JPH02179450A - Testing tool - Google Patents

Abstract

PURPOSE:To shorten a time for analysis by bringing a developing layer and a ground layer thereof into contact with each other without bonding them. CONSTITUTION:A test piece 5a of a laminate formed by laying a reagent layer 3 and a developing layer 4 for developing a specimen uniformly on one surface of a light-transmitting substrate 2 is surrounded to be held by a holding member 8 composed of first and second holding member pieces 6 and 7. the two layers 3 and 4 are integrated with and fixed to each other by a pressure of the member 8 without using a bonding force of a bonding layer as an intermediary. Accordingly, a time required for the specimen developed on the developing layer 4 to permeate into the reagent layer 3 is shortened, the permeability of a gas being necessary for color development reaction is also improved, the speed of the reaction is increased and thus a time required for analysis is shortened.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to a test device for measuring and analyzing components in a specimen, such as the amount of sugar in blood or urine.

<Prior Art> A multilayer analysis sheet for liquid sample analysis is known as a sheet that can easily quantify specific components in a specimen (Japanese Patent Publication No. 61347/1983).

This multilayer analytical sheet consists of 1 or 2 layers containing predetermined reagents in a binder, such as gelatin, on a light-transparent hydrophobic support.
The above reagent layer is coated and formed, and a liquid sample spreading layer made of a hydrophilized fabric is adhered onto the reagent layer.

Further, an analytical function supporting layer such as a color shielding layer or a light reflecting layer may be provided between the reagent layer and the developing layer.

According to such a multilayer analysis sheet, when a sample (blood or serum) is dropped onto the liquid sample development layer, the sample is spread uniformly on the liquid sample development layer, and then the sample permeates into the reagent layer. , a specific component in the sample (e.g. glucose)
reacts with the reagent and develops a color, which is emitted from the support side,
The specific component is quantified based on the intensity of the reflected light.

However, this multilayer analysis sheet connects the reagent layer (or analysis function auxiliary layer) and liquid sample development layer via a separately provided adhesive layer (0.5 to 15p thick) such as gelatin.
Another disadvantage is that the reaction time (the time from dropping the sample until it develops color) is long because the gelatin in the reagent layer adheres and bonds in a wet state. This will be explained in detail as follows.

The principle behind the color development of the reagent layer in blood sugar level measurement is as shown in the formula below: glucose in the sample and oxygen taken in from the outside air are decomposed into gluconic acid and hydrogen peroxide by the enzyme glucose oxidase (GOD). Hydrogen peroxide is decomposed into water and oxygen atoms by an enzyme called peroxidase (POD), and these oxygen atoms react with chromogens to form pigments.

GOD Glucose +02-11 Gluconic acid 1820□0D H20□--[0]+820 [01+Chromogen-Dye However, in conventional multilayer analysis sheets, the reagent layer and the liquid sample development layer are bonded together as described above. Since the 02 is bonded to the reagent layer, the permeability of the 02 is poor, and the 02 from the atmosphere enters the reagent layer.
The feed rate of is slowed down, and as a result, the reaction rate of the reaction shown above is slowed down.

Additionally, if the reagent layer and liquid sample spreading layer are combined, the permeability and permeability of the sample will be poor (and the time it takes for the sample to reach the reagent layer and permeate after being dropped is also long). Become.

Note that these problems are particularly noticeable when the reagent layer and the liquid sample spreading layer are bonded together via the adhesive layer.

<Problems to be Solved by the Invention> An object of the present invention is to eliminate the drawbacks of the prior art described above and to provide a test device with a short reaction time.

〈Means for solving problems〉 Such objects are achieved by the present invention as described below.

That is, the present invention provides a test device for analyzing components in a specimen, which comprises a reagent layer and a developing layer for uniformly spreading the specimen on one side of a light-transmitting support. This test device is characterized in that these are integrally fixed by the pressure of a holding member so that they are on the outside.

Moreover, it is preferable that the holding member is a test device having a structure having a holding part that holds the peripheral edge of the laminate from the support to the spreading layer, and having an opening inside the holding part.

Preferably, the holding member is a test device in which an annular convex portion surrounding the opening is formed on a side of the holding portion that contacts the support body and/or a side that contacts the spreading layer. .

According to the present invention with such a configuration, since the developing layer and the layer below it are not bonded, the time required for the dropped sample to reach and permeate the reagent layer is short, and the color reaction is This improves the permeability of gases such as oxygen, which are necessary for gases, and ensures smooth supply of gases such as oxygen. As a result, the rate of color reaction is accelerated and the time required for analysis is shortened.

<Example> Hereinafter, the test device of the present invention will be described in detail with reference to preferred examples shown in the accompanying drawings.

FIG. 1 is a cross-sectional front view showing an example of the configuration of the test device of the present invention. As shown in the figure, the test device 1a has a reagent layer 3 formed on one side of a support 2, and a spreading layer 4 on the reagent layer 3.
It has a test piece 5a made of a laminate having a configuration in which
The periphery of the test piece 5a is held between a holding member 8 consisting of a pair of first and second holding member pieces 6 and 7.

The support 2 has a plate shape with a thickness of about 20 to 200 mm, for example, and is made of a light-transmitting material, preferably a transparent material.

Specific constituent materials of the support 2 include polyethylene terephthalate, cellulose ester (cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.), polycarbonate of bisphenol A, polymethyl methacrylate, polyvinyl chloride, polypropylene, polystyrene, polyvinyl. Examples include various resins such as alcohol, glass, and the like. Alternatively, sheets made of two or more of the above materials may be laminated.

The reagent layer 3 has a composition in which a predetermined reagent is contained (dispersed) in a binder such as gelatin, polyvinyl alcohol, polvinylpyrrolidone, agarose, sodium polyvinylbenzenesulfonate, polyurethane, or polyvinylpropionate. consists of things.

The coating method for forming the reagent layer, etc. can be any suitable method used in the industry, such as dip coating or extrusion coating.If necessary, two or more layers can be applied at the same time. It is also possible to form by coating, and by changing the composition and coating conditions, it is possible to obtain an arbitrary dry film thickness, for example, 1 to 50 tiles.

The reagent in the reagent layer 3 is appropriately determined depending on the component to be quantified in the specimen. For example, when quantifying glucose in a specimen, the reagents include glucose oxidase, peroxidase, and a chromogen.

In the illustrated example, there is only one reagent layer 3, but a plurality of reagent layers each containing a reagent having a different composition may be laminated.

The spreading layer 4 is located on the outer surface of the test piece 5a, spreads the dropped sample uniformly on the spreading layer, and supplies a substantially constant amount of the sample or components in the sample per unit area to the reagent layer 3. It has the effect of

This spreading layer 4 is made of a non-fibrous or fibrous porous material.

Examples of non-fibrous porous materials include a dispersion in which a porous material such as plush polymer (general name: Menplan filter), diatomaceous earth, microcrystalline material (for example, microcrystalline cellulose (trade name: Avicel of FMC Corporation)) is dispersed in a binder. ,
Examples include porous aggregates made of microspherical beads of glass or resin bonded to each other.

Examples of the fibrous porous agent include woven fabrics, non-woven fabrics, aggregates of short fibers, etc. Among them, the hydrophilized fabric disclosed in Japanese Patent Publication No. 61-61347 is particularly suitable. preferable.

Such a spreading layer 4 is placed on the reagent layer 3, but the two layers 3.4 are in contact with each other without having substantial bonding force by themselves.

That is, an adhesive layer made of a composition having an adhesive function, such as gelatin, is interposed between the reagent layer 3 and the spreading layer 4, so that both layers 3.
4, or by pressing the spreading layer 4 onto the reagent layer 3 in a wet state, and gelatin or the like in the reagent layer 3 binds both layers 3.
4 is excluded.

With this configuration, the time required for the sample developed on the development layer 4 to reach and permeate the reagent layer 3 is shortened, and the permeability of gases such as oxygen necessary for color reaction is also reduced. This increases the reaction rate and reduces the time required for analysis.

In this way, the spreading layer 4 does not have a bonding force with the reagent layer 3 below it, so simply being placed on the reagent layer 3 will cause misalignment, and the reagent layer 3 and the spreading layer 3 may be misaligned. A gap is likely to be created between the two. Therefore, as shown in FIG. 1, the periphery of the test piece 5a is held by the holding member 8, and the spreading layer 4 is pressed and fixed against the reagent layer 3.

This holding member 8 includes first holding member pieces 6 that fit together and hang together.
and a second holding member piece 7, and when both holding member pieces 6 and 7 are fitted, the respective clamping parts 61 and 71 press the peripheral edge of the test piece 5a with a predetermined pressure. There is.

Furthermore, approximately central portions of the first and second holding member pieces 6 and 7 (inside the holding portions 61.71) have openings 62, respectively.
and 72 are formed.

The opening 62 of the first retaining member piece 6 is inserted into the support body 2 by the analyzer.
It is provided as a window for measuring the intensity of coloration of the reagent layer 3 by projecting and receiving light from the side. On the other hand, the open lower portion 2 of the second holding member piece 7 is provided as a space for supplying the specimen to the developing layer 4.

The shapes of these openings 62 and 72 include circular, oval, and regular polygonal shapes.

FIG. 3 is a perspective view showing another example of the structure of the holding member.

The first holding member piece 6 shown in the figure has a structure in which an annular convex portion 63 surrounding an opening 62 is formed on the inside of the holding portion 61 (the side that contacts the support body 2).

The convex portion 63 is an annular portion formed along the peripheral edge of the circular opening 62, and preferably has a flat top.

Further, unlike the illustrated example, two or more annular convex portions having different diameters may be provided concentrically.

Note that the height of the convex portion 63 is preferably about 0.05 to 0.31 IIm.

Further, although not shown, the second holding member piece 7 can also be formed to have a similar shape to the convex portion 63.

In this way, by providing the annular convex portion on one or both of the first and second holding member pieces 6.7, it is possible to reliably prevent the spreading layer 4 and the reagent layer 3 from shifting or floating,
Various parts of the spreading layer 4 (particularly within the open lower part 2) can be bonded to the reagent layer 3 with uniform pressure. As a result, it is possible to prevent unevenness in coloring, thereby reducing measurement errors.

Note that when annular convex portions are formed on both the first and second holding member pieces 6 and 7, their inner and outer diameters are different, and one is shaped so that it fits inside the other. is preferred.

The constituent materials of the first and second holding member pieces 6.7 are not particularly limited, and include various resins such as polyethylene, polypropylene, vinyl chloride, polystyrene, and ABS, or metals such as aluminum and stainless steel. can be mentioned.

Furthermore, in FIGS. 1 and 3, the first holding member piece 6 and the second holding member piece 7 are shown as being constructed separately, but in the present invention, both the holding member pieces are connected,
Alternatively, it may be formed integrally. For example, one side of both the holding member pieces 6.7 may be connected to each other, and this connecting portion may be made flexible so that both holding member pieces can rotate around the connecting portion.

FIG. 2 is a cross-sectional front view showing another example of the configuration of the test device of the present invention. The test device 1b shown in the figure is different from the test device 1a in the configuration of the test piece 5b, but the other configurations are the same.

The test piece 5b has a reagent layer 3 formed on one side of the support 2, and an analysis function auxiliary layer 9 formed on the reagent layer 3.
The structure is such that the development layer 4 is placed on the analysis function auxiliary layer 9. Note that the support 2, the reagent layer 3, and the developing layer 4
This is the same as described above.

For example, when the sample is whole blood, the analysis function auxiliary layer 9 in this reagent piece 5b filters blood cells, which are components that interfere with color detection, and also blocks colored light from the filtered material (blood cells) side. It has the function of shielding or reflecting light from the support side.

The structure of this analytical function auxiliary layer 9 is such that fine particles of titanium oxide, barium sulfate, aluminum, etc. are dispersed in a binder such as gelatin.

The content of the fine particles in the analytical function auxiliary layer 9 is 10 to 90
It is preferable to set it to about wt%.

Further, the thickness of the analytical function auxiliary layer 9 is preferably about 2 to 20 mm.

Note that it is preferable to use the fine particles having a particle size of about 0.1 to 3 mm.

According to such a test device 1b, the spreading layer 4 comes into contact with the analysis function auxiliary layer 9 below it without having any bonding force.

In addition, in the present invention, in addition to or in place of the analysis function auxiliary layer 9, a layer for another purpose may be provided.

In addition, the test device of the present invention can be used for specimens (e.g., whole blood,
It can be applied not only to the analysis of sugar content in body fluids such as serum, urine, and saliva, but also to the analysis of uric acid, GOT, GPT, etc., and can also be applied to other fields such as the analysis of food and environmental samples. is also possible.

Experimental Example (Example of the Present Invention) A solution having the composition shown in Table 1 below was coated on a transparent support made of PET film with a thickness of 0.1 mm and dried to form a reagent layer with a dry film thickness of about 10 μs. .

Next, this was cut into a size of 1.5 cm x 1.5 cm, and a spread layer of the same size made of a knitted fabric impregnated with a surfactant was placed on the reagent layer, and these were assembled into the configuration shown in FIG. A test device A was obtained by sandwiching the sample between a holding member (mount) consisting of a pair of holding member pieces with convex portions (made of polypropylene) and bringing them into close contact.

(Comparative Example) A solution having the same composition was applied onto the same support as in the present invention example, and a similar developing layer was layered on the solution in an undried state.

Next, a load of approximately 50 g/cm'' was applied for 5 minutes to press the reagent layer and the spreading layer together, allowing the gelatin (binder) in the reagent layer to penetrate into the knitted fabric of the spreading layer, and then drying this. The reagent layer and developing layer were adhered and imaged.

Thereafter, this was cut into 1.5 cm x 1.5 cm sizes to obtain test device B.

Table 1 Composition of coating liquid for reagent layer For the above test devices A and B, sample containing glucose (
The reaction rate was investigated using serum). The detailed procedure is as follows.

First, glucose was dissolved in physiological saline to a concentration of 5000 mg/dl, and heat treated at 37° C. for 2 hours to isomerize the solution, which was used as a glucose stock solution.

Next, the glucose concentration of a commercially available standard serum (manufactured by Bio-Rad, Liphocheck Biochemical Control Serum Engineering) was measured using a commercially available measurement kit (Glucose Test Wako, manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.), and based on this measurement value, the standard Add glucose stock solution to serum, glucose concentration is 100.20 respectively.
0.300 and 400 mg 7 dl were added to prepare four types of serum samples No. 1, 2.3 and 4.

Prepare four test devices A and B, and place the four serum sample numbers on the spreading layer of each test device.
, 1 to 4 were dropped in 10 μρ portions, and the light was projected and received on the support side, and the intensity of reflected light at a wavelength of 565 nm was measured at 30 second intervals.

Note that light was projected onto the support from two directions at an angle of 45° with respect to the support surface, and light was received in a direction perpendicular to the support surface.

In addition, for measurement, an instant multi-measurement spectrometer MC manufactured by Tenba Denshi Co., Ltd.
PD-200 was used.

The measurement results obtained in this way are shown in Figure 4 (example of the present invention).
and shown in the graph of FIG. 5 (comparative example). In addition, the fourth
The figure and the graph in FIG. 5 show the change over time of the logarithmically converted value of the relative reflection intensity after coloring with respect to the reflection light intensity before serum drop, which was measured in advance.

As shown in FIG. 4, in the test device A of the present invention, the difference in glucose concentration among serum samples Nos. 1 to 4 was 30.
It is recognized from about seconds, and clearly appears after 60 seconds.

On the other hand, as shown in Fig. 5, in the test device B of the comparative example, the difference in glucose concentration between serum samples No. 1 to 4 is still small at 60 seconds, and reaches its current level at about 90 to 120 seconds. .

As described above, it can be seen that the test device A of the present invention shows coloration according to the glucose concentration earlier than the test device B of the comparative example, and can perform sugar content analysis in a shorter time.

A test prepared in the same manner as the above-mentioned inventive examples and comparative examples except that a layer containing 49 wt % of titanium oxide powder with an average particle size of 0.3- in gelatin was formed on the reagent layer as an analytical function auxiliary layer. For instruments C and D, the same measurements as above were performed using whole blood instead of standard serum.
The same results as above were obtained, and it was confirmed that the test device C of the present invention having the analysis function auxiliary layer was also capable of conducting analysis in a shorter time compared to the test device of the comparative example.

<Effects of the Invention> As described above, according to the test device of the present invention, the spreading layer and the layer below it do not have substantial bonding force (they are in contact with each other, so penetration of the sample into the reagent layer and The color reaction is promoted, and the time required for analysis can therefore be shortened.

[Brief explanation of the drawing]

FIG. 1 and FIG. 2 are sectional front views each showing an example of the configuration of the test device of the present invention. FIG. 3 is a perspective view showing an example of the configuration of a holding member used in the test device of the present invention. FIG. 4 and FIG. 5 are graphs showing changes over time in the coloring state (relative reflection intensity) of the test devices of the present invention example and the comparative example in the experimental examples, respectively. Explanation of symbols 18% lb...Test device 2...Support 3...Reagent layer 4...Development layers 5a, 5b...Test piece 6...First holding member piece 7... Second holding member piece 61.71...Campling part 62.72...Opening 3...Convex part 8...Holding member 9...Analysis function auxiliary layer

Claims (3)

[Claims] (1) A test device for analyzing components in a specimen, which includes a reagent layer and a developing layer for uniformly developing the specimen on one side of a light-transmitting support, with the developing layer on the outside. A test device characterized in that these are integrally fixed by pressing a holding member in such a manner. (2) The test device according to claim 1, wherein the holding member has a structure in which the holding member has a holding part that holds the peripheral edge of the laminate from the support to the spreading layer, and has an opening inside the holding part. (3) The holding member has an annular convex portion surrounding the opening formed on a side of the holding portion that contacts the support body and/or a side that contacts the spreading layer. test equipment.