JP2002162397A - Dry type analyzing element - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an analyzing device which allows simple and quick measurement with a small amount of analyte and agglutination reaction, coagulation reaction, and the like are measured. SOLUTION: A dry type analyzing element is provided where a hydrophilic polymer layer is provided on an impervious supporting body, on which a development layer comprising a porous material is provided with a spacer in between.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an analytical instrument which can quickly and easily analyze a small amount of a sample in the field of clinical analysis and the like.

[0002]

2. Description of the Related Art Methods of diagnosing human diseases by analyzing blood, urine, etc. as specimens have been used for a long time.

[0003] This analysis method is roughly classified into a wet method and a dry method. In a dry method using a dry analysis element or the like, an analysis utilizing an agglutination reaction or a coagulation reaction may not be performed in some cases.

[0004] In the wet method, a sample and a necessary reagent are put in a container, a detection reaction is performed in a solution, and measurement is performed.

[0005]

The problems of the wet method are that a large amount of sample is required and that the method is not simple and quick.
That is, about 0.1 to 0.5 ml is required for each analysis item. When analyzing a plurality of analysis items, a considerable amount of sample is required. It will be a burden. In addition, since each reagent is added to each container, it takes time and effort, and the size of the entire analyzer becomes large.

On the other hand, in the dry method, there are cases where it is not possible to measure the agglutination reaction and the coagulation reaction.

An object of the present invention is to provide an analytical instrument which can easily and quickly measure a small amount of a sample and can measure an agglutination reaction, a coagulation reaction and the like.

[0008]

A major feature of the dry analysis element lies in the development layer. In other words, the sample supplied to the dry analytical element is spread in a plane without substantially unevenly distributing the components contained in the sample in the developing layer, and is supplied to the layer below it at a substantially constant rate per unit area. By doing so, the stability was ensured, and the development of this spreading layer made it possible to complete the dry analytical element.

Usually, a porous material can be used for the spreading layer having the above function. The porous material may be fibrous or non-fibrous. As the fibrous material, for example, filter paper, nonwoven fabric, woven fabric, knitted fabric, glass fiber filter paper and the like can be used. Examples of non-fibrous materials include membrane filters made of cellulose acetate and the like described in JP-A-49-53888, JP-A-49-53888, and JP-A-55-90859.
No. (corresponding US Pat. No. 4,258,001),
No. 8-70163 (corresponding US Pat. No. 4,486,53)
Any of the continuous void-containing granular structure layers composed of inorganic or organic fine particles described in 7) and the like may be used, but fibrous materials are preferred.

However, the inventors of the present invention have studied that the analysis using the agglutination reaction and the coagulation reaction is performed using a dry analytical element. Was found.

The inventors of the present invention have conducted intensive studies on the reason why a conventional dry analytical element cannot analyze a measurement system including an agglutination reaction and the like. It was found that it was not carried out smoothly and quantitativeness could not be secured. Then, a hydrophilic polymer layer and a developing layer are provided in this order on the support, and a large gap is provided by separating the hydrophilic polymer layer and the developing layer by interposing a spacer between the hydrophilic polymer layer and the developing layer. The present inventors have found that the measurement can be performed even if it includes a coagulation reaction, and have completed the present invention.

That is, the present invention relates to a dry analytical element in which a hydrophilic polymer layer is provided on a water-impermeable support, and a developing layer made of a porous material is provided thereon via a spacer. It is.

In the dry analysis element of the present invention, the supplied sample is developed in the developing layer and transferred to the hydrophilic polymer layer. Since the hydrophilic polymer layer is separated by a space from the developing layer and the spacer, the hydrophilic polymer layer supplied with the sample dissolves or is as if it had melted without being restricted by the movement of the developing layer. And quantitative agglutination and coagulation reactions are obtained.

[0014]

DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows an example of the structure of a dry analytical element according to the present invention. In this dry analysis element, a reagent layer 2 in which a reagent necessary for analysis is contained in a hydrophilic polymer layer is provided on a water-impermeable support 1, and a developing layer 4 is provided thereon via a spacer 3. ing.

As the water-impermeable support, a known water-impermeable transparent support which has been used in a dry analytical element can be used. More specifically, the thickness is about 50 μm made of polyethylene terephthalate, polycarbonate of bisphenol A, polystyrene, cellulose ester (eg, cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.).
A transparent film of m to 1 mm, preferably about 80 μm to about 300 μm can be used.

If necessary, a known undercoat layer or adhesive layer may be provided on the surface of the support to enhance the adhesion to the hydrophilic polymer layer.

It is preferable that the hydrophilic polymer layer is provided on the support, and that no other layer besides the undercoat layer and the adhesive layer is interposed therebetween. The hydrophilic polymer that is the matrix forming the hydrophilic polymer layer is soluble in known water used in conventional dry chemistry analysis elements,
Various swellable and hydrophilic polymers can be used.
Specific examples include polyvinyl alcohol, polyacrylamide, polyvinylpyrrolidone, polymethylvinyl ether, copolymers of polyacrylamide and polyvinylpyrrolidone, methylcellulose derivatives, starch / acrylate graft copolymer crosslinked products, crosslinked polyacrylic acid, gelatin ( Examples thereof include, but are not limited to, acid-treated gelatin, deionized gelatin, etc., gelatin derivatives (eg, phthalated gelatin, hydroxyacrylate-grafted gelatin, etc.), carboxymethyl starch, and the like. It is preferable that the hydrophilic polymer layer is transparent in principle. The thickness (when dried) of the hydrophilic polymer layer is about 2 to 100 μm, preferably 5 to 50 μm.
About μm is appropriate.

The spacer separates the hydrophilic polymer layer from the developing layer, and has a shape similar to the outer edge of the developing layer or about 5 to 10% smaller than the outer edge of the developing layer. In some cases, the support rod is passed or a net is stretched. The frame is usually in a continuous shape, but may be in an intermittent shape as required. When a developing layer support member such as a support rod or a net is also provided in the frame, it is preferable to take care that the support member does not contact the hydrophilic polymer layer. The distance of the support member from the hydrophilic polymer layer is about 150 to 600 μm at the center, preferably 3 μm.
About 00 to 550 μm is appropriate. As the size of the spacer, the outer diameter (the length of a side in the case of a square, the diameter in the case of a circle) is about 1 to 15 mm, usually about 4 to 12 mm,
Height is about 150-600μm, usually 300-500μ
About m is appropriate. FIGS. 2A to 2D show examples of the shape of the spacer when the dry analysis element is circular. FIG.
The spacer (a) is a frame only. This spacer is formed of a square rod, but may be a round rod. In this way, when the spacer is only a frame, the peripheral edge of the developed layer is bonded to the spacer, or a spacer is provided with a retaining piece so that the developed layer is bent and does not contact the hydrophilic polymer layer. It is preferable to fix and spread the spreading layer on the spacer. The spacer shown in FIG. 2B is an example in which a cross-shaped support rod is provided inside a circular frame. The cross-shaped support rod is thinner than the frame, further thinned toward the center, and separated from the hydrophilic polymer layer. The spacer in FIG. 2C is an example in which a net is stretched in a circular frame. The periphery of the net is fixed to the frame. In the spacer of FIG. 2D, a small circle support rod is provided inside a circular frame, and four portions between the small circle and the circular frame are connected by connecting rods. The spacer may be made of a hydrophobic material such as nylon, polyethylene, Teflon (registered trademark), or polystyrene, but is preferably subjected to a hydrophilic treatment.

The spacer may be adhered to the hydrophilic polymer layer or may simply be placed. The adhesive for bonding can be selected from the hydrophilic polymers exemplified for the hydrophilic polymer layer.

The developing layer is a layer for developing the sample and supplying it to the hydrophilic polymer layer, and can also function as a reflector at the time of colorimetry. Further, it has a function of preventing the supplied sample from scattering at the time of transportation or the like. In this deployment layer,
A woven fabric and a knitted fabric such as polyester can be selected. The thickness of the spreading layer is about 150 to 400 μm, preferably about 150 to 360 μm.

The dry analytical element of the present invention basically comprises a spreading layer, a spacer, a hydrophilic polymer layer and a support, and does not include other layers. However, a functional layer, for example, a water-absorbing layer of a dry analytical element, can be provided between the hydrophilic polymer layer and the support.

The shape of the dry analysis element is not particularly limited, and examples thereof include a square, a hexagon, and a circle. The size of the dry analytical element is the diameter in the case of a circle, 4
In the case of a square, the side length is about 1 to 15 mm, usually 4 to 1
It may be about 2 mm.

In the present invention, the target analyte is not particularly limited. Analytical methods such as enzymes, lipids, inorganic ions, metabolites, proteins, etc., which are usually measured in the field of clinical tests, various biological components such as globulin, antigens, antibodies, drugs, hormones, tumor markers, DNA, RNA, etc. If it is established, it can be analyzed.

The dry analytical element of the present invention preferably contains all the reagents necessary for analysis. This reagent may be the same as a known dry analytical element. All the reagents necessary for the analysis are indispensable reagents, and other reagents are added or deleted as appropriate. As a rule, all of the reagents are applied, dried, or contained in the hydrophilic polymer layer. As a method for containing a reagent, there are a method in which the solution is contained in a reagent solution or a hydrophilic polymer solution and applied to a support, a method in which the solution is sprayed or spotted on the support, a method using a spin coater, and the like. For the drying of the reagent, heat drying, drying under reduced pressure, freeze drying, etc. are performed depending on the thermal stability of the reagent. On the other hand, the reagent may be added together with the specimen (at the front, back, simultaneously) without being contained in advance.

The supply amount of the specimen may be about 5 to 100 μl, usually about 5 to 70 μl.

Incubation conditions may be the same as when a known dry analytical element is used.

After the reaction is completed, the side light is on the upper side, the lower side,
It may be from any of the horizontal directions, and may be either reflection photometry or transmission photometry. When photometry is performed from above the support, the photometry is performed after the development layer is removed. Photometry can also usually use a prism or a mirror.

[0028]

EXAMPLES Example 1 Preparation of Dry Analysis Element 1 180 μm colorless and transparent polyethylene terephthalate (support)
Apply the reagent solution on the carrier
And dried to form a reagent layer. Carboxymethyl starch 5.81 g / m²  Colloidal gold-labeled anti-human hemoglobin antibody 0.21 g / m²

The above analysis element has a diameter of 1 as a spacer.
A plastic sheet of 0.3 mm thickness punched out of a circle of 0.5 cm is adhered with a double-sided tape of 0.05 mm, and then a tricot knitted fabric made by knitting a 36 gauge spun polyester yarn with a double-sided tape of 0.05 mm. Glued. This analytical element is cut into a 12 × 13 mm chip,
A dry analytical element 1 of Example 1 was set in a slide frame described in JP-A-57-6352.

Comparative Example 1 Preparation of Dry Analysis Element 2 Colorless and transparent polyethylene terephthalate of 180 μm
Apply the reagent solution on the carrier
And dried to form a reagent layer. Carboxymethyl starch 5.81 g / m²  Colloidal gold-labeled anti-human hemoglobin antibody 0.21 g / m²

Next, water was supplied to the entire surface of the reagent layer at a supply amount of about 30 g / m ² to wet the surface, and a tricot knitted fabric in which a polyester spun yarn equivalent to 50 denier was knitted with 36 gauge was lightly lightened. The layers were laminated under pressure and dried.

In this way, an integrated multilayer analytical element having a colorless and transparent film of polyethylene terephthalate, a reagent layer and a developing layer in this order was produced. Next, this analytical element was cut into chips of 12 × 13 mm.
A dry analytical element 2 of Comparative Example 1 was accommodated in a slide frame described in Japanese Patent No. 3452.

[Measurement Example 1] Measurement of human hemoglobin Exocell. INC, Hum
An Hemoglobin AO (Hb) was diluted with a 0.2 M aqueous solution of ammonium chloride (pH 6.8) containing 6% polyethylene glycol 6000, and then diluted with 0,100,2.
70 μL of 50, 500, 1000 ng / ml series was spotted. After incubating each analytical element at 37 ° C. for 5 minutes, the reflection density was measured at 540 nm from the support side.

FIG. 3 shows the calibration curve. In the figure, black circles indicate Example 1 and white circles indicate Comparative Example 1.

As shown in the calibration curve, the dry analytical element 1 of Example 1 can secure the slope of the calibration curve four times or more as compared with the dry analytical element 2 of Comparative Example 1, and can perform quantitative measurement of Hb. It has been shown.

[0036]

The dry analytical element of the present invention has both the advantages of the wet method and the dry method, and can perform measurement quickly and easily using a small amount of sample. It has the advantage that agglutination and coagulation reactions can also be measured.

[Brief description of the drawings]

FIG. 1 is a cross-sectional view showing an example of the structure of a dry analysis element of the present invention.

FIG. 2 is a plan view and a sectional view showing an example of the shape of a spacer.

FIG. 3 is a graph showing the results of measurement of the relationship between hemoglobin concentration and coloring density using the dry analysis element of the present invention.

[Explanation of symbols]

 REFERENCE SIGNS LIST 1 support 2 reagent layer (hydrophilic polymer layer) 3 spacer 4 spreading layer

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Kazuya Kawasaki 3--11-46 Izumi, Asaka-shi, Saitama Prefecture Inside Fujisha Shin Film Co., Ltd. (72) Yoshikazu Amano 3--11-46, Izumi-zu, Asaka-shi, Saitama No. Fujisha Shin Film Co., Ltd. (72) Inventor Masato Nagata 6-2-10 Ginza, Chuo-ku, Tokyo Inside Godoshu Seiki Co., Ltd. (72) Inventor Toru Tanaka 6-2-10 Ginza, Chuo-ku, Tokyo F-term (for reference) in Godo Shusei Co., Ltd. 2G042 AA01 BD19 CA10 DA10 FA14 FB07 FC02 2G045 AA13 AA16 CA25 CA26 CB03 DA51 FA11 FB03 GC10 GC11

Claims (2)

[Claims] 1. A dry analytical element in which a hydrophilic polymer layer is provided on a water-impermeable support, and a developing layer made of a porous material is provided thereon via a spacer. 2. The dry analytical element according to claim 1, wherein the hydrophilic polymer layer is a reagent layer containing a reagent necessary for analysis.