JPH0954085A - Dry testing tool for blood sample - Google Patents

Abstract

PROBLEM TO BE SOLVED: To easily measure hemoglobin A1 C and A simultaneously by providing, on a support, a plurality of parallel structural bodies in each of which a reagent layer containing a hemolyzer or a modifier is provided on a solid antibody layer on the light transmitting part of the support, and surrounding both antibody layers with a remaining-fluid absorbing layer. SOLUTION: A support is provided with a light transmissible part, such as a through hole, and solid antibody layers are stacked on the top of the light transmissible part. Two structural bodies each provided with a reagent layer containing a hemolyzer and/or a modifier are provided in parallel on the support, and a remaining-fluid absorbing layer is provided around two solid antibody layers. Anti-hemoglobin(Hb) A and anti-HbA1 C, are held in solid phase in the respective antibody layers. A blood sample is dropped onto both of the reagent layers by equal amounts, hemolyzed, and modified, and subsequently a developing solution is dropped and developed. HbA containing HbA1 C. is trapped in the solid anti-HbA layer, and HbA in the solid anti-HbA1 C, layer. Since the Hb is colored, the amount of HbA and HbA1 C. is determined through the observation of the degree of coloring of each solid layer, and the ratio between them is calculated.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a clinical test,
The present invention relates to a test device for simultaneously and simply measuring hemoglobin A _1c (abbreviated as HbA _1c ) and hemoglobin A (abbreviated as HbA) in whole blood.

[0002]

2. Description of the Related Art So-called glycosyl hemoglobin, in which sugar is bound to hemoglobin in blood, is used for diagnosis or follow-up of diabetes because its concentration reflects the average blood sugar concentration in the past 1-2 months. Widely used as a suitable index.

Conventionally, the measurement of glycosyl hemoglobin has been carried out by an electrophoresis method, a mini-column method, an HPLC method, etc. Among them, in the field of clinical examination, the HPLC method is mainly used in view of required time and separability. Has become. However, recently, a method of measuring by an immune reaction, a so-called immunoassay method has been used.

Since this immunoassay method utilizes the specificity of an antibody for glycosyl hemoglobin, unstable (Reybile type) glycosyl hemoglobin, carbamylated hemoglobin, acetaldehyde hemoglobin, and acetylated hemoglobin which are problematic in the HPLC method are obtained. Accurate and rapid measurement is possible without being affected by the above, and sample processing capacity is high.

[0005] Further, there is also known a disposable test device which simplifies this immunoassay method and allows visual determination of hemoglobin (hemoglobin A, S and C) in blood (“Screening” 3 , 67).
~ 76 (1994)). However, this determines the difference between hemoglobins A, S, and C, not the difference between hemoglobin and glycosyl hemoglobin.

The conventional HPLC method and immunoassay method require a relatively large-scale analyzer, and the measurement is performed in a hospital laboratory or a laboratory center. In contrast,
Although the above-mentioned currently available disposable test devices can be measured by the patient themselves, they require hemolysis as a pretreatment and can measure various hemoglobins as described above. However, glycosyl hemoglobin cannot be measured. When the pretreatment is performed by the patient himself / herself, the problem of contamination arises in addition to the complexity of the operation.

When quantifying glycosyl hemoglobin, hemoglobin A which is a specific glycosyl hemoglobin in hemoglobin A (hereinafter abbreviated as HbA).
It has been clinically generalized to use the ratio of _1c (hereinafter, abbreviated as HbA _1c ) to HbA as an index, and it is displayed as a normal ratio, and about 4 to 6% is a standard for its normal value. There is. If a kit that can easily and quickly measure these HbA _1c values without the need for pretreatment is developed, its merit will be extremely large.

When performing an immunoassay, HbA _1c
The specific antibody against HbA has an epitope at a site having a different chemical structure from HbA, and the site is HbA.
Not fully exposed in _1c . Therefore, in order to perform an immunoassay, in many cases, the epitope must be exposed by denaturing HbA _1c , and the denaturing operation was also a major technical issue.

[0009]

SUMMARY OF THE INVENTION The present invention does not require a large-scale analyzer, and does not require pretreatment, and can be operated by a simple operation even when a small or medium-sized hospital, a practitioner or the patient himself is at home.
It is an object of the present invention to provide a dry test device capable of simultaneously measuring bA _1c .

[0010]

The above problems can be solved by a test device having the following structure. That is, the present invention provides the following configurations i) i on one support having a through hole or a light transmissive portion for enabling optical measurement.
i) ie; i) laminated directly above the through hole or light transmissive portion,
A solid-phased antibody layer (or a solid-phased adsorbent layer), ii) a reagent layer, which is located on the solid-phased antibody layer (or the solid-phased adsorbent layer) and contains a hemolytic agent and / or a denaturant,
Of the solid-phased antibody layer (or the solid-phased adsorbent layer) having two residual liquid absorption layers, each structure being arranged in parallel on a support. It is a dry test device for blood samples that is installed so as to surround it. Hereinafter, the test tool of this type of aspect is defined as aspect 1.

Further, a solid-phased antibody layer (or a solid-phased adsorbent layer) is provided directly on one support having a through hole or a light-transmitting portion for enabling optical measurement. One is retained, and further one reagent layer containing a hemolytic agent and / or a denaturant is retained on the immobilized antibody layer (or immobilized adsorbent layer), and the residual liquid absorption layer is fixed. There is also provided a dry test device for a blood sample, which is installed so as to surround the phased antibody layer (or the solid phase adsorbent layer). Hereinafter, the test device of this type of aspect is defined as aspect 2.

The present invention can be applied to the measurement of various antigens in blood, but is particularly suitable for the measurement of hemoglobin A and A _1c , and a specific embodiment relating to the present invention will be specifically described later with reference to the drawings. explain.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention is based on the principle that a sample is hemolyzed / denatured in a reagent layer and trapped by a solid-phased antibody layer (or a solid-phased adsorbent layer). A development layer may be provided between the derivatized antibody layer (or the solid-phased adsorbent layer). Providing the development layer is particularly useful because the binding reaction between the sample denatured in the reagent layer and the antibody, the labeling substance, etc. in the reagent layer described later is performed in the development layer.

The inventors have already disclosed in Japanese Patent Application No. 7-113765 a test device which has solved the above-mentioned conventional problems. The invention according to the present application occupies less space due to the different structure as compared with the previous invention, and the liquid uniformly advances (from the top to the bottom) due to the action of gravity, resulting in liquid unevenness when developing the developing liquid. It has the advantage of being less likely to occur.

[0015]

Embodiments of the present invention will be described below with reference to the accompanying drawings. FIG. 1 is a sectional view showing a typical embodiment of the present invention. 2 to 9 are a plan view and a sectional view showing a specific embodiment of the present invention. In FIGS. 2 to 9, only the residual liquid absorption layer is covered with a moss, but this is to make the drawings easier to see, and the structure is exactly the same as in FIG. As can be understood from the drawings, the test device according to the present invention essentially has a layer having liquid transfer ability stacked to form one large layered structure that spreads the liquid from top to bottom. It is in.

FIG. 1 FIG. 1 shows a typical view of a dry test device for blood samples according to the present invention. As can be seen from the figure, aspect 1
In the second aspect, two layered structures are provided in parallel on the support, and in the second aspect, one layered structure is provided on the support. When stacking the constituents that form each layer as shown in the figure, make sure that the layers do not shift and that the layers are in close contact with each other.
It is advisable to provide a frame to hold down the laminate. In the figure, a cover having a through hole is installed on the upper part, and the cover and the support are bonded together while pressing the laminate. The material of the cover may be the same as that forming the support. Specific examples of each component will be described later.

FIG. 2 FIG. 2 is a cross-sectional view of a dry test device for a blood sample in which anti-HbA antibody and anti-HbA _1c antibody are immobilized in each immobilized antibody layer. When the same amount of blood sample is dropped onto the reagent layer A and the reagent layer B, the hemolyzing agent and the denaturing agent simultaneously modify the hemolysis. Subsequently, when the developing solution is added to the reagent layer A and the reagent layer B, the denatured hemoglobin is developed, and HbA containing HbA _1c is trapped in the immobilized antibody layer on which the anti-HbA antibody is immobilized, and HbA
HbA is present in the immobilized antibody layer on which the _1c antibody is immobilized.
Only _1c is trapped. Other substances (excess blood components and HbA other than HbA _1c in the solid-phased antibody layer on which the anti-HbA _1c antibody is solid-phased)
Is absorbed by the residual liquid absorption layer around the immobilized antibody layer. Since hemoglobin is colored, the degree of coloring of each of the solid-phased antibody layer on which the anti-HbA antibody is solid-phased and the solid-phased antibody layer on which the anti-HbA _1c antibody is solid-phased is determined by the support. Observe from the through-hole or the light-transmissive part. As for the observation method, there is no problem with the conventional method such as visual observation or using a colorimeter. After the observation, the amounts of HbA and HbA _1c are obtained from each antibody layer, and the respective values are used to calculate the ratio of HbA _1c to HbA.

Here, the materials for the respective constituent components may be any known ones, but the concrete examples are as follows. Support: Film, sheet, or plate-shaped plate mainly composed of polystyrene, polyester, cellulose acetate, etc. Remaining liquid absorption layer: Mainly composed of cellulose, cellulose derivative (cellulose acetate, nitrocellulose, etc.), glass fiber, etc. Filter paper or continuous porous membrane with sufficient pore volume Hemolytic agent; saponin, surfactant, etc. Denaturing agent; thiocyanate, protease, etc. Reagent layer; same material as residual liquid absorption layer Adsorbed denaturant Antibody layer (or adsorbent layer); Immobilized antibody or adsorbent on the same material as residual liquid absorption layer Development solution: Contains small amounts of proteins, salts, and surfactants Buffer solution

The denaturant in the reagent layer is added so that the anti-HbA _1c antibody exposes the sugar moiety of HbA _1c , which is the antigen, so that the antibody can easily attack. Among the two structures, HbA is the antigen. In that case, it is not necessary to expose the sugar moiety, and therefore it is not necessary to add a denaturant to the reagent layer.

An example of a dry test device for a blood sample, which further contains a labeling substance in the reagent layer, is shown below. When a labeled substance is used, a mechanically and optically clear label can be measured, so that the measurement accuracy becomes high. ● FIG. 3 In FIG. 3, labeled anti-HbA antibody is contained as a labeling substance in both reagent layers, and anti-HbA _1c antibody and anti-HbA antibody are separately immobilized in separate immobilized antibody layers. It is sectional drawing of the dry test device for blood samples which has been phased. However, the labeled anti-HbA antibody and the immobilized anti-HbA antibody have different epitopes for HbA. When the same amount of blood sample is dropped onto the reagent layer A and the reagent layer B, the hemolyzing agent and the denaturing agent simultaneously modify the hemolysis, and the modified HbA and HbA _1c react with the labeled anti-HbA antibody in the reagent layer. The complex of HbA-labeled anti-HbA antibody and HbA _1c -labeled anti-Hb
A complex of A antibody is formed. However, this labeled anti-Hb
The antibody A does not have the glycosyl hemoglobin sugar side as an epitope, but has the hemoglobin side epitope as an epitope. When the developing solution is subsequently added to the reagent layer A and the reagent layer B, both complexes are trapped in the immobilized antibody layer in which the anti-HbA antibody is immobilized, and the complex of HbA _1c -labeled anti-HbA antibody is Only the anti-HbA _1c antibody is trapped in the solid phase immobilized antibody layer. here,
Since the labeled anti-HbA antibody and the immobilized anti-HbA antibody have different epitopes for HbA, two antibodies can simultaneously bind to one HbA. However, even if the epitopes are different, it may be difficult for the two antibodies to bind simultaneously due to steric hindrance or the like, and therefore it is necessary to be careful in selecting the antibody. Other substances are absorbed in the residual liquid absorption layer around the immobilized antibody layer. From the through-holes or the light-transmitting portion of the support, the coloring degree of all HbA in the immobilized antibody layer on which the anti-HbA antibody is immobilized is measured by a measuring device such as a colorimeter, while the anti-HbA _1c antibody is detected. The label of the solid-phased antibody layer that has been solid-phased is measured by a device that reads the label. After the observation, the amounts of HbA and HbA _1c in each antibody layer are obtained, and the ratio of HbA _1c to HbA is calculated using each value. Here, the support, the residual liquid absorption layer, the hemolytic agent, the denaturing agent, and the developing solution are the same as those in FIG. 2, and the other materials include the following. Labeling means for labeled antibodies: metal colloids (gold, silver, copper), fine particles such as latex and colored products thereof, dyes (hematoxylin, malachite green, rhodamine B, galocyanine, neal blue A, azure C, xylene cyanol, etc.) , Fluorescent dyes (fluorescein, rhodamine, etc.), luminescent substances (acridinium, luciferase, etc.)

FIG. 4 shows that the labeled boronic acid derivative is contained in the reagent layer in one reagent layer and not in the other reagent layer.
FIG. 4 is a cross-sectional view of a dry test device for blood specimen in which bA antibody is immobilized on both immobilized antibody layers. Among the boronic acid derivatives, dihydroxyboryl derivatives (for example, xylene cyanol aminophenylboronic acid (Xlenecy)
anol Aminophenylboronic a
cid)) is a substance that chemically binds to two cis-type hydroxyl groups in a sugar, and Hb in order to specifically bind to a sugar.
A and HbA _1c can be distinguished. When the same amount of blood sample is dropped onto the reagent layers A and B, the hemolyzing agent and the denaturing agent simultaneously modify the hemolyzed blood, and in the reagent layer A, the modified HbA _1c and the labeled boron in the reagent layer are added. The acid derivative reacts to form the HbA _1c -boronic acid derivative complex. Here, since the dihydroxyboryl derivative has a small molecular weight unlike an antibody or the like, it can bind to a sugar that makes the face slightly visible from hemoglobin without modifying the hemoglobin. There is no particular need to add a denaturing agent. Of course, you can put it in. Subsequently, the developing solution is added to the reagent layers A and B.
In addition, the HbA _1c -boronic acid derivative complex has anti-H
The bA antibody is trapped in the solid-phased antibody layer that has been solid-phased. Here, this anti-HbA antibody is not an antibody having the sugar side of glycosyl hemoglobin as an epitope but an antibody having the hemoglobin side as an epitope. Eventually, HbA on one of the immobilized antibody layer containing _{HbA 1c} is, the other immobilized antibody layer _{HbA 1c} - boronic acid derivative complex and HbA is trapped. Other substances are absorbed in the residual liquid absorption layer around the immobilized antibody layer. The amount of total hemoglobin A in one immobilized antibody layer is determined from the through hole or the light-transmitting portion of the support, and the amount of HbA _1c is determined by measuring the label in the other immobilized antibody layer. afterwards,
The ratio of HbA _1c to HbA is calculated using each value. Here, the types of labels applied to the support, the residual liquid absorption layer, the hemolytic agent, the denaturing agent, the developing solution, and the boron derivative are the same as those in FIG.

FIG. 5 FIG. 5 shows that the labeled anti-HbA _1c antibody was contained in one reagent layer, the labeled anti-HbA antibody was contained in the other reagent layer, and the anti-HbA antibody was immobilized in both reagents. FIG. 3 is a cross-sectional view of a dry test device for a blood sample that is solid-phased in each phased antibody layer. Here, the labeled anti-HbA antibody and the immobilized anti-HbA antibody HbA
Due to the different epitopes for HbA, two antibodies can bind to one HbA at the same time. However, even if the epitopes are different, it may be difficult for the two antibodies to bind simultaneously due to steric hindrance or the like, and therefore it is necessary to be careful in selecting the antibody. In addition, the labeled anti-HbA _1c antibody does not use the sugar side of glycosylhemoglobin as an epitope, but the hemoglobin side as an epitope. Furthermore, the immobilized anti-HbA antibody is HbA _1c.
The epitope that binds to is HbA _1c and labeled anti-HbA
_It recognizes an epitope at a site different from the site to which the _1c antibody is bound. When the same amount of the blood sample is dropped onto the reagent layer A and the reagent layer B, the hemolyzing agent and the denaturing agent simultaneously change the hemolysis and denature the Hb in the reagent layer A.
A _1c and the labeled anti-HbA _1c antibody react to form a complex, and in the reagent layer B, a complex of HbA-labeled anti-HbA antibody and a complex of HbA _1c -labeled anti-HbA antibody are formed. To do. When the developing solution is subsequently added to the reagent layer A and the reagent layer B, all of the above-mentioned three kinds of complexes are trapped in the solid-phased antibody layer on which the anti-HbA antibody is solid-phased. Other substances are absorbed in the residual liquid absorption layer around the immobilized antibody layer. The amount of HbA and HbA _1c is determined by measuring the label in each immobilized antibody layer from the through-holes or light-transmitting portion of the support. After that, using each value, H
Calculate the ratio of HbA _{1c to} bA. Here, the support, the residual liquid absorption layer, the hemolytic agent, the denaturing agent, and the developing solution are the same as those in FIG.

FIG. 6 FIG. 6 shows that the labeled anti-HbA _1c antibody was contained in one reagent layer, the labeled anti-HbA antibody was contained in the other reagent layer, and the ion-exchange resin was used for both. FIG. 3 is a cross-sectional view of a dry test device for blood sample that is solid-phased in each phased adsorbent layer. When the same amount of the blood sample is dropped into the reagent layers A and B, the hemolyzing agent and the denaturing agent simultaneously modify the hemolyzed blood, and in the reagent layer A, the modified HbA _1c is labeled with anti-HbA labeled with anti-HbA.
_{The 1c} antibody reacts to form an HbA _1c -labeled anti-HbA _1c antibody complex, and in the reagent layer B, HbA and HbA _1c
A labeled anti-HbA antibody reacts with HbA-labeled anti-Hb
A HbA _1c -labeled anti-HbA antibody complex is formed with the A antibody complex. When the developing solution is subsequently added to the reagent layer A and the reagent layer B, the above complex is trapped in each of the solid-phased adsorbent layers in which the ion-exchange resins are solid-phased.
Other substances are absorbed by the residual liquid absorption layer around the solid-phased adsorbent layer. HbA and HbA by measuring the respective labels from the through-holes or light-transmitting parts of the support
_Calculate the amount of _1c . Then, using each value, the ratio of HbA _1c to HbA is calculated. Where the support,
The residual liquid absorption layer, hemolytic agent, denaturing agent, and developing solution are the same as those in FIG. Examples of ion exchange resins include diethylaminoethyl cellulose and carboxymethylated polyvinyl alcohol, and the above solid-phase adsorption is obtained by adsorbing and fixing these ion exchange resins on the same material as the residual liquid absorption layer. It is a drug layer.

Up to this point, the test device has two reagent layers and two immobilized antibody layers (or immobilized adsorbent layers). However, there is also the above-mentioned aspect 2 "a test device of a type in which one immobilized antibody layer (or immobilized adsorbent layer) and one reagent layer are held on a support". This type of test device saves the trouble of dropping blood samples in the same amount in two reagent layers. Also, when manufacturing the test device, only half the steps of the first aspect are required. However,
In Aspect 1, HbA _1c was independently trapped in one immobilized antibody layer, but in Aspect 2, HbA _1c and HbA _1c were also trapped.
Since HbA and HbA _1c are visually the same as well as being trapped in a single immobilized antibody layer, it is necessary to label HbA _1c mechanically and optically to distinguish it from HbA. There is. After applying the sample, the amount of HbA on one side is judged from the overall coloration and the amount of HbA _1c on the other side is judged from the label. The following is an example of the test tool of the second embodiment including one reagent layer and one solid-phased antibody layer (or solid-phased adsorbent layer).

[FIG. 7] FIG. 7 shows a dry test for a blood sample in which the labeled anti-HbA _1c antibody is contained in the reagent layer and the anti-HbA antibody is immobilized in the immobilized antibody layer. It is sectional drawing of a tool. When a blood sample is dropped on the reagent layer, HbA _1c, which is denatured at the same time as hemolysis by the action of a hemolytic agent and a denaturing agent, and the binding portion (sugar portion) is denatured to expose the labeled anti-HbA _1c antibody in the reagent layer. React to form a complex. Continuing the addition of developing solution to the reagent layer, _{HbA 1c} - labeled anti _{HbA 1c} antibody complex and HbA is Yuki is expanded, anti-HbA antibody immobilized to HbA to immobilized antibody layer has an _{HbA 1c} - the complex of labeled _{anti-HbA 1c} antibody is trapped. Here, the anti-HbA antibody is an antibody having an epitope on the hemoglobin side, rather than an epitope on the sugar side of glycosyl hemoglobin. Other substances are absorbed in the residual liquid absorption layer next to the immobilized antibody layer. From the through-holes or the light-transmitting portion of the support, the degree of coloring of hemoglobin in the immobilized antibody layer is measured with a colorimeter, and the label is measured with a device for reading the label. After the observation, the amounts of HbA and HbA _{1c in} the antibody layer were calculated, and the respective values were used to determine HbA _1c for HbA.
Calculate the ratio of Here, the support, the residual liquid absorption layer, the hemolytic agent, the denaturing agent, the developing solution, and the labeling means for the labeled antibody are the same as those shown in FIG.

If the labeling concentration is measured by a colorimeter different from that for measuring the degree of coloring of hemoglobin in the immobilized antibody layer, the size of the device becomes large. However, since hemoglobin is red, it is possible to measure the labeling concentration with a colorimeter by using a dye having a wavelength different from that of red as the labeling substance (so-called dual wavelength measurement), and miniaturization of the measuring device is expected. . The same applies hereinafter.

FIG. 8 shows that a labeled boronic acid derivative is contained in the reagent layer,
FIG. 3 is a cross-sectional view of a dry test device for a blood sample in which an anti-HbA antibody is immobilized on a solid-phased antibody layer. When a blood sample is dropped onto the reagent layer, it is simultaneously denatured by the action of the hemolytic agent and the denaturing agent, and in the reagent layer, HbA _1c which is denatured and the sugar moiety is exposed reacts with the labeled boronic acid derivative to form a complex. To form. For the same reason as the test device shown in FIG. 3, no denaturant is required. When the developing solution is subsequently added to the reagent layer, the HbA and the complex are developed and reach the immobilized antibody layer on which the anti-HbA antibody is immobilized. After arrival, Hb
Both A and the complex are trapped in the immobilized antibody layer on which the anti-HbA antibody is immobilized. Where this anti-HbA
The antibody does not have the sugar side of glycosyl hemoglobin as the epitope, but has the hemoglobin side as the epitope. Other substances are absorbed in the residual liquid absorption layer next to the immobilized antibody layer. From the through-holes or the light-transmitting portion of the support, the degree of coloring of hemoglobin in the immobilized antibody layer is measured with a colorimeter, and the label is measured with a device for reading the label. After the observation, the amounts of HbA and HbA _{1c in} the antibody layer are obtained, and the ratio of HbA _1c to HbA is calculated using each value. Here, the support, residual liquid absorption layer, hemolytic agent, denaturing agent, developing solution, labeling substance, and boronic acid are the same as those in FIG.

[FIG. 9] FIG. 9 is a cross section of a dry test device for a blood sample in which a labeled anti-HbA _1c antibody is contained in a reagent layer and an ion exchange resin is immobilized on a solid phase adsorbent layer. It is a figure. When a blood sample is dropped on the reagent layer, it is simultaneously denatured by hemolysis and denaturing agents, and in the reagent layer, the denatured HbA _1c with exposed sugar chains reacts with the labeled anti-HbA _1c antibody to form a complex. Form the body. When the developing solution is subsequently added to the reagent layer, the complex and HbA are developed and reach the solid-phased adsorbent layer. After the arrival, both the HbA _1c- derived complex and HbA are trapped by the action of the ion exchange resin. Other substances are absorbed in the residual liquid absorption layer next to the immobilized antibody layer. The amount of HbA in the solid-phased antibody layer is judged from the overall coloring, and the amount of HbA _1c is judged from the label from the through-holes or the light-transmitting part of the support. Then, using each value, HbA for HbA
Calculate the ratio of _1c . Here, the support, the residual liquid absorption layer,
The hemolytic agent, denaturing agent, developing solution, labeling means, and ion exchange substance are the same as those in FIG.

[0029]

EFFECTS OF THE INVENTION By using the dry test device according to the present invention, HbA and HbA _1c can be simultaneously measured by a simple operation even when the patient is at home without using a large-scale analyzer and pretreatment. Further, since the liquid is not chromatographed in the lateral direction, the liquid flows uniformly and the unevenness is eliminated, which leads to improvement in reliability of the measurement result.

[0030]

[Brief description of drawings]

FIG. 1 is a sectional view of a typical embodiment of a test device according to the present invention. FIG. 2 is a cross-sectional view of a dry test device for a blood sample in which anti-HbA antibody and anti-HbA _1c antibody are immobilized on each immobilized antibody layer. FIG. 3 shows that a labeled anti-HbA antibody is contained as a labeling substance in both reagent layers, and the anti-HbA _1c antibody and the anti-HbA antibody are immobilized in separate immobilizing antibody layers, respectively. FIG. 3 is a cross-sectional view of a dry test device for a blood sample that is present. FIG. 4 shows that one of the reagent layers contained the labeled boronic acid derivative in the reagent layer and the other reagent layer did not contain the anti-HbA antibody in both solid phase immobilized antibody layers. It is sectional drawing of the dry-type test device for blood samples which has been made into. FIG. 5 shows that the labeled anti-HbA _1c antibody was contained in one reagent layer, the labeled anti-HbA antibody was contained in the other reagent layer, and the anti-HbA antibody was immobilized on both of the immobilized antibody layers. It is sectional drawing of the dry test device for blood samples which has been phased. FIG. 6 shows that the labeled anti-HbA _1c antibody is contained in one reagent layer, the labeled anti-HbA antibody is contained in the other reagent layer, and the ion-exchange resin is contained in both solid-phase adsorbent layers. FIG. 3 is a cross-sectional view of a solid-state dry test device for blood sample.
FIG. 7 is a cross-sectional view of a dry test device for a blood sample in which the labeled anti-HbA _1c antibody is contained in the reagent layer and the anti-HbA antibody is immobilized in the immobilized antibody layer. FIG.
FIG. 3 is a cross-sectional view of a dry test device for a blood sample in which a labeled boronic acid derivative is contained in a reagent layer and an anti-HbA antibody is immobilized on a solid-phased antibody layer. FIG. 9 is a cross-sectional view of a dry test device for blood samples in which a reagent layer contains a labeled anti-HbA _1c antibody and an ion exchange resin is immobilized on a solid-phase adsorbent layer.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shigeru Doi 57 57 Higashikujo Nishiamita-cho, Minami-ku, Kyoto-shi, Kyoto Prefecture Kyoto Daiichi Kagaku Co., Ltd.

Claims (14)

[Claims] 1. On a support having a through-hole or a light-transmissive portion for enabling optical measurement, each of the following constitutions i and ii, ie; i) the through-hole or light-transmitting: A solid-phased antibody layer (or a solid-phased adsorbent layer) laminated on the solid portion, ii) a hemolytic agent positioned on the solid-phased antibody layer (or the solid-phased adsorbent layer), and / Or a reagent layer containing a denaturant,
Of the solid-phased antibody layer (or the solid-phased adsorbent layer) having two residual liquid absorption layers, each structure being arranged in parallel on a support. Are arranged around the
Dry test device for blood samples. 2. An anti-hemoglobin A (abbreviated as HbA) antibody and an anti-hemoglobin A _1c (abbreviated as HbA _1c ) antibody are immobilized in each immobilized antibody layer. The dry test device for a blood sample according to claim 1. 3. The reagent layer further comprises a labeling substance.
The dry test device for a blood sample according to claim 1 or 2. 4. A labeled anti-HbA antibody is contained as a labeling substance in both reagent layers, and the anti-HbA _1c antibody and the anti-HbA antibody are immobilized in separate immobilization antibody layers, respectively. The dry test device for a blood sample according to claim 3, wherein the labeled anti-HbA antibody and the immobilized anti-HbA antibody have different epitopes for HbA. 5. One of the reagent layers contains a labeled boronic acid derivative, and the other reagent layer does not contain a labeled boronic acid derivative.
The dry test device for a blood sample according to claim 3, wherein the antibody is immobilized on both of the immobilized antibody layers. 6. The dry test device for a blood sample according to claim 5, wherein the boronic acid derivative is a dihydroxyboryl derivative. 7. A labeled anti-HbA _1c antibody is contained in one reagent layer, a labeled anti-HbA antibody is contained in the other reagent layer, and an anti-HbA antibody is immobilized on both of the immobilized antibody layers. The dry test device for a blood sample according to claim 3, wherein the labeled anti-HbA antibody and the immobilized anti-HbA antibody are phased and have different epitopes for HbA. 8. A labeled anti-HbA _1c antibody is contained in one of the reagent layers, a labeled anti-HbA antibody is contained in the other of the reagent layers, and an ion exchange resin is contained in both of the immobilized adsorbent layers. The dry test device for a blood sample according to claim 3, wherein the dry test device is immobilized. 9. A solid-phased antibody layer (or solid-phased adsorbent layer) is provided on a single support having a through hole or a light-transmitting portion to enable optical measurement. In addition, one reagent layer containing a hemolytic agent and / or a denaturant is further laminated on the solid-phased antibody layer (or the solid-phased adsorbent layer), and the residual liquid absorption layer is a solid phase. A dry test device for blood samples, which is installed so as to surround the immobilized antibody layer (or solid-phase adsorbent layer). 10. The blood according to claim 9, wherein the reagent layer contains a labeled anti-HbA _1c antibody, and the anti-HbA body is immobilized on the immobilized antibody layer. Dry test device for specimens. 11. The method according to claim 9, wherein the reagent layer contains a labeled boronic acid derivative, and the anti-HbA antibody is immobilized on the immobilized antibody layer. Dry test device for blood samples. 12. The dry test device for a blood sample according to claim 11, wherein the boronic acid derivative is a dihydroxyboryl derivative. 13. The reagent layer contains a labeled anti-HbA _1c antibody and a labeled anti-HbA antibody labeled with a label different from the above-mentioned label, and the ion exchange resin is a solid phase adsorbent. The dry test device for a blood sample according to claim 9, wherein the dry test device is immobilized on a layer. 14. The development layer according to claim 1, further comprising a spreading layer provided between the reagent layer and the immobilized antibody layer (or the immobilized adsorbent layer). Dry test device for blood samples.