JP4255121B2 - Measuring apparatus and measuring method - Google Patents

Description

  The present invention relates to a measuring apparatus and a measuring method that have a blood dropping section that can separate blood cells and that can measure a specific component in blood quickly, simply, and accurately at a medical site. More specifically, the present invention relates to a measuring apparatus and a measuring method capable of measuring glycated albumin in blood quickly, simply and inexpensively with high accuracy.

2. Description of the Related Art Conventionally, when measuring components in blood, a method of measuring using a large automatic analyzer after making plasma or serum components by centrifugation or the like is often used. However, in facilities such as small and medium-sized hospitals and clinics where the number of specimens is small, it is difficult to install and use automatic analyzers. Measurements are often outsourced to external laboratory facilities.
Methods using test specimens that can be used in clinical practice have also become widespread, but there are problems such as differences in the reading visually and individual control of reactions at room temperature. As a result of confirmation by the present inventors, when the reaction time becomes longer, the reagent or the sample evaporates during the reaction, and when an attempt is made to detect a macromolecule such as a protein using an enzyme reaction, it takes time to color at room temperature. A problem was observed that it took over or hardly colored.
Therefore, there has been a demand for a measuring device that does not require a complicated operation at the clinical site, can quickly obtain a highly reliable value, and is small in size and can be installed anywhere.

  The present invention uses blood as a sample, has a blood cell separable part, and uses a test tool that can measure a specific component in blood accurately, quickly, simply, and inexpensively at a clinical site, reaction control, detection, It is an object of the present invention to provide a small-sized measuring apparatus capable of providing timely measurement results accurately by calculation and a measuring method using the same.

As a method for separating blood cells, there is a method using a blood cell separation membrane other than the centrifugation operation. In a general blood cell separation membrane, plasma does not pass through the membrane unless external force such as pressurization or decompression is applied. According to the study by the present inventors, it has been found that no plasma component can be obtained by simply placing whole blood on the membrane. Obviously, if a suction device is used, it will pass through the membrane, but this will increase the size and complexity of the device, and it will be difficult to make it a device that can be handled easily at the clinical site.
Furthermore, the present inventors can detect a specific component in plasma by a test piece holding an enzyme / reagent, and control not only low molecules in blood but also macromolecules by controlling the enzyme reaction on the test piece by heating and cooling. It was found that the enzyme reaction can be easily performed on the components.
Further, it is found that the specimen and reagent are prevented from evaporating and absorbing moisture by covering the test piece, and the reaction product of the test piece can be easily measured by measuring reflected light, and the present invention is completed based on these findings. It came to.
That is, the present invention has the following configuration.
(1) A measurement device comprising a test piece in which a test piece capable of measuring a specific component in blood and a blood cell separable portion connected by a capillary tube is housed, and a temperature control unit for temperature-controlling a reaction on the test piece, A measuring apparatus comprising: a detection unit that detects a reactant on a test piece; and a calculation unit that calculates a concentration of a specific component based on a detection result.
(2) The measuring apparatus according to (1), wherein the test piece includes an enzyme that reacts with a specific component in blood.
(3) The measuring device according to (1) or (2), wherein the temperature control unit controls the temperature of the reaction on the test piece independently for each test piece.
(4) The measuring device according to any one of (1) to (3), wherein the detecting unit is an independent detecting unit for each test piece.
(5) Any of the above (1) to (4), wherein a transparent cover is disposed on the upper surface of the test piece, and a detection unit for detecting a reactant on the test piece is disposed on the cover side. A measuring device according to the above.
(6) The measuring apparatus according to any one of (1) to (5), wherein the test tool is stored in a shielded space.
(7) The measuring device according to any one of (1) to (6), wherein the detection unit includes a light source and means for detecting reflected light of light emitted from the light source.
(8) The measuring apparatus according to any one of (1) to (7), wherein the calculation unit that calculates the concentration of the specific component in the blood performs the following steps.
1) detecting the reflected light intensity of the reactant 2) associating the detected reflected light intensity with the concentration of the component in the blood 3) calculating the ratio of the component to be obtained from the obtained concentration (9) The measuring apparatus according to any one of (1) to (8), wherein the specific component is glycated albumin.
(10) A method for measuring a specific component in blood including the following steps.
1) Blood is dropped on a test tool in which a test piece capable of measuring a specific component in blood and a blood cell separable part are connected by a capillary tube, and the measurement tool is accommodated in a measuring device and reacted under temperature-controlled conditions. A step of forming an object on a test tool, 2) a step of detecting a reaction product by a detection unit arranged for each test piece, and a step of associating and calculating the detected reaction product with the concentration of a component in blood.

  By using the measuring apparatus and the measuring method of the present invention, plasma can be easily separated simply by dropping whole blood into a dropping part at a medical site, and a specific component in plasma, particularly glycated albumin, can be quickly obtained. It becomes possible to measure easily and inexpensively with high accuracy.

Hereinafter, the configuration and preferred embodiments of the present invention will be described in more detail.
<Construction materials of test tools>
As the blood cell separable part that can be used in the present invention, a blood cell separation membrane can be used. The material of the blood cell separation membrane is not particularly limited, and examples thereof include polymer materials such as cellulose, polyethersulfone, polypropylene, polyethylene, nylon, polysulfone, and polyamide, and glass fibers. For convenience, a commercially available polyethylene membrane or polysulfone membrane can be used. As blood passes through the blood cell separation membrane, blood cells are separated from the blood.

As a test piece that can be used in the present invention, any test piece may be used as long as it is a sheet-like material, for example, a filter paper-like body mainly composed of paper, nonwoven fabric, cellulose derivative, glass fiber, or the like. Can be used. In addition, as a property thereof, any material may be used as long as it has water absorbency and can retain reagents and enzymes. The thickness of the test piece may be about 0.1 mm to 2 mm, and more preferably about 0.2 mm to 1 mm. The area of the test strip, may be any area of about 0.01 cm ² 2 cm ² in order to measure one sample, 0.05 cm ² 1 cm ² is more preferable.

Moreover, as a material for the support, known materials may be used, but specifically,
Films, sheets, and plate-shaped products mainly composed of high molecular plastics such as polystyrene, polyester, polyethylene terephthalate, and cellulose acetate are preferable. Optically, the support is used as a reflective layer during reaction measurement, so that it does not transmit light. Desirable ones such as white polyethylene terephthalate (PET) are desirable.

  The cover material may also be a known material. Specifically, a film or a sheet-like material mainly composed of polyester, cellulose acetate, polycarbonate, polyethylene terephthalate, polystyrene or the like is preferable, and optically for measurement. It is preferable to use a light-transmitting material that has little interference with the irradiation light. For example, a transparent PET film is preferably used.

<Preparation of test piece and reagent used>
In order to retain the reagent and enzymes on the test piece, a liquid reagent having the required composition is prepared, and the test piece is placed in the liquid, for example, at room temperature, preferably refrigerated, preferably for 0.1 minute to 1 day. May be soaked for about 1 minute to 8 hours and dried.
The drying method may be normal pressure or reduced pressure, and the temperature may be, for example, about 4 ° C. to 60 ° C. for about 1 minute to 2 days, but preferably about 4 ° C. to about 30 ° C., where enzymes are not easily deactivated. . More preferably, a test paper can be produced more efficiently and safely by applying cold air at a low temperature of 13 to 17 ° C. and a low humidity of 10% or less.

  As the protease that can be used in the present invention, any protease may be used as long as it is a protease that acts on albumin to excise a glycated amino acid or a peptide containing a glycated amino acid. Further, the enzyme may be a purified product or a non-purified product as long as the desired activity is expressed.

  Preferred examples of proteases that can be used in the present invention include, for example, proteases derived from animals such as trypsin and chymotripsin, proteases derived from plants such as papain and bromelain, and microorganisms. Examples include protease.

The caseinfoline method was used as a method for measuring protease activity. The definition of activity was defined as 1 U for color development corresponding to 1 μg of tyrosine at 37 ° C. for 1 minute.
Further, regarding the use of the protease in the present invention, it is of course possible to use the protease alone, but other endoproteases or other exoproteases may be used simultaneously.

  Examples of enzymes that act on glycated amino acids that can be used in the present invention include dehydrogenases, kinases, oxidases, and the like that act by recognizing the ketoamine structure of glycated amino acids, but oxidases that are available in large quantities at the lowest cost are preferred.

In the case where a reagent containing an enzyme that acts on protease and glycated amino acid is held on a test piece, the concentration of each enzyme in the reagent solution containing these enzymes is preferably 125 U / ml or more and 0.25 mU or more. In order to hold the reagent on the test piece, it is sufficient to immerse a test piece of 5 cm × 5 cm in 1 ml of the reagent having the above concentration. In this case, if all the reagents are absorbed, the protease content per 1 cm ^{2 of the} test piece is 5 U / The amount of enzyme acting on glycated amino acid is 10 mU or more. The protease concentration may be any amount as long as the protease content per 1 cm ² of the test piece is 5 U / ml or more, but is preferably 10 KU or less per 1 cm ^{2 of} the test piece in view of the increase in background and cost.

  The pH of the liquid reagent containing the protease and the enzyme acting on the glycated amino acid may be selected so that the reaction proceeds efficiently in consideration of the optimum pH of the enzyme used.

  In addition, a test piece containing a protease and an enzyme that acts on a glycated amino acid is easy to detect if a coloring reagent that changes the reaction into a color is held at the same time.

  As the protein coloring reagent, any protein coloring reagent may be used as long as it uses a known method for measuring protein. For example, when the protein is albumin, bromocresol green (BCG), bromocresol purple (BCP) ), An albumin-specific dye such as bromophenol blue, or a coloring reagent using an albumin antibody may be used.

<Preparation of test tool>
Examples of the test tool comprising a test piece capable of measuring a specific component in blood of the present invention and a blood cell separable part connected to the test piece by a capillary tube include, for example, a specimen dropping part capable of separating blood cells, Capillary tube and air vent that can suck a certain amount of sample from the dripping part, test piece containing reagents that react with specific components in blood, prevent evaporation of reagents from the test piece, and interfere with optical detection It is preferable to use a test tool that is configured to increase the efficiency of optical measurement by disposing a transparent cover that is not used and an optical reflective layer below the test piece.

  The capillary tube may be formed on the test device by a known method. For example, three films having a thickness of 0.01 mm to 1 mm, a width of 0.5 mm to 10 cm, and a length of 1 mm to 10 cm are used. There is a method of forming a capillary tube by forming a groove having a width of 0.01 mm to 1 cm in a film and sandwiching the film with a film having no groove from above and below. The films can be bonded together by a known technique, and can be easily bonded with a double-sided tape that does not interfere with optical measurement. At this time, a blood dripping hole having a diameter of 0.05 mm to 8 cm and a circular vent hole having a diameter of 0.05 mm to 5 mm are opened in the upper film, and a blood dripping hole is provided between the upper film and the middle film. A structure in which a blood cell separation membrane is disposed at a position and a test piece is disposed in the middle of a capillary groove is desirable.

  An example of a more specific configuration is shown in FIG. 5 is a cover material made of an upper film, 6 is an intermediate plate made of a middle film, and 7 is an optical reflector that also serves as a support made of a lower film. The cover material 5 is provided with one blood dropping hole 12 and the same number of vent holes 3 as the test piece 1. In the middle plate 6, for example, four grooves are formed radially from the position corresponding to the blood dropping hole, corresponding to the number of test pieces. Reference numeral 1 denotes a test piece, which is located on the capillary tube in a path between the blood dropping hole and the vent hole. The capillary tube 4 is formed by an optical reflection plate that also serves as a groove of the intermediate plate 6, the cover material 5, and the support. 8 is a Peltier element, and 9 is a temperature control device using the Peltier element. Reference numeral 10 denotes a detection unit.

  In the test tool configured as described above, an operation until a specific component of blood reacts with an enzyme on a test piece and is detected will be described. When the blood to be measured is dropped onto the blood cell separation membrane 2 from the blood dropping hole 12, the blood passes through the blood cell separation membrane 2 disposed immediately below by the suction force of the capillary, where the blood cells are separated and removed. Only the remaining components are guided to the test piece 1 arranged in the flow direction of the capillary 4. When the blood component reaches the test piece 1, it reacts with the enzyme on the test piece to generate a reaction product. The reaction product is measured by an optical measuring device provided on the top of each test piece.

  The upper film is provided with a vent hole 3 at a position corresponding to the tip of the capillary tube, so that ventilation is performed from the capillary tube prior to the flow of liquid to the test piece. By disposing the vent hole at a predetermined position from the test piece, it is possible to control the amount of liquid distributed to the test piece. That is, if the distance from the test piece to the vent hole is shortened, the distribution amount is small, and conversely, if the distance from the test piece is increased, the distribution amount can be increased.

  It is desirable to provide a cover 5 on the upper surface of the test piece in order to prevent evaporation of reagents from the test piece. Further, it is desirable that the cover is a transparent cover so as not to interfere with the measurement when the reactant on the test piece is optically measured. For example, a transparent PET film is preferably used.

  A combination of the test piece and the specimen will be described. The configuration of FIG. 2 can measure a plurality of components mainly using a plurality of types of test pieces for one sample of blood. For example, a test piece containing an enzyme that reacts with glycated albumin for one blood, a test piece containing a reagent that reacts with albumin, and a test piece containing a glycated amino acid are set in a test tool. It can measure simultaneously and the ratio of glycated albumin in blood can be calculated from a measurement result. Further, as another configuration, for example, a plurality of blood separable portions can be provided, and one component or a plurality of components can be measured for each.

<Detection means>
It is most convenient to detect the reaction by irradiating the test piece colored with the reagent and detecting the reflected light, but other methods may be used. For example, a UV lamp, a halogen lamp, a tungsten lamp, a light emitting diode (LED), a laser, or the like can be used as a light source for irradiation, but it is preferable to use an LED that can be easily incorporated into a small measuring apparatus and generates little heat.
The light irradiation angle may be any, but detection of reflected light is preferably 45 ° or perpendicular to the detection surface. Detection can be easily performed by using a photodiode, a commercially available integrating sphere, or the like.

  When detecting a plurality of test strips, the test strips can be moved and detected by one set of detection units, or the detection units can be prepared by the number of test strips. However, it is desirable to arrange an independent detection unit for each test piece in the case where the optimum wavelength is different or from the viewpoint of control of the measurement position and in the case of a small measurement apparatus such as the present invention.

  Optical detection can be performed from the side of the cover by placing a cover on the top of the test piece, but can also be performed from below the test piece. In this case, a light transmissive filter is also disposed as a support under the test piece, and a light reflecting plate is also disposed as a cover above the test piece. When providing the temperature control apparatus mentioned later, providing in the upper part of a cover is desirable.

  Furthermore, in order to raise the efficiency of optical measurement, the structure which has arrange | positioned the optical reflection layer in the test piece lower part is preferable. As the optical reflection layer, a light-impermeable layer, for example, white PET or the like is desirable.

  The detected reflected light may be converted to the glycated protein concentration by comparing with the sensitivity of the glycated protein having a known concentration. In general, since the sensitivity is constant depending on the lot of the test paper, the sensitivity at a glycated protein concentration whose concentration is known for each lot may be measured in advance so that it can be converted based on these.

<Temperature control means>
In order to perform the measurement using the measuring apparatus of the present invention, a sample is dropped on the appropriate lower part of the blood of the test tool, and the color development of the test piece due to the reaction between the aspirated sample and the reagent is optically measured. Although the test piece is usually dry, the reagent component dissolves due to the moisture of the specimen, and the reaction proceeds automatically.
The temperature of the reaction using the test piece is usually room temperature. However, in the case of an enzyme, particularly an enzyme using a polymer as a substrate, the reaction and reproducibility are improved by carrying out the reaction at a constant temperature with a heat retaining or heating function. .

  For example, a temperature control device can be provided below the filter. Since the reaction on the test piece is mainly an enzyme reaction, the reaction proceeds more efficiently and can be measured with high accuracy by appropriately controlling the temperature. In particular, when a plurality of test pieces are arranged in the test tool and the reaction temperatures of the test pieces are different, it is desirable to perform independent temperature control for each test piece. As such temperature control means, a known heating or cooling device can be used, but it is desirable to use a Peltier element as a small temperature control device with high accuracy.

  In the measurement apparatus of the present invention, the space for storing the test tool is a reaction part and a detection part, so that the influence of diffuse reflection of light from the outside and the influence of changes in temperature or humidity are avoided, and more accurate measurement is performed. It is desirable to be shielded. For example, in a metal box whose inside is painted black, the measuring device can have a display unit and an input unit on the outside, and a detection unit, a reaction unit, and a control unit on the inside. Accordingly, it is preferable that the temperature and humidity around the stored test tool are controlled except when the test tool is taken in and out, and it is shielded from light from the outside.

  The term “small” as used in the present invention means a size that can be easily carried manually. Accordingly, the vertical and horizontal heights are each desirably 20 cm or less, and more specifically, 20 cm in length, 15 cm in width, and 10 cm or less in height are preferable. The weight is 3 kg or less, more preferably 1 kg or less.

  As a sample that can be used for the measurement of the present invention, whole blood, serum, or plasma can be used when the substance to be measured is present in serum, for example, when measuring glycated albumin or the like. For the purpose of measurement, it is preferable to use whole blood.

<Measurement device>
FIG. 1 is a block diagram showing a schematic configuration of a measuring apparatus according to the present invention.
In FIG. 1, reference numeral 180 denotes a test tool, which is housed in the detection unit 160 of the measurement apparatus and performs measurement. Reference numeral 18 denotes a blood cell separable portion, which is connected to the test piece 20 by the capillary 15. The test piece 20 contains a reagent and an enzyme and is impregnated with a reagent necessary for reacting with a specific component in blood to produce a reaction product. For example, when measuring glycated albumin, it is impregnated with protease, ketoamine oxidase, peroxidase, 4-aminoantipyrine, N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-toluidine sodium (TOOS), etc. Has been.

The blood is dropped on the blood cell separable portion 18, moves through the capillary 15, and reaches the test piece 20. Here, it reacts with the reagent to produce a reaction product. And the reflected light of this product is measured from the cover 25 side of the test piece upper part. Reference numeral 30 denotes a light emitting element, which irradiates predetermined light with a reaction product. For example, in the case of glycated albumin, light having a wavelength of about 570 nm is irradiated. A light receiving element 40 detects the reflected light intensity of light irradiated from the light emitting element 30 and reflected by the test piece 20. The change of the reaction product in the test piece 20 can be detected by this reflected light intensity.
Reference numeral 50 denotes an A / D converter that converts a detection signal (analog signal) output from the light receiving element 40 according to the reflected light intensity into a digital signal. Reference numeral 170 denotes a control unit that controls the operation of the entire apparatus. The control unit 170 includes, for example, a microprocessor, and is detected by a program unit 100 that stores a control program and the like, a data storage unit 60 that stores measurement data and the like, and a temperature sensor 150. A temperature control unit is provided that gives instructions to the heating / cooling unit according to temperature. Reference numeral 110 denotes a clock unit necessary for the control program to issue an instruction for timing of measurement and data storage.
Reference numeral 130 denotes an input unit, which includes, for example, a key for instructing power on / off, a key for instructing output of measurement data, and the like. Reference numeral 90 denotes a display unit such as a liquid crystal display, and 140 denotes a power supply unit for supplying power to the entire apparatus.
Next, although the Example of this invention is described in detail, this invention is not limited at all by this.

<Preparation of glycated albumin measuring reagent test piece>
The glycated albumin measurement reagent test piece was immersed in a 0.4 mm thick, 50 mm × 50 mm filter paper (Whatman's chromatographic filter paper) at room temperature for 5 minutes in a reagent with the enzyme concentration changed in the following composition, and then Air-dried at 37 ° C. for 20 minutes to prepare a glycated albumin measuring reagent test piece. After drying, it was treated at room temperature for 2 hours, sealed in a light-shielding manner with a desiccant and stored in a refrigerator until use.
50 mM Tris buffer, pH 7.5
2000 U / ml protease type XXVII (manufactured by Sigma)
4 mM 4-aminoantipyrine (4-AA; manufactured by Dojindo Laboratories)
2 mM N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-toluidine
(TOOS: Doujin Chemical Laboratory Co., Ltd.)
4000 mU / ml ketoamine oxidase (Asahi Kasei)
4U / ml peroxidase (Roche)

<Preparation of glycated amino acid measuring reagent test piece>
The glycated amino acid measurement reagent test piece was prepared and stored in the same manner as the glycated albumin detection reagent test piece using the reagent having the following composition.
50 mM Tris buffer, pH 7.5
4 mM 4-aminoantipyrine (4-AA; manufactured by Dojindo Laboratories)
2 mM N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-toluidine
(TOOS: Doujin Chemical Laboratory Co., Ltd.)
4000 mU / ml ketoamine oxidase (Asahi Kasei)
4U / ml peroxidase (Roche)

<Preparation of test specimen for albumin measurement reagent>
The test specimen for albumin measurement reagent was prepared and stored in the same manner as the preparation of the test specimen for glycated albumin measurement reagent, using reagents having the following composition.
50 mM citrate buffer, pH 4.0
1% Brij 35 (Wako Pure Chemical Industries)
0.03% Bromocresol Green (Wako Pure Chemical Industries, Ltd.)

<Preparation of test tool>
The configuration of the test tool will be described with reference to FIG. Four test pieces 1 prepared in Example 1 were cut out to have a diameter of 4 mm. For the blood cell separation membrane 2, a filter made of polysulfone was used, and one piece having a diameter of 10 mm was cut out. The test tool is configured in a state where three films are stacked.
The upper film is the cover material 5, the middle film is the middle plate 6, and the lower film is the optical reflector 7 that also serves as a support. The cover material 5 is a transparent PET film (30 mm × 40 mm) having a thickness of 0.2 mm, provided with one blood dropping hole 12 having a diameter of 8 mm and the same number of ventilation holes 3 having a diameter of 1 mm as the test piece 1. . An intermediate plate 6 is disposed under the cover material. The intermediate plate is a PET film (30 mm × 40 mm) having a thickness of 0.4 mm, and a groove having a width of 1 mm corresponding to the number of test pieces is formed in the blood dropping hole. Four are formed radially from the corresponding positions. Further, a circular holding portion (not shown) is provided at a position corresponding to the test piece and the blood cell separation membrane. The position of the test piece was on the capillary tube and arranged so as to be a path between the blood dropping hole and the vent hole.
Further, a support body and an optical reflection plate 7 are disposed below the intermediate plate 6. This is made of a white PET film (30 mm × 40 mm) having a thickness of 0.6 mm, and the capillary 4 is formed in such a manner that an intermediate plate is sandwiched together with the cover material 5 of the transparent film. These three films are bonded with a double-sided tape.
In addition, a temperature control device 9 including a Peltier element 8 is provided below the test tool, and each test piece can be controlled to a desired temperature.

<Measurement of glycated albumin>
The glycated albumin ratio was measured using whole blood samples of 5 healthy subjects and 5 diabetic patients. The glycated albumin concentration, glycated amino acid concentration, and albumin concentration were converted by separately measuring a calibrator for Lucika GA (manufactured by Asahi Kasei Co., Ltd.). A digital fiber sensor 11 (manufactured by Keyence Corporation) using a light emitting diode was used as the detection unit 10, and the reflected light after the sample was dropped was measured for 10 minutes and recorded.
For the specimen, the glycated albumin concentration, the glycated amino acid concentration, and the albumin concentration were determined, and the ratio of glycated albumin was calculated from these values. The results are shown in the left column of Table 1.
From this, it can be seen that unnecessary substances such as blood cells in whole blood are removed by the blood cell separation membrane, so that glycated albumin can be measured directly from a whole blood sample using the test tool of the present invention.
Furthermore, after the same whole blood sample was centrifuged, the results (right column in Table 1) measured with an automatic analyzer using Lucika GA reagent (manufactured by Asahi Kasei) were in good agreement with the measurement apparatus of the present invention. It is clear that an accurate measurement is possible.

<Effect of transparent film cover on reagent evaporation>
Two types of test tools, a normal type in which the upper surface of the test piece was covered and an open type in which a hole with a diameter of 3 mm was formed in the upper surface of the test piece, were prepared by the procedure of Example 2 without using a blood cell separation membrane.
As a sample, Lucica GA calibrator H (Asahi Kasei Co., Ltd.) was used, and 8 μL was directly sucked into a capillary tube at room temperature in an analytical balance and absorbed by a test piece. The moment of absorption was defined as 0 seconds, and thereafter, the weight was measured every 30 seconds until 600 seconds, and the remaining weight% from the weight at 0 seconds was calculated. The results are shown in FIG. As is clear from this, the test piece without the cover evaporates as much as 40% in 10 minutes, whereas the test piece with the cover evaporates only 10%.
Therefore, it was clarified that by covering the upper surface of the test piece, the evaporation of the reagent during measurement can be reduced by 30%, and more accurate measurement can be performed.

<Influence of measurement temperature>
A calibrator for Lucika GA (manufactured by Asahi Kasei Co., Ltd.) was used as a specimen, reaction temperatures were set to 30 ° C., 40 ° C., 45 ° C., and 50 ° C., and changes in reflectance over time were measured. The results are shown in FIG. As is clear from this, it has been clarified that the reaction rate on the test piece is accelerated as the temperature increases from 30 ° C. to 45 ° C. during measurement, and stable measurement by temperature control is possible.

  The present invention can measure a blood sample quickly, conveniently, and accurately at a clinical site, and can be suitably used in the field of clinical diagnosis.

It is a block diagram which shows schematic structure of the measuring apparatus of this invention. It is a perspective view which shows a part of structure of the measuring apparatus of this invention. It is a graph which shows the influence of the cover in the measurement of the glycoalbumin of this invention based on Example 4 of this invention. It is a graph which shows the influence of the measurement temperature in the measurement of the glycoalbumin of this invention based on Example 5 of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Test piece 2 Blood cell separation membrane 3 Ventilation hole 4 Capillary tube 5 Cover material 6 Middle plate 7 Optical reflecting plate which served also as support 8 Peltier element 9 Temperature control part 10 Detection part 11 Sensor 12 Blood drop hole 15 Capillary 18 Blood cell separation part DESCRIPTION OF SYMBOLS 20 Test piece 30 Light emitting element 40 Light receiving element 50 A / D conversion part 60 Data storage part 70 Calculation part 80 Display control part 90 Display part 100 Program 110 Clock part 120 Temperature control part 130 Input part 140 Power supply part 150 Heating / cooling part Temperature sensor 160 Detection Unit 170 Control Unit 180 Test Tool

Claims (12)

A glycated albumin measuring apparatus comprising a test piece capable of measuring glycated albumin in blood and a test tool of (1) below, a temperature control unit for controlling the temperature of the reaction on the test piece, and a reaction product on the test piece. An apparatus for measuring glycated albumin, comprising: a detecting section for detecting; and a calculating section for calculating a concentration of glycated albumin based on a detection result.
(1) A test tool including a blood cell separable portion connected to a test piece by a capillary and a vent hole located at the tip of the capillary. The measuring device according to claim 1, wherein the test piece includes an enzyme that reacts with glycated albumin in blood. The measuring apparatus according to claim 1, wherein the temperature control unit controls the temperature of the reaction on the test piece independently for each test piece. The measuring device according to claim 1, wherein the detecting unit is an independent detecting unit for each of the test pieces. The measuring apparatus according to claim 1, wherein a transparent cover is disposed on an upper surface of the test piece, and a detection unit that detects a reactant on the test piece is disposed on the cover side. . The measuring apparatus according to claim 1, wherein the test tool is stored in a shielded space. The measuring device according to claim 1, wherein the detection unit includes a light source and means for detecting reflected light of light emitted from the light source. The test piece is a test piece containing an enzyme that acts on protease and glycated amino acid, and the test piece is 1 cm. ² The glycated albumin measuring apparatus according to any one of claims 1 to 7, which is a test piece having a protease content and an enzyme content acting on a glycated amino acid of 5 U / ml or more and 10 U / ml or more, respectively. The measuring apparatus according to any one of claims 1 to 8, wherein a calculation unit that calculates the concentration of glycated albumin in blood performs the following steps.
(1) A step of detecting the reflected light intensity of the reactant (2) A step of associating the detected reflected light intensity with a concentration of a component in blood (3) A step of calculating a ratio of a component to be obtained from the obtained concentration A method for measuring glycated albumin in blood, comprising the following steps.
(1) Blood is dropped on a test piece capable of measuring glycated albumin in blood and the test tool shown in (A) below , the measurement tool is stored in the measuring device, and the reaction product is tested under temperature-controlled conditions. (2) A step of detecting the reaction product by a detection unit arranged on each test piece (3) A step of associating the detected reaction product with the concentration of a component in blood and calculating
(A) A glycated albumin test tool comprising a blood cell separable part connected to a test piece by a capillary, and a vent located at the tip of the capillary The test piece is a test piece containing an enzyme that acts on protease and glycated amino acid, and the test piece is 1 cm. ² The method for measuring glycated albumin according to claim 10, which is a test piece having a protease content and an enzyme content acting on glycated amino acids of 5 U / ml or more and 10 U / ml or more, respectively. A glycated albumin ratio measuring apparatus containing the test tool of (2) below, in which the test piece of (1) below and a blood cell separable part are connected by a capillary tube, and a temperature control unit for controlling the temperature of the reaction on the test piece An apparatus for measuring a glycated albumin ratio, comprising: a detection unit for detecting a reactant on a test piece; and a calculation unit for calculating a concentration of a specific component based on a detection result. ;
(1) A test piece containing a protease and an enzyme that acts on a glycated amino acid, the test piece 1 cm ² Test pieces having a protease content and an enzyme content acting on glycated amino acids of 5 U / ml or more and 10 U / ml or more,
(2) A test tool comprising the test piece of (1), a blood cell separable portion connected to the test piece by a capillary, and a vent hole located at the tip of the capillary.

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