking at least two upper and lower sheets, and either one of the upper and lower planar sheets has a gap or a recess into which a sample collecting part of a sheet-like carrier having at least a sample collecting part can be inserted in one direction. Specimen collection that allows the upper and lower planar sheets to be contacted in an airtight state after insertion and loading of the specimen collection part of the sheet-like carrier having the specimen collection part, and that the evaporation of water components in the specimen collection part of the sheet-like carrier can be reduced A device for preventing transpiration of water components in a specimen collecting part of a sheet-like carrier having a part.
7). 7. The transpiration prevention device according to item 6 above, wherein at least one surface of the flat sheet has a transparent portion at which the sample collection portion can be visually observed.
8). On the lower plate sheet of the water component-impermeable member, an intermediate plate sheet having a gap or a recess capable of inserting and loading the sample collection portion of the sheet-like carrier in one direction is adhered, and the sample collection portion of the sheet-like carrier is attached. The transpiration prevention device according to 6 or 7 above, which is composed of an upper plate sheet that can be brought into airtight contact with the middle plate after insertion and loading.
9. 9. The transpiration prevention device according to 8 above, wherein the bonding is performed with a double-sided tape.
10. 10. The transpiration prevention device according to item 8 or 9 above, wherein the upper plate sheet that is hermetically contactable with the middle plate is a suction cup system.
11. The sheet-like carrier having at least the sample collection part and the means for covering the sample collection part used in the method according to any one of the preceding items 1 to 5 or the transpiration prevention device according to any one of the preceding items 6 to 10 are combined. Reagent kit.

  Since the means of the present invention can achieve the intended effect at a very low cost and easily, it has great value as a means for detecting a large amount of biochemical substances. By simply setting the evaporation prevention kit of the present invention at the tip of the test strip, it is possible to prevent the evaporation of collected biochemical samples such as saliva, and the time from sample collection to analysis can be freely selected Is possible. As a result, it is possible to analyze any number of saliva samples at any time with one analyzer main body, and it is not necessary to prepare a plurality of expensive analyzer main bodies.

  According to the first aspect of the present invention, a biochemical substance is collected at least in a specimen collecting portion of a sheet-like carrier having a specimen collecting section, and then at least the specimen collecting section is impervious to the water component in order to prevent evaporation of water components in the specimen. A method for measuring the concentration and / or activity of a biochemical substance, which is coated with a coating made of a conductive material, and then the coating is released to bring the sample collection part into contact with the reagent and lead to a sample detection reaction. It is.

The sample of the present invention is intended for saliva, particularly amylase in saliva in the examples. However, the present invention is not limited to this, and a wide range of biochemical substances can be measured or tested using at least a sheet-like carrier having a sample collection part. It can be used. In addition to saliva, it can also be applied to urine, sputum, nasal discharge, tears, sweat, gingival crevicular fluid, stool, waste liquid, extract and the like.
The sample is collected in the sample collection unit. The collection is performed by a capillary phenomenon, suction, dripping, tongue licking of the collection unit, and the like in the collection unit, but is not particularly limited. The sample collection unit is made of a sheet-like member such as water-absorbing paper, water-absorbing cotton, woven fabric, or non-woven fabric, and can hold at least water components in the portion. Immediately after the collection, the specimen collection part is covered with a coating made of the water component-impermeable member of the present invention to prevent evaporation of moisture.

  The water component impervious member used for the coating of the present invention can be widely used as long as it is inexpensive, retains ease of form processing, and prevents moisture permeation. For example, as plastic materials, for example, polyester [polyethylene terephthalate (PET), polyethylene naphthalate (PEN), polybutylene terephthalate (PBT), etc.], olefin resin [polyethylene (PE), polypropylene (PP), ethylene-propylene, etc. Copolymer, ethylene-vinyl acetate copolymer (EVA) and other olefin resins having an α-olefin as a monomer component], polyethersulfone (PES) (polyethersulfone), polysulfone, polyvinylchloride (PVC) ), Polyphenylene sulfide (PPS), amide resins [polyamide (nylon), wholly aromatic polyamide (aramid), etc.], polyimide (PI), polyamideimide, polyetherimide (PEI), polyesterimide, methacrylate Resin [polymethyl methacrylate (PMMA) etc.], styrene resin [polystyrene, acrylonitrile-styrene copolymer (AS resin), acrylonitrile-butadiene-styrene copolymer (ABS resin) etc.], polycarbonate (PC), Polyacetal, polyarylene ether (such as polyphenylene ether), polyphenylene sulfide, polyarylate, polyaryl, polyurethanes, polyether ketones [polyether ether ketone (PEEK), polyether ketone ketone, etc.], polyacrylic acid esters (polyacrylic) Acid butyl, polyethyl acrylate, etc.) and epoxy resins. These materials (plastic materials) can be used alone or in combination of two or more.

  The covering means of the present invention includes 1) covering the sample collecting part with a suction-type flat sheet, 2) covering the sample collecting part with an adhesive-type flat sheet, and 3) covering the sample collecting part with a cap. 4) For example, it is preferable to place the entire sheet-like carrier having the specimen collecting part in an airtight bag or container.

  Covering the sample collection part with a suction-type flat sheet means that the surface layer of one of the flat sheets to be covered has a cell suction function, and this cell suction function makes contact with the opposing flat sheet to increase the adsorption force. Demonstrate and the sample collection part is loaded between both planar sheets. The planar sheet having the cell suction function is composed of, for example, a base material layer and a cell suction layer, and may have a separator or a protective layer in order to ensure the virginity of the cell suction layer. As a sheet of the cell suction cup, for example, there is a ZEON Kasei ZEON AL sheet. This uses PET film (50μ of easy adhesion type) as the base material and acrylic resin (acrylic emulsion is foamed) as the foam layer. The thickness of the acrylic resin is 300 ± 50μ, 100 ± 30μ. Etc. The standard of the size of the cell suction cup “foam diameter” is mainly in the range of 20 to 100 μm, but is not promised.

  Covering the specimen collection part with an adhesive-type flat sheet means that one surface layer of the flat sheet to be coated has an adhesive layer, and this adhesive layer is in contact with the opposing flat sheet for adsorption. The specimen collecting part is loaded between the two flat sheets by exerting force. The pressure-sensitive adhesive layer is not particularly limited as long as it can be easily peeled off and maintains adhesiveness. For example, an acrylic pressure-sensitive adhesive, a rubber-based pressure-sensitive adhesive, a silicone-based pressure-sensitive adhesive, a urethane-based pressure-sensitive adhesive, and a polyester-based pressure-sensitive adhesive layer Known pressure-sensitive adhesives such as pressure-sensitive adhesives, styrene-diene block copolymer-based pressure-sensitive adhesives, vinyl alkyl ether-based pressure-sensitive adhesives, polyamide-based pressure-sensitive adhesives, fluorine-based pressure-sensitive adhesives, improved creep characteristics, and radiation-curable pressure-sensitive adhesives Can be appropriately selected and used. An adhesive can be used individually or in combination of 2 or more types.

In the present invention, the planar sheet has a gap or a recess into which at least the specimen collection part of the sheet-like carrier can be inserted and loaded in one direction, and the specimen collection part loaded in the gap or the depression is the surface of the covering part of the planar sheet There is enough space to prevent contact. The specimen collecting portion of the loaded sheet-like carrier is optimally sized and shaped to fit in the gap without any gap. After inserting and loading the sample collecting part of the sheet-like carrier having the sample collecting part, the upper and lower planar sheets can be contacted in an airtight state, thereby reducing transpiration of water components in the sample collecting part of the sheet-like carrier.
In order to provide the gap or recess, the intermediate plate having the gap or recess is bonded to the lower plate which is a planar sheet and / or the sheet to be coated (upper plate) which is a planar sheet by known means. Can be achieved. This bonding means can be easily achieved by using a double-sided adhesive sheet (tape), for example. Using a plurality of these intermediate plates, providing a difference in the gap portion, providing a step in the non-sample collection portion and the sample collection portion of the sheet-like carrier to be loaded, and making the sample collection portion a deeper gap or recess makes the sample collection portion more Can be secured.

In the present invention, the planar sheet is generally a planar sheet formed by superimposing at least two upper and lower sheets of a sheet serving as a lower plate and a covering sheet (upper plate). The end fixing type or two separate sheets may be used. In the folding type, one sheet can be folded at substantially the center, and when in use, the folded sample collecting part is covered so as to sandwich the sheet-like carrier having the sample collecting part, and the upper and lower two sheets are overlapped. At this time, the intermediate plate is bonded to the overlapping flat sheet portion, and the sample collection portion is covered with a gap or a recess formed by the intermediate plate without any gap.
With the fixed end type, one end is fixed in a state where two upper and lower sheets are overlapped. When in use, the stacking is released, and the sample collecting part is loaded on one of the upper and lower planar sheets. Thereafter, the specimen collection part is covered with a flat sheet. Two separate sheets refer to two separate planar sheets separated in advance. In these cases as well, a void or a recess can be easily provided by an intermediate plate as described above.

  The planar sheet has at least one transparent portion on at least one surface where the specimen collecting portion can be visually observed. This transparent part prevents the transpiration of water components in the specimen collecting part of the sheet-like carrier by bringing the specimen collecting part of the sheet-like carrier having the specimen collecting part into contact with the upper and lower planar sheets in an airtight state after insertion and loading. Thus, it is possible to check the state from the outside of the sample collection unit. In a system for detecting by a change in the color of the sample collection unit, the measurement can be performed without removing the planar sheet.

  The planar sheet as described above is constructed as follows, for example. An intermediate plate sheet having a gap capable of inserting and loading the sample collecting part of the sheet-like carrier in one direction is adhered on the lower plate sheet of the water-impermeable member, and the sample collecting part of the sheet-like carrier is inserted and loaded. It is composed of an upper plate sheet that can be hermetically contacted with the middle plate later. The water component-impermeable member is as described above, and the airtight contact is a sucker method or an adhesive method. In the case of the adhesive method, a simple double-sided pressure-sensitive adhesive tape is used for adhesion, and a flat sheet member other than the sample collection unit and the coating are adhered and adhered to prevent moisture permeation from the sample unit. is there.

  Covering the specimen collection part with a cap means that the covering means is a cap formed of a water component-impermeable member, and the part including the specimen collection part after specimen collection is closely attached so as to cover the cap. The By the covering with the cap, transpiration of the water component in the specimen collecting part is prevented.

  Placing the entire sheet-like carrier having the specimen collecting part in an airtight bag or container means that the specimen is collected after the specimen is collected by using an airtight bag or container formed of a water-impermeable member for the covering means. The entire sheet-like carrier including the part is hermetically loaded into an airtight bag or container. By this loading, the transpiration of the water component in the specimen collecting part is prevented more than a certain amount.

  The sheet-like carrier having at least the specimen collection part as described above, and means for covering the specimen collection part used in the method described above or the transpiration prevention device can be provided in combination as a reagent kit.

  Hereinafter, the present invention will be described by way of examples, but the present invention is not limited thereto.

Example 1
FIG. 1 is a representative example of an evaporation preventing device. It is an example of the evaporation prevention instrument comprised by laminating | stacking two or more planar sheets containing at least 1 or more adhesive material. In the figure, 1 is a stopper, 2 is a special film, 3 is a middle plate, 4 is a double-sided tape (black), and 5 is a lower plate. In the figure, W = means an outline of the horizontal length, that is, a horizontal length of about 50 mm, and l = means an outline of the vertical length, that is, about 50 mm and 58 mm. In this embodiment, the device is composed of four layers of sheets, and there are U-shaped gaps between the three middle plates and the four double-sided tapes. The lower plate is bonded. As a result, the formed U-shaped gap can be loaded with a sheet-like carrier having a specimen collection part. In order to cover the sample collection portion more densely, it can be achieved by providing a recess at the center of the flat sheet instead of being U-shaped and covering only the sample collection portion (FIG. 1-2). Four special films, which are coatings, are provided on the uppermost part to achieve dense coating. The special film has a cell sucker or a peelable pressure-sensitive adhesive layer that can be closely adhered to at least three intermediate plates. Each of these sheets is fixed to each other with, for example, one stopper, and as shown in FIG. 2, when the sheet-like carrier is loaded, two special films can be rolled up. It should be noted that the gap in the part covering the specimen collecting part has a sufficient gap to avoid contact with the two special films even when at least two special films are covered. Table 1 illustrates the thickness of each sheet, but this thickness can be freely adjusted according to the thickness of the sheet-like carrier. Adjusted.
FIG. 3 shows a state in which a sheet-like carrier is loaded in the transpiration prevention device, and a specimen collecting part is provided at the tip of the sheet-like carrier. The transpiration prevention device uses a U-shaped transparent member that can confirm this part, and this part can be visually confirmed in the covering state of the specimen collecting part.
FIG. 5 shows that the covering is a cap method. If an integral-molded cover method (evaporation prevention cap) or an integral-molded bag is used, another mode in which it is not necessary to get used to wearing and an effect that saliva does not adhere can be expected.

(Example 2)
An anti-transpiration device constituted by laminating flat sheets was prepared, and its anti-evaporation effect was evaluated. Saliva was used as the specimen, and a salivary amylase system was used as the measurement system. Collect saliva from one subject and drop 25 μL onto a saliva collection strip (an example of a sheet-like carrier with a specimen collection section) to indicate “with transpiration prevention device” and “without transpiration prevention device (control)” And in a constant temperature room, each was stored under conditions of 26 ° C. and 50% humidity. Measurements are 7 points after 0, 0.5, 1, 2, 3, 6, 12 hours for “with transpiration prevention device”, and 0, 0.5, 1 hour after for “without transpiration prevention device”. A total of 3 points were measured, and each point was measured 3 times. The initial value of the salivary amylase activity value was 40 kU / L, but in the control, the salivary amylase activity value rapidly increased after the start of the experiment, and increased by 35% compared to the initial value after 1 hour. On the other hand, the saliva amylase activity value was relatively stable in the case of “with transpiration prevention device”, and the effect of preventing evaporation was recognized. FIG. 4 illustrates the results. In the figure, ◯ indicates a case with a transpiration prevention device, and △ indicates a control.

  In the above experiment, in order to analyze the α-amylase activity present in the saliva of a subject, saliva can be obtained by attaching a covering that covers the saliva collection part in a device that collects saliva using capillary action. If you use an evaporation prevention device to prevent evaporation of the sample, you can prevent evaporation of the collected saliva sample simply by setting a transpiration prevention device with a simple structure at the tip of the test strip, and the time from saliva collection to analysis Can be freely selected. As a result, it is possible to analyze any number of saliva samples at any time with one analyzer main body, and it is not necessary to prepare a plurality of expensive analyzer main bodies.

  It can be used in the health / medical field as a measurement kit used for measuring biochemical substance activity such as amylase activity, particularly as a measurement kit for measuring amylase activity in specimens such as saliva of a subject.

Composition of transpiration prevention equipment Composition of transpiration prevention equipment Concept of use of transpiration prevention equipment Appearance of transpiration prevention device Temporal changes in salivary amylase activity in transpiration prevention devices and controls Concept of transpiration prevention device (integrated molding cover method (evaporation prevention cap))

Explanation of symbols

1 Stopper 2 Special Film 3 Middle Plate 4 Double-sided Tape (Black)
5 Lower plate

Claims (11)

  In order to collect biochemical substances at least in the sample collection part of the sheet-like carrier having the sample collection part, and then prevent the evaporation of the water component in the sample, at least the sample collection part is made of a water component-impermeable member. A method for measuring the concentration and / or activity of a biochemical substance, in which the coating is opened and then the coating is released to bring the specimen collection part into contact with the reagent, thereby leading to a specimen detection reaction. The method for measuring the concentration and / or activity of a biochemical substance according to claim 1, wherein the covering means is selected from the following.
1) Cover the specimen collection part with a suction cup type flat sheet.
2) Cover the specimen collection part with an adhesive-type flat sheet.
3) Cover the specimen collection part with a cap.
4) Place the entire sheet-like carrier having the specimen collection part in an airtight bag or container. 3. The method according to claim 2, wherein a sheet carrier is sandwiched between at least two upper and lower planar sheets. The method according to claim 2 or 3, further comprising a transparent portion on at least one surface of the planar sheet, through which at least the specimen collection portion can be visually observed. The method according to any one of claims 2 to 4, wherein the sheet portion corresponding to the specimen collecting portion of the planar sheet has a gap or a recess. It is a planar sheet formed by stacking at least two upper and lower sheets, and either one of the upper and lower planar sheets has a gap or a recess into which a sample collecting part of a sheet-like carrier having at least a sample collecting part can be inserted in one direction. Specimen collection that allows the upper and lower planar sheets to be contacted in an airtight state after insertion and loading of the specimen collection part of the sheet-like carrier having the specimen collection part, and that the evaporation of water components in the specimen collection part of the sheet-like carrier can be reduced A device for preventing transpiration of water components in a specimen collecting part of a sheet-like carrier having a part. The transpiration prevention device according to claim 6, further comprising a transparent portion on which at least the specimen collection portion can be visually observed on at least one surface of the flat sheet. On the lower plate sheet of the water component-impermeable member, an intermediate plate sheet having a gap or a recess capable of inserting and loading the sample collection portion of the sheet-like carrier in one direction is adhered, and the sample collection portion of the sheet-like carrier is attached. The transpiration prevention device according to claim 6 or 7, comprising an upper plate sheet that can be brought into airtight contact with the middle plate after insertion and loading. 9. The anti-transpiration device according to claim 8, wherein the adhesion is performed with a double-sided tape. The transpiration prevention device according to claim 8 or 9, wherein the upper plate sheet that can be hermetically contacted with the middle plate is a suction cup system. A sheet-like carrier having at least a sample collection part and a means for covering the sample collection part used in the method according to any one of claims 1 to 5 or a transpiration prevention device according to any one of claims 6 to 10 are combined. Reagent kit.