JPH09133673A - Instrument and method for measuring liquid sample - Google Patents

Abstract

PROBLEM TO BE SOLVED: To quickly and easily analyze a liquid sample for a specific component with accuracy with an inexpensive simple structure. SOLUTION: An instrument for measuring liquid sample is provided with at least one bendable section 2 and a reagent layer 4 containing a reagent which reacts to a specific component and generates a reaction product on a substrate 3 which can be bent together with the section 2. When the specific component contained in a liquid sample is measured, the substrate 3 is bent in a U-shape together with the section 2 with the reagent layer 4 inside so that a sample supplying section 6 can be formed of a capillary tube chamber 5 and its opening section between facing surfaces 3a and 3b of the substrate 3.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a liquid sample measuring tool and a liquid sample measuring method, and more particularly to a liquid sample measuring tool and a liquid sample measuring method for measuring a specific component in a liquid sample.

[0002]

2. Description of the Related Art Many measuring tools have been developed as means for rapidly and easily measuring a specific component in a liquid sample such as blood, urine, saliva, sweat and spinal fluid. In particular, the dry measurement tool does not require preparation of a reagent, and enables specific components in a liquid sample to be measured by a simple operation such as dropping a sample to be measured on the tool.

In the field of clinical tests, when a specific component in a liquid sample is, for example, glucose, lactic acid, uric acid, cholesterol, etc., an assay method using an enzyme reaction with high substrate specificity has been conventionally used. Has been. Examples of conventional dry measuring tools include, for example, Japanese Examined Patent Publication No. 49-33.
The invention disclosed in Japanese Patent Laid-Open No. 800-15075
The invention disclosed in Japanese Patent Laid-Open No. 1-18065 and the invention disclosed in Japanese Patent Laid-Open No. 4-188065 are known.

[0004]

[Problems to be Solved by the Invention]
The diagnostic agent for detecting body fluid contents of the invention disclosed in JP-A-33800 uses a water resistance test film in which a detection system using an oxidase and a peroxidase is dispersed in a polymer and applied to a plastic member. There is.

In this known water resistance test film, it is possible to measure the produced coloring dye from the sample supply side of the film by contacting the sample with the reagent layer for a certain period of time and then wiping it off with absorbent cotton or the like. And However, this wiping operation caused individual differences due to time and strength. Further, when the liquid sample is blood of another person, the wiping operation may cause an infection such as a disease.

Then, the invention disclosed in Japanese Patent Laid-Open No. 2-150751 was proposed as a measuring tool which does not require a wiping operation. The device and method for measuring a specific component in a liquid sample, which is the invention disclosed in this publication, has been filed by the present applicant.

The tool for measuring a specific component in the liquid sample uses a porous film as a support for the reagent layer, and the reagent layer is placed on the porous film. However, this has a problem in that in the sample holding layer arranged above the reagent layer, a difference occurs due to the permeability of the liquid sample, and bubbles are generated between the reagent layer and the sample holding layer.

Therefore, a measuring tool has been proposed in which the liquid sample is not dropped on the sample holding layer. That is JP-A-4-18806.
A liquid sample measuring tool and a measuring method according to the invention disclosed in Japanese Patent Laid-Open No. The invention disclosed in this publication is also an invention filed by the present applicant, and this liquid sample measuring tool is provided with a support and a cover at an interval that induces a capillary phenomenon to form a capillary chamber. It is characterized by.

However, this also has a problem that the structure of the capillary chamber is complicated, and moreover, it is not disclosed in Japanese Patent Laid-Open No. 188065.
Since the cover used in the invention described in the publication is a molded product, it also causes a cost increase.

An object of the present invention is to solve the problems of the conventional liquid sample measuring technique, and to provide a low cost and simple structure,
It is an object of the present invention to provide a liquid sample measuring tool and a liquid sample measuring method capable of measuring a specific component in a liquid sample quickly, simply and accurately.

[0011]

The present invention is a liquid sample measuring tool, which is configured as follows in order to solve the above-mentioned technical problems. That is, the liquid sample measurement tool of the present invention includes at least one bendable portion, a support that can be bent at this bendable portion, and a support that is disposed on this support and that reacts with a specific component to generate a reaction product. And a reagent layer containing possible reagents. <Specific Configuration of Invention of Liquid Sample Measuring Tool> The liquid sample measuring tool of the present invention comprises the above-mentioned essential components, but the specific components are also realized in the following cases. The specific components thereof, when a specific component in the liquid sample reacts with the reagent layer to generate a reaction product, in order to capture the reaction product by an optical change,
The support has a through hole at a position other than the bendable portion, and a light transmissive film having the reagent layer fixed thereto is arranged so as to cover the through hole.

Further, in the liquid sample measuring device of the present invention, in addition to the above-mentioned features, it is preferable that the light transmissive film is composed of a porous film. Further, in the liquid sample measuring tool of the present invention, when measuring a specific component in a liquid sample, the support is bent into a substantially U shape at the bendable portion so that the reagent layer is located inside, and It is characterized in that the sample supply section is formed by the capillary chamber and the opening thereof between the facing surfaces of the support.

Further, the present invention is a liquid sample measuring method,
In order to solve the above-mentioned technical problem, it is configured as follows. That is, the present invention comprises a support having at least one bendable portion, which can be bent at the bendable portion, and a reagent layer which is disposed on the support and shows an optical change by reacting with a specific component. A method for measuring a specific component in a liquid sample using a liquid sample measuring tool provided, wherein the support is bent into a substantially U shape at the bendable portion so that the reagent layer is located inside, A sample supply part is formed between the facing surface of the support and a capillary chamber and its opening, a liquid sample is supplied from the sample supply part to the capillary chamber, and the liquid sample that moves in the capillary chamber by a capillary phenomenon is It is characterized in that a specific component in a liquid sample is measured by measuring the amount of the reaction product substance produced by reacting by touching the reagent layer to produce a reaction product substance. <Specific Structure of Invention of Liquid Sample Measuring Method> The liquid sample measuring method of the present invention comprises the above-mentioned essential components, but the specific components are also realized in the following cases. The specific component is characterized in that when a specific component in the liquid sample reacts with the reagent layer to generate a reaction product, the reaction product is captured by an optical change.

In the liquid sample measuring method of the present invention, in addition to the above-mentioned characteristics, the optical change in the reagent layer is the transmittance, and for that reason, a part of the support is formed in the light-transmitting part. It is characterized by

Further, in the liquid sample measuring method of the present invention, the optical change in the reagent layer is the reflectance, so that part of the support is formed in the light-reflecting portion. .

Furthermore, at the same time as analyzing the optical change in the reagent layer, the supply of blood is detected by the optical change in the capillary chamber, and for that purpose, the sample adjacent to at least the reagent layer in the support is detected. It is characterized in that a part of the supply portion side is formed as a light transmissive portion.

According to the liquid sample measuring tool and the liquid sample measuring method having such a configuration, when the specific component in the liquid sample is measured using the liquid sample measuring tool, the reagent layer is positioned inside from the bendable portion. Bend this measuring tool in a substantially horizontal U shape. As a result, the sample supply section is formed between the facing surface of the support and the capillary chamber and its opening.

When the liquid sample is supplied to the capillary chamber from the sample supply unit, the liquid sample moves in the capillary chamber by the capillary phenomenon and comes into contact with the reagent layer. As a result, the reagent layer reacts with the specific component in the liquid sample and exhibits a change in the amount of the reaction product. The change in the amount of the reaction product can be measured by an appropriate means, and the measurement result can be confirmed by, for example, outputting it to a display, a printer or the like.

[0019]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, a liquid sample measuring tool of the present invention will be described in detail with reference to the embodiments shown in the drawings together with a liquid sample measuring method. First, FIG. 1 shows a liquid sample measuring tool 1 according to an embodiment of the present invention. The liquid sample measuring tool 1 of this embodiment basically includes at least one bendable portion 2, a support 3 that can be entirely bent at this bendable portion, and a support 3 that is disposed on the support 3 and has a specific component. And a reagent layer 4 that reacts with (in this embodiment, the change in the amount of the reaction product is detected by an optical change).

In this liquid sample measuring tool 1, when measuring a specific component in a liquid sample, as shown in FIGS. 2 and 3, the support 3 can be bent so that the reagent layer 4 is located inside. Bend it into a horizontal U shape. As a result, in the liquid sample measuring tool 1 in the ready-to-use state, the capillary chamber 5 is formed between the facing surfaces 3a and 3b of the bent support body 3, and the sample supply portion 6 is formed at the open portion. .

As is apparent from FIGS. 2 and 3, the sample supply unit 6 has both ends 7a and 7b of the support 3 along the planned bending lines 8a and 8b in the bending direction of the bendable portion 2. Is bent slightly to the opposite side, and the support 3 is bent at the bendable portion 2
When bent in a V shape, both ends thereof are configured to spread out in an eight shape. This makes the suction of the liquid sample into the capillary chamber 5 more effective.

Various materials can be used as the support 3 constituting the liquid sample measuring tool 1, but polyethylene terephthalate film, polystyrene film and cellulose acetate film are preferable. Further, if the support 3 is made of a light-transmissive material, the optical change in the reagent layer 4 in the capillary chamber 5 can be easily measured, and thus the ease of measurement can be obtained. At the same time, not only can it be surely and easily confirmed that the liquid sample, for example, blood, has been supplied from the sample supply unit 6, but also the time from the supply of blood to the measurement can be fixed and automatically performed. It becomes possible to analyze.

The support 3 which constitutes the liquid sample measuring tool 1 has a diameter of 1 to 1 at a position apart from the bendable portion 2.
It is preferable to form the 10 mm through hole 9. In this case, preferably, the light transmissive film 10 to which the reagent layer 4 is fixed is arranged so as to cover the through hole 9. Such a configuration is particularly effective, for example, when the specific components of blood are glucose, uric acid, and cholesterol, and when an oxidase is used.

Various materials such as a polyethylene terephthalate film and a polystyrene film can be used as a suitable material for the light transmitting film 10, but a porous film is preferable. The reason is
This is because such a porous film is air permeable and can easily supply a sufficient amount of oxygen necessary for the oxidation reaction from the air to the reagent layer.

However, even if the specific components of blood are glucose, uric acid, and cholesterol, for example, when oxidase is not used, for example, when dehydrogenase is used,
Even if the through-hole 9 having a diameter of 1 to 10 mm is formed in the light-transmitting support 3, the light-transmitting film 10 to which the reagent layer 4 is fixed does not need to be disposed so as to cover the through-hole 9.

When a porous film is used as the light transmissive film 10, various porous films can be used. For example, to give some examples, Nuclepore (manufactured by Nuclepore), Celgard (manufactured by Hoechst Celanese), Cyclopore (manufactured by Whatman), Omnipore (manufactured by Millipore), GORE-TEX (manufactured by GORE-TEX), Filter Light (manufactured by MEMTEC) ) And other commercially available porous films.

Further, the optical change in the reagent layer 4 may be the transmittance, and in this case, the support 3 at the position where the reagent layer of the support 3 is arranged needs to be a light transmitting portion. . As the light transmitting portion, in the embodiment shown in FIG. 2, a through hole 9 is formed in the support 3 and the light transmitting film 10 closes the through hole 9. Further, the optical change in the reagent layer 4 may be reflectance. However, in that case, it is necessary to make at least the portion of the support 3 a light-reflecting portion.

As described above, when the optical change in the reagent layer 4 is used as the transmittance or the reflectance, if another light transmitting portion is formed at an appropriate position of the support 3, the optical change in the capillary chamber 5 will be suppressed. The analysis and the supply of the liquid sample can also be detected by an optical change in the capillary chamber 5.

As an example, when the liquid sample is blood, it can be detected that blood is supplied to the capillary chamber 5 at the absorption wavelength of hemoglobin. When it becomes possible to detect the supply of the liquid sample as described above, the detection enables a time start as an auto-start function, and the optical change in the reagent layer 4 is accurately measured after a fixed time, for example, 1 minute. be able to.

In this case, the position where the optical change in the capillary chamber 5 is measured is a position which is close to the reagent layer 4 in distance as shown in FIG. 2, and therefore the light transmitting portion therefor is located on the sample supply side. A relatively small through hole 11 in the support 3 of
Can be formed and the hole 11 is closed with a light-transmitting film 12. The reason for detecting the supply of the liquid sample to the capillary chamber 5 at a position close to the reagent layer 4 in this way is that the start of the reaction in the reagent layer 4 and the detection of the supply of blood are almost simultaneous. It should be.

The reagent layer 4 contains an enzyme capable of measuring a specific component and a dye precursor. For example, when the specific component is glucose, glucose oxidase, peroxidase and 4-aminoantipyrine, N-ethyl-N-
It contains various dye precursors such as (2-hydroxy-3-sulfopropyl) -3,5-dimethylaniline.

Further, the reagent layer 4 can contain insoluble particles that shield light, so that it can be measured from the porous film side even in the analysis of a sample containing a coloring component such as blood. This makes it possible to hide a coloring component such as hemoglobin and obtain an accurate measurement result.

As the insoluble particles, light-reflective white particles such as titanium dioxide, zinc oxide, barium sulfate and magnesium oxide are preferable, but any substance can be used as long as it is a substance which does not dissolve in a liquid sample and shields light. It may be one.

Examples thereof include fluorocarbon, polystyrene latex particles, calcium carbonate, talc, aluminum powder, dextran, acrylic polymer particles, polystyrene latex particles and the like.

A hydrophilic polymer may be added to the reagent solution in order to enhance the coating performance of the reagent solution onto the light-transmitting film 10. Examples of the hydrophilic polymer include hydroxypropyl cellulose, methyl cellulose, sodium alginate, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, modified gelatin, agar and the like.

Further, in order to maintain the shape of the reagent layer 4 and improve the permeability, an emulsion emulsion adhesive or latex particles may be added to these hydrophilic polymers. For example, as the emulsion emulsion adhesive, a vinyl polypropionate copolymer commercially available under the trade name of propiophane (manufactured by BASF) can be used. The coating performance and reactivity can also be improved by adding a surfactant, a plasticizer, a stabilizer, etc. to the reagent solution.

Further, the reagent layer 4 in the liquid sample measuring tool of the present invention may be composed of two layers, a reaction layer containing an enzyme, a dye precursor and the like and a separation layer containing insoluble particles such as titanium dioxide. It is possible.

[0038]

EXAMPLES Next, examples of the liquid sample measuring tool of the present invention will be shown, but the present invention is not limited thereto.

Quantification of glucose in blood <Example of tool for measuring liquid sample> Porous film made of polypropylene as a light-transmitting film, thickness 25 μm
m of Celgard (trademark of Hoechst Celanese film) was coated with a reagent solution having the following composition with a wet thickness of 100 μm and dried at 40 ° C. for 30 minutes. (Reagent solution composition) Glucose oxidase (manufactured by Toyobo) 10 ku Peroxidase (manufactured by Toyobo) 10 ku 4-aminoantipyrine (manufactured by Kishida Chemical Co., Ltd.) 50 mg N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3,5-dimethyl Aniline (Dojindo) 100 mg Propiophan 5D (BASF) 300 mg 0.1 M borate buffer (pH 7.0) 8 ml 0.1 MTES buffer (pH 7.0) 2 ml 50 wt% titanium dioxide aqueous solution 1 g The reagent layer 4 adhered to the flexible film is cut into 7 mm × 7 mm together with the porous film, and the through hole 9 of 4 mm is cut.
And a light-reflective white polyethylene terephthalate film (thickness 188 μm) as the support 3 cut into 10 mm × 30 mm and thermocompression-bonded using a thermoplastic resin. However, the porous film and the support 3 were adhered to each other, and the porous film covered the 4 mm through hole 9. Then, as shown in FIG. 1, a fold line was formed as the bendable portion 2.

As a result, the liquid sample analysis tool 1 was obtained, and the reagent layer 4 was placed inside and bent in a U-shape as shown in FIG. 2 to form a gap (capillary chamber 5) between the facing surfaces of the support 3. Two
It was fixed so as to be 00 μm. <Example of Liquid Sample Measuring Method> About 10 μl of blood samples having the same hematocrit value but different glucose concentrations were supplied from the sample supply unit 6 to the liquid sample measuring tool 1 according to the above-mentioned embodiment, and after 1 minute, The reflectance of the reagent layer 4 at the measurement wavelength λ = 640 nm is measured from below through the through hole 9 using a color difference meter (manufactured by Nippon Denshoku Industries Co., Ltd., Σ-90).
It was measured at point (A).

From this reflectance, according to the simplified form of the Kubelka-Munk equation: K / S = (1-R) ² / 2R K: constant S: scattering coefficient R: reflectance / 100 / S value was determined.

Since the K / S value correlates with the molar concentration, it is possible to convert the concentration by preparing a calibration curve. If a dedicated measuring instrument is used, the reflectance of the capillary chamber at the measurement wavelength λ = 600 nm is measured at point (B), and the reaction time is started. Further, the reflectance of the reagent layer at the measurement wavelength λ = 640 nm can be measured at point (A) through the through hole 9 of the support 3. However, at the point (B), it is necessary to make the support 3 light-transmissive.

[0043]

As described above, according to the liquid sample measuring tool of the present invention, a reagent that reacts with a specific component in a liquid sample to produce a reaction product on a support that can be bent at a bendable portion. By arranging and configuring the reagent layer containing the liquid sample, it is possible to provide a liquid sample measuring tool that measures a specific component in the liquid sample at a low cost and has an extremely simple structure.

Further, according to the liquid sample measuring method of the present invention, a reagent layer containing a reagent that reacts with a specific component in a liquid sample to generate a reaction product is arranged on a support that can be bent at a bendable portion. When measuring a specific component in a liquid sample using the liquid sample measuring tool, the support is bent into a U-shape at the bendable part with the reagent layer inside, and the capillary chamber and its opening are provided between the facing surfaces of the support. By forming the sample supply part with the part to measure the specific component of the liquid sample, the specific component in the liquid sample can be measured quickly and easily.

[Brief description of the drawings]

FIG. 1 is a plan view showing a liquid sample measurement tool according to an embodiment of the present invention.

FIG. 2 is a cross-sectional view showing the liquid sample measurement tool shown in FIG. 1 when it is folded in a ready-to-use state.

FIG. 3 is a perspective view showing the liquid sample measurement tool shown in FIG. 2.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 Liquid sample measuring tool 2 Bendable part 3 Supports 3a, 3b Confronting surface of a support bent in a U shape 4 Reagent layer 5 Capillary chamber 6 Sample supply part 7a, 7b Ends 8a, 8b of support Bending Scheduled line 9 Through hole (for reagent layer) 10 Light transmissive film (for reagent layer) 11 Through hole (for auto start) 12 Light transmissive film (for auto start) A Position for measuring optical change of reagent layer B Position to measure optical changes in the capillary chamber for autostart

Claims (9)

[Claims] 1. A support comprising at least one bendable part, which can be bent at this bendable part, and a support which is disposed on this support and can react with a specific component in a liquid sample to generate a reaction product. A liquid sample measuring tool comprising a reagent layer containing a reagent. 2. When the specific component in the liquid sample reacts with the reagent layer to generate a reaction product, the support is placed at a position other than the bendable portion so as to capture the reaction product by an optical change. The liquid sample measuring tool according to claim 1, wherein a light-transmissive film having a through-hole formed in the through-hole and having the reagent layer fixed thereto is disposed so as to cover the through-hole. 3. The liquid sample measuring tool according to claim 2, wherein the light transmissive film is a porous film. 4. When measuring a specific component in a liquid sample,
The support is bent into a substantially U-shape at the bendable portion so that the reagent layer is located inside, and a capillary chamber and a sample supply portion are formed at the open portion between the facing surfaces of the support. The liquid sample measuring tool according to claim 1, 2 or 3. 5. A support comprising at least one bendable part, which can be bent at this bendable part, and a support which is disposed on this support and can react with a specific component in a liquid sample to generate a reaction product. A method for measuring a specific component in a liquid sample using a liquid sample analysis tool comprising a reagent layer containing a reagent, wherein the support is provided at the bendable portion so that the reagent layer is located inside. Bending into a substantially U shape, forming a sample chamber with the capillary chamber and its opening between the facing surfaces of the support, supplying a liquid sample from the sample supplying unit to the capillary chamber, and causing the capillary chamber to undergo the capillary phenomenon. A liquid sample characterized in that a liquid sample which moves due to contact with the reagent layer reacts to produce a reaction product, and a specific component in the liquid sample is measured by measuring the amount of the reaction product produced. Sample measurement method. 6. A liquid sample measuring method, characterized in that when a specific component in the liquid sample reacts with the reagent layer to generate a reaction product, the reaction product is captured by an optical change. 7. The liquid sample measurement according to claim 6, wherein the optical change in the reagent layer is a transmittance, and therefore a part of the support is formed in a light transmissive part. Method. 8. The liquid sample measuring method according to claim 6, wherein the optical change in the reagent layer is a reflectance, and thus a part of the support is formed in a light-reflecting part. . 9. An optical change in the reagent layer is measured, and at the same time, the supply of blood is detected by an optical change in the capillary chamber, so that the sample adjacent to at least the reagent layer on the support is detected. 9. The liquid sample measuring method according to claim 6, wherein a part of the supply section side is formed as a light transmissive section.