JP3298836B2 - Sample analysis tool - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a dry sample analyzer used for analyzing a target component in a sample such as blood or urine.

[0002]

2. Description of the Related Art Conventionally, in the fields of biochemical analysis and clinical examination, various samples have been analyzed, and these samples can be analyzed in a short time, in a large amount and collectively. Has been requested.

[0003] In response to such demands, a number of dry sample analysis tools (also referred to as "test films") have been developed in which a reagent is preliminarily contained in a porous membrane formed of paper or plastic material or the like. .

According to such a dry sample analysis tool,
Compared with wet (liquid) analysis in which the reagent and the target component are reacted in a liquid, the operation of adding the reagent, the operation of reacting with the sample, and the like can be simplified, so that the analysis can be performed quickly and easily. . This dry sample analysis tool is actually used for analysis of glucose, ketone bodies, proteins and the like in blood, urine, saliva and the like.

[0005] However, in the above-mentioned dry sample analysis tool, when whole blood is used as a sample to analyze a specific component in plasma, there is a problem that red blood cells hinder the analysis. For example, when analysis is performed using a coloring reagent, the color of the coloring reagent may not be accurately measured because the red blood cells themselves are colored. For this reason, it is necessary to remove red blood cells by centrifugation or the like before performing analysis.
However, when the amount of the sample is small, it is difficult to separate components by centrifugation, and in a dry-type sample analysis tool in which the easiness of the analysis operation is advantageous, an operation such as the centrifugation treatment is unnecessary. It is desirable.

[0006] In order to solve such a problem, there have been proposed various sample analysis tools as shown below.

Japanese Patent Publication No. 49-33800 discloses a water resistance test film in which a polymer containing a coloring reagent is applied to a plastic film, and this is used as a reagent layer. In this test film, only the plasma component of the whole blood sample moves to the reagent layer and reacts with the coloring reagent, so that it is not necessary to centrifuge the whole blood sample. But,
Red blood cells remain on the sample supply surface of the test film,
At the time of measurement, it is necessary to wipe red blood cells from the sample supply surface, which complicates the operation.

Japanese Patent Publication No. 53-21677 discloses a multilayer test film in which a reagent layer and a sample developing layer are laminated in this order on a liquid-impermeable, light-permeable support layer. According to this test film, since the measurement is performed from the support layer side opposite to the sample development layer, there is no need to wipe the sample. However, since the test film has a laminated structure in which the reagent layer is sandwiched between other layers, when the reagent requires oxygen, the oxygen may be insufficient.

To solve this problem of oxygen deficiency, Japanese Patent Application Laid-Open No. 60-82859 discloses a test film having an oxygen-permeable protein-impermeable light-blocking layer between a reagent layer and a sample developing layer. Is disclosed. However, this test film has a complicated structure.

[0010] In the sample analysis device disclosed in Japanese Patent Application Laid-Open No. 60-205364, a reagent layer and a sample development layer are laminated, and the reagent layer and the support layer are formed of a porous and hydrophobic oxygen supply. They are stacked via layers. However, since the oxygen supply layer needs to hold a sufficient amount of oxygen, it needs to have a certain thickness or more, and depending on the thickness, the light transmittance from the support layer side may be insufficient. . In this case, measurement must be performed from the sample supply side, but an operation of removing the sample such as wiping is required.

The sample analysis device disclosed in Japanese Patent Application Laid-Open No. 63-101775 has a porous reagent layer and a support having through holes. According to this sample analysis device, sufficient oxygen is supplied to the reagent layer from the through hole, and the measurement is performed from the side opposite to the sample supply side, so that it is not necessary to wipe the sample. Furthermore, if this sample analysis tool is combined with a measuring device, automatic detection becomes possible,
Accuracy and simplicity in measurement can be significantly improved. However, in this sample analysis device, two-wavelength measurement is indispensable in order to eliminate the influence of red blood cells, and the structure of the measuring device and the method of correcting the measured value are complicated.

[0012] The sample analysis tool disclosed in Japanese Patent Application Laid-Open No. Sho 63-172962 includes a plurality of reagent layers, a sample supply region, a detection region, an absorption region, and the supply region and the absorption region on a support layer. It has a carrier band to connect. However, since this sample analysis tool has a complicated laminated structure and the gaps of the transport band and the like require considerable accuracy, the analysis accuracy may not be stable.

On the other hand, recently, as shown below, a sample analysis tool using a porous membrane whose average pore diameter changes in the thickness direction has been proposed. The change in the average pore diameter is called asymmetry.

Japanese Patent Application Laid-Open No. 3-56856 discloses a sample analysis tool in which an asymmetric porous membrane whose average pore diameter continuously decreases in the thickness direction is laminated on a support layer. According to this sample analysis tool, the blood sample
Red blood cells are separated when moving in the porous membrane in the thickness direction by capillary action. However, in this sample analysis device, since the sample is moved in the thickness direction to separate red blood cells, there is a limit to the distance between the sample introduction section and the analysis section, and there is a possibility that the effect of red blood cells cannot be sufficiently eliminated. is there.

[0015] In addition, Japanese Patent Application Laid-Open Nos. Hei 6-47953 and Hei 7-72145 disclose sample analyzing tools having the asymmetric porous membrane. A complicated layered structure is required, and for the same reason as described above, the effects of red blood cells may not be sufficiently eliminated.

[0016]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a sample analyzing tool which has a simple structure and can quickly and easily analyze a target component in a sample with high accuracy.

[0017]

In order to achieve the above object, a sample analysis device of the present invention provides a sample analysis device in which a sample is placed in a porous layer having a sample introduction portion and an analysis portion in a direction perpendicular to a thickness direction thereof. In addition, the porous layer moves by capillary action, and the average pore diameter of the sample introduction section of the porous layer is larger than the average pore diameter of the analysis section of the porous layer.

In the sample analysis device of the present invention, the sample moves in the porous layer in a direction perpendicular to the thickness direction thereof,
During this movement, the components are separated, so that the length of the sample introduction section and the analysis section can be increased regardless of the layer thickness. Therefore, for example, when whole blood is used as a sample, the sample analysis device of the present invention does not complicate its structure.
Further, even if the thickness of the porous layer is further reduced, the influence of red blood cells can be sufficiently eliminated. Further, in the sample analysis device of the present invention, if the average pore diameter in the thickness direction of the porous layer is made uniform, the components are not separated in the thickness direction. Measurement can be performed from any surface.

In the sample analysis device of the present invention, the analysis means is not particularly limited, and for example, means such as optical means and electrochemical means can be applied. Examples of the optical means include measurement of transmitted light, measurement of reflected light, and measurement of fluorescence intensity. Examples of the electrochemical means include a method of measuring a current change and a potential change based on a redox reaction of a sample or a reagent.

In the sample analysis device of the present invention, it is preferable that the pore size distribution of the porous layer is a pore size distribution in which the average pore size decreases continuously or discontinuously along the moving direction of the sample. In addition, it becomes discontinuously small,
For example, it refers to a case where the size gradually decreases.

In the sample analysis device of the present invention, the average pore size of the sample introduction portion of the porous layer is 3.0 to 100.
The diameter is preferably in the range of 0 μm, more preferably 10.
The range is from 0 to 50.0 μm. The average pore size of the analysis part of the porous layer is preferably in the range of 0.01 to 10.0 μm, more preferably 0.05 to 1.0 μm.
It is in the range of μm.

In the sample analysis device of the present invention, it is preferable that the porous layer has a reagent disposing portion. If the reagent is arranged on the porous layer, when the sample moves in the porous layer by capillary action, the reagent and the target component in the sample can contact and react in the reagent disposing portion. Because.

The location of the reagent placement section is not particularly limited, and is appropriately determined depending on the target component, the type of reagent, and the like. In the sample analysis tool of the present invention, the analysis section also serves as the reagent placement section. Is preferred. Further, in addition to the analysis unit, a reagent placement unit may be provided between the sample introduction unit and the sample introduction unit and the analysis unit.

[0024] In the sample analysis device of the present invention, it is preferable that the porous layer includes two or more reagent disposing portions in series along the moving direction of the sample. According to this, when a reagent composed of two or more types of components and a reagent that cannot mix the respective components before reacting with the sample is used, when the sample component moves in the porous layer, This is because each of the reagents can be brought into contact with the target component in the sample in the order of the reagent disposing portions.

[0025] In the sample analysis device of the present invention, the porous layer may include two or more reagent disposing portions arranged in parallel with the moving direction of the sample and two or more analysis portions arranged in parallel with the moving direction of the sample. It is preferable to provide a part. According to this, the same sample can be simultaneously analyzed (hereinafter, referred to as “multi-analysis”) for a plurality of analysis items.

The number of the analysis items in the sample analysis device of the present invention is not particularly limited, but is usually 2 to 10 items.

[0027] In the sample analysis device of the present invention, it is preferable that a chromogenic reagent is disposed in the reagent disposing portion.

[0028] In the sample analysis device of the present invention, it is preferable that the chromogenic reagent is arranged by at least one method selected from the group consisting of a printing method, a coating method, an impregnation method and a spraying method.

When the immunoassay is applied to the sample analysis device of the present invention, it is preferable that a labeled antibody for the analyte and a carrier-immobilized antibody for the analyte are arranged in the reagent disposing portion. According to this sample analysis device, the conjugate of the labeled antibody, the analyte and the immobilized antibody can be separated from the unbound labeled antibody by the porous layer, and the conjugate The analyte can be measured by measuring the label of or the label of the unbound labeled antibody.

In the sample analysis device of the present invention, it is preferable that the label is at least one label selected from the group consisting of a radioactive label, a fluorescent label, a chromogenic label, and a specific wavelength light absorbing label.

In the sample analysis device of the present invention, when a sample is analyzed by the electrochemical means, it is preferable that an electrode is arranged in the analysis section. The electrode is not particularly limited and is appropriately determined depending on the type of reaction system between the reagent and the target component, and examples thereof include a hydrogen peroxide electrode, an oxygen electrode, and a pH electrode.

In the sample analysis device of the present invention, it is preferable that the porous layer is disposed on a support layer. According to this, a sample analyzer having a sufficient strength can be obtained regardless of the strength of the porous layer.

In the sample analysis device of the present invention, it is preferable that the support layer has a through hole for supplying oxygen. According to this, when the reaction system between the reagent and the target component requires oxygen, sufficient oxygen can be supplied.

[0034] In the sample analysis device of the present invention, when the sample is whole blood, the porous layer has a region of an average pore size through which red blood cells cannot pass between the sample introduction part and the analysis part. Is preferred. According to this, for example, plasma and red blood cells can be separated, and the influence of red blood cells can be eliminated.

It should be noted that the sample analysis device of the present invention is not limited to the analysis of blood components except for the red blood cells, and the red blood cells may be analyzed in some cases.

Further, in the analytical device of the present invention, the specimen to be analyzed is not limited to the whole blood as long as it can move in the porous layer by capillary action.
Examples include urine, cerebrospinal fluid, saliva, plasma, serum and the like.

[0037]

Next, an embodiment of the present invention will be described. Note that the present invention is not limited to the embodiments described below.

(Embodiment 1) FIG. 1 shows an example of a sample analysis tool of the present invention. FIG. 1 (a) is a perspective view schematically showing the sample analysis tool, and FIG. 1 (b) is a perspective view of I in FIG. 1 (a).
FIG. 1C is an enlarged sectional view schematically showing a portion B in FIG. 1B.

As shown in FIG. 1A, the sample analysis device 1 has a configuration in which a rectangular plate-shaped porous layer 11 is laminated on a support layer 12. A part of the one end surface (the left surface in the figure) of the porous layer 11 is a circular sample introduction part 13 and the other end surface (the right surface in the figure).
Is a circular analysis unit 14. Further, the analysis unit 14 also serves as a reagent placement unit. As shown in FIG. 1C, the average pore diameter of the porous layer 11 is continuously reduced along the arrow A direction. FIG.
As shown in (b), the support layer 12 has a through-hole 18 at a position corresponding to the reagent disposition portion 14 of the porous layer 11.

The sample analysis device 1 can be usually manufactured by bonding and laminating the support layer 12 and the porous layer 11 with an adhesive or ultrasonic vibration welding. The lamination method is not particularly limited, and besides this, for example, lamination may be performed using a double-sided tape or the like. Further, a porous layer may be formed directly on the support layer.

The size of the sample analysis device 1 is appropriately determined depending on the type of analysis and the like.
00.0 mm, overall width 1.0-30.0 mm, overall thickness 100.0-5000.0 μm.

The material for forming the porous layer 11 includes, for example, polysulfone, polyamide, polyester, polyolefin, cellulose and the like. Among them, polysulfone is preferred because of ease of production and moderate rigidity. Is preferred. The material is not limited to one type, and two or more types may be used in combination. Further, other materials may be contained as long as they do not interfere with the sample analysis device of the present invention.

The porous layer 11 may be manufactured using, for example, the above-described materials, or a commercially available porous film, for example, a porous film manufactured by Memtech Japan Co., Ltd. may be used.

For example, in a case where whole blood is used as a sample and red blood cells are analyzed for plasma components, if the porous layer 11 includes a region having an average pore diameter equal to or less than the maximum diameter of red blood cells, the porous layer 11 may have an average pore diameter. The pore size is not particularly limited, but is preferably, for example, in the range shown below. The porous layer 11
The average pore diameter of the sample introduction part 13 is usually 3.0 to 10
0.0 μm, more preferably from 10.0 to
The range is 50.0 μm. The average pore size of the analysis part 14 of the porous layer 11 is usually 0.01 to 10.0 μm.
m, more preferably 0.05 to 1.0 μm
m.

The total length of the porous layer 11 is appropriately determined according to the set distance between the sample introduction section 13 and the analysis section 14. In the present embodiment, the total length is usually 10.0 to 100.
0 mm, more preferably 20.0 to 5 mm.
It is in the range of 0.0 mm. Further, the overall thickness is usually in the range of 100.0 to 2000.0 μm, more preferably in the range of 100.0 to 1000.0 μm, and the overall width is in the range of 1.0 to 30.0 mm, more preferably. Is
It is in the range of 1.0 to 20.0 mm.

The size of the sample introduction section 13 is not particularly limited, and is usually a circle having a diameter of 2.0 to 20.0 mm. In this sample analysis device 1, a part of the surface of the porous layer 11 is used as it is as the sample introduction part 13. In addition, for example, a porous material such as filter paper may be provided on a part of the surface of the porous layer. May be arranged, and the specimen may be introduced through this.

The size of the analysis section 14 is usually a circle having a diameter of 1.0 to 20.0 mm, but is not limited thereto.

The distance between the sample introduction unit 13 and the analysis unit 14 is appropriately determined according to the type of the sample and the target component.
Usually, it is in the range of 10.0 to 100.0 mm.

The shape of the reagent disposing portion 14 is not particularly limited, and examples thereof include a circular shape, an elliptical shape, and a band shape. Among them, the band shape is preferable. In the case of the band shape, the length direction is preferably substantially perpendicular to the direction of arrow A. Usually, the length of the strip-shaped reagent disposition portion (the length in the vertical direction) is 1.0.
２20.0 μm, and its width (length in the direction of arrow A) is 1.0-20.0 μm.

The reagent to be placed in the reagent placement section 14 is not particularly limited, and is appropriately determined depending on the type of the target component and the like. Examples of the components of the reagent include enzymes such as glucose oxidase and peroxidase, buffer substances such as phosphates and carbonates, coloring agents such as tetramethylbenzine and o-tolidine, and high-potentials such as hydroxypropylcellulose and polyvinylpyrrolidone. Molecular polymers and the like. These various components can be used as the reagent by combining them according to the target components.

The method for disposing the reagent in the reagent disposing section 14 includes, for example, a printing method, a coating method, an impregnation method, as described above.
Although a spraying method and the like can be mentioned, a printing method is particularly preferable because the reagent can be arranged in a spot shape. The placement of the reagent in the reagent placement unit 14 by the printing method can be performed, for example, by applying a small amount of reagent to the same location using an inkjet method and repeating the operation of drying the reagent a plurality of times. .

In this embodiment, the reagent placement section 14 is provided at the same location as the analysis section 14,
As mentioned above, this is not a limitation. Further, when a reagent is not required, for example, when the target component itself shows light absorbency at a specific wavelength, the reagent placement section may not be provided.

The material for forming the support layer 12 is as follows.
For example, besides paper and glass, polyethylene terephthalate (PET), polyvinyl chloride, acrylic resin, acrylonitrile-butadiene-styrene copolymer (ABS)
And other plastics can be used. Among them, PET, polyvinyl chloride, acrylic resin, ABS, and the like are preferable in terms of strength, dimensional stability, and the like. As for the plastic, those molded into a film, a plate or the like are preferable. In addition, the said material is not limited to any one kind, and may use two or more kinds together.

For example, when the measurement is performed by optical means from the support layer 12 side, it is preferable that at least a portion of the support layer 12 corresponding to the analysis section 14 is light-transmissive. In this case, it is preferable to use a transparent material such as polystyrene, acrylic resin, or polyethylene terephthalate as the material. Note that the material is not particularly limited as long as the support layer 12 has a through hole 18 at a position corresponding to the analysis unit 14 so that light irradiation can be performed.

The thickness of the support layer 12 is usually 10.0
To 4000.0 μm, preferably 30.
The range is from 0 to 1000.0 μm. Further, the total length and the total width are appropriately determined according to the size of the porous layer.

The location of the through-holes 18 in the support layer 12 is not particularly limited as long as it corresponds to the reaction site between the target component of the porous layer 11 and the reagent. It is provided in the place where it does.

The size of the through-hole 18 is appropriately determined according to the size of the reagent disposing portion 4, and usually has a diameter of 1 mm.
２０20 mm, preferably 1-10 m in diameter
m.

Next, the method of using the sample analyzing device 1 will be described with reference to an example in which whole blood is used as the sample 15 and the target component in the plasma is analyzed. In this example, the sample analyzing device 1 has a region in which a color-forming reagent is disposed in the analysis section 14 and a region having an average pore diameter smaller than the maximum diameter of red blood cells between the sample introduction section 13 and the analysis section 14 of the porous layer 11. Having.

That is, first, the whole blood sample 15 is dropped into the sample introduction section 13 of the sample analysis tool 1. The drop of the whole blood sample 15 is caused by an arrow A in the porous layer 11 due to the capillary phenomenon.
Move in the direction. During this movement, the red blood cells stop on the way, and only the plasma component reaches the analyzer 14. Then, the target component in the plasma reacts with the coloring reagent to form a color. This color development is measured with an optical device such as a densitometer. For example, the analyzer 14 is irradiated with incident light 16a from a densitometer, and the reflected light 16b is detected by a detector 17 of the densitometer, and the color development is measured. Then, if there is a calibration curve or the like, it is used to calculate the amount of the target component in the plasma from the measured values.

(Embodiment 2) FIG. 2 shows an example of a sample analysis tool provided with two reagent placement sections in series along the moving direction of the sample. Unless otherwise indicated, this sample analysis device has the same configuration as the sample analysis device of the first embodiment, and the same reference numerals in FIG. 2 denote the same parts as in FIG. .

As shown in FIG.
The first reagent disposing unit 25 is provided between the sample introducing unit 13 and the analyzing unit 24, and the analyzing unit 24 also serves as the second reagent disposing unit 24.

The sample analysis device 2 is useful when two or more types of reagents are used and they cannot be mixed in advance. Examples of such reagents include enzyme-substrate reagents and the like, for example, trypsin and its substrate-based reagents, and the substrate is usually a chromogenic substrate that generates a color product by an enzyme reaction. it can.

The distance between the first reagent disposition portion 25 and the second reagent disposition portion 24 is not particularly limited as long as there is no risk that the reagents are mixed with each other, but is usually preferably in the range of 1.0 to 20.0 mm. .

The size of the sample analysis device 2 is usually 10.0 to 100.0 mm in overall length and 1.0 to 30.0 mm in overall width.
mm, and the total thickness is 100.0 to 5000.0 μm.

Next, the method of using the sample analysis device 2 will be described with reference to an example in which whole blood is used as a sample and a trypsin inhibitor in plasma is analyzed. In this example, trypsin is placed in the first reagent placement section 25, and its chromogenic substrate (eg, benzoyl-arginine-p-nitroanilide) is placed in the second reagent placement section (analysis section) 24, respectively. You.

That is, first, a whole blood sample is dropped into the sample introducing section 13. The sample moves in the direction of arrow A in the porous layer 11 due to capillary action. During this movement, the sample component that has reached the first reagent placement section 25 first dissolves the trypsin reagent, and the trypsin reagent moves with the sample component. During this movement, trypsin
It is inhibited depending on the amount of trypsin inhibitor in plasma. Next, these are used as the second reagent placement unit (analysis unit) 24.
, Where uninhibited trypsin reacts with the chromogenic substrate to develop color. This color development is measured in the same manner as in the first embodiment. Then, using a calibration curve or the like, the trypsin activity was determined from the measured values, and further,
From this activity value, the amount of trypsin inhibitor in plasma is determined.

In the present embodiment, the second reagent placement section 24 is located at the same place as the analysis section 24, but the present invention is not limited to this. The first and second reagent placement units may be placed between the analysis unit 24 and the first reagent placement unit 25.
May be arranged at the same location as the sample introduction unit 13.

Further, in this embodiment, an example in which the reagent disposing portions are provided at two places is shown, but the present invention is not limited to this, and two or more A part can be provided.

If reagents for each analysis item are placed in a plurality of reagent placement sections and the reaction of these reagents is measured, multiple analysis can be performed for one sample.

(Embodiment 3) FIG. 3 shows an example of a sample analysis tool for multi-analysis. Unless otherwise indicated, this sample analysis device has the same configuration as the sample analysis device of the first embodiment, and the same reference numerals in FIG. 3 denote the same parts as in FIG.

As shown in the figure, in the sample analyzing device 3, the first analyzing section 34 is arranged in parallel with the moving direction A of the sample.
And a second analyzer 35, wherein the first analyzer 34 also serves as the first reagent placement unit 34, and the second analyzer 35 also serves as the second reagent placement unit 35. Arrow C in the figure is
The arrow indicates the direction in which the sample moves to the first analyzer 34, and the arrow D indicates the direction in which the sample moves to the second analyzer 35.

Reagents are arranged in the analysis sections 34 and 35 according to the analysis items.

In the sample analysis device 3, the overall size is appropriately determined according to the number of analysis items. In this embodiment, since there are two analysis items, the total length is usually 10.0 to 100.0 mm, and the overall width is 1.0 to 30.0 m.
m, and the total thickness is 100.0 to 5000.0 μm.

The distance between the first analysis unit 34 and the second analysis unit 35 is not particularly limited as long as it does not affect the detection of each analysis item, but is usually 1.0 to 20.0.
mm.

Next, this multi-analyte sample analysis device 3
The method of using is described using an example in which whole blood is used as a sample and a target component in plasma is analyzed by two types of reaction systems. In this example, reagents of different reaction systems are arranged in the first reagent arrangement section 34 and the second reagent arrangement section 35, respectively.

That is, first, a whole blood sample is added to the sample introduction section 33 of the sample analysis tool 3. For example, the sample may be added in a band shape in the direction perpendicular to the arrow direction A, or may be dropped in a spot shape according to the number of analysis items. Then, the sample moves in the porous layer 11 in the arrow C direction and the arrow D direction, and reacts with each reagent in the first reagent placement section and the second reagent placement section,
Color develops. The color-developed sample analysis tool 3 is provided to an optical measuring instrument capable of performing multi-analysis, and each of the first analysis section 34 and the second analysis section 35 is irradiated with light, and each color is subjected to the first analysis. The measurement is performed in the same manner as in the embodiment.

In this embodiment, an example is shown in which two analysis sections are provided. However, the sample analysis tool of the present invention is not limited to this. An arrangement can be provided.

(Embodiment 4) Next, an example of a sample analysis device using an immunoassay will be described. The sample analysis tool has the same configuration as the sample analysis tool of the first embodiment shown in FIG. 1 unless otherwise specified.

In this sample analysis device, a sample introduction section and an analysis section are provided on the surface of the porous layer, and the sample introduction section also serves as a reagent placement section. A labeled antibody and a carrier-immobilized antibody are arranged in the reagent disposing portion.

The antibody can be prepared by a conventional method.
Immobilization of the antibody on a carrier and labeling of the antibody,
It can be performed by a conventional method.

The carrier is not particularly limited.
For example, beads, latex (microcapsules) and the like can be used. The size of the carrier is appropriately determined depending on the pore size distribution of the porous layer and the like.
１10.0 μm, preferably with a diameter of 0.0
It is in the range of 2 to 1 μm.

As the label, various labels as described above can be used. For example, as the specific wavelength light absorbing label,
Latex, colloidal gold and the like can be mentioned. The latex can bind to the antibody by a carbodiimide bond, and the colloidal gold can bind to the antibody by physical adsorption. The size of the label is not particularly limited, but is usually in the range of 0.01 to 1 μm.

When the label is a color label, for example, it is preferable to further provide a reagent placement section at the same location as the analysis section, and to place a detection reagent for detecting the color label. The detection reagent is not particularly limited as long as it is a reagent system capable of detecting the label.

Next, an example of a method of using the sample analysis device using the immunization method will be described with reference to FIG. In FIG. 4, the same portions as those in FIG. 1 are denoted by the same reference numerals.

In this example, whole blood is used as the sample 44, and the target component in the plasma is the target antigen 43. And
The sample introduction part also serves as a reagent placement part, in which the carrier-immobilized antibody 41 and the labeled antibody 42 are placed. Further, a region having an average pore diameter smaller than the maximum diameter of the carrier of the immobilized antibody 41 is provided between the sample introduction part and the analysis part of the porous layer 11.

That is, first, as shown in FIG. 4A, a sample 44 containing the target antigen 43 is dropped into the sample introduction section, and as shown in FIG. A conjugate in which the labeled antibody 42 and the immobilized antibody 41 are bound to the antigen 43 is generated. The sample containing the sample moves in the direction of arrow A in the porous layer 11. In the course of this movement, the conjugate and the labeled antibody 42 not bound to the antigen 43 are separated according to the pore size of the porous layer 11, and as shown in FIG. Only the labeled antibody 42 moves to the analysis section. Then, the analyzer 16 is irradiated with the incident light 16a, and the reflected light is measured. Thus, the amount of the unbound labeled antibody 42 is measured, and the amount of the antigen can be determined based on the measured value.

In this example, the unbound labeled antibody 42
Was measured, but the labeled antibody 42 bound to the immobilized antibody 41 via the antigen 43 may be measured. in this case,
In the porous layer 11 of the sample analysis device, a region where the conjugate is not permeated and accumulates is defined as an analysis unit, and measurement is performed here.

In the present embodiment, each antibody is arranged in one reagent disposing portion as described above, but is not limited thereto. For example, the antibodies may be arranged in series along the moving direction of the sample. May be provided and arranged separately.

(Embodiment 4) Although the sample analysis device of the above-described embodiment has been described centering on analysis by optical means, as described above, the sample analysis device of the present invention is an electrochemical device. Means are also applicable. The sample analysis tool using the electrochemical means has the same structure as the sample analysis tool using the optical means of Embodiment 1 and the like unless otherwise specified.

In the sample analysis tool using such an electrochemical means, it is preferable that electrodes are arranged in the analysis section, and that the analysis section also serves as a reagent arrangement section. The location of the electrode is not particularly limited as long as it can be measured, and may be any of the upper surface and the lower surface of the porous layer.If the electrode has a support layer, it may be between the porous layer and the support layer. .

The above-mentioned electrode usually comprises a working electrode, a counter electrode and a terminal portion. The material for forming the electrode is, for example,
There are silver chloride and the like. The electrode can be formed using the above-described material by a screen printing method, a photolithography method, or the like.

Usually, the outer diameter of the working electrode is 1.0 to 10.0 mm, and the outer diameter of the counter electrode is 1.0 to 1 mm.
0.0 mm, the gap width between the two electrodes is 0.5 to 10.0 m
m. The length of the entire electrode including the terminal portion is usually 10.0 to 100.0 mm. The shape of the electrode is not particularly limited.

The type of the electrode is appropriately determined depending on the type of the target component or the reagent. For example, a hydrogen peroxide electrode, an oxygen electrode, a pH electrode or the like as described above can be used.

When the porous layer and the support layer are analyzed by electrochemical means, they do not need to be light-transmissive and may be colored.

The reagent to be placed in the reagent placement section is
It can be appropriately determined according to the type of the target component, and for example, the above-mentioned enzymes, buffer substances and the like can be used.

For example, when the target component for analysis is glucose, glucose oxidase can be used as the reagent, and a reaction for generating hydrogen peroxide in the presence of water and oxygen is catalyzed as shown below.

Glucose + oxygen + water → gluconic acid +
hydrogen peroxide

The glucose can be measured, for example, by measuring the amount of the generated hydrogen peroxide or the amount of consumed oxygen. The amount of hydrogen peroxide can be determined, for example, by using the hydrogen peroxide electrode and measuring a current generated by oxidizing the hydrogen peroxide by the electrode and a change amount thereof. Further, the oxygen amount can be obtained by using the oxygen electrode and measuring a current generated by reducing oxygen by the electrode and a change amount thereof. Alternatively, the glucose can be measured by measuring a pH change due to the production of gluconic acid by a voltage change of a pH electrode.

[0099]

As described above, the sample analysis device of the present invention can quickly and easily measure a target component in a sample accurately without requiring a complicated laminated structure. Therefore, if this sample analysis tool is applied to the field of clinical medicine, for example, for blood tests, urine tests, immunological tests, etc., a large number of samples can be analyzed in a short time and collectively, and the efficiency of various diagnoses can be improved. Can greatly contribute to

[Brief description of the drawings]

1 (a) is a perspective view of an embodiment of the sample analysis device of the present invention, (b) is a cross-sectional view of (a), (c) is B of (b).
It is an expanded sectional view of a part.

FIG. 2 is a plan view of another embodiment of the sample analysis device of the present invention.

FIG. 3 is a plan view of still another embodiment of the sample analysis device of the present invention.

FIGS. 4A to 4C are cross-sectional views showing an analysis process in still another embodiment of the sample analysis device of the present invention.

[Explanation of symbols]

 1, 2, 3 Sample analysis tool 11 Porous layer 12 Support layer 13, 33 Sample introduction section 14 Analysis section or reagent placement section 15, 44 Sample 16a Incident light 16b Reflected light 17 Detection section 18 Through hole 24 Analysis section or second section Reagent placement section 25 First reagent placement section 34 First analysis section or first reagent placement section 35 Second analysis section or second reagent placement section 41 Immobilized antibody 42 Labeled antibody 43 Antigen

────────────────────────────────────────────────── ─── Continuation of front page (56) References JP-A-3-56856 (JP, A) JP-A-6-88816 (JP, A) JP-A-1-302161 (JP, A) JP-A-1- 158350 (JP, A) JP-A-10-78433 (JP, A) JP-A-10-78432 (JP, A) JP-A-6-74853 (JP, A) JP-A-7-72145 (JP, A) (58) Field surveyed (Int. Cl. ⁷ , DB name) G01N 33/48-33/98

Claims (15)

(57) [Claims] A sample moves in a porous layer having a sample introduction part and an analysis part in a direction perpendicular to a thickness direction thereof by capillary action, and an average pore diameter of the sample introduction part of the porous layer is: A sample analysis tool that is larger than the average pore size of the analysis section of the porous layer. 2. The sample analysis tool according to claim 1, wherein the pore size distribution of the porous layer is such that the average pore size decreases continuously or discontinuously along the moving direction of the sample. 3. An average pore diameter of the sample introduction part of the porous layer is in a range of 3.0 to 100.0 μm, and an average pore diameter of the analysis part of the porous layer is 0.01 to 10.0 μm. The sample analysis device according to claim 1 or 2, wherein the sample analysis device has a range. 4. The sample analysis tool according to claim 1, wherein the porous layer includes a reagent disposing portion. 5. The analyzer according to claim 4, wherein the analyzer also serves as a reagent arranging unit.
The sample analysis tool according to the above. 6. The sample analysis tool according to claim 4, wherein the porous layer includes two or more reagent disposing portions in series along the moving direction of the sample. 7. The method according to claim 4, wherein the porous layer comprises two or more reagent disposing portions arranged in parallel with the moving direction of the sample and two or more analyzing portions arranged in parallel with the moving direction of the sample. The sample analysis tool according to any one of the preceding claims. 8. The sample analysis tool according to claim 4, wherein a chromogenic reagent is arranged in the reagent arrangement section. 9. The sample analysis tool according to claim 8, wherein the chromogenic reagent is arranged by at least one method selected from the group consisting of a printing method, a coating method, an impregnation method and a spraying method. 10. The sample analysis tool according to claim 4, wherein the labeled antibody for the analyte and the carrier-immobilized antibody for the analyte are arranged in the reagent disposing section. 11. The sample analysis device according to claim 10, wherein the label is at least one label selected from the group consisting of a radioactive label, a fluorescent label, a chromogenic label, and a specific wavelength light absorbing label. 12. The sample analysis tool according to claim 1, wherein an electrode is arranged in the analysis section. 13. The device for analyzing a sample according to claim 1, wherein the porous layer is disposed on the support layer. 14. The sample analysis device according to claim 13, wherein the support layer has a through hole for supplying oxygen. 15. The sample analysis tool according to claim 1, wherein the porous layer has a region having an average pore diameter through which red blood cells cannot pass between the sample introduction section and the analysis section.