JPH08285849A - Convenient measuring method and apparatus - Google Patents

Abstract

PURPOSE: To obtain an accurate and inexpensive convenient measuring apparatus suitable as a disposable measuring apparatus in which the measurement can be carried out quickly and conveniently by single step operation without requiring an independent B/F separation step. CONSTITUTION: The convenient measuring apparatus comprises a test specimen sampling section 12 including a spacer 20 having a through hole 20a for introducing a test specimen and a liquid permeable membrane disposed while closing the upstream side of the through hole 20a, and a reagent section 14 for retaining a soluble reagent in a porous material disposed downstream. The convenient measuring apparatus further comprises a decision section 16 having at least one reactive region composed of a porous material fixed with a hydrophilic substance disposed downstream, long upstream and downstream sheets 28, 30 disposed farther downstream, and a long absorption sheet 26 composed of a liquid absorbing material held between both sheets, wherein a slit 38 is made in the absorption sheet 26.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to the measurement of an analyte in a liquid test sample by utilizing a specific binding reaction between an analyte and an affinity substance having an affinity for the analyte. The present invention relates to a measuring device that is simple and quick, and a measuring method using the measuring device.

[0002]

2. Description of the Related Art Measurement of trace substances in a living body using a specific binding reaction is frequently carried out for many purposes such as diagnosis of various diseases and judgment of therapeutic effect. Recently, it is often carried out not only by medical institutions but also by unskilled persons at home. For the measurement of in-vivo trace substances using these specific binding reactions, a highly sensitive and highly precise precision measurement method and a simple measurement method that is easy to operate and obtains results in a short time can be selected, depending on the purpose of use. It can be used properly according to the type. In particular, the simple measurement method can be performed easily by simple operation without using a reaction device or measurement equipment that requires specialized knowledge or technology, so that a sufficient diagnosis can be made even with semi-quantitative or qualitative measurement. It is a very convenient method and is widely used for pregnancy diagnosis.

Presently, as a simple measuring method utilizing an immunological reaction which is one of the specific binding reactions, an agglutination reaction or an aggregating inhibition reaction using latex particles as a carrier, and an enzyme immunoassay method using an enzyme as a labeling substance. (EIA), a simple immunological measurement method using a colloidal non-metal or colored latex particles as a labeling substance is widely used. Further, in recent years, as a method widely used in a simple immunological measurement method, a method using a porous reaction membrane in which a substance having an affinity for an analyte is immobilized on a membrane of a porous material is used. It has been known. Unlike those EIAs that utilize a substrate for an enzyme in the coloring process, most of these methods often fix a coloring substance directly to an antibody.
By using this method, the operation becomes simpler and the result can be obtained in a shorter time, as compared with the conventional aggregation reaction or aggregation inhibition reaction using latex particles, EIA and the like.

As a measuring method (apparatus) using a porous reaction membrane, Japanese Patent Publication No. 3-504166 and Japanese Patent Laid-Open No. 3-1.
There is a simple immunological measurement method using chromatography disclosed in Japanese Patent No. 76659. In these methods, a porous reaction membrane is used as a chromatographic strip, and the reaction is carried out while a reagent set on a specific portion of the surface is moved together with a test sample according to the principle of chromatography. However, with these methods,
It requires a time for the test sample and the reagent to move over a certain distance of the chromatograph strip, and has a drawback that the reaction time is relatively long.

While these measuring devices use the method of flowing the test sample along the chromatographic strip of the porous reaction membrane, the test sample is flowed (passed) in the direction of filtration through the porous reaction film. Devices and methods are also known, and specific examples include the following. Japanese Patent Application Laid-Open No. 57-63454 discloses an apparatus including a porous reaction membrane and a liquid absorption layer, which allows a test sample to pass through the porous reaction membrane and then be absorbed in the liquid absorption layer, and a signal generating system is used. Discloses a method of generating a signal on a porous reaction membrane and measuring the signal. In Japanese Patent Laid-Open No. 1-214760, after passing a test sample through a porous reaction membrane, colored latex particles having another affinity substance fixed thereto are supplied to and passed through the porous reaction membrane, and further washed. It discloses a method of performing the measurement depending on the presence or absence of colored spots on the porous reaction film after that.

[0006] In these methods, the operation of adding a solution other than the test sample to the apparatus and the washing operation are required, so that the measurement operation is complicated, and therefore, it takes a considerable time to obtain the measurement result. When the test sample contains insoluble contaminants, etc., it has a drawback that the contaminants accumulate on the porous reaction membrane and interfere with the measurement. On the other hand, in JP-A-62-500121, a porous material holding a soluble reagent is placed on the upper part of a porous reaction membrane on which an affinity substance is fixed, and the soluble reagent is dissolved by adding a test sample. Disclosed is a reaction apparatus in which a porous reaction membrane having an affinity substance immobilized thereon is passed through and a liquid is absorbed in a liquid absorption zone bonded to a lower portion. In this device, since the soluble reagent is incorporated in the reaction device in advance, the operation of adding the reagent solution is omitted, and the porous body containing the soluble reagent has a function as a filter. It is possible to prevent the accumulation of impurities and the like on the top. However, in order to observe the results of the reaction, it is necessary to remove the porous body containing the soluble reagent from the device,
There is a drawback in the complexity of the operation.

A method for adhering a solution on a porous reaction film to carry out a reaction is also known, and the following is exemplified. In Japanese Unexamined Patent Publication (Kokai) No. 4-161853, gold colloid is used as a soluble reagent, and a porous and elastic material having a function of depositing a soluble reagent and absorbing a test sample liquid under a porous reaction membrane on which an affinity substance is immobilized is used. Disclosed is a method of placing a member having properties and observing a signal by separating the container after completion of the reaction. Further, since the signal is unclear in this method, JP-A-4-232861 discloses a method using an enzyme-labeled antibody and a substrate as a color-developing method. However, in these methods, the operation of separating the container is indispensable for observing the signal, and the complexity of the operation has not been eliminated.

Various measuring devices are known in which a part of the device is not separated or opened when observing a signal. As such a device, for example, a device for flowing a test sample along a chromatographic strip of a porous reaction membrane is disclosed in the above-mentioned JP-A-3-176659 and JP-A-HEI-3.
In addition to the devices disclosed in Japanese Patent Publication No. 504166, a device disclosed in Japanese Patent Publication No. 63-500330 is a device for passing a test sample in a direction in which it is filtered by a porous reaction membrane.

In the apparatus disclosed in Japanese Patent Publication No. 63-500330, an upper structure designed to drop the reaction solution only on the reaction zone at the center of the filter and a reaction zone.
Disclosed is a reactor comprising a lower structure designed so that the reaction solution concentrated in the filter flows horizontally toward the periphery inside the filter and is absorbed by an absorber adjacent to the upper part or the lower part of the peripheral part. ing. In this device, the lower part of the filter is an enclosure and the window is open, so that the signal can be read from the bottom through the window. Although this device has the convenience that the signal can be observed only by turning the reaction device over after the reaction is completed, the reaction device itself is not designed for simple and quick measurement in one step. In other words, in this device, it is necessary to carry out operations such as filtration of test specimens and immunological reactions in another reaction container, and then attach the container to the receiving port of the reaction device and inject the reaction liquid. However, the simple measuring device has a drawback that the operation is complicated and it takes time to obtain a result. Further, since it is necessary to move the reaction liquid concentrated in the reaction zone limited to the center of the filter to the peripheral portion of the porous reaction membrane by the principle of chromatography,
There is a drawback that the measuring device must be manufactured strictly and the device itself becomes complicated.

When a result is obtained by a simple operation in one step using a simple measuring device, it is necessary to pay particular attention to sufficient B / F separation (for example, in an antigen-antibody reaction, an antigen and an antibody are bound to each other to cause the binding). It is necessary to physically separate the bound type: Bound, B from the unbound free type: Free, F). That is, it is necessary to remove the unreacted reagent remaining on the determination part by various methods so that the reagent such as unreacted labeled antibody does not interfere with the visual determination. Therefore, for example, Japanese Patent Laid-Open No. 1-21447
According to the method disclosed in Japanese Patent Laid-Open No. 60, it is necessary to wash the porous reaction membrane with water, a buffer solution or the like, if necessary, and this method makes the operation complicated. In addition, in the simple measurement method using the principle of chromatography, an extra test sample containing the analyte is supplied to cause the reaction on the porous reaction membrane to be followed by washing for B / F separation. Therefore, it takes a considerable time for measurement. Further, the method of moving the test sample in the vertical direction generally has an advantage that the reaction time is shorter than that in the horizontal direction, but as described above, it is necessary to remove a part of the device to visually judge the result. Many of them require that the judgment surface be pulled out as a unit, which has the drawback that blood or urine used as a test sample may come into contact with a hand or a part of the body during the operation. It is a thing.

In order to solve such a problem, the inventors of the present invention have disclosed, as a device capable of performing a sufficient B / F separation and a quick measurement by a simple operation in one step, as disclosed in JP-A-6-2.
A simple measuring device disclosed in Japanese Patent No. 73419 is proposed. In this device, a porous reaction membrane that fixes an affinity substance is used as a determination unit, and a porous material coated with a soluble reagent is placed on this, and the solution that has passed through the porous reaction membrane is absorbed by the absorbent material. Has a configuration. In this device, the test sample first passes through a porous material coated with a soluble reagent,
It is configured to reach the porous reaction membrane in the state where the soluble reagent is dissolved, and is absorbed through the liquid impermeable sheet downstream of the porous reaction membrane so that it passes through the porous reaction membrane efficiently. It is configured to contact the material. Further, the liquid impermeable sheet and the absorbent are not placed on the reaction area where the affinity substance is fixed in the porous reaction film and the lower surface around the reaction area, and the measurement result is obtained from the observation window on the back side of the device. It is designed for observation. According to this device, since the measurement result can be observed from the observation window, it is not necessary to separate or release the device in the measurement operation. Furthermore, since the soluble reagent is applied to the porous material arranged on the porous reaction membrane, the measurement can be performed by a one-step operation of adding the test sample to the device. As a result, the measurement is completed in a shorter time than the apparatus using the above-mentioned principle of chromatography. In addition, the porous material coated with the soluble reagent exhibits the function as a filter, and the process in which the test sample passes through the porous reaction film also exhibits the function as a filter. The influence of foreign substances in the test sample of is prevented.

[0012] As described above, although the measuring operation has become considerably simple, it is still necessary to measure a fixed amount of the test sample, and there is still room for improvement in simplification. Also,
Although the time required for measurement is shorter in a device in which the test sample passes through the porous reaction membrane than in a device using the principle of chromatography, results can be obtained in a shorter time, such as in mass screening, or a larger number can be obtained in a certain time. It is required to further reduce the time required for the measurement in order to obtain the result of the sample. Furthermore, the conventional device requires not only a case for holding various members such as a porous material coated with a soluble reagent, a porous reaction film, a liquid impermeable sheet, and an absorbent, but also the device. Since it is not easy to assemble, it is not satisfactory in terms of mass productivity and cost when it is used for a disposable use, and improvement is desired in terms of the manufacturing process and manufacturing cost of the device.

[0013]

The simple measurement requires accurate and short-time results to be obtained even when operated by an unskilled person, which results in quickness and simplicity of measurement, reduction in product cost, and the like. It is a big issue. Although there are many simple measuring devices and methods using a porous reaction membrane at present, as described above, it is difficult to manufacture inexpensively because of a long measuring time, multiple operation steps, and a complicated container structure. None of them satisfy the low cost, simplicity, and certainty required for simple measurement, such as the image being unclear. One of the factors that influence the speed and convenience described above is how easily, quickly and reliably B / F separation can be performed. In other words, there is a demand for the development of a simple measurement device and method that do not require B / F separation as a special independent process. By solving these problems, it is expected that the simple measurement will be able to obtain accurate results in a shorter time with a simpler operation.

The present invention does not require B / F separation as an independent step in the measurement operation, enables quick and simple measurement in one step operation, has high measurement accuracy, and is easy to manufacture. It is an object of the present invention to provide a simple measuring device which is inexpensive and suitable as a disposable measuring device.

[0015]

In order to solve the above-mentioned problems, the inventors of the present invention can obtain accurate results in a short time with a simple operation in one step, and an inexpensive and easy-to-manufacture measuring device. As a result of the research on, it is composed of a porous material on which an affinity substance is immobilized, preferably a determination part having a porous reaction membrane, and a porous material holding a soluble reagent which is solubilized by the addition of a test sample upstream thereof. A reagent part, and a test sample collecting part having a spacer having a through hole for introducing a test sample upstream thereof and a liquid-permeable membrane arranged to cover the upstream surface of the through hole, The absorption part, which is placed downstream of the judgment part and absorbs unreacted and excess test samples, has a structure in which an absorption sheet is sandwiched between liquid-impermeable sheets, and is long with respect to the judgment part and reagent part. In addition, A slit for facilitating the flow of ventilation and test specimens bets, by visibly constituting the determination unit further from the back (downstream) was found to be able to achieve the object.

That is, the present invention is a measuring device for measuring an analyte in a liquid test sample, which comprises a spacer having a through hole for introducing the test sample and an upstream side of the through hole. And a test sample collecting section having a liquid permeable membrane arranged in a porous manner, and a soluble reagent which is arranged downstream of the test sample collecting section facing the through hole and which is dissolved and released upon contact with the test sample is porous. At least one reaction region formed by immobilizing an affinity substance capable of binding to the analyte or soluble reagent, which is arranged downstream of the reagent part, on a porous material A determination part, and a long upstream sheet and a downstream sheet made of a liquid impermeable material, which are arranged downstream of the determination part, and a long absorption sheet made of a liquid absorbent material and sandwiched between the two sheets. And near the edge The absorbent sheet further has a through-hole corresponding to the downstream side of the determining section, and an absorbent sheet in contact with the determining section. A slit extending in the direction is formed, the upstream sheet is formed with a through hole corresponding to the through hole of the absorbent sheet, and the downstream sheet is
There is provided a simple measuring device characterized in that at least a portion corresponding to a through hole of the absorbent sheet is transparent.

Further, in the simple measuring device of the present invention, the slit of the absorbent sheet may communicate with the atmosphere outside the device, and the test sample collecting portion has a space inside the through hole formed in the spacer. Therefore, it is preferable to have a function of collecting a fixed amount of test sample.

Further, in the simple measuring device of the present invention, the judgment part may have a plurality of reaction zones holding different affinity substances, and the reaction part is fixed to the porous material of the judgment part. The affinity substance thus obtained is preferably an antibody or an antigen, the soluble reagent retained in the porous material of the reagent part is an antibody or an antigen, and the antibody or antigen is a labeling substance for enabling detection. The labeling substance is preferably selected from the group consisting of colloidal metals, colloidal non-metals, dyes and latex particles, and the porous material of the determination part and / or reagent part is preferably Are preferably selected from the group consisting of cellulose, cellulose derivatives, nitrocellulose, porous synthetic polymers, glass fiber filters, nonwovens and cloths. The liquid absorbent material forming the absorbent sheet of the absorbent section is preferably selected from the group consisting of cellulose, a cellulose derivative, a porous synthetic polymer, a glass fiber filter, a nonwoven fabric, a particle absorbent and a cloth, It is particularly preferable that the porous material of the determination part is nitrocellulose, and the liquid absorbent material forming the porous material and the absorbent sheet of the reagent part is cellulose.

The measuring method of the present invention is a measuring method for measuring an analyte in a liquid test sample by using the simple measuring device of the present invention, which contains the analyte. A test sample that seems to be dropped or dropped onto the test sample collecting part of the simple measuring device, or by immersing the test sample collecting part in the test sample, a fixed amount of the test sample is put in the through hole of the spacer. After collecting, by flowing into the porous material of the reagent portion, the soluble reagent retained in the porous material of the reagent portion is eluted and passed through the reagent portion together with the test sample to flow into the determination portion, By directly or indirectly binding the affinity substance and the soluble reagent fixed in the reaction region of the porous material of the determination part,
A detectable signal is formed in the reaction zone, and the test sample that has passed through the determination part is rapidly absorbed by the absorption sheet by being accelerated by the slit formed in the absorption sheet of the absorption part, and the downstream sheet A measurement method for observing the signal from a transparent portion to measure an analyte in a liquid test sample is provided.

In the measuring method of the present invention, it is preferable that the test sample is a body fluid or a solution obtained by diluting or extracting and diluting a biological component.

The simplified measuring device of the present invention and the measuring method using this measuring device will be described in detail below.

The measurement in the present invention utilizes a specific binding reaction of an analyte, and a substance having an affinity for the analyte, that is, an affinity substance which specifically binds to the analyte, and an analysis This is a measurement using a soluble reagent that is a labeled substance that is a specific binding substance for the target substance or the affinity substance. The device of the present invention is
It has a determination part having a reaction region formed by immobilizing an affinity substance on a porous material, and a reagent part formed by holding a soluble reagent in the porous material so that it can be eluted, and a test sample collecting part (described later in detail). Hereinafter, a liquid test sample that is expected to contain the analyte is collected in the sampling unit), the test sample is caused to flow into the reagent unit, the soluble reagent is eluted with the test sample, and the determination unit together with the test sample. Then, in the determination part, a signal is generated by the specific binding reaction of the analyte, the soluble reagent and the affinity substance, and the analyte is measured. Regarding this measuring method, Japanese Patent Application Laid-Open No. 6-27341
This is described in detail in Japanese Patent No. Although the test sample that has passed through the determination unit is absorbed by the absorption unit, in the present invention, the flow of the test sample and the flow of the test sample are absorbed by the absorption sheet that makes the absorption unit long and absorbs the test sample. Since the slit that promotes ventilation is formed, the test sample can be quickly absorbed in the absorbing part and the liquid flow of the test sample can be promoted, whereby extremely quick and simple measurement can be performed. This point will be described in detail later.

In the measurement utilizing various specific binding reactions, the measurement results of various analytes are judged by comparison with their physiological concentrations.

As the analyte to be measured by the simple measuring device of the present invention, specifically, various proteins or peptides functioning as antigens or antibodies, nucleic acids, effector molecules, receptor molecules, enzymes, coenzymes, inhibitors, Sugar chains and lipids or compounds having these, lectins and other biological substances, drugs, drug metabolites, etc.,
More specifically, human chorionic gonadotropin (hC
G), luteinizing hormone (LH), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), human placental lactogen (hPL), beta-2 microglobulin (β
2m), ferritin, HBs antigen, estrogen, HB
s antibody, HCV antibody, HIV antibody, rubella antibody, IgE antibody and the like.

The test sample means a body fluid or biological component expected to contain such an analyte, and a solution obtained by diluting or extracting and diluting these.
Specifically, blood, plasma, serum, urine, saliva, lymph,
Vaginal secretions, milk, nipple discharge, semen, intracystic fluid, fecal extract, tissue extract, and the like, and solutions obtained by diluting or extracting and diluting these with water, physiological saline, etc. are mainly mentioned.

As described above, the affinity substance is a substance which specifically reacts with the analyte (specific binding substance) or a substance which binds to the analyte via the specific binding substance. Such affinity substances include various proteins or peptides that function as antigens or antibodies, nucleic acids, lectins, sugar chains or compounds having these, biotin, avidin, enzymes, enzyme substrates, enzyme inhibitors, receptors, ligands, drugs, drugs. Metabolites, derivatives of analytes, and the like, and more specifically, human chorionic gonadotropin (hCG), luteinizing hormone (LH), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), human Examples thereof include placenta lactogen (hPL), beta-2 microglobulin (β2m), ferritin, HBs antigen, estrogen and the like, and antibodies against these. In the present invention, antibody, antigen, nucleic acid, avidin, biotin are preferably used, and antibody or antigen is particularly preferable.

The soluble reagent is a substance obtained by labeling a substance that specifically binds to an affinity substance, a substance that specifically binds to an analyte, or a substance that binds to an analyte via a specific binding substance. . Specific binding substances include various proteins or peptides that function as antigens or antibodies,
Examples include nucleic acids, lectins, sugar chains or compounds having these, biotin, avidin, enzymes, enzyme substrates, enzyme inhibitors, receptors, ligands, drugs, drug metabolites, and analyte derivatives, and more specifically humans. Chorionic gonadotropin (hCG), luteinizing hormone (LH), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), human placental lactogen (hPL),
Beta-2 microglobulin (β2m), ferritin, H
Examples include Bs antigen, estrogen and the like, and antibodies against them. In the present invention, the specific binding substance used as a soluble reagent is an antibody, an antigen, a nucleic acid, avidin,
Biotin is preferred, and antibody or antigen is particularly preferred.

The soluble reagent used in the present invention is labeled by binding such a specific binding substance to a labeling substance by physical adsorption such as hydrophobic bond or chemical bond such as covalent bond. is there. Examples of the labeling substance used in the present invention include enzymes or substrates, fluorescent compounds, radioactive labels, chemiluminescent compounds, colloidal metals, colloidal nonmetals, dyes, latex particles, chromogens, catalysts, liposomes, microorganisms and the like. Of these, enzymes, fluorescent compounds, colloidal metals, colloidal nonmetals, dyes, latex particles and the like are preferably used. Particularly preferred labeling substances are colloidal metals, colloidal non-metals, dyes and latex particles, and dyes are particularly preferable.

In the soluble reagent, the amount of the labeling substance labeled with the specific binding substance varies depending on the type of the soluble reagent and the labeling substance used. For example, an anti-hCG antibody is used as the specific binding substance and dispersed in the labeling substance. When a dye is used, the final concentration of the antibody solution to which the disperse dye solution is added is 0.3 to 30 for 0.5 mg / ml of the antibody.
It is preferred to use mg / ml disperse dye. Further, the preparation of the soluble reagent in this case is performed by, for example, incubating the disperse dye and the antibody at 56 ° C. for 30 minutes, and
After ice-cooling for 1 minute, centrifugation was performed at 22,000 × g for 20 minutes, and albumin / saccharose / phosphate buffered saline (PBS) was added to the resulting precipitate to suspend it, whereby a disperse dye-labeled antibody can be obtained.

In the present invention which utilizes the specific binding reaction of the analyte, the affinity substance and the soluble reagent, a sandwich type reaction and a competitive type reaction can be used as the specific binding reaction. The sandwich type is a form in which the analyte and the affinity substance fixed to the porous material of the determination part and the soluble reagent retained in the porous material of the reagent part are bound to each other, and The intensity of the signal increases proportionally. On the other hand, the competitive type is a form in which the binding of the affinity substance fixed to the porous material of the determination part is made to compete between the analyte and the soluble reagent retained in the porous material of the reagent part. Yes, the intensity of the signal decreases in proportion to the analyte concentration. The presence or amount of the analyte can be known by comparing the signal intensity obtained by each method with the signal intensity of the standard substance.

Examples of the affinity substance and the soluble reagent used in the present invention include anti-hCG antibody as the affinity substance and dye-labeled anti-hCG antibody as the soluble reagent when the analyte is hCG. When the analyte is AFP, the affinity substance is anti-AFP
Antibodies, soluble reagents are dye-labeled anti-AFP antibodies; if the analyte is CEA, affinity substances are anti-CEA antibodies, soluble reagents are dye-labeled anti-CEA antibodies; analytes are estrogen In some cases, the affinity substance is an anti-estrogen antibody, the soluble reagent is a dye-labeled estrogen derivative, and when the analyte is an anti-HIV antibody, the affinity substance is an HIV.
Examples of the antigen peptide and the soluble reagent include dye-labeled anti-human immunoglobulin antibodies;

Hereinafter, the simple measuring device of the present invention and the measuring method of the present invention using the simple measuring device will be described in more detail with reference to the preferred examples shown in the accompanying drawings. The simple measuring device of the present invention is, of course, not limited to the examples described below.

FIG. 1 shows an exploded perspective view of an example of the simple measuring device of the present invention, and FIG. 2 shows a schematic sectional view thereof.
In the following description, the upper part of the drawing will be referred to as upper and the lower part will be referred to as lower. As shown in FIG. 1 and FIG. 2, the simple measuring device 10 of the present invention basically includes a (test sample) collecting unit 12, a reagent unit 14, a determining unit 16, and a long measuring unit from the top in the drawings. The absorber 18 is included. In such a simple measuring device 10, the test sample is added to the collection unit 12, passes through the reagent unit 14 downward, flows into the determination unit 16, passes through this downward, and is finally absorbed by the absorption unit 18. It
In addition, a signal indicating the measurement result is formed in the determination unit 16 and the measurement result is obtained by detecting the signal from the lower surface.

In the simple measuring device 10 of the illustrated example, the sampling part 12 comprises a spacer 20, a liquid-permeable membrane 22 arranged on the upper surface of the spacer 20, and a seal 24 arranged under the spacer 20. Composed.

The spacer 20 is a rectangular parallelepiped member in which a through hole 20a for introducing a test sample (hereinafter referred to as a sample) is formed, and the sample passes through the membrane 22 and the through hole 20a.
To be collected. Here, in the simple measuring device 10 of the present invention, the addition of the sample is basically performed by applying the sample to the sampling unit 12, immersing the sampling unit 12 in the sample, dropping the sample into the sampling unit 12, or the like. . Therefore, in the present invention, the amount of the sample to be sampled can be defined by the volume of the through hole 20a, and it is possible to sample a fixed amount of sample easily, and more simple and reliable measurement can be realized.
That is, the device described in Japanese Patent Application Laid-Open No. 6-273419, which is a device through which a sample passes through a porous reaction membrane as a filtration membrane, requires an operation for measuring a certain amount of a test sample with an instrument or device such as a pipette. On the other hand, the present invention is the sampling unit 1
A fixed amount of the sample can be measured even by simply applying or immersing the sample in 2, and the measurement operation is very simple. In the apparatus of the illustrated example, the reagent portion 14 is housed in the seal 24 and inserted and arranged in the through hole 20a. Therefore, the volume of the through hole 20a is equal to that of the reagent portion 14
It is necessary to take into consideration the volume etc.

In the simple measuring device 10 of the illustrated example, as a preferred mode, the spacer 20 is provided with an air vent 20b which communicates the side surface thereof with the through hole 20a. By having the air vent 20b, it is possible to more smoothly and surely collect a certain amount of the sample in the through hole 20a, and it is possible to realize simpler and more reliable measurement.

There is no particular limitation on the external shape of the spacer 20, and in addition to the rectangular parallelepiped shown in the figure, the shape, such as a columnar (elliptic column) shape, a polygonal column such as a hexagonal column, or the shape shown in FIG.
It may be appropriately determined in consideration of productivity, usability, and the like. However, the spacer 20 is required to have a thickness and a size that can secure the required strength even when the through hole 20a (further, the air vent 20b) is formed and can form the through hole 20a having a sufficient volume. Therefore, the spacer 20 preferably has a thickness (up and down direction) of about 0.5 to 5 mm. The through hole 20a is not limited to the cylindrical shape shown in the figure, and various shapes can be used, but the cylindrical shape is preferable in terms of productivity, uniformity of the flow of the sample, and the like. Further, the size may be appropriately determined according to the amount of the sample required for measurement, but in consideration of the thickness of the spacer 20, if the spacer is a cylinder, the diameter of the circle is 3 to.
It is about 15 mm.

The material for forming the spacer 20 is not particularly limited, but it is necessary to allow the sample collected in the through hole 20a to flow into the reagent portion 14 without leaking. Therefore, it is necessary to secure sufficient liquid impermeability until the measurement is completed, and various liquid impermeable materials and low liquid permeability materials are preferably used. Specifically, it is preferably selected from expanded polyethylene, synthetic rubber, various plastics such as vinyl chloride and acrylic, expanded polystyrene and the like, and expanded polyethylene is particularly preferred.

A liquid-permeable film 22 is attached to the upper surface of the spacer 20 to cover the upper surface of the through hole 20a. This membrane 2
2 also acts as a contaminant removal filter that removes foreign matters such as suspended matter and precipitates contained in the sample, and more preferably tissue debris and cell clumps. The membrane 22 having such an action has a high sample permeation rate, has a filtering performance to the extent that impurities can be removed at a required level, and has a strength necessary to hold a fixed amount of the sample. It may be formed of various materials that satisfy the requirements. Specific examples include woven fabrics, non-woven fabrics, meshes, membranes and the like made of rayon, cellulose, cellulose derivatives and the like. The method of attaching the film 22 to the spacer 20 is not particularly limited, and any known method depending on the forming material of the spacer 20 and the film 22, such as a method using a double-sided tape or a method using an adhesive, can be used. . In addition to the attachment, a fixing member may be used to fix the film 22 on the upper surface of the spacer 20 to cover the through hole 20a.

In the simple measuring device 10 of the illustrated example, as a preferred mode, the sampling section 12 has a seal 24 for holding the reagent section 14. In the illustrated example, the seal 2
4 has a bottom surface shape similar to the bottom surface of the spacer 20, and has a hollow convex portion 2 having a shape substantially the same as that of the through hole 20a in the central portion.
4a is a liquid impermeable member. Also, the convex portion 2
In the center of 4a, a hole 24b for allowing the sample to pass is formed. In the seal 24, the protrusion 24a is formed in the through hole 20a.
And is fixed to the spacer 20, and the reagent portion 14 is inserted into the convex portion 24a. By having the seal 24, the reagent portion 14 is held at a predetermined position and communicates with the through hole 20a, and the collected sample efficiently flows into the central portion of the reagent portion 14 to provide a uniform liquid in the surface direction. The flow can be realized.

As the material for forming the seal 24, various liquid-impermeable materials can all be used, and various polyesters, polyethylene, polypropylene, vinyl chloride,
It is preferably selected from nylon and the like, and polyester is particularly preferable.

The present invention is not limited to the one having such a seal 24, but the through hole 20a of the spacer 20.
A holding part such as a step may be provided to hold and fix the reagent part 14 here, or may be fixed to the spacer 20 so as to cover the bottom surface of the through hole 20a, or fixed to the determination part 16. The reagent unit 14 may be held by the above.

A reagent section 14 is arranged below the sampling section 12. The reagent portion 14 is made of a porous material that holds the above-mentioned soluble reagent. In the illustrated example, the reagent portion 14 is made of a disk-shaped porous material. Is stored and retained in the through hole 20 from below.
It is arranged in a state of being inserted in a.

Such a reagent part 14 elutes the soluble reagent held by the porous material by the inflow of the sample and supplies it to the sample, and further removes foreign matters and impurities which could not be removed by the membrane 22, It is possible to prevent foreign matters from flowing into the determination unit 16. Therefore, the determination unit 1 based on impurities
The effect on 6 can be prevented, the signal can be made clear, and more accurate measurement can be performed. There is no particular limitation on the porous material that can be used for the reagent portion 14, and cellulose,
Cellulose derivatives, woven fabrics made of porous synthetic polymers and the like, non-woven fabrics and membranes, glass fiber filters, various filter papers are preferably exemplified, but cellulose,
Those using a cellulose derivative or a porous synthetic polymer are preferably used, and those using cellulose are particularly preferable. Here, as described above, the reagent unit 14 needs to have a function of filtering the sample and rapidly dissolving the soluble reagent held by the inflow of the sample, and thus the compatibility with the analyte or the soluble reagent is required. In consideration of this, it is preferable to select the material, size, etc. of the porous material. The shape of the reagent portion 14 is not limited to the disc shape shown in the figure,
From the viewpoint of the uniformity of the sample liquid flow and the like, it is preferable that it has a disc shape.

As a method for holding the soluble material in the porous material so that the soluble material can be eluted by the inflow of the sample, a method of impregnating the porous material with the solution containing the soluble reagent and then drying the solution is exemplified. Here, the drying is preferably performed by freeze-drying because the soluble reagent to be retained can be quickly and uniformly distributed. Further, when the reagent part 14 is prepared by holding the soluble reagent in the porous material, it is preferable to add salt, sugar, protein or the like to freeze-dry in order to enhance the stability, and the soluble reagent due to the inflow of the sample. A surfactant can be added as a dispersant to enhance the solubility of the.

Below the reagent section 14, a judgment section 16 is arranged. The determination unit 16 is formed by immobilizing the above-mentioned affinity substance on a porous material, and preferably has a reaction region in which the affinity substance is immobilized on a film of the porous material (porous film), and is porous. It is a reaction film. In the simple measuring device 10 of the present invention, the mixture of the sample and the soluble reagent flows into the determination unit 16 to cause the above-mentioned sandwich-type or competitive-type specific binding reaction, and the reaction in which the affinity substance is immobilized. A signal is generated in the area. It should be noted that this signal is a signal corresponding to the labeling substance labeled with the above-mentioned soluble reagent, such as discoloration, coloring, luminescence, and the like.

As the porous material, woven cloth, non-woven cloth or membrane made of porous synthetic polymer such as cellulose, cellulose derivative, nitrocellulose, nylon, etc. as a raw material,
Although glass fiber filters and various filter papers are preferably exemplified, those using cellulose, cellulose derivatives and porous synthetic polymers are preferable, and those using nitrocellulose are particularly preferable. What is necessary is that these porous materials have pores that communicate from the front surface to the back surface, and that the unreacted soluble reagent has a sufficient pore size to pass through the pores that communicate with each other. That is. Usually, the pore size is preferably 1 to 50 μm, particularly 3 to 10 μm.
m is preferred.

The affinity substance is bound to the porous material by physical adsorption such as hydrophobic bond or chemical bond such as covalent bond by a method such as heating after applying the affinity substance to the porous film. Fixed. The immobilization method may be an ordinary method of immobilizing an affinity substance, that is, all known methods of immobilizing the above-mentioned specific binding substance on a holding material such as a porous material are available, and for example, a porous material When nitrocellulose is used, it can be carried out by applying a necessary amount of a solution containing an affinity substance, drying it at about 37 ° C., and then blocking the unbound portion with albumin or the like. Alternatively, it can be carried out by coating the nitrocellulose membrane with albumin in advance, applying a necessary amount of a solution containing an affinity substance, and drying at about 37 ° C.

The amount of the affinity substance immobilized on the porous material varies depending on the type of the substance to be analyzed, the type of the affinity substance used, etc., but is preferably about 10 to 200 μg / cm ² . Also,
The shape of the affinity substance to be immobilized on the porous material on the plane (that is, the reaction area = the shape of the signal) is a cross, parallel lines, a circle, a double circle, a star, a triangle, or a simple illustration. It is preferable that the shape is such that it can be easily discriminated.

Here, when the signal can be visually confirmed, the measurement result may be visually provided. Although the shape of the signal in the reaction zone is as described above, a semi-quantitative determination can be made by binding a certain concentration of standard substance to a part of the signal and comparing the signal intensity of the analyte with the signal intensity of the standard substance. It is possible. Alternatively, a fixed amount of labeling substance may be applied. Furthermore, it is also possible to quantify by measuring the signal intensity of the signal of the reaction part using an appropriate optical instrument. For example, if the analytes of varying concentrations are bound as dots in the reaction part of the porous reaction membrane as a standard substance, and the antibodies to the analytes are bound on the same reaction membrane with dots of the same diameter for analyte measurement, By comparing the signal intensities, semiquantitative or simple quantification can be performed.

There may be a plurality of reaction regions in which the affinity substance is fixed to the porous material. The measuring apparatus of the present invention having the above-described configuration can bring the sample mixed with the soluble reagent into contact with the reaction zone of the determination unit 16 in a uniform state, so that the signal intensity of each portion in the reaction zone is uniform. The reaction under the same conditions can occur in a plurality of reaction zones. Therefore, in addition to signals for analyte measurement, multiple signals to control the measurement are set on the reaction zone, or multiple reaction zones with different sensitivities are formed to compare the signal intensities and semiquantitative and then quantitative determination is performed. It is possible, and it is also possible to adopt a configuration in which a plurality of different types of measurements can be performed simultaneously by forming a plurality of reaction zones with different affinity substances immobilized. Further, since the signal intensity on the reaction zone is uniform, there is an advantage that the design of the reaction zone (signal) can be freely selected as described above.

An absorber 18 is arranged below the determination unit 16. The absorbing portion 18 is basically formed by sandwiching a long absorbing sheet 26 made of a liquid absorbing material between a liquid impermeable upper sheet 28 and a lower sheet 30 and having one end in the longitudinal direction. A laminated body such as the determination unit 16 is placed near the part. In the present invention, the absorption sheet 26
The term “long length” and the like means extending in one lateral direction with respect to the vertical direction of the flow of the sample from the sampling unit 12 to the determination unit 16.

In the illustrated example, the absorbent sheet 26 has a substantially rectangular shape, and the upper sheet 28 and the lower sheet 30 have a rectangular shape having substantially the same size as the absorbent sheet 26, and the absorbent sheet 26 is entirely sandwiched. Furthermore, in a preferred embodiment, the upper surface of the upper sheet 28 is covered with an opaque upper color sheet 32, while the lower surface of the lower sheet 30 is also covered with an opaque lower color sheet 34, which enhances the design of the simple measuring device 10. ing.

The absorbing sheet 26 absorbs the sample (mixture with the soluble reagent) collected by the collecting section 12 and passed through the reagent section 14 and the judging section 16. That is,
The flow of the sample that has passed through the determination unit 16 is changed from the vertical direction to the horizontal direction (in the illustrated example, a large flow is to the right) by the absorption unit 18. In the simple measuring device 10 of the present invention, the sample that has passed through the determination unit 16 is prevented from flowing out by absorbing the sample by permeating the sample into the absorption sheet 26, and at the same time, from the sampling unit 12 The liquid flow of the sample passing through the determination unit 16 and reaching the absorption sheet 26 is accelerated to shorten the measurement time.

As described above, the determination unit 16 and the like are the absorption unit 18
Placed near one end of the. Therefore, the through-hole 36 for observation is formed in the absorption sheet 26 at a position corresponding to the reaction area of the determination unit 16. In the conventional device that allows a sample to pass through the porous reaction membrane (determination unit 16), the absorbing member is usually placed directly below the porous reaction membrane, but in this structure, the reaction region is hidden by the absorbing member or the like. As a result, the signal resulting from the reaction cannot be directly observed from the back surface of the measuring device. On the other hand, the absorption sheet 2
By having such an observation through hole 36 in 6, it is possible to directly observe the determination unit 16 (reaction area) from the lower surface side of the simple measurement device 10, and, as in the conventional device, a part of the device. It is possible to confirm the signal and obtain the measurement result without performing operations such as removing and disassembling the sample, and it is also possible to prevent the sample from touching the hand or the like during the measurement.

In addition, since the observation through-hole 36 (and the communication through-hole 28a formed in the upper sheet 28) is provided, a space having the lower sheet 30 as the lower surface is provided below the reaction area of the determination unit 16. Can be formed. Having this space results in that the sample is once stored in this space and then absorbed by the absorption sheet 26, and the analyte, soluble reagent, and affinity substance in the sample are in contact with each other for a necessary time, and The reaction can proceed to produce a signal, and more accurate measurement results can be obtained.

In the simple measuring device 10 of the present invention, the absorbent sheet 26 is provided with a slit 38 which is connected to or close to the through hole 36 for observation and which vertically penetrates the absorbent sheet 26 in the longitudinal direction of the absorbent sheet 26. Is formed to extend. In the illustrated example, the slit 38 is formed to be connected to the observation through hole 36, and its end (the end on the opposite side of the observation through hole 36) communicates with the slit 38 and the atmosphere through the top sheet 28. A through hole 40 for bleeding air may be formed. In the simple measuring device 10 of the present invention, the absorption portion 18 (absorption sheet 26) is elongated, and further, by having such a slit 38 or further air bleeding means, air bleeding for the flow of the sample, the absorption sheet By promoting the lateral flow of the sample liquid in 26 and the absorption of the sample into the absorption sheet 26, the liquid flow of the sample from the collection unit 12 through the determination unit 16 to the absorption sheet 26 is accelerated, Achieved even shorter measurement time. According to the device disclosed in the above-mentioned JP-A-6-273419, the measurement time is shorter than the devices disclosed in JP-A-3-176659 and JP-A-3-504166 which use the principle of chromatography. However, there is a limit to the shortening of the measurement time because the absorber has not been devised to accelerate the liquid absorption rate. The simple measuring device 10 of the present invention has a gap and a slit 38 under the determination unit 16 so as to accelerate the liquid absorption speed, and the measurement time is further shortened.

The shape of the observation through hole 36 is not particularly limited, but a circular shape is preferable from the viewpoint of production and the flow of the reaction solution. Further, the size thereof is preferably large enough for observing the reaction zone of the judging section 16 and smaller than the judging section 16, and specifically, the diameter of the circle is 2 to 12
mm is preferred. Further, the size of the slit 38 is not particularly limited, and may be appropriately determined according to the size of the absorbent sheet 26 and the like. According to the study by the present inventors, the length of the slit 38 is usually preferably about 5 to 100 mm depending on the length of the absorbent sheet 26 in the longitudinal direction (hereinafter referred to as length) and the like. Although the width depends on the length of the absorbent sheet 26 in the lateral direction (hereinafter referred to as width), it is preferably 1 to 3 mm in consideration of measurement time and the like. The slit 38
Is not limited to those extending in the same direction as the longitudinal direction of the absorbent sheet 26, and may extend obliquely with respect to the longitudinal direction of the absorbent sheet 26, and may be bent or bent as necessary. You may have.

The through hole 40 for bleeding air may have a circular shape with a diameter of about 1 to 12 mm, like the through hole 36 for observation. In the present invention, the through hole 40 (and
Corresponding to this, the through holes 28 formed in the top sheet 28
b) is not an indispensable constituent element. For example, the slit 38 may be formed to be longer so as to penetrate the absorbent sheet 26, and may be opened to the atmosphere from the end face of the absorbent portion 18 in the lateral direction. . Alternatively, as shown in FIG. 3A, the through hole 40 and the like may not be formed and the structure may not be opened to the atmosphere. In this case, the length of the slit 38 is preferably 1 mm or more, more preferably 2 mm or more. Although there is a limit, the larger the communication between the slit 38 and the outside, the shorter the measuring time tends to be. However, even if the slit 38 and the outside are not in direct communication, as shown in the later embodiment, A sufficiently quick measurement is possible.

Further, in the present invention, the slit 38
Is not limited to the one formed by being connected to the observation through hole 36, and the slit 38 may be formed in the vicinity of the observation through hole 36 as shown in FIG. At this time, the distance between the slit 38 and the observation through hole 36 is preferably 3 mm or less, and more preferably 2 mm or less.

The size of the absorbent sheet 26 is not particularly limited, and may be a size capable of sufficiently absorbing the sample depending on the amount of the sample collected in the collecting section 12 and the material forming the absorbent sheet 26. . The thickness of the absorbent sheet 26 is
Since it has a large influence on the absorbency of the sample and the volume of the space formed below the reaction zone of the determination unit 16, an appropriate thickness is required, and 0.05 to 1.5 mm is preferable, and 0.1 to 1.5 mm is particularly preferable. 1.2 mm is preferred. Further, according to the study by the present inventors, in order to allow the absorbent sheet 26 to absorb the sample well and to realize a quick measurement, the distance from the slit 38 to the end of the absorbent sheet 26 in the width direction (that is, The width of the absorbent sheet 26 from the slit 38 at the slit 38 forming portion) is 1 mm or more, preferably 2 mm or more.
Therefore, it is preferable to determine the widths of the absorbent sheet 26 and the slit 38 in consideration of this point.

The material of the absorbent sheet 26 is not particularly limited, and various liquid absorbents can be used. For example, various materials such as cellulose, cellulose derivatives, porous synthetic polymers, various synthetic fibers, etc. can be used. Examples thereof include woven fabrics, non-woven fabrics, membranes, and materials obtained by processing a particle absorbent such as a methyl methacrylate-vinyl acetate copolymer system into a sheet shape. Among them, those using cellulose, cellulose derivatives and porous synthetic polymers are preferable, and those using cellulose are particularly preferable. Further, by selecting the material of the absorption sheet 26, the absorption rate of the sample can be adjusted. For example, if the absorption rate is increased, more rapid measurement is possible, and conversely, by decreasing the absorption rate, the contact time between the affinity substance and the analyte or the like in the reaction zone of the determination unit 16 is lengthened. The measurement sensitivity can be changed as a result.

In the simple measuring device 10 of the present invention, the absorbing part 18 is constructed by sandwiching the absorbing sheet 26 made of such a liquid absorbing material between the liquid impermeable upper sheet 28 and the lower sheet 30. Have. In the illustrated example, the absorbent sheet 26, the upper sheet 28, and the lower sheet 30 are provided by the adhesive applied to the upper sheet 28 and the double-sided tape 42 arranged below the absorbent sheet 26.
And are glued together.

The upper surface sheet 28 arranged above the absorbent sheet 26 has a communication through hole 28a corresponding to the above-mentioned observation through hole 36 of the absorbent sheet 26 through which a sample flows into the absorbent sheet 26, and the same air vent. A through hole 28b corresponding to the through hole 40 for use is formed. Such a top sheet 28
By having the above, it is possible to prevent the unreacted and excess samples from being absorbed by the absorbent sheet 26 without passing through the reaction region of the determination unit 16 and the space portion formed thereunder. On the other hand, the lower surface sheet 30 prevents the specimen reaching the observation through hole 36 (the space portion) from flowing out downward. Here, the simple measuring device 10 of the present invention
In the above, since the signal generated in the determination unit 16 is detected from the lower sheet 30, the lower sheet 30 is transparent at least in the portion corresponding to the observation through hole 36 of the absorption sheet 26. Further, by making the absorbent sheet 26 entirely sandwiched between the upper sheet 28 and the lower sheet 30, it is possible to prevent the sample from touching the hand and the like, which is preferable in terms of hygiene.

As the material for the upper sheet 28 and the lower sheet 30, all known liquid impermeable materials can be used, and specifically, polyethylene, polyester, polypropylene, vinyl chloride, aluminum film, Examples include non-woven fabric impregnated with an adhesive. Further, this sheet may be formed by stacking a plurality of materials. Further, the method of adhering the absorbent sheet 26 to the upper sheet 28 and the lower sheet 30 is not particularly limited, and any known method using a double-sided tape or an adhesive can be used. When the double-sided tape is used, it is preferable to pull out the double-sided tape corresponding to the observation through-hole 36, the slit 38, and the through-hole 40 of the absorbent sheet 26 as illustrated. In addition, when observing a signal in the present invention, it is possible to measure and quantify with an optical device in addition to visual observation. preferable. The upper sheet 28 and the lower sheet 30 used in the present invention are preferably made of a material such as vinyl chloride having a high light transmittance, a smooth surface, and a uniform liquid wettability (or water repellency).

By the way, in the simple measuring device 10 of the illustrated example, the width of the half-strength of the absorption sheet 26 on the observation through-hole 36 side (upstream side in the sample flow direction) is narrower than that of the top sheet 28 and the like. ing. That is, the upper sheet 28 and the lower sheet 30 are directly adhered to each other in this portion. With such a configuration, when the sample flows out (liquid leakage) from the end surface of the absorbing part 18 to the outside, and the sample is applied to the sampling part 12, or the sampling part 12 is dipped in the sample, the sample is collected. This is more preferable because it is possible to suitably prevent the invasion of the sample (liquid invasion) from the end surface of the absorbing section 18. Here, the width of the absorbent sheet 26 may be made narrower than that of the upper sheet 28 or the like as a whole, but normally, the simplified measuring device 1 of the present invention is used.
For the absorbent section 18 of 0, a sheet in which a large number of members of the absorbent section 18 are continuously formed in the width direction is formed for each member, and the sheets are laminated and adhered in a predetermined order as shown in the drawing. After that, the sheet is cut and used as the absorbing portion 18 of each simple measuring device 10. Therefore, if the width of the absorbent sheet 26 is narrowed as a whole, the productivity of the absorbent section 18 and thus the simple measuring device 10 is significantly reduced. It is preferable that the region to some extent has a width similar to that of the top sheet 28 and the like. The region where the width of the absorbent sheet 26 is narrowed on the upstream side in the flow direction may be appropriately determined according to the amount of the sample to be collected, the liquid absorption capacity of the absorbent sheet 26, the productivity, and the like.

The top sheet 28 and the bottom sheet 3
It is more preferable that 0 is made slightly longer than the absorption sheet 26, and the upper-side sheet 28 and the lower-side sheet 30 are directly adhered to each other at the end portion on the observation through-hole 36 side (upstream side). It is possible to prevent liquid leakage and liquid invasion of the sample.

As a preferred mode, the simplified measuring device 10 of the illustrated example is configured to emit a sign for notifying the end of measurement. This signature is determined, for example, by adding the dye 44 to an appropriate position of the absorption sheet 26 and at an appropriate position of the absorption unit 18 (the flow of the sample and the speed of the above-mentioned specific binding reaction). ), A window for observing the end sign for visually observing the dye can be formed. With such a configuration, the sample is absorbed by the absorption sheet 26 after passing through the determination unit 16, and the sample 44 is dissolved and colored while the sample penetrates the absorption sheet in the longitudinal direction of the slit 38, Next, when the colored sample reaches the window for observing the end signature, the end signature is indicated. Further, the generation of such a signature may utilize coloring or discoloration accompanied by a chemical reaction.

In the simplified measuring device 10 of the illustrated example, in order to prevent the absorption sheet 26 and the double-sided tape 42 from being visible to the user and to improve the design of the device, the upper surface of the upper surface sheet 28 is preferably a The upper surface color sheet 32 that is opaque and has an arbitrary color or pattern is covered with the lower surface sheet 30, and the lower surface of the lower surface sheet 30 is also covered with the same lower surface color sheet 34. On the top color sheet 32,
Through-holes 32a corresponding to the through-holes 40 for bleeding the absorbent sheet 26 are provided in the lower color sheet 34.
The through hole 34a corresponding to the observation through hole 36 of No. 6 is further provided on both sheets with the windows 32b and 34 for observing the end sign.
b are formed respectively. This top color sheet 32
The lower surface color sheet 34 and the lower surface color sheet 34 are the upper surface sheet 28 and the lower surface sheet 30 by a known method such as an adhesive or a double-sided tape.
Adhered to.

It is to be noted that the upper surface color sheet 32 and the lower surface color sheet 34 may not be provided and the upper surface sheet 28 and the lower surface sheet 30 may be colored to have the same effect.

It is preferable that the absorbing portion 18 of the simple measuring device 10 of the present invention as described above is constructed so as to be flexible at least in the longitudinal direction thereof so that the self-shape can be held. With such a configuration, the operation of collecting a sample is easy, and it is preferable in terms of handleability.

Although the absorbing portion 18 in FIG. 1 has a rectangular shape, the present invention is not limited to this, and various shapes such as an elliptical shape, a rhombic shape and the like shown in FIG. It is possible.

The simple measuring device 10 of the present invention as described above is
Basically, after producing the absorbing portion 18 as described above,
A predetermined position on the upper surface of the absorbing portion 18, that is, the absorbing sheet 26
The determination section 16 is laminated and adhered so that the observation through-hole 36 and the reaction area coincide with each other. It is manufactured by stacking the sampling part 12 accommodating the reagent part 14 on the convex part 24 a of the seal 24 so as to be in agreement, and adhering it to the upper surface of the absorbing part 18. Alternatively, after fixing the determination unit 16, the reagent unit 14 is placed at a predetermined position on the determination unit 16, and the sampling unit 1 is placed so that the reagent unit 14 is housed in the convex portion 24a.
It is produced by placing 2 and adhering it to the upper surface of the absorbing portion 18. Therefore, the device of the present invention is disclosed in
The devices disclosed in Japanese Patent Laid-Open No. 166 and Japanese Patent Application Laid-Open No. 6-273419 do not require an indispensable case, and thus have high flexibility, easy sample collection operation, and high productivity. It can be offered to the market simply and inexpensively.

The measurement using the simple measuring apparatus 10 of the present invention, that is, the measuring method of the present invention is basically performed as follows. First, the sample is applied to the sampling part 12, the sampling part 12 is immersed in the sample, and the through hole 20 of the sampling part 12 is provided.
The sample is collected by dropping the sample on a. Here, in the device of the present invention, the through hole 2 of the spacer 20 is used.
Due to the action of 0a and the air bleeding 20b, a fixed amount of sample can be collected easily and quickly. It should be noted that foreign matter and impurities larger than the pores of the membrane 22 contained in the sample are
It is removed by the membrane 22. Needless to say, it is preferable to keep the collecting unit 12 upward after collecting the sample.

The sample collected by the sampling unit 12 is the reagent unit 1
4 for observing the absorbent sheet 26, which is formed below the determination unit 16 after passing through the determination unit 16 and eluting the soluble reagent when being mixed therewith The space formed by the through holes 36 and the like is reached. As a result, the analyte, the soluble reagent, and the affinity substance come into contact with each other to cause the above-mentioned sandwich-type or competitive-type specific binding reaction, and a signal is generated in the reaction region in which the affinity substance is immobilized. Here, in addition to the film 22 described above, foreign substances and impurities contained in the sample are further removed by the porous material of the reagent section 14, so that the signal generated in the determination section 16 is very clear and accurate. Further, by having the space, the analyte, the soluble reagent and the affinity reagent can be in contact with each other for a sufficient time, and the reaction is surely proceeded.

On the other hand, when the specimen that has flowed into the space formed below the determination portion 16 comes into contact with the absorption sheet 26, it is impregnated into the absorption sheet 26 and absorbed. Here, in the simple measuring device 10 of the present invention, the absorption sheet 26 has an observation through-hole 36.
Since the slit 38 and the through-hole 40 that are connected to the absorption sheet 26 are formed, the liquid flow of the sample in the absorption sheet 26 is promoted, and the liquid flow of the sample is preferably exhausted, and the absorption of the sample into the absorption sheet 26, That is, the flow of the sample from the collection unit 12 to the determination unit 16 is quick. Therefore, the signal generation to the determination unit 16 is performed quickly, and the measurement can be completed in a shorter time than in the conventional case.

After the measurement is completed, this signal is confirmed from the lower surface of the simple measuring device 10 through the through hole 34a of the lower surface color sheet 34 and the through hole 36 for observation of the absorbing sheet 26 to obtain the measurement result.

The simple measuring device of the present invention and the measuring method using the same have been described above in detail. However, the present invention is not limited to the above-mentioned examples, and various types of devices can be used without departing from the scope of the present invention. Of course, improvements and changes may be made.

[0079]

EXAMPLES The present invention will be described below with reference to specific measurement examples, but the present invention is not limited thereto.

[Example 1 (Measurement of hCG)] The simple measuring device 10 shown in FIGS. 1 and 2 is as follows.
Was prepared and hCG was measured.

(A) (Test sample) Preparation of sampling section 12 A through hole 20a having a diameter of 9 mm is formed in the approximate center of a plate-like foamed polyethylene having a width of 12 mm, a length of 18 mm and a thickness of 2 mm, and further, a width of 3 mm. The air vent 20b was formed, and the spacer 20 as shown in FIG. 1 was produced. Also, a portion of a polyester sheet having a width of 12 mm and a length of 15 mm corresponding to the through hole 20a was extruded, and then a hole having a diameter of 6 mm was opened at the center thereof to form a convex portion 24a to obtain a sheet 24. Furthermore, a non-woven fabric made of rayon having a width of 12 mm and a length of 18 mm was prepared to form a liquid-permeable membrane 22. Fabricated spacer 20
The film 22 is attached to the upper surface of the sheet using an adhesive, and the convex portion 24a is inserted into the through hole 20a from below the spacer 20 to stack and adhere the sheet 24, as shown in FIGS. The sampling unit 12 was produced.

(B) Purification of Anti-hCG Monoclonal Antibody Two kinds of antibodies obtained by the cell fusion method using polyethylene glycol (immunity experimental manipulation method VII 2211, 1978), each recognizing different antigenic determinants on hCG, were obtained. Two hybridoma strains producing a monoclonal antibody were cultivated, and immunoglobulin G fractions were respectively obtained from 10 L (liter) of each of the obtained culture supernatants by the ammonium sulfate precipitation method.
Each was purified by affinity chromatography using a protein A sepharose column to obtain antibodies A and B. The obtained antibodies A and B were dialyzed against physiological saline to obtain 250 mg of antibody A and 245 mg of antibody B, respectively.

(C) Preparation of reagent part 14 1 g of a disperse dye (KAYARON RED VIOLET manufactured by Nippon Kayaku Co., Ltd.) was suspended in 10 mL (milliliter) of distilled water, and 1600 ×
Centrifuge at 10g for 10 minutes and add 22,000 xg to the supernatant.
After 10 minutes of centrifugation at 10 ° C., 10 mL of distilled water was added to the obtained precipitate and suspended to obtain a disperse dye for labeling. 0.8 mg prepared by diluting 0.5 mL of this disperse dye for labeling with physiological saline
0.5 mL of the above-mentioned antibody A was added thereto at a temperature of 56 ° C. for 30 minutes. After ice-cooling for 20 minutes, 5 mL of glycine buffer was added and the mixture was centrifuged at 22000 xg for 20 minutes. The obtained precipitate was suspended in 5 mL of glycine buffer and then centrifuged again at 22000 xg for 20 minutes. It was separated and washed. 1.25 mL of 2.5% bovine serum albumin (hereinafter referred to as BSA) / 10% saccharose / phosphate buffered saline (hereinafter referred to as PBS) was added to the obtained precipitate.
Was added and suspended to obtain a disperse dye-labeled anti-hCG antibody (hereinafter referred to as dye-labeled antibody A). 0.03 mL of the obtained dye-labeled antibody A was dispensed into a circular filter paper (No. SM-45 manufactured by Azumi Filter Paper Co., Ltd.) cut into a circle having a diameter of 8 mm to impregnate the filter paper,
Lyophilization was performed to produce the reagent part 14 to which the soluble reagent was applied.

(D) Preparation of determination part 16 A nitrocellulose membrane (manufactured by Toyo Roshi Kaisha, Ltd.) having a pore size of 5 μm was dipped in 0.5% BSA, pre-coated and washed with a physiological saline solution, and then with a physiological saline solution. Apply 4 mg / mL of the diluted antibody B above, dry at 37 ° C.,
The determination unit 16 is used.

(E) Preparation of absorbing section 18 A sheet of cellulose filter paper (N0.1 filter paper manufactured by Whatman) having a width of 12 mm, a length of 118 mm and a thickness of 0.18 mm is formed on a sheet of observation through holes 36, slits 38 and air vents. The through-holes 40 for water are formed and the absorbent sheet 26 as shown in FIG. 1 is manufactured. The observation through-hole 36 and the through-hole 40 have a diameter of 6 mm, the slit 38 has a width of 1 mm and a length of 84 mm (from the inside of both holes to the inside), and the observation through-hole 36 is an end of the absorbent sheet 26 in the longitudinal direction. It was formed at a position 4 mm from the part.
In addition, the observation through hole 36, the slit 38, and the through hole 4
The formation position of 0 is the center of the absorbent sheet 26 in the width direction.
Further, as shown in FIG. 1, both sides in the width direction on the observation through-hole 36 side (upstream side) of the absorbent sheet 26 were narrowed by 1 mm (total of 2 mm on both sides) over 78 mm. Furthermore, from the upstream end of such an absorbent sheet 26, a length of 16 mm
1 μL of Food Red No. 2 was added dropwise to the position as a dye for emitting a sign indicating the end of measurement. Such an absorbent sheet 26 has a width of 12 mm, a length of 120 mm and a thickness of 0.
The upper sheet 28 and the lower sheet 30 made of a 075 mm polyester transparent sheet were sandwiched and adhered. In addition, in the upper surface sheet 28, communication through holes 28 a and through holes 28 b having the same diameter are formed at positions corresponding to the observation through holes 36 and the through holes 40 of the absorbent sheet 26. Further, the absorbent sheet 26 and the upper surface sheet 28 are bonded to each other with an adhesive, the absorbent sheet 26 and the lower surface sheet 30 are bonded to each other with a double-sided tape 42 having holes drilled in the same manner as the absorbent sheet 26, and the adhesive sheet 26 is bonded to the absorbent sheet 26. The upper sheet 28 and the lower sheet 30 were projected by 2 mm from the upstream end.
Furthermore, the width 12 mm and the length 104 mm are made of white polyester and have a through hole 32 a of the same diameter at a position corresponding to the air vent through hole 40 of the absorbent sheet 26, and 5 mm from the upstream side.
An upper surface color sheet 32 having a window 32b for observing an end sign having a diameter of 3 mm at the position is laminated on the upper surface of the upper surface sheet 28 so that the downstream end faces are aligned and bonded with an adhesive. It is made of white polyester with a length of 120 mm, and has a through hole 34a of the same diameter at a position corresponding to the through hole 36 for observation of the absorption sheet 26, and a window 34b for observing an end sign having a diameter of 3 mm at a position 21 mm from the upstream side. The formed lower surface color sheet 34 is laminated on the lower surface of the lower surface sheet 30 and adhered with an adhesive to form the absorbing portion 18.

(F) Manufacture of the simple measuring device 10 The judgment part 16 manufactured in (d) above is laminated on the upper surface of the absorbent part 18 manufactured in (e) above in accordance with the observation through holes 36 of the absorbent sheet 26. Then, the periphery was adhered with an adhesive. Furthermore, the reagent part 14 prepared in (c) above is inserted into the protrusion 24a of the seal 24, and the sampling part 12 prepared in (a) above is inserted.
Were laminated so that the reagent part 14 corresponded to the through holes 36 for observation, and adhered to the upper sheet 28 to fabricate the simple measuring device 10 of the present invention shown in FIGS. 1 and 2.

(G) Measurement of hCG hCG standard product (according to 3rd IS 75/537) was diluted with urine of a healthy man to obtain 1,000, 500, 20.
HCG standard solutions of 0, 100 and 50 mIU / mL were prepared.
The sampling unit 12 of the simple measurement device 10 prepared in (f) above was dipped in each hCG standard solution and taken out after about 5 seconds, and the sampling unit 12 was placed upward, and the end sign was the window 32b (3).
4b), the measurement was terminated, and the color development in the determination unit 16 (antibody B-immobilized membrane) was visually determined from the back side of the measuring device. Those with coloration were judged as positive (+), and those without coloration were judged as negative (-). The results are shown in Table 1.

Table 1 (hCG measurement results) ─────────────────────────────── hCG concentration mIU / ml 0 50 100 200 500 1000 ─────────────────────────────── Judgment − ＋＋＋＋＋＋ ───────── ────────────────────── End sign occurrence time (seconds) 21 32 22 25 28 25 ─────────────── ────────────────

[Example 2 (LH measurement)] Reagent part 14
And the simple measuring device 10 shown in FIGS. 1 and 2 was produced in the same manner as in Example 1 except that the determining unit 16 was changed to the following, and LH was measured.

(A) Purification of Anti-LH Monoclonal Antibody Two types of monoclonal antibodies obtained by the cell fusion method using polyethylene glycol (immunity experimental manipulation method VII 2211, 1978), each recognizing different antigenic determinants on LH, were obtained. Two hybridoma strains producing a monoclonal antibody were cultured, and 10 L of each obtained culture supernatant was purified in the same manner as in Example 1 above to obtain antibodies C and D. The obtained antibodies C and D were dialyzed against physiological saline to obtain 205 mg of antibody C and 255 mg of antibody D, respectively.

(B) Preparation of Reagent Part 14 1 g of disperse dye (KAYARON RED VIOLET manufactured by Nippon Kayaku Co., Ltd.) was suspended in 10 mL of distilled water and centrifuged at 1000 × g for 10 minutes. After centrifugation at g for 10 minutes, 10 mL of distilled water was added to the resulting precipitate to suspend it to give a disperse dye for labeling. 0.5 mL of disperse dye for labeling,
2 mg / mL of the antibody C prepared by diluting with physiological saline was added to 0.2 wt.
5 mL was added and incubated at 56 ° C. for 30 minutes.
After ice-cooling for 20 minutes, add 5 mL of PBS and add 22,000.
After centrifugation at xg for 20 minutes, the obtained precipitate was suspended in 5 mL of distilled water and again centrifuged at 22,000 xg for 20 minutes to wash. 1.25 mL of 7.5 was added to the resulting precipitate.
% Bovine serum albumin / 20% saccharose / glycine buffer was added and suspended to obtain a disperse dye-labeled anti-LH antibody (hereinafter referred to as dye-labeled antibody C). Filter paper obtained by cutting the dye-labeled antibody C into a circle having a diameter of 8 mm (No. S manufactured by Azumi Filter Paper Co., Ltd.
M-45) was dispensed in 0.03 mL aliquots to impregnate each with M-45) and freeze-dried to produce reagent part 14.

(C) Preparation of determination part 16 A nitrocellulose membrane (manufactured by Toyo Roshi Kaisha, Ltd.) having a pore size of 5 μm was immersed in 0.5% BSA for precoating and washed with a physiological saline solution, and then the reaction zone was obtained. Range (8
An antibody solution was applied in two spots with a diameter of 2 mm to a square (mm mm) to prepare a determination part 16. That is, one was coated with 2 mg / mL of the antibody D diluted with physiological saline in the same manner as in (d) of Example 1, and dried at 37 ° C. The other is LH50mIU / mL as a positive control
The amount of the anti-mouse IgG antibody corresponding to the color development was applied in the same manner and dried.

(D) LH measurement LH standard product (UCB, Luteinizing Hormone 1st IRP
68/40) was diluted with urine of a healthy man to prepare LH standard solutions 200, 100 and 50 mIU / mL LH standard solutions. The sampling unit 12 of the simple measuring device 10 is dipped in this solution, taken out after about 5 seconds, placed with the sampling unit 12 upward, and the measurement is finished when the end sign appears in the window 34b (32b), and the judgment is made. Color development in the part 16 (antibody D-immobilized membrane) was visually judged from the back side of the simple measurement device 10. Judgment was made such that no coloration was observed or the coloration was weaker than that of the positive control, negative (-), and those having the same or strong coloration were positive (+). Table 2 shows the results.

Table 2 (Measurement of LH) ────────────────────────── LH concentration 0 50 100 200 mIU / ml ───── ───────────────────── Judgment − ＋＋＋ ──────────────────────── ─── End sign occurrence time (seconds) 35 23 25 26 ──────────────────────────

[Example 3 (Effect of presence or absence of slit)]
A simple measuring device was produced in exactly the same manner as in Example 1 except that the absorbing sheet 26 of the absorbing portion 18 did not have the slit 38 and the through hole 40 for venting air. This simple measuring device and the simple measuring device 1 of the present invention produced in Example 1
0 in the same manner as in (g) of Example 1 and 100
The hCG measurement was carried out using 0 mIU / mL hCG standard solution. The measurement was performed 10 times for each device, and it was examined whether the presence or absence of the slit had an influence on the reaction time (end sign appearance time). As shown in the results in Table 3, it was confirmed that the reaction time (end sign appearance time) was significantly shortened by providing the slit.

Table 3 (Effect of presence or absence of slit) ────────────────────────────────── Yes No No Yes ───────────────────────────────── Judgment ＋ ＋ ─────────── ─────────────────────── End sign appearance time (seconds) 59.8 28.8 Standard deviation 5.5 3.8 Coefficient of variation (%) 9.2 13.2 Significant difference (no slit) + ^* ───────────────────────────────── ^*: There is a significant difference of 0.1% risk rate (+) To

[Embodiment 4 (Study of slit width)] A simple measuring apparatus was prepared in exactly the same manner as in Embodiment 1 except that the absorbent sheet 26 of the absorbent section 18 did not have the slit 38 and the through hole 40 for venting air. It was made. Further, a simple measuring device was produced in exactly the same manner as in Example 1 except that the width of the slit 38 was changed to 2 mm, 3 mm and 5 mm (invention product).
Using these simple measuring devices and the simple measuring device 10 of the present invention produced in Example 1, hCG was measured using 1000 mIU / mL hCG standard solution in exactly the same manner as in (g) of Example 1. went. Five measurements were performed for each device, and it was examined whether the slit width affects the reaction time (end sign appearance time). Table 4
As shown in the results, the reaction time (the time of appearance of the end sign) was hardly affected by the width of the slit.

Table 4 (Slit width examination) ───────────────────────────────────── No slit width 1mm 2mm 3mm 5mm ──────────────────────────────────── Judgment ＋＋＋＋＋＋ ──── ───────────────────────────────── End sign Appearance time (seconds) 58.0 25.6 30.2 29.6 35.8 Standard deviation 4.7 4.5 2.8 4.4 8.7 Coefficient of variation (%) 8.1 17.6 9.3 14.9 24.3 Significant difference (no slit) ＋ ^* ＋ ^* ＋ ^* ＋ ^* ──────────────────────── ───────────── *: Significantly different (+) with a risk rate of 0.1%

[Embodiment 5 (Slit length study)] A simple measuring apparatus was prepared in exactly the same manner as in Embodiment 1 except that the lengths of the slits 38 of the absorbent sheet 26 of the absorbent section 18 were changed to 20 mm and 100 mm. (Invention product) Using these simple measuring devices and the simple measuring device 10 of the present invention produced in Example 1, hCG was measured using 1000 mIU / mL hCG standard solution in exactly the same manner as in (g) of Example 1. went. Five measurements were performed for each device to examine whether the slit length affects the reaction time (end sign appearance time). As shown in the results in Table 5, the reaction time (end signature appearance time) was hardly affected by the slit length.

Table 5 (Slit length examination) ─────────────────────────────────── Slit length 84mm 20mm 100mm ────────────────────────────────── Judgment + ＋ ＋ ──────────── ─────────────────────── End sign Appearance time (seconds) 26.8 29.8 28.0 Standard deviation 4.7 9.5 2.9 Coefficient of variation (%) 17.5 31.9 10.4 Significant difference (vs. 84mm) None None ──────────────────────────────────

[Embodiment 6 (Effect of presence or absence of slit)]
The slit 38 of the absorbent sheet 26 has a length of 8 mm and a width of 2 mm.
(See FIG. 3A), through holes 40, 28 for venting air
A simple measuring apparatus as shown in FIG. 4 was produced in exactly the same manner as in Example 1 except that it had no b or the like and was not opened to the atmosphere (invention product). Using such a simple measuring device of the present invention, in the same manner as in (g) of Example 1, 100
The hCG measurement was carried out using 0 mIU / mL hCG standard solution. The measurement was performed 10 times, and compared with the measurement results obtained by the measuring device having neither the slit 38 nor the air vent through hole 40 measured in the third embodiment, the slit was measured in the device having no air vent through hole 40. It was examined whether the presence or absence of the item affects the reaction time (the time when the end sign appears). As shown in Table 6, the through holes 4 for venting air
It was confirmed that even in the apparatus having no 0, the reaction time (the time of appearance of the end sign) was remarkably shortened by installing the slit, similarly to the result obtained in Example 3.

Table 6 (Effect of presence or absence of slit) ────────────────────────────────── Presence or absence of slit ───────────────────────────────── Judgment + ＋ ────────────── ──────────────────── End signature appearance time (seconds) 59.8 32.4 Standard deviation 5.5 6.5 Coefficient of variation (%) 9.2 20.1 Significant difference (no slit) + ^* ─ ──────────────────────────────── *: Significantly different (+) at a risk rate of 0.1% From the results, the effect of the present invention is clear.

[0103]

As described above in detail, according to the present invention, the B / F as an independent step in the measurement operation is performed.
It does not require separation and can perform quick and simple measurement in one step operation, and also has high measurement accuracy and is inexpensive.
It is possible to realize a simple measuring device that is most suitable for disposable applications.

[Brief description of drawings]

FIG. 1 is a schematic exploded perspective view of an example of a simple measuring device of the present invention.

FIG. 2 is a schematic sectional view of the simple measurement device shown in FIG.

FIG. 3A and FIG. 3B are schematic perspective views showing another example of an absorbent sheet used in the simple measuring device of the present invention.

FIG. 4 is a schematic exploded perspective view of another example of the simple measurement device of the present invention.

[Explanation of Codes] 10 Simple Measuring Device 12 Sample Specimen Collection Part 14 Reagent Part 16 Judgment Part 18 Absorption Part 20 Spacer 20a Through Hole 22 Membrane 24 Seal 26 Absorption Sheet 28 Upper Surface Sheet 30 Lower Surface Sheet 32 Upper Color Sheet 34 Lower Color Sheet 36 Through hole for observation 38 Slit 40 Through hole 42 Double-sided tape

Claims (12)

[Claims] 1. A measuring device for measuring an analyte in a liquid test sample, comprising: a spacer having a through hole for introducing a test sample; and a spacer covering the upstream side of the through hole. A test sample collecting part having a liquid permeable membrane, which is arranged to face the through hole downstream of the test sample collecting part, and a soluble reagent which is dissolved and released by contact with the test sample in a porous material. A determination unit having at least one reaction region formed by holding a reagent unit held therein and a affinity substance capable of binding to the analyte or soluble reagent, which is arranged downstream of the reagent unit, on a porous material. And, arranged downstream of the determination unit, a long upstream sheet and a downstream sheet made of a liquid impermeable material, having a long absorbent sheet made of a liquid absorbent material and sandwiched between the two sheets, The judgment area is near the edge Further, the absorbent sheet has a through-hole corresponding to the downstream side of the determination section, and the absorbent sheet extends in one direction continuously or close to the plane of the through-hole. Existing slits are formed, the upstream sheet is formed with through holes corresponding to the through holes of the absorbent sheet, and the downstream sheet is transparent at least at a portion corresponding to the through holes of the absorbent sheet. And a simple measuring device. 2. The simple measuring device according to claim 1, wherein the test sample collecting section has a function of collecting a predetermined amount of test sample by a space in a through hole formed in the spacer. 3. The determination unit has a plurality of reaction zones holding affinity substances different from each other.
Simple measurement device described in. 4. The simple measuring device according to claim 1, wherein the affinity substance fixed to the porous material of the determination part is an antibody or an antigen. 5. The soluble reagent retained in the porous material of the reagent part is an antibody or an antigen, and the antibody or antigen is labeled with a labeling substance to enable detection. The simple measurement device according to any one of 1. 6. The simple measuring apparatus according to claim 5, wherein the labeling substance is selected from the group consisting of colloidal metal, colloidal non-metal, dye and latex particles. 7. The porous material of the judgment part is selected from the group consisting of cellulose, cellulose derivative, nitrocellulose, porous synthetic polymer, glass fiber filter, non-woven fabric and cloth. Simple measurement device described. 8. The porous material of the reagent part is selected from the group consisting of cellulose, cellulose derivatives, nitrocellulose, porous synthetic polymers, glass fiber filters, non-woven fabrics and cloths. Simple measurement device described. 9. The liquid absorbent material forming the absorbent sheet of the absorbent part is selected from the group consisting of cellulose, cellulose derivatives, porous synthetic polymers, glass fiber filters, non-woven fabrics, particle absorbents and cloths. Item 1
8. The simple measuring device according to any one of 8. 10. The porous material of the determination part is nitrocellulose, and the liquid absorbent material forming the porous material and the absorbent sheet of the reagent part is cellulose.
The simple measurement device according to any one of 1. 11. A method for measuring an analyte in a liquid test sample by using the simple measuring device according to claim 1, which seems to contain the analyte. A certain amount of the test sample is collected in the through hole of the spacer by dropping or sprinkling the test sample to be collected on the test sample collecting part of the simple measuring device or by immersing the test sample collecting part in the test sample. Then, the soluble reagent retained in the porous material of the reagent part is eluted by flowing into the porous material of the reagent part to pass through the reagent part together with the test sample and flow into the determination part. Directly or indirectly binds the affinity substance fixed to the reaction zone of the porous material and the soluble reagent to form a detectable signal in the reaction zone and passes the determination zone. The test sample was The absorption is accelerated by the slit formed in the absorption sheet of the absorption section and quickly absorbed by the absorption sheet, and the signal is observed from the transparent portion of the downstream sheet to measure the analyte in the liquid test sample. Measuring method. 12. The measuring method according to claim 11, wherein the test sample is a body fluid or a solution obtained by diluting or extracting and diluting a biological component.