JP6652974B2 - Test fixture for transmitted light measurement and component measuring device set - Google Patents

Description

  The present invention relates to a test device and a component measuring device set, and more particularly to a test device and a component measuring device set capable of suppressing deterioration over time while suppressing a decrease in reaction rate.

  2. Description of the Related Art Conventionally, a component measuring device for measuring a predetermined component in a body fluid such as blood or urine has been widely used (for example, see Patent Documents 1 and 2). As a component measuring device of this kind, a colorimetric component measurement is performed by injecting a body fluid such as blood into a body fluid measuring chip having a coloring reagent and optically measuring the degree of coloration when the coloring reagent reacts with the body fluid. An apparatus is known (for example, see Patent Document 3).

  When a component in a body fluid such as blood or urine is measured using such a component measuring device, as shown in FIG. 17, a color-forming reagent layer includes a lower layer P made of an enzyme (for example, peroxidase) and a dye ( For example, there is a technique in which an upper layer Q made of a redox-based coloring reagent) is different from each other (for example, see Patent Document 4).

WO 2014/049704 Japanese Patent No. 4999007 Japanese Patent Publication No. Hei 10-505676 Japanese Patent No. 54406633

  However, when the enzyme and the dye are supported on different layers, it takes time for body fluids such as blood and urine to dissolve the dye in the upper layer Q and for the solution containing the dye to dissolve and react with the enzyme in the lower layer P. However, there is a problem that the reaction speed becomes slow.

  On the other hand, when the reagents in the color forming reagent are uniformly applied in a mixed state (in the case of a uniform structure), particularly in a wet state, before the body fluid such as blood or urine is injected. However, there is a problem that the color develops and deteriorates with time. The present inventors have conducted intensive studies on such a problem of aging, and have found that the cause is that the dye in the coloring reagent reacts with another reagent (mediator or enzyme) in the coloring reagent. Was.

  Therefore, an object of the present invention is to provide a test tool and a component measuring device set that can suppress deterioration over time while suppressing a decrease in reaction rate.

A test device according to a first aspect of the present invention is a test device used for measuring a predetermined component in a body fluid, wherein the test device reacts with a predetermined component in the body fluid or reacts with a predetermined component in the body fluid. Has a reagent layer structure including a first reagent that promotes the reaction and a second reagent that reacts with the reaction of the first reagent, wherein the reagent layer structure is such that the first reagent and the second reagent are It is characterized by having a reagent separation layer arranged separately from each other in the layer.
As a result, the diffusion distance can be shortened while separating the first reagent and the second reagent from each other. Thus, compared to the case of a uniform structure having the same thickness, a reduction in the reaction rate can be suppressed, Deterioration can be suppressed.

  As one embodiment of the present invention, the reagent separation layer includes a granular first granular portion including the first reagent, and a granular second granular portion including the second reagent, and the first granular portion includes: It is preferable that the second granular portion is disposed separately from the second granular portion.

  As one embodiment of the present invention, it is preferable that the reagent layer structure is a multilayer structure in which the reagent separation layers are stacked.

  As one embodiment of the present invention, the first granular portion in the first reagent separation layer of the multilayer structure is adjacent to the second granular portion in the second reagent separation layer formed on the first reagent separation layer. Preferably, the second granular portion in the first reagent separation layer is adjacent to the first granular portion in the second reagent separation layer.

  As one embodiment of the present invention, the first granular portion is applied so as to be adjacent only to the second granular portion, and the second granular portion is applied so as to be adjacent only to the first granular portion. Is preferred.

  As one embodiment of the present invention, it is preferable that the reagent layer structure further includes a single-grain layer composed of only one of the first granular portion and the second granular portion.

  As one embodiment of the present invention, the one-grain layer is such that the one granular portion is adjacent to the other granular portion of the first granular portion and the second granular portion in the reagent separation layer. It is preferable to be laminated on the reagent separation layer.

  As one embodiment of the present invention, it is preferable that the one granular portion is composed of only the first reagent or the second reagent.

  As one embodiment of the present invention, it is preferable that the first reagent contains at least one of an enzyme and a mediator, and the second reagent is a dye.

  As one embodiment of the present invention, the test device includes a supply port to which a bodily fluid is supplied, and a flow channel having the supply port formed at one end, and a component measuring device for measuring a predetermined component in the bodily fluid. It is preferable that the reagent layer structure is provided in the flow channel.

  A component measuring device set according to a second aspect of the present invention includes a body fluid measuring chip as a test device according to the first aspect of the present invention, and the body fluid measuring chip is mounted thereon, and measures a predetermined component in the body fluid. And a component measuring device.

  Advantageous Effects of Invention According to the present invention, it is possible to provide a test tool and a component measuring device set that can suppress deterioration over time while suppressing a decrease in reaction rate.

It is a top view showing the body fluid measuring chip concerning one embodiment of the present invention. FIG. 2 is a sectional view taken along line II in FIG. 1. FIG. 2 is a plan view showing an example of a reagent separation layer constituting the reagent layer structure in FIG. 1. FIG. 4 is a plan view illustrating a modification of the reagent separation layer in FIG. 3. FIG. 2 is a perspective view illustrating an example (a three-dimensional mosaic structure) of the reagent layer structure in FIG. 1. It is a top view showing the blood glucose meter set concerning one embodiment of the present invention. It is a longitudinal cross-sectional view which shows the blood glucose meter and the blood glucose measurement chip of the blood glucose meter set shown in FIG. 5 is a graph showing a reaction solution spectrum of Experimental Example 1, wherein the vertical axis represents absorbance and the horizontal axis represents wavelength (nm). 5 is a graph showing the spectrum of the reaction solution of Experimental Example 2, wherein the vertical axis represents absorbance and the horizontal axis represents wavelength (nm). It is a graph which shows the reaction solution spectrum of Experimental example 3, and a vertical axis | shaft shows an absorbance and a horizontal axis shows a wavelength (nm). 5 is a graph showing the spectrum of the reaction solution of Experimental Example 4, wherein the vertical axis represents absorbance and the horizontal axis represents wavelength (nm). FIG. 3 is a plan view (electron micrograph) showing a reagent layer structure of Comparative Example 1. FIG. 2 is a plan view (electron micrograph) showing a reagent layer structure of Example 1. It is a graph which shows the reaction rate (color-forming rate) of each reagent layer structure (Comparative example 1, Comparative example 2, Example 1), a vertical axis | shaft is absorbance and a horizontal axis is time after spotting (ms: millisecond). Is shown. It is a graph which shows the change of the amount of increase in absorbance per 0.6 second of each reagent layer structure (Comparative example 1, Comparative example 2, Example 1), a vertical axis | shaft shows a change in absorbance, and a horizontal axis shows time after spotting ( (ms: milliseconds). It is a graph which shows the change of the amount of increase in absorbance of each reagent layer structure (Comparative Example 1, Example 1) after irradiation with a fluorescent lamp for 3 hours. It is a graph which shows the change of the light absorbency increase amount after the heat acceleration test (60 degreeC, 3 days) of each reagent layer structure (Comparative Example 1, Example 1). FIG. 9 is a perspective view showing a two-layer structure of Comparative Example 2. FIG. 3 is a diagram for explaining a pitch P of droplets. FIG. 9 is a diagram for explaining a reagent layer structure of Comparative Example 3. It is a figure for explaining the reagent layer structure of Examples 2-4. It is a graph which shows the light absorbency (coloring amount) after 25 seconds of Comparative Example 3 and Examples 2-4, and a vertical axis | shaft shows light absorbency.

(Test tool)
Hereinafter, an embodiment of a test device according to the present invention will be described with reference to FIGS. In the drawings, common members are denoted by the same reference numerals.

  The test device of the present invention is not particularly limited as long as it is used to measure a predetermined component in a body fluid, and can be appropriately selected depending on the purpose.For example, a body fluid measurement chip, a test paper, and the like Is mentioned.

<< Body fluid measurement chip >>
Hereinafter, a body fluid measurement chip as a specific example of the test device will be described in detail.
FIG. 1 is a plan view showing a body fluid measurement chip according to one embodiment of the present invention. FIG. 2 is a sectional view taken along line II in FIG.

As shown in FIGS. 1 and 2, the body fluid measurement chip 100 according to the present embodiment includes a supply port 10, a flow path 20, and a reagent layer structure 30, and can be attached to a component measurement device described later.
Hereinafter, details of each member of the body fluid measurement chip 100 and the characteristic portion configured by each member in the present embodiment will be described.

<< supply port 10, flow path 20, and reagent layer structure 30 >>
As shown in FIGS. 1 and 2, the body fluid measurement chip 100 includes a first base 1 forming a bottom surface, a second base 2 forming a top surface, and a first base 1 and a second base 2. Adhesive portions 3 and 4 are provided between the base materials 2 and provided at both ends in the width direction orthogonal to the chip thickness direction.
As described above, the first base material 1 and the second base material 2 are sandwiched between the first base material 1 and the second base material 2 with the spacers (not shown) between the first base material 1 and the second base material 2. By bonding the second substrate 2, a gap of a predetermined size is formed between the first substrate 1 and the second substrate 2. The gap having the predetermined size serves as a flow path 20 having the supply port 10 formed at one end, so that the bodily fluid can flow into the bodily fluid measurement chip 100. Further, a reagent layer structure 30 is provided in the channel 20. In FIGS. 1 and 2, the reagent layer structure 30 is formed on the first base material 1, but is not limited thereto, and is provided in the flow path 20 without closing the flow path 20. It should just be done.

  The material of the first substrate 1 is not particularly limited and can be appropriately selected depending on the purpose (irradiation / reception of light). For example, polyethylene terephthalate (PET), polymethyl methacrylate, polystyrene, cyclic polyolefin And transparent organic resin materials such as cyclic olefin copolymers and polycarbonates; and transparent inorganic materials such as glass and quartz.

  The material of the second base material 2 is not particularly limited and can be appropriately selected depending on the purpose (irradiation and reception of light). For example, a transparent organic material such as a hydrophilic-treated polyester film (3M hydrophilized film) can be used. And a resin material.

  The thickness of the bonding portions 3 and 4 is appropriately adjusted so that the distance of the flow path 20 in the chip thickness direction becomes a desired value. For example, a spacer having an arbitrary thickness is arranged between the first base material and the second base material and then bonded or fused, or as a bonding member for bonding the first base material and the second base material. A double-sided tape having a function may be used.

-Reagent layer structure 30-
The reagent layer structure 30 includes at least a first reagent that reacts with a predetermined component in a body fluid or promotes a reaction with a predetermined component in a body fluid, and a second reagent that reacts with the reaction of the first reagent. And, if necessary, other components. As a combination of the first reagent and the second reagent, for example, a “combination having different pHs and losing the activity when the reagents having the different pHs are mixed” can be considered.

The layer structure of the reagent layer structure 30 is not particularly limited as long as it has a reagent separation layer, and is appropriately selected depending on the purpose.
The reagent layer structure 30 may have a single-layer structure consisting of only a reagent separation layer, or may have a multilayer structure having at least one reagent separation layer. A multilayer structure of two or more layers (that is, a multilayer structure in which all layers are reagent separation layers) is preferable.

--- Reagent separation layer ---
In the present specification, the “reagent separation layer” means “a layer in which a first reagent and a second reagent are arranged separately from each other in a layer”. For example, as shown in FIG. 3 and FIG. 4, the reagent separation layers 30a and 30b include a portion A containing the first reagent (but not containing the second reagent) and a portion A containing the second reagent (where the first reagent is used). And B) (not included) are disposed separately from each other.

  Here, the part A (see FIGS. 3 and 4) containing the first reagent (but not containing the second reagent) is composed of a plurality of first granular parts and contains the second reagent (however, the first reagent ) (See FIGS. 3 and 4) may be composed of a plurality of second granular portions.

  The average particle size of the first granular portion and the second granular portion is not particularly limited, and can be appropriately selected depending on the purpose.

Further, in the embodiment shown in FIG. 5, the reagent separation layer 30 includes a granular first granular portion X including the first reagent (but not including the second reagent) and a second reagent (the first granular portion X includes the first reagent). It is preferable that a second granular portion Y (containing no reagent) is provided, and the first granular portion X and the second granular portion Y are arranged separately.
Here, as shown in FIG. 5, the reagent layer structure 30 is such that the first granular portion X1 in any reagent separation layer (first reagent separation layer) of the reagent layer structure 30 is located on the first reagent separation layer. It is more preferable that the second granular portion Y2 in the second reagent separation layer formed adjacent to the second granular portion Y2 in the first reagent separation layer be adjacent to the first granular portion X2 in the second reagent separated layer. .
In FIG. 5, the first granular portion X is applied so as to be adjacent only to the second granular portion Y, and the second granular portion Y is applied so as to be adjacent only to the first granular portion X. That is, the first granular portion X is entirely covered with the second granular portion Y, and the periphery of the second granular portion Y is entirely covered with the first granular portion X. Here, the three-dimensional structure in FIG. 5 is similar to the crystal structure of sodium chloride (sodium chloride type structure).
Thus, when a sample (body fluid) is injected into such a three-dimensional structure, the sample three-dimensionally goes around the gaps around the first granular part and the second granular part, and diffuses uniformly while dissolving the reagent. Therefore, a reduction in the reaction rate can be suppressed as compared with the case where the same thickness is used for a uniform structure.

--- First reagent ---
The first reagent is not particularly limited and may be appropriately selected depending on the purpose as long as it reacts with a predetermined component in a body fluid or promotes a reaction with a predetermined component in a body fluid. Examples of the first reagent include enzymes and mediators. , Hydroperoxide, and the like. These may be used alone or in combination of two or more.

--- Enzyme ---
The enzyme plays a role of, for example, reacting with a predetermined component (glucose) in a body fluid (blood, urine) to extract electrons from glucose.
The enzyme is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include glucose dehydrogenase (GDH), glucose oxidase (GOD), peroxidase (POD), cholesterol oxidase, cholesterol esterase, lipoprotein lipase, and the like. Glycerol oxidase, phospholipase D, choline oxidase, and the like. These may be used alone or in combination of two or more.

−−− Mediator −−−
The mediator promotes, for example, (i) a reaction between a first reagent (enzyme) and a predetermined component (glucose) in a body fluid (blood, urine), and (ii) once receives an electron extracted from glucose by the enzyme. It plays a role of handing over to pigments.
The mediator is not particularly limited and can be appropriately selected depending on the purpose. For example, 5-methylphenazinium methyl sulfate (PMS), 1-methoxy-5-methylphenazinium methyl sulfate ( mPMS), NAD, FAD, PQQ, potassium ferricyanide, and the like.
These may be used alone or in combination of two or more.

--- Second reagent--
The second reagent is not particularly limited as long as it reacts with the reaction of the first reagent, and can be appropriately selected depending on the purpose. For example, the above-described mediator, dye (color former), and fluorescent substance , Transition metal salts, sodium nitrite, and the like.

−−− Mediator −−−
The mediator plays a role of, for example, once receiving an electron generated by a reaction between an enzyme and glucose and transferring it to a dye (reacting with the reaction of the first reagent).

--- Dye--
The dye (coloring agent) plays a role of, for example, receiving an electron or hydrogen peroxide generated by the reaction between the enzyme and glucose to form a color (reacting with the reaction of the first reagent).
The dye is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include tetrazolium salts (eg, WST-4, WST-1, WST-5, MTS, MTT, tetrazolium salt (A)) , O-tolidine, m-tolidine, benzidine, tetramethylbenzidine, O-methylbenzidine, O-dianisidine, 2,7-diaminofluorene, and those obtained by oxidatively coupling a coupler with a Trinder reagent. These may be used alone or in combination of two or more.
The chemical formula of the tetrazolium salt (A) is shown below.

Where X = Na.

--- Sodium nitrite ---
For example, in blood glucose measurement, it is originally desired to observe only color development based on the reaction of glucose. However, when blood lyses, hemoglobin reacts with a dye and develops color (pseudo color development). Here, since the sodium nitrite has a property of converting hemoglobin into another substance which does not react with a dye called methemoglobin, by including the sodium nitrite in the second reagent, it is possible to convert hemoglobin into methemoglobin. , Pseudo color development can be prevented.
Since sodium nitrite has the property of deactivating an enzyme, no reaction occurs when combined with the enzyme. Therefore, it is necessary to include the second reagent separated from the first reagent containing the enzyme.

--- Transition metal salt ---
When the dye is a tetrazolium salt, a transition metal salt (transition metal ion) and a tetrazolium salt may be subjected to a chelate reaction to form a chelate complex to form a color. The transition metal salt is not particularly limited as long as it generates ions in an aqueous liquid (eg, water, buffer, blood, body fluid), and can be appropriately selected depending on the purpose. Cobalt chloride, bromide, sulfate and organic acid salts are preferred.

-Manufacturing method of reagent layer structure 30-
The method for producing the reagent layer structure includes at least a coating step and a drying step, and includes other steps as necessary.

−−Coating process−−
The application step is a step of separately applying a first coating liquid containing a first reagent and a second coating liquid containing a second reagent.
The coating method is not particularly limited and may be appropriately selected depending on the purpose. Examples thereof include an inkjet method, a screen printing method, and a gravure printing method.
These may be used alone or in combination of two or more.

−−Drying process−−
The drying step is a step of drying the applied coating liquid.
The drying temperature and drying time in the drying step are not particularly limited, and can be appropriately selected depending on the purpose.

When the reagent layer structure 30 has a three-dimensional structure (FIG. 5), for example, it can be manufactured as follows.
First, two types of solutions (for example, a “dye solution (first solution)” and a “solution of a mixture of a mediator and an enzyme (second solution)”) are prepared. Next, two types of droplets having substantially the same particle size (the first droplet derived from the first solution, the first droplet derived from the first solution, the first solution and the second solution) are printed by a printing technique such as an inkjet method, a screen printing method, or a gravure printing method. A second droplet derived from the second solution) is prepared, and the first droplet and the second droplet are dropped one by one on the substrate. At this time, the first droplet is applied in one row, the second droplet is applied in one row with a shift of half the particle size in the droplet arrangement direction, and the first droplet is shifted in half the particle size in the droplet arrangement direction. The operation of applying one line is repeated to apply and form the first reagent separation layer (see FIG. 5).
Next, after drying the formed first reagent separation layer, a second reagent separation layer is applied and formed on the first reagent separation layer in the same manner.
Further, the formed second reagent separation layer is dried, and after drying, a third reagent separation layer is similarly formed on the second reagent separation layer to form a three-dimensional three-dimensional structure (FIG. 5). Make it. When forming the upper layer (for example, the second reagent separation layer), the upper layer (for example, the first reagent separation layer) is stacked by being shifted by half the particle size in the vertical and horizontal directions with respect to the droplet arrangement direction. By repeating the above, it is possible to suppress a decrease in the reaction rate and deterioration over time.
Here, by using a printing technique, the fluidity of the droplet can be controlled.
The “solution” in this specification includes not only a case where a solute is completely dissolved in a solvent but also a case where a part or all of a solute is dispersed without being dissolved. In addition, the solution usually has a high viscosity and may be in a gel state.

Although the above-described three-dimensional structure (FIG. 5) is manufactured using two types of solutions (droplets), the present invention is not limited to this. Solution (first solution) "," mediator solution (second solution) ", and" enzyme solution (third solution) ". The pattern using three types of solutions (droplets) is as follows: (i) a first pattern in which each layer separates and arranges three types of solutions (droplets) according to a certain rule, and Each layer is separated by two kinds of solutions (droplets) out of three kinds of solutions (droplets), arranged and arranged in a second pattern, (iii) three kinds of solutions (droplets) are fixed A third pattern in which a layer (three types of droplet layers) that is separated and arranged according to the following rule and a layer (two types of droplet layers) that separates and sorts two types of solutions (droplets) It is preferable that three types of droplets (particles) are applied so as not to contact the same type of droplets (particles).
Specific examples of the (ii) second pattern include, for example, a first reagent separation layer prepared using a first solution (droplets) and a second solution (droplets); A second reagent separation layer formed on the separation layer and formed using the second solution (droplets) and the third solution (droplets); and a third solution ( And a third reagent separation layer formed using the first solution (droplet) and the first solution (droplet). When forming the upper layer (for example, the third reagent separation layer), the upper layer (for example, the second reagent separation layer) is stacked by being shifted by half the particle size in the vertical and horizontal directions with respect to the droplet arrangement direction. By repeating the above, it is possible to further suppress the reduction of the reaction rate and the deterioration over time.

Hereinafter, the pitch (interval) of the droplet when the droplet lands on the base material in the coating step will be described in detail.
The pitch P of the droplets (FIG. 18) is not particularly limited as long as the first reagent and the second reagent are arranged separately from each other in the reagent separation layer of the resultant test device. Although it can be appropriately selected according to the purpose, (r1 + r2) / 2 to (r1 + r2) × 2 is preferable, and about r1 + r2 is more preferable. Here, r1 indicates the radius of the sphere assuming that the first droplet (reference numeral 200 in FIG. 18) including the first reagent is spherical, and r2 indicates the second droplet (including the second reagent). The reference numeral 300 in FIG. 18 indicates the radius of the sphere when it is assumed to be spherical.
If the pitch of the droplets is less than (r1 + r2) / 2, the first droplet and the second droplet are excessively overlapped with each other, and the reaction does not occur because the enzyme is deactivated. If the pitch of the droplets exceeds (r1 + r2) × 2, the gap (dead space) between the first droplet and the second droplet is too large, and the time required for the reaction is long. Sometimes it becomes.
When the application method is an ink jet method, the droplets are usually spherical, and the droplets after application are circular, but when the application method is, for example, a screen printing method, the droplets after application are It can be rectangular. As described above, when the droplet after application is rectangular, a length 0.5 times the length of one side of the rectangle can be used instead of the radii r1 and r2 of the circle.

<Test paper>
Hereinafter, a test paper as a specific example of the test device will be described in detail.
The test paper includes a base material and a reagent layer structure (same configuration as the above-described reagent layer structure 30) formed on the base material.

<<< base material >>>
The material of the base material is not particularly limited and may be appropriately selected depending on the purpose as long as the material has a certain degree of rigidity. For example, polyether sulfone, cyclic polyolefin, cyclic polyolefin copolymer, polystyrene, polyethylene terephthalate And plastics such as polyvinyl chloride, polyvinyl acetate, polypropylene, and celluloid; ceramics; metals; These may be used alone or in combination of two or more.

(Component measurement device set)
Next, a component measuring device set as one embodiment of the present invention will be described.
The component measuring device set as one embodiment of the present invention includes a body fluid measuring chip which is an example of the test device of the present invention described above, and a component measuring device to which the body fluid measuring chip is attached and which measures a predetermined component in the body fluid. Is provided.
Hereinafter, a component measuring device set including a transmission-type component measuring device that measures light transmitted through a reaction product of a body fluid and a coloring reagent will be described, but the present invention is not limited thereto. A component measurement device set including a reflection-type component measurement device that measures light reflected from the reactant may be used.

  FIG. 6 shows a blood glucose meter set 500 as a component measuring device set according to an embodiment of the present invention. The blood glucose meter set 500 includes a blood glucose meter 110 as a component measuring device and a blood glucose measuring chip 100a as a body fluid measuring chip 100. The blood glucose measuring chip 100a is attached to the tip of the blood glucose meter 110. The blood glucose meter 110 includes a display 111 that displays a measurement result and operation details, a power button 112 for instructing activation and termination of the blood glucose meter 110, an operation button 113, a removal lever 114 for removing the blood glucose measurement chip 100a, It has. The display 111 is composed of a liquid crystal or an LED.

  FIG. 7 is a longitudinal sectional view separately showing the tip of the blood glucose meter 110 of the blood glucose meter set 500 and the blood glucose measurement chip 100a. In order to mount the blood glucose measurement chip 100a on the blood glucose meter 110, a mounting portion 22 having an opening 21 formed at the tip of the blood glucose meter 110 is provided, and a mounting hole 23 for mounting the blood glucose measurement chip 100a is defined inside the blood glucose meter 110. I do. Further, inside the blood glucose meter 110, a predetermined component of a body fluid (mainly, blood is described as an example in the present embodiment) collected by the blood glucose measurement chip 100a (in the present embodiment, a blood glucose level is mainly described as an example). Is provided with an optical measuring unit 24 for measuring. The blood glucose meter 110 further includes a processing unit 25 that processes a signal obtained from the measurement light to calculate a blood glucose level, and an eject pin 26 that removes the blood glucose measurement chip 100a in conjunction with the removal lever 114 (FIG. 6). ing. Hereinafter, each configuration will be described.

  At the time of measurement, the blood glucose measurement chip 100a is mounted in the mounting hole 23. The mounting work is performed manually by the user. Although not shown, an appropriate lock mechanism or the like for fixing the blood glucose measuring chip 100a at a predetermined position in the mounting hole 23 is preferably installed in order to minimize variations in the mounting position caused by manual operation.

  The optical measurement unit 24 includes an irradiation unit 31 that irradiates light to a reaction product of the coloring reagent and the blood, and a light receiving unit 32 that receives light transmitted through the reaction product as measurement light. In the present embodiment, a light emitting diode (LED) is used for the irradiation unit 31, but a halogen lamp, a laser, or the like may be used. As the light receiving unit 32, for example, a photodiode (PD) is used. The light receiving section 32 may be any as long as it can convert the received light into a predetermined signal, and may be a CCD, a CMOS, or the like.

  In the present embodiment, the irradiation unit 31 includes a first light emitting element 51 that emits light having a first wavelength and a second light emitting element 52 that emits light having a second wavelength different from the first wavelength. Including. Here, the first wavelength is a wavelength for detecting the degree of color development according to the blood sugar amount, and is in a wavelength band of, for example, 600 to 900 nm. The second wavelength is a wavelength for detecting the concentration of red blood cells in blood, and is in a wavelength band of, for example, 510 to 590 nm.

  The arrangement of the irradiation unit 31 and the light receiving unit 32 and the positional relationship between them will be described. Inside the blood glucose meter 110, a first space 41 and a second space 42 are formed. The irradiation unit 31 is arranged in the first space 41, and the light receiving unit 32 is arranged in the second space 42. When the blood glucose measurement chip 100a is not mounted on the blood glucose meter 110, the first space 41 and the second space 42 face each other with the mounting hole 23 interposed therebetween (see FIG. 7). In a state where the blood glucose measurement chip 100a is mounted on the blood glucose meter 110, the first space 41 and the second space 42 are located at positions where the reagent layer structure 30 on the blood glucose measurement chip 100a is held. They face each other. In order to reduce the energy loss of the irradiation light 33, the irradiation unit 31 is preferably arranged at a position where the irradiation light 33 can be irradiated perpendicularly to the bottom surface of the blood glucose measurement chip 100a.

Although not shown, when using a halogen lamp that irradiates the irradiation unit 31 with white light, a method of providing a spectral filter to extract only a specific wavelength as the irradiation light 33 may be used.
In addition, a method including a condenser lens is also preferable in order to effectively carry out irradiation with low energy.

  Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.

(Experimental example 1)
First, glucose dehydrogenase (GDH) 2U / μL as an enzyme, 5-methylphenazinium methylsulfate (PMS) 10 mM as a mediator, 2-benzo-thiazolyl-3- (4-carboxy-2) as a dye An aqueous solution containing 50 mM of -methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) and 250 mM of MES buffer (pH 6.5) was prepared. The absorbance of (PMS / 0h) and the sample (PMS / 22h) after being left in a light-shielded state at room temperature for 22 hours were measured using an ultraviolet-visible spectrophotometer. FIG. 8 shows the measurement results.
Next, 33.3 nkat (2U) of glucose dehydrogenase (GDH) as an enzyme, 1 mM of 1-methoxy-5-methylphenazinium methylsulfate (mPMS) as a mediator, and 2-benzo-thiazolyl- as a dye Contains 50 mM of 3- (4-carboxy-2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) and 250 mM of MES buffer (pH 6.5) An aqueous solution was prepared, and the absorbance of the sample immediately after the preparation (mPMS / 0h) and that of the sample (mPMS / 22h) left for 22 hours in a light-shielded state at room temperature were measured using an ultraviolet-visible spectrophotometer. FIG. 8 shows the measurement results.
From FIG. 8, the absorbance spectrum of sample PMS / 22h has a peak near 650 nm as compared with the absorbance spectrum of sample PMS / 0h, and the absorbance spectrum of sample mPMS / 22h is the absorbance spectrum of sample mPMS / 0h. It can be seen that the vicinity of 650 nm is raised as compared with. This indicates that the dye has developed color.
From the above, it was found that when a solution containing an enzyme, a mediator and a dye was left in a light-shielded state at room temperature for 22 hours, unintended color formation occurred and stability was low.

(Experimental example 2)
50 mM 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) as dye and MES buffer An aqueous solution containing 250 mM (pH 6.5) was prepared, and the absorbance of a sample immediately after preparation (WST4-0h) and that of a sample (WST4-22h) after being left in a light-shielded state at room temperature for 22 hours were measured with an ultraviolet-visible spectrophotometer. It measured using. FIG. 9 shows the measurement results.
From FIG. 9, the absorbance spectrum of sample WST4-22h was almost the same as the absorbance spectrum of sample WST4-0h. This indicates that the dye hardly developed color.
From the above, it was found that even if the solution containing only the dye was left in a light-shielded state at room temperature for 22 hours, little color was generated and the stability was high.

(Experimental example 3)
First, 10 mM of 5-methylphenazinium methyl sulfate (PMS) as a mediator and 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [4- (2- An aqueous solution containing 50 mM of sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) and 250 mM of MES buffer (pH 6.5) was prepared, and a sample immediately after the preparation (PMS / WST4-0h) was shielded from light at room temperature. The absorbance with the sample (PMS / WST4-22h) after standing for 22 hours at was measured using an ultraviolet-visible spectrophotometer. FIG. 10 shows the measurement results.
Next, 10 mM of 1-methoxy-5-methylphenazinium methyl sulfate (mPMS) as a mediator and 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [ An aqueous solution containing 50 mM of 4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) and 250 mM of MES buffer (pH 6.5) was prepared, and a sample immediately after the preparation (mPMS / WST4-0h) The absorbance of the sample (mPMS / WST4-22h) after being allowed to stand for 22 hours in a light-shielded state at room temperature was measured using an ultraviolet-visible spectrophotometer. FIG. 10 shows the measurement results.
From FIG. 10, the absorbance spectrum of sample PMS / WST4-22h has a peak near 650 nm as compared with the absorbance spectrum of sample PMS / WST4-0h, and the absorbance spectrum of sample mPMS / WST4-22h is Compared to the absorbance spectrum of mPMS / WST4-0h, it can be seen that around 650 nm is raised. This indicates that the dye has developed color.
From the above, it was found that if a solution containing a mediator and a dye was left in a light-shielded state at room temperature for 22 hours, unintended color formation occurred and stability was low.

(Experimental example 4)
First, glucose dehydrogenase (GDH) 2U / μL as an enzyme and 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl as a dye ] An aqueous solution containing 50 mM of 2H-tetrazolium (WST-4) and 250 mM of MES buffer (pH 6.5) was prepared, and the sample immediately after the preparation (GDH / WST4-0h) was left in a room light-shielded state for 22 hours. The absorbance with the subsequent sample (GDH / WST4-22h) was measured using an ultraviolet-visible spectrophotometer. FIG. 11 shows the measurement results.
From FIG. 11, it can be seen that the absorbance spectrum of sample GDH / WST4-22h has a peak near 650 nm as compared to the absorbance spectrum of sample GDH / WST4-0h. This indicates that the dye has developed color.
From the above, it was found that when a solution containing an enzyme and a dye was left in a light-shielded state at room temperature for 22 hours, unintended color development might occur.

  The present inventors have conducted experiments of Experimental Examples 1 to 4, and as a result of intensive research, for the first time, the dye and the mediator or the enzyme should be separated instead of mixed (instead of having a uniform structure). Thus, the present inventors have arrived at the present invention.

(Comparative Example 1)
<Preparation of reagent sheet with uniform structure>
2 U / μL of glucose dehydrogenase (GDH) as an enzyme, 1 mM of 1-methoxy-5-methylphenazinium methylsulfate (mPMS) as a mediator, and 2-benzo-thiazolyl-3- (4-carboxy) as a dye An aqueous solution containing 100 mM of 2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) and 250 mM of PIPES buffer (pH 7.5) was prepared. Next, the prepared solution was placed on a polyethylene terephthalate (PET) film (manufacturer: Toray Industries, Inc., trade name: Lumirror T60) and an inkjet printer (manufacturer: microjet, trade name: Lab-Jet500 Bio) was applied. It was applied and dried at 25 ° C. overnight to prepare a reagent sheet of uniform structure (thickness: 30 μm). FIG. 12 shows a micrograph of the uniform structure surface of the prepared uniform structure reagent sheet.
<Preparation of sample>
A high glucose solution (40 g / dL) was added to the blood to prepare a sample having a glucose concentration of 300 mg / dL.
<Absorbance measurement after sample spotting>
The prepared sample 3 mm ³ was spotted on the prepared reagent sheet, and the absorbance was measured using an ultraviolet-visible spectrophotometer. The results are shown in FIGS.
<Absorbance measurement after light irradiation>
The absorbance of the reagent sheet immediately after the preparation (initial) and the reagent sheet after being left under an indoor fluorescent lamp (average of 750 Lx) for 3 hours (after irradiation with fluorescence) were measured using an ultraviolet-visible spectrophotometer. FIG. 16A shows the average absorbance ratio (%) of the initials after light irradiation.
<Absorbance measurement after thermal acceleration test>
The absorbance of the reagent sheet immediately after preparation (initial) and the reagent sheet after the heat acceleration test (60 ° C., 3 days) were measured using an ultraviolet-visible spectrophotometer. FIG. 16B shows the average absorbance ratio (%) after the thermal acceleration test with respect to the initial.

(Comparative Example 2)
Preparation of a sample and measurement of absorbance after spotting of the sample were performed in the same manner as in Comparative Example 1, except that a reagent sheet having a two-layer structure was prepared instead of preparing a reagent sheet having a uniform structure in Comparative Example 1. The evaluation results are shown in FIGS.

<Preparation of a two-layered reagent sheet>
An aqueous solution (first solution) containing 2 U / μL of glucose dehydrogenase (GDH) as an enzyme and 1 mM of 1-methoxy-5-methylphenazinium methylsulfate (mPMS) as a mediator was prepared. Next, 100 mM of 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) as a dye was added. And an aqueous solution (second solution) containing 250 mM (pH 7.5) of PIPES buffer. The first liquid was applied on a polyethylene terephthalate (PET) film using an inkjet printer to form a first layer, and dried at 25 ° C. for 10 minutes. After drying, the second liquid was applied on the first layer using an inkjet printer, and dried at 25 ° C. overnight to prepare a two-layer structured reagent sheet (thickness: 30 μm).

(Example 1)
In Comparative Example 1, preparation of a sample, measurement of absorbance after spotting of a sample, measurement of light after irradiation were performed in the same manner as in Comparative Example 1 except that a reagent sheet having a three-dimensional mosaic structure was prepared instead of preparing a reagent sheet having a uniform structure. Was measured, and the absorbance after the heat acceleration test was measured. The evaluation results are shown in FIGS.

<Preparation of 3D mosaic structure reagent sheet>
First, an aqueous solution (first solution) containing 2 U / μL of glucose dehydrogenase (GDH) as an enzyme and 1 mM of 1-methoxy-5-methylphenazinium methylsulfate (mPMS) as a mediator was prepared. Next, 100 mM of 2-benzo-thiazolyl-3- (4-carboxy-2-methoxyphenyl) -5- [4- (2-sulfoethylcarbamoyl) phenyl] -2H-tetrazolium (WST-4) as a dye was added. And an aqueous solution (second solution) containing 250 mM (pH 7.5) of PIPES buffer.
The prepared first liquid and second liquid are put into respective storage units of an ink jet printer (manufacturer: MicroJet, trade name: Lab-Jet500 Bio), and applied on a polyethylene terephthalate (PET) film. A first layer was formed and dried at 25 ° C. for 10 minutes. After drying, similarly to the first layer, the second layer was formed on the first layer so as to have a predetermined laminated state (FIG. 5), and dried. After drying, similarly to the second layer, a third layer is formed on the second layer so as to have a predetermined laminated state (FIG. 5), dried, and a three-dimensional mosaic structure reagent sheet (FIG. 5, film thickness 30 μm) ) Was prepared. FIG. 13 shows an electron micrograph of the surface of the prepared three-dimensional mosaic structure reagent sheet.

  14 and 15, the reaction rate of the two-layered reagent sheet of Comparative Example 2 was lower than that of the uniform-structured reagent sheet of Comparative Example 1, whereas the three-dimensional mosaic-structured reagent sheet of Example 1 was compared. It was found that a decrease in the reaction rate can be suppressed as compared with the uniform-structure reagent sheet of Example 1.

As shown in FIG. 16A, when left under a fluorescent lamp for 3 hours, compared with the uniform structure reagent sheet of Comparative Example 1 (about 1.5 times the absorbance of the original before fluorescence irradiation), 3 of Example 1 The dimensional mosaic structure reagent sheet (about 1.3 times the absorbance of the original before fluorescence irradiation) was suppressed in color development, and it was found that deterioration over time could be suppressed.
From FIG. 16B, when the heat acceleration test (60 ° C., 3 days) was performed, the uniform structure reagent sheet of Comparative Example 1 (approximately 1.4 times the absorbance of the original before the heat acceleration test) was obtained. The three-dimensional mosaic structure reagent sheet of Example 1 (approximately 1.2 times the absorbance of the original before the thermal acceleration test) was suppressed in color development, indicating that deterioration over time could be suppressed.

(Comparative Example 3)
<Preparation of reagent sheet with uniform structure>
4 U / μL of glucose dehydrogenase (GDH) as an enzyme, 1 mM of 1-methoxy-5-methylphenazinium methyl sulfate (mPMS) as a mediator, 50 mM of tetrazolium salt (A) as a dye, and 200 mM of nickel ion And an aqueous solution containing 350 mM sodium nitrite. Next, the prepared solution was placed on a polyethylene terephthalate (PET) film (manufacturer: Toray Industries, Inc., trade name: Lumirror T60: 1 in FIG. 19A) and an inkjet printer (manufacturer: microjet, trade name: Lab) -Jet500 Bio) (solid coating: FIG. 19A) and dried at 25 ° C. overnight to prepare a uniform structure reagent sheet (film thickness 12 μm).
The concentrations of the reagents in the reagent sheet having the uniform structure are as follows.
Glucose dehydrogenase (GDH): 4 U / μL
1-methoxy-5-methylphenazinium methyl sulfate (mPMS): 1 mM
Tetrazolium salt (A): 50 mM
Nickel ion: 200 mM
Sodium nitrite: 350 mM
<Preparation of sample>
A high-concentration glucose solution (40 g / dL) was added to water to prepare a sample having a glucose concentration of 400 mg / dL.
<Absorbance measurement after sample spotting>
The prepared specimen 3 mm ³ was spotted on the prepared reagent sheet, and the absorbance was measured 25 seconds after spotting using a UV-visible spectrophotometer. The results are shown in FIG.

(Example 2)
In Comparative Example 3, the preparation of the sample and the measurement of the absorbance after the sample was spotted were performed in the same manner as in Comparative Example 3 except that a reagent sheet having a multilayer structure (two-step coating) was prepared instead of preparing a reagent sheet having a uniform structure. went. FIG. 20 shows the evaluation results.

<Preparation of laminated structure reagent sheet (two-step coating)>
First, an aqueous solution (first solution) containing 2 U / μL of glucose dehydrogenase (GDH) as an enzyme was prepared. Next, an aqueous solution containing 1 mM of 1-methoxy-5-methylphenazinium methyl sulfate (mPMS) as a mediator, 50 mM of a tetrazolium salt (A) as a dye, 200 mM of nickel ions, and 350 mM of sodium nitrite ( Second liquid) was prepared.
The prepared first liquid and second liquid are put into respective storage units of an ink jet printer (manufacturer: MicroJet, trade name: Lab-Jet500 Bio), and applied on a polyethylene terephthalate (PET) film. A first layer was formed (400A in FIG. 19B) and dried at 25 ° C. for 10 minutes. After drying, similarly to the first layer, a second layer is formed on the first layer so as to be in a predetermined laminated state (400B in FIG. 19B), dried, and a laminated structural reagent sheet (two-step coating: film thickness) 10 μm).
The concentration of each reagent in the laminated structure reagent sheet (two-step coating) is as follows.
Glucose dehydrogenase (GDH): 2 U / μL
1-methoxy-5-methylphenazinium methyl sulfate (mPMS): 1 mM
Tetrazolium salt (A): 50 mM
Nickel ion: 200 mM
Sodium nitrite: 350 mM

(Example 3)
Preparation of a specimen and measurement of absorbance after specimen spotting were performed in the same manner as in Comparative Example 3, except that a laminated structure reagent sheet (three-step coating) was prepared in Comparative Example 3 instead of producing a uniform structure reagent sheet. went. FIG. 20 shows the evaluation results.

<Preparation of laminated structure reagent sheet (three-step coating)>
First, an aqueous solution (first solution) containing 2 U / μL of glucose dehydrogenase (GDH) as an enzyme was prepared. Next, an aqueous solution containing 1 mM of 1-methoxy-5-methylphenazinium methyl sulfate (mPMS) as a mediator, 50 mM of a tetrazolium salt (A) as a dye, 200 mM of nickel ions, and 350 mM of sodium nitrite ( Second liquid) was prepared.
The prepared first liquid and second liquid were put into respective storage units of an ink jet printer (manufacturer: MicroJet, trade name: Lab-Jet500 Bio), and applied onto a polyethylene terephthalate (PET) film. A first layer was formed (400A in FIG. 19B) and dried at 25 ° C. for 10 minutes. After drying, similarly to the first layer, the second layer was formed on the first layer so as to have a predetermined laminated state (400B in FIG. 19B), and dried. After drying, a third layer (a single-grain layer composed of only the first granular portion composed of the first liquid) is formed on the second layer so as to be in a predetermined laminated state (400C in FIG. 19B), and dried. A laminated structure reagent sheet (three-step coating: film thickness 11 μm) was prepared. Here, the first granular portion in the third layer is stacked on the second layer (reagent separation layer) so as to be adjacent to the second granular portion made of the second liquid in the second layer (reagent separation layer). .
The concentrations of the respective reagents in the laminated structure reagent sheet (three-step coating) are as follows.
Glucose dehydrogenase (GDH): 3 U / μL
1-methoxy-5-methylphenazinium methyl sulfate (mPMS): 1 mM
Tetrazolium salt (A): 50 mM
Nickel ion: 200 mM
Sodium nitrite: 350 mM

(Example 4)
Preparation of a specimen and measurement of absorbance after specimen spotting were performed in the same manner as in Comparative Example 3, except that a laminated structure reagent sheet (four-step coating) was prepared instead of producing a uniform structure reagent sheet in Comparative Example 3. went. FIG. 20 shows the evaluation results.

<Preparation of reagent sheet with laminated structure (four-step coating)>
First, an aqueous solution (first solution) containing 2 U / μL of glucose dehydrogenase (GDH) as an enzyme was prepared. Next, an aqueous solution containing 1 mM of 1-methoxy-5-methylphenazinium methyl sulfate (mPMS) as a mediator, 50 mM of a tetrazolium salt (A) as a dye, 200 mM of nickel ions, and 350 mM of sodium nitrite ( Second liquid) was prepared.
The prepared first liquid and second liquid are put into respective storage units of an ink jet printer (manufacturer: MicroJet, trade name: Lab-Jet500 Bio), and applied on a polyethylene terephthalate (PET) film. A first layer was formed (400A in FIG. 19B) and dried at 25 ° C. for 10 minutes. After drying, similarly to the first layer, the second layer was formed on the first layer so as to have a predetermined laminated state (400B in FIG. 19B), and dried. After drying, a third layer (a single-grain layer composed of only the first granular portion made of the first liquid) was formed on the second layer so as to have a predetermined laminated state (400C in FIG. 19B), and dried. After drying, a fourth layer (a single-grain layer composed of only the first granular portion composed of the first liquid) is formed on the third layer so as to be in a predetermined laminated state (400D in FIG. 19B), and a laminated structure reagent sheet is formed. (Four-step coating: film thickness 12 μm) was produced. Here, the first granular portion in the third layer is laminated on the second layer (reagent separating layer) so as to be adjacent to the second granular portion made of the second liquid in the second layer (reagent separating layer). . Further, the first granular portions in the fourth layer are stacked so as to be alternate with the first granular portions in the third layer (400D in FIG. 19B).
The concentration of each reagent in the laminated structure reagent sheet (four-step coating) is as follows.
Glucose dehydrogenase (GDH): 4 U / μL
1-methoxy-5-methylphenazinium methyl sulfate (mPMS): 1 mM
Tetrazolium salt (A): 50 mM
Nickel ion: 200 mM
Sodium nitrite: 350 mM

From FIG. 20, it can be seen that in Comparative Example 3 (solid coating: FIG. 19A) as a control, the enzyme was inactivated and almost no color was formed, whereas the control had a reagent separation layer (first and second stages). Examples 2 to 4 were found to develop color.
FIG. 20 shows that in Examples 2 to 4, as in Examples 3 and 4, as the amount of the enzyme increased, the color development speed increased, and as a result, the absorbance increased.

  The present invention relates to a test device and a component measuring device set, and more particularly to a test device and a component measuring device set capable of suppressing deterioration over time while suppressing a decrease in reaction rate.

1: First base material 2: Second base material 3: Adhesive part 4: Adhesive part 10: Supply port 20: Flow path 21: Opening part 22: Mounting part 23: Mounting hole 24: Optical measuring part 25: Processing part 26 : Eject pin 30: reagent layer structure 30a: reagent separation layer 30b: reagent separation layer 31: irradiation part 32: light receiving part 33: irradiation light 41: first space 42: second space 51: first light emitting element 52 : Second light emitting element 100: body fluid measurement chip 100a: blood glucose measurement chip 110: blood glucose meter 111: display 112: power button 113: operation button 114: detachment lever 200: first droplet 300: second droplet 400A: First-stage application state 400B: Second-stage application state 400C: Third-stage application state 400D: Fourth-stage application state 500: Blood glucose meter set (component measuring device)
A: Part containing the first reagent (but not containing the second reagent) B: Part containing the second reagent (but not containing the first reagent) P: Lower layer made of enzyme Q: Upper layer made of dye X: 1st granular part X1: 1st granular part X2 in a 1st reagent separation layer: 1st granular part Y in a 2nd reagent separation layer: 2nd granular part Y1: 2nd granular part Y2 in a 1st reagent separation layer: 2nd Second granular part r1: radius of first droplet (circle) r2: radius of second droplet (circle) in reagent separation layer

Claims (10)

A transmitted light measurement test device used to measure a predetermined component in a body fluid,
It has a reagent layer structure including a first reagent that reacts with a predetermined component in the body fluid or promotes a reaction with the predetermined component in the body fluid, and a second reagent that reacts with the reaction of the first reagent. And
The reagent layer structure has a reagent separation layer in which the first reagent and the second reagent are arranged separately from each other in a direction along the layer in the layer ,
The reagent separation layer includes a granular first granular portion including the first reagent and a granular second granular portion including the second reagent,
A test device for transmitted light measurement, wherein the first granular part and the second granular part are arranged separately.   The test device for transmitted light measurement according to claim 1, wherein the reagent layer structure has a multilayer structure in which the reagent separation layers are stacked. The first granular portion in the first reagent separation layer of the multilayer structure is adjacent to the second granular portion in the second reagent separation layer formed on the first reagent separation layer,
The test device for transmitted light measurement according to claim 2, wherein the second granular portion in the first reagent separation layer is adjacent to the first granular portion in the second reagent separation layer.   The said 1st granular part is apply | coated so that it may adjoin only the said 2nd granular part, The said 2nd granular part may be apply | coated so that it may adjoin only the said 1st granular part. 4. The test device for transmitted light measurement according to 1.   The transmitted light measurement method according to any one of claims 1 to 4, wherein the reagent layer structure further includes a single-grain layer including only one of the first and second granular portions. Test equipment.   The one-grain layer is laminated on the reagent separation layer such that the one granular portion is adjacent to the other granular portion of the first granular portion and the second granular portion in the reagent separation layer. Item 6. A test device for transmitted light measurement according to Item 5.   The test device for transmitted light measurement according to claim 5, wherein the one granular portion is composed of only the first reagent or the second reagent.   The test device for transmitted light measurement according to any one of claims 1 to 7, wherein the first reagent contains at least one of an enzyme and a mediator, and the second reagent is a dye. The test device for transmitted light measurement includes a supply port to which a body fluid is supplied, and a flow path having the supply port formed at one end, and a body fluid mountable on a component measuring device for measuring a predetermined component in the body fluid. Measuring tip,
The test device for transmitted light measurement according to any one of claims 1 to 8, wherein the reagent layer structure is provided in the flow channel. A body fluid measurement chip as a transmitted light measurement test device according to claim 9, and the body fluid measurement chip is attached,
A component measuring device for measuring a predetermined component in the body fluid.