JPH0339652A - Testing means - Google Patents

Abstract

PURPOSE:To accelerate coloration reaction and to shorten the time required for analysis and measurement by constituting a carrier to carry a reagent of knitted and woven materials. CONSTITUTION:The testing means 1a to be used mainly for serum, urine, saliva, etc., has a test piece 5a of the laminate on which the reagent carrier 3 to carry the reagent on one surface of a base 2 and the circumference thereof is crimped by a holding member 8 consisting of 1st, 2nd holding members 6, 7. The time when a dropping specimen penetrates over the entire part of a reagent layer is short if this carrier 3 is constituted of the knitted and woven materials. The penetratability of the gas required for the coloration reaction is then improved and the supply is smoothly executed. The rate of the coloration reaction is, therefore, increased and the time required for the analysis is shortened. The production of the testing means to a uniform thickness is possible and the spread of the specimen is sufficient and uniform. The degree of the coloration is stable and the unequal coloration is obviated. The measurement with the higher accuracy is, therefore, possible.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to a test device for measuring and analyzing the amount of components 1 in a specimen, such as cholesterol and sugar in blood or urine.

<Prior Art> A multilayer analysis sheet for liquid sample analysis is known as a device that can easily quantify specific components in a specimen (Japanese Patent Publication No. 61-F31347).

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 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, in this multilayer analysis sheet, since the reagent layer is made by coating a coating solution containing a hydrophilic binder such as gelatin on a support and drying it, the reaction time (time from dropping the sample to color development) is short. The shortcoming is that the length is long and errors may occur in the measurement. 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 (COD), 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.

COD Glucose + O, --- Gluconic Acid Ten HiO OD H, O As shown in the formula below, cholesterol ester in the sample is decomposed into cholesterol and free fatty acids by collosterol esterase (COE).This cholesterol and the cholesterol in the sample are converted into Δ4-cholestenone by cholesterol oxidase (COD). is decomposed into hydrogen peroxide, and this hydrogen peroxide is decomposed into water and oxygen atoms by peroxidase (POD).
This oxygen atom reacts with the chromogen to form a pigment.

COE − Cholesterol ester −−−→Cholesterol I role + free fatty acid OD Cholesterol 10□−−−−−−→ Δ 4-Cholestenone 10, O□OD H*Os −111 [0]+H*0 [01 + Chromogen - Dye However, in conventional multilayer analysis sheets, the reagent layer is formed by applying and drying a coating solution, so the permeability of 02 is poor, and the supply rate of 02 from the atmosphere to the reagent layer is low. becomes slower, and as a result, the reaction rate of the reaction shown above becomes slower.

Furthermore, the permeability and permeability of the specimen in the reagent layer are poor, and it takes a long time for the dropped specimen to permeate the entire thickness of the reagent layer.

Moreover, if the permeability of the specimen into the reagent layer is poor, the spread throughout the reagent layer will be insufficient or non-uniform, and sufficient coloration will not be obtained or coloration will be uneven.

In addition, 1. Since the reagent layer is formed by coating, it is difficult to accurately specify the film thickness after drying (dry film thickness). Therefore, the film thickness varies due to subtle differences in coating conditions (i.e.
If the content of the reagent changes), the degree of coloration will vary accordingly, making it impossible to obtain a product with stable detection accuracy.

Moreover, in a single multilayer analysis sheet, there may be unevenness in the thickness of the reagent layer, resulting in uneven coloring (thick areas are dark colored, thinner areas are light colored). ), which causes measurement errors.

Furthermore, a multilayer analysis sheet having such a configuration is not suitable for analyzing cholesterol in a sample. That is, the reagent layer is
Hydrophilic binders such as gelatin account for most of the binders, but cholesterol is oily and exists in the sample as minute oil globules, making it poorly compatible with the reagent layer and difficult to penetrate.

<Problems to be Solved by the Invention> An object of the present invention is to eliminate the various drawbacks of the prior art described above, and to provide a test device that has a short reaction time and improved measurement accuracy.

Means to solve problems〉 Such objects are achieved by the present invention as described below.

That is, the present invention is a test device for analyzing components in a specimen by reaction with a reagent, wherein the carrier supporting the reagent is made of a woven or knitted fabric.

Moreover, it is preferable that the carrier is a test device having hydrophilic properties.

Moreover, it is preferable that the thickness of the carrier is 50 to 500 J and 11 mm.

According to the present invention having such a configuration, since the carrier supporting the reagent layer is made of a woven or knitted fabric, the time required for the dropped sample to permeate the entire reagent layer is short, and The permeability of gases such as oxygen necessary for color reactions is improved, and their supply is performed smoothly. As a result, the rate of color reaction is accelerated and the time required for analysis is shortened.

In addition, carriers made of woven or knitted fabrics can be manufactured to have a uniform thickness, and the spread of the specimen is sufficient and uniform, so the degree of coloration is stable and there is no unevenness in coloration. Measurements can be taken.

Further, since a carrier made of woven or knitted fabric easily permeates regardless of the nature of the component to be placed in the specimen, it can be used even when analyzing oily components such as cholesterol.

<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 a test device of the present invention. The test device 1a shown in the figure is a test device mainly used for specimens such as serum, urine, saliva, etc., and has a laminated structure in which a reagent carrier 3 carrying a reagent is placed on one side of a support 2. This test piece has a test piece 5a made of a solid body, and the periphery of the test piece 5a is held between a holding member 8 made 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 500 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 carrier 3 is made of woven or knitted fabric.

Here, woven or knitted fabrics include woven fabrics, m-type fabrics, or similar fabrics, and do not include nonwoven fabrics or filter paper.

As the textile structure, all kinds of practically available types, such as plain weave, twill weave, and satin weave, can be used.

Further, there is no particular limitation on the composition of the knitted fabric, and examples thereof include horizontal knitting (flat knitting), warp knitting (tricot knitting), circular knitting, and flat knitting.

The reagent carrier 3 made of such a woven or knitted fabric has higher permeability to the specimen or its components and to gases such as oxygen necessary for the coloring reaction, compared to conventional reagent layers formed by coating and drying a coating liquid. Are better. In particular, it easily penetrates even oily components such as cholesterol, so it is suitable for use as a test tool for detecting such components.

In addition, the reagent carrier in the present invention is preferably one that has regular weaving or knitting, such as a plain woven fabric or a plain knitted fabric.The reason is that the sample permeates uniformly into the reagent carrier 3
This is because uneven coloring can be prevented by keeping the amount of specimen per unit area of the reagent carrier constant.

Examples of fibers constituting such woven or knitted fabrics include natural fibers such as cotton, kapok, flax, hemp, ramie silk, and wool; chemical fibers such as nylon, Tetoron, rayon, acetate, vinylon, acrylic, and polyethylene terephthalate; Alternatively, a blend of two or more of these natural and chemical fibers (for example, a blend of nylon and Tetron) can be used.

Further, although the thickness of these fibers depends on the type thereof, it is usually preferably about 0.1 to 1.0 denier.

It is preferable that the reagent carrier 3 made of such a woven or knitted fabric has hydrophilic properties.
This is because diffusion is promoted.

In such cases, the woven or knitted fabric constituting the reagent carrier 3 is hydrophilic in itself (for example, made of fibers such as cotton, silk, nylon, etc.), or the woven or knitted fabric is hydrophilic. Examples include those that have been treated.

Hydrophilic treatment may be carried out by a simple method such as thoroughly washing a commercially available woven or knitted fabric to remove glue or other processing agents, but preferably by applying a surfactant as described below to the woven or knitted fabric. Good.

Application of a surfactant can be achieved, for example, by immersing the washed fabric in a surfactant solution (e.g. 0.05-5%) or by spraying a similar surfactant solution on the fabric. After that, drying is performed.

As for the type of surfactant, any of water-soluble nonionic (nonionic) cationic, anionic, and amphoteric surfactants can be used. Particularly preferred are nonionic surfactants such as alkylaryl ethers or fatty acid esters of polyoxyethylene or polyglycerin, sorbitan esters, and the like.

Other methods for making woven or knitted fabrics hydrophilic include using an aqueous solution of a hydrophilic polymer such as polyvinyl alcohol, or an aqueous solution of a hydrophilic polymer containing fine powder such as titanium oxide or barium sulfate, or a wetting agent such as glycerin or polyethylene glycol. It is also possible to wet the woven or knitted fabric and then dry it.

In this case, the concentration of the hydrophilic polymer solution is between 0.1 and 5.
It is preferable to set it to about Owt%.

The thickness of the reagent carrier 3 is preferably 50 to 500M, more preferably 100 to 400M, and the thickness is 50JA.
If it is less than 11, the amount of supported reagent is small, so the degree of coloration is reduced. In addition, if the thickness exceeds 500 μm, if the amount of sample is small, the sample may not penetrate the entire thickness of the reagent carrier 3, or it will take time to penetrate, making it difficult to obtain uniform coloration and rapid reaction. Become.

Such a reagent carrier 3 supports a reagent to be described later.

This reagent 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 addition, when quantifying total cholesterol in a sample, cholesterol esterase, cholesterol oxidase,
Contains peroxidases and chromogens.

As a method for supporting the reagent on the reagent carrier 3, for example, the reagent carrier 3 may be immersed in a liquid containing a reagent (for example, 0.1 to 10% chromogen), or a similar liquid may be sprayed on the reagent carrier 3. It is preferable to wet the material and then dry it.

Further, the amount of the reagent in the reagent carrier 3 varies depending on the type of reagent, and the preferable range is illustrated in Table 1 below.

Table 1 Such a reagent carrier 3 may be simply placed on the support 2 (having no binding force with the support), but
It may be bonded or bonded (having bonding strength) to the support 2.

In addition, in the illustrated example, the reagent carrier 3 has only one layer, but
It may also be a stack of a plurality of reagent carriers each containing a different reagent composition.

As shown in FIG. 1, the test piece 5a is held by a holding member 8 around the test piece 5a, and the reagent carrier 3 is pressed and fixed to the support 2.

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 holding member piece 6 is provided as a window through which the analyzer emits and receives light from the support 2 side to measure the intensity of coloring of the reagent carrier 3. On the other hand, the opening 72 of the second holding member piece 7 is provided as a space for supplying the sample to the reagent carrier 3.

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

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 FIG. 1, 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 or For example, both holding member pieces 6 may be integrally formed.
7 may be connected to each other, and by making this connecting portion flexible, both holding member pieces may be configured to 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 ib shown in the figure is a test device mainly used for whole blood, and the configuration of the test piece 5b is the same as that of the test device 1.
This is different from that in a.

Hereinafter, the differences will be explained, and the explanation of similar matters will be omitted.

The test piece 5b has a reagent carrier 3 placed on one side of the support 2 (
The analytical function support layer 4 is further placed (or adhered) on the reagent carrier 3.

Note that the support 2 and reagent carrier 3 are the same as those described above.

The analytical function support N4 in this reagent piece 5b filters blood cells, which are components that interfere with color detection, for example, when the sample is whole blood, and also filters colored light from the filtered material (blood cells) side. It has the function of shielding or reflecting light from the support side.

Specific examples of the analysis function auxiliary layer 4 include filter members such as membrane filter 1 filter paper, woven fabric, and glass fiber.

Note that in the present invention, in addition to or in place of the analysis function auxiliary layer 9, a layer for another purpose (for example, a development layer) may be provided.

In the test devices 1a and 1b described above, as another example of the structure of the holding member, an opening 62.7 is provided on the inside of the holding part 61 and/or 71 (the side that contacts the support 2 or the reagent carrier 3).
An example of this is a structure in which an annular convex portion surrounding the 2 is formed.

In this way, by providing the annular convex portion on one or both of the first and second holding member pieces 6.7, the mutual misalignment of the support 2, the reagent carrier 3, and the analytical function auxiliary layer 4 can be prevented. It is possible to reliably prevent floating or floating, and to join them 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 test device of the present invention is not limited to a structure in which a test piece is held between holding members as in the illustrated example of the structure, but also a test device in which a test piece is used alone, and a reagent carrier is attached to a strip-shaped support. It may be in any form, such as a

Further, if the reagent carrier 3 itself W has a predetermined rigidity, the support 2
It is not necessarily necessary to provide.

The test device of the present invention is used for specimens (e.g., whole blood, serum,
Not only analysis of cholesterol and sugar content in body fluids such as urine and saliva, but also uric acid.

It can be applied to the analysis of GOT, GPT, etc., and can also be applied to other fields such as the analysis of food and environmental samples.

Experimental example> (Example of the present invention) A reagent carrier shown below was placed on a transparent support made of a PET film with a thickness of 1 mm, and a filter member was placed on top of the transparent support, and these were placed with a pair of protrusions. A test device A was obtained by sandwiching and closely adhering a holding member (mount) made of a holding member piece (made of polypropylene).

The sizes of the support, reagent carrier, and filter member were all 1.5 cm×1.5 ca.

[Reagent carrier] Material: 0. Knitted fabric using ID yarn Thickness: 2404 Manufacturing method: The above knitted fabric piece was impregnated with a liquid containing a reagent and a surfactant and having the composition shown in Table 2 below, and dried at 40°C for 60 minutes. A reagent carrier was obtained.

Table [Filter material] Material: Menplan filter (FM-300 manufactured by Fuji Photo Film Co., Ltd.) Thickness: P (Comparative example) The following table was applied as a coating liquid for forming a reagent layer on the same support as in the present invention example. A solution having the composition shown in No. 3 was applied, and a similar filter member was placed on top of the solution in an undried state.

Next, a load of approximately 50 gf/ca+" was applied for 5 minutes to press the reagent layer and the filter member together, allowing some of the gelatin (binder) in the reagent layer to penetrate into the filter member. Dry and adhere the reagent layer and filter part.
It became law. In addition, the dry film thickness of the reagent layer was 6 to 8.

Thereafter, this was cut into a size of 1.5 cm x 1.5 cm to obtain a test device B.

Test devices A and B shown in the table above were colored using a sample containing cholesterol (human whole blood), and the reaction rate was investigated. The detailed procedure is as follows.

First, low-density lipoprotein (LDL) containing cholesterol was fractionated from human plasma to obtain a concentrated LDL solution with a known cholesterol concentration, which was used as a stock solution.

Next, the total cholesterol concentration of human whole blood was measured using a commercially available measurement kit (Determiner TC"555" manufactured by Kyowa Medex), and based on this measurement value, the cholesterol stock solution was added to the human whole blood, and the cholesterol concentration
．． 200, 300 and 400 mg/di, and four types of blood samples No. 1, 2.3 and 4 were prepared.

Prepare four test devices A and B, drop 10μ of each of the four blood samples No. 1 to 4 onto the filter member of each test device, project light from the support side, After receiving the light, the intensity of the reflected light with a wavelength of 560 nm is reduced to 1
Measurements were taken at 0 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 Otsuka Electronics Co., Ltd.
PD-200 was used.

The measurement results obtained in this way are shown in Figure 3 (example of the present invention).
and shown in the graph of FIG. 4 (comparative example). In addition, the third
The figure and the graph in FIG. 4 show changes over time in logarithmically converted values of the relative reflection intensity after coloring with respect to the reflection light intensity before blood dripping, which was measured in advance.

As shown in FIG. 3, in the test device A of the present invention, the difference in cholesterol concentration among blood samples Nos. 1 to 4 was recognized from about 10 seconds and clearly appeared at 40 seconds.

On the other hand, as shown in FIG. 4, in the test device B of the comparative example, the difference in cholesterol concentration among blood samples Nos. 1 to 4 was still small at 40 seconds, and slightly appeared at about 50 seconds.

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

Furthermore, when the reagent carrier of Test Device A and the reagent layer of Test Device B were observed, the former was uniformly colored over the entire area of the reagent carrier, but the latter had spotty uneven coloration.

<Effects of the Invention> As described above, according to the test device of the present invention, since the carrier supporting the reagent is composed of a woven or knitted fabric, "the penetration of the specimen or its components into the reagent carrier and the penetration of the specimen into the reagent carrier" Since the supply of oxygen and the like becomes active, the color reaction is promoted, and therefore the time required for analysis and measurement can be shortened.

In addition, compared to conventional reagent layers formed by coating and drying, the uniformity of coloration is improved, allowing for more accurate analysis and
Measurement becomes possible.

[Brief explanation of drawings]

FIG. 1 and FIG. 2 are sectional front views each showing a configuration example of a test device of the present invention. FIG. 3 and FIG. 4 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 La, 1b...Test device 2...Support 3...Reagent carrier 4...Analysis function auxiliary layer 5a, 5b...Test piece 6...First holding member piece 7. ...Second holding member piece 61.71...Holding part 62.72...Opening 8...Holding member 1G, 1 IG "1G, 3 Wooden edge I 11 Doshimei 1a Terama (Kike)

Claims (3)

[Claims] (1) A test device for analyzing components in a specimen by reaction with a reagent, characterized in that a carrier supporting the reagent is made of a woven or knitted fabric. (2) Claim 1, wherein the carrier is hydrophilic.
Test equipment described in . (3) The test device according to claim 1 or 2, wherein the carrier has a thickness of 50 to 500 μm.