JPH08160040A - Determining device for occult blood of feces - Google Patents

Abstract

PURPOSE: To obtain a determining device for occult blood of feces which has a relatively simple construction and utilizes an immunoassay enabling simplification of an operation for determination of the occult blood of feces over all processes thereof. CONSTITUTION: This device has a feces collecting appliance 2 and a buffer solution container 3 and the feces collecting appliance 2 is provided with a feces collecting rod 5, while the buffer solution container 3 is provided with a determining part 11. By breaking through a closing material 12 by operating a closing member piercing member 13 from the outside, in the determining part 11, a buffer solution wherein a sample of feces is dissolved is made to flow into the determining part 11. Solids therein are removed by a separating filter 15 and the presence or absence of occult blood of feces is determined in a developing layer 16 according to an immuno-chromatographic method.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a simple fecal occult blood determining apparatus for determining the presence or absence of fecal occult blood using an immunization method.

[0002]

2. Description of the Related Art Fecal occult blood is produced by digestive system diseases resulting from contamination of blood in the stool in the digestive tract. Particularly, the correlation with diseases such as colorectal cancer has been confirmed, and it is a very important and widely used test as an index for early diagnosis of colorectal cancer, which is increasing in number in Japan.

Currently used fecal occult blood test methods include:
There are two main methods. The first method is called a conventional chemical occult blood reaction (hereinafter referred to as “chemical method”), which is a nonspecific reaction for detecting peroxidase-like activity of hemoglobin (hereinafter referred to as Hb). Is. This method is based on a mechanism in which a chromogen (such as guaiac butter or orthotolidine) is oxidized in the presence of a peroxidase-like activity-retaining substance and hydrogen peroxide solution to develop a color. Therefore, in this method, since the drug is a peroxidase-like activity-retaining substance, food containing Hb, and the like are positive, medication and dietary restriction were required.

A simple measuring system is disclosed as an inspection device based on the present chemical method, and some of them are widely used. For example, Japanese Examined Patent Publication No. 2-45826 and Japanese Patent Laid-Open No. Sho 5
In JP-A-9-168370, JP-A-62-266463, and JP-A-62-153573, a small amount of a stool sample is applied to a card containing a test paper impregnated with guaiac fat or the like, and then peroxidized. A system is disclosed in which hydrogen water is dropped to check the color appearance. In these systems,
Whether a suitable amount of stool sample is applied on a part of the guaiac fat attached to the card type body and hydrogen peroxide solution is dripped on the part on the back side of the application part to see if the guaiac fat in the dripping part develops color. Determine with. It is advantageous that it can be performed without opening the stool sample application part, that it can be handled hygienically, and that it has a simple structure, but the drawbacks derived from the above-mentioned measurement principle have not been solved.

The second method has been recently developed to solve the above-mentioned drawbacks of the chemical method, and is a method based on an immune reaction (hereinafter referred to as "immunization method"). Since this method is a method based on an antigen-antibody reaction using an anti-human Hb antibody, it has high specificity and does not require dietary restrictions such as chemical methods. The determination is performed by a general immunoserologic test method such as a latex agglutination method or an enzyme-labeled immunoassay, but the most widely used method at present is the latex agglutination method.

In the latex agglutination method, a stool sample solution and a latex reagent are dropped on a judgment plate and mixed, and judgment is made based on the presence or absence of an agglutination image. In principle, the specificity is higher than that of the chemical method, but skill in determining the agglutination method is required. Further, the test system based on the immunoassay is provided as a kit including a judgment plate, a latex reagent, a stirring rod, a stool collection instrument, a solution, etc., but has a complicated structure and is as simple as the above chemical method. The system has not been developed.

[0007] The simplification of these immunization methods is to carry out a step unique to the immunization method, which is not present in the chemical method, that is, a step of dissolving a stool sample in a buffer solution or the like, and a non-dissolved solid content in the dissolved solution after being removed by filtration. It is done about the process of doing. That is, for these operations, a stool collection device capable of relatively quantitative and hygienic handling is disclosed, for example, Japanese Utility Model Publication No. 3-44363 and Japanese Utility Model Publication No. 2-13977.
Japanese Utility Model Publication No. 1-66071 Japanese Utility Model Publication No. 1-85
In Japanese Patent No. 668 and Japanese Utility Model Application Laid-Open No. 64-42454, a collection rod for collecting feces is attached to the lid of a container containing a buffer solution, and the lid is set in the buffer container after the collection of feces. Thereby, the stool at the tip of the collecting rod is dispersed in the buffer solution. In addition, a filter mechanism is installed in the dropping portion to remove insoluble solids when the droplet is dropped onto the determination plate from the dropping portion after being shaken well. After dropping onto the judgment plate, the operation is carried out according to the usual method of the above-mentioned latex agglutination method.

The simplification of the fecal occult blood measurement system by the immunization method is considered to be achieved by the integration of the above steps and the antigen-antibody reaction, but the technique has not been realized so far.

[0009]

In recent years, with the spread of the concept of self-care, a simple test system based on an immune reaction has become available as a general test drug at pharmacies and the like. For example, for pregnancy diagnostics, a simple test system using an immunochromatography method has been established. The biggest problem in using the immunochromatography method for fecal occult blood test is that the stool sample lysis and filtration steps are difficult to be incorporated into the system.

An object of the present invention is a fecal occult blood test apparatus utilizing an immune reaction, which has a relatively simple structure, but the operation is simplified over all steps of the fecal occult blood determination operation, and is unskilled. An object of the present invention is to provide a fecal occult blood determining apparatus that enables a person to easily and accurately determine the presence or absence of fecal occult blood.

[0011]

The inventions set forth in claims 1 to 3 have been accomplished to achieve the above object,
A stool collection tool having a stool collection stick for holding the collected stool attached to the outer surface thereof, and a lid connected to the stool collection stick, and a buffer container closed by the lid of the stool collection tool. The fecal occult blood determination device is provided with each of the following features.

That is, according to the first aspect of the invention, a buffer solution container containing a buffer solution, a lid attached to the buffer solution container so as to close the buffer solution container, and a lid body connected to the lid body. And a stool collection device having the stool collection rod. A determination unit is provided on the side wall of the buffer solution container. The determination unit is a closed member provided on the inner wall of the buffer container so as to close the inside of the determination unit to prevent the buffer solution from flowing into the determination unit, and the closed state by the closed member is operated by an external operation. A closed state opening member attached to the buffer solution container so that it can be opened, and a stool sample inside the determination section to remove the solid content in the buffer solution in which the stool sample that has flowed in when the closed state is opened And a separation filter provided in.
Furthermore, in the determination part, a spreading layer is arranged so that the fecal sample solution whose solid content has been removed by the separation filter is supplied. An anti-hemoglobin antibody-sensitized carrier is placed on the spreading layer and the anti-hemoglobin antibody is immobilized. Further, in order to observe the presence or absence of occult blood and the end of the determination from the outer surface of the buffer solution container, a determination window and an end window are formed on the side wall of the buffer solution container so that the spreading layer can be partially observed. There is.

In the fecal occult blood determining apparatus according to the second and third aspects of the present invention, a buffer solution container containing a buffer solution, a lid body and a lid body attached to the buffer solution container so as to close the buffer solution container. And a stool collection tool having a stool collection rod connected to the. Also, in the inventions according to claims 2 and 3, the determination portion is provided on the side wall of the buffer solution container.

According to the second and third aspects of the present invention, the judging section is provided on the inner wall of the buffer solution container so as to close the judging section in order to prevent the buffer solution from flowing into the judging section. The material and the closed state opening member attached to the buffer container so that the closed state by the closed material can be opened by an external operation, and the stool sample that has flowed in when the closed state is opened is dissolved. In order to remove the solid content in the buffer solution,
And a separation filter provided in the determination unit.

In the invention described in claims 2 and 3,
An immune filter is arranged in the determination unit. This immune filter is arranged on the opposite side of the closure material with the separation filter interposed therebetween so that the fecal sample solution in which the solid content is removed by the separation filter is supplied.

According to the second aspect of the invention, in the immunofilter, the anti-hemoglobin antibody-sensitized carrier is movably set by the buffer solution. Further, the immunofilter is configured to have pores through which an anti-hemoglobin antibody-sensitized carrier can pass but a product of an antigen-antibody reaction between the anti-hemoglobin antibody-sensitized carrier and hemoglobin cannot pass.

Further, in the invention described in claim 3,
In the immunofilter, the anti-hemoglobin antibody-sensitized carrier is movably installed by the buffer solution, and the second anti-hemoglobin having a different recognition site from the anti-hemoglobin sensitized by the anti-hemoglobin antibody-sensitized carrier is further provided. The antibody is immobilized so that it cannot be moved, and has a pore through which the anti-hemoglobin antibody-sensitized carrier can pass.

[0018]

When determining the fecal occult blood using the fecal occult blood determining apparatus according to the invention described in claims 1 to 3, first, the stool sample of the stool collection device is pierced into the stool sample and pulled out, Collect a stool sample. After that, the stool collection rod is inserted into the buffer solution container, and the stool collection device is attached to the buffer solution container.
As a result, the stool sample is dissolved in the buffer solution in the buffer container,
A buffer solution is prepared in which the stool sample is dissolved. Claims 1-3
In the invention described in (1), the closing material is attached to the inner wall of the buffer solution container in order to prevent the buffer solution in which the stool sample is dissolved from flowing into the determination unit. Therefore, as it is, the buffer solution in which the fecal sample is dissolved does not flow into the determination unit.

In the invention described in claims 1 to 3, the stool sample is dissolved by externally operating the closed state opening member provided in the determination part to open the closed state by the closing material. The dissolution liquid flows into the determination unit. The buffer solution in which the stool sample is dissolved is filtered by the separation filter in the determination unit to remove the solid content. Next, the buffer solution in which the stool sample is dissolved and the solid content is removed is supplied to the spreading layer in the invention described in claim 1 and to the immune filter in the invention described in claims 2 and 3. .

According to the first aspect of the invention, the presence or absence of fecal occult blood is determined by the immunochromatographic method in the spreading layer, as will be apparent from the description of the examples given below.
Similarly, in the invention described in claims 2 and 3, the presence or absence of fecal occult blood is determined by the immune filter according to the immune filter method.

Further, in the invention described in claim 1, since the judgment window and the end window are formed on the side wall of the buffer solution container,
The determination can be easily made from the outside, and the end of the determination can be confirmed. In the inventions described in claims 2 and 3, if the buffer solution container is transparent, the determination can be easily performed from the outside, and the end of the determination can be confirmed.

As described above, in the inventions according to claims 1 to 3, after the stool sample is collected using the stool collection device and the stool collection device is attached to the buffer container, the closed state opening member is provided. The presence or absence of fecal occult blood can be easily determined by operating the. That is, since the determination unit is provided on the inner wall of the buffer solution container, it is possible to reliably and easily perform from the collection of a stool sample to the determination of fecal occult blood, while having a relatively simple structure.

[0023]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be clarified by describing non-limiting embodiments of the present invention with reference to the drawings.

First Embodiment FIGS. 1 (a) and 1 (b) are a sectional view and a longitudinal sectional view for explaining a fecal occult blood determining apparatus according to an embodiment of the present invention. The fecal occult blood determination device 1 of the present embodiment is composed of a fecal collection device 2 and a buffer solution container 3. Less than,
Each member will be described in detail.

The stool collection device 2 has a lid 4 and a stool collection rod 5 connected to the lid 4. In the present embodiment, the lid 4 and the stool collecting rod 5 are integrally formed of synthetic resin, but after the lid 4 and the stool collecting rod 5 are formed as separate members, they are fixed by an adhesive or the like. It may have been done.

The lid 4 is made of a cylindrical member opened downward, and has an internal thread 4a formed on the inner peripheral surface of the cylindrical side wall. On the other hand, the stool collecting rod 5 is connected to the center of the lower surface of the top plate of the lid body 4 and is composed of an elongated columnar member. A stool collection part 6 is formed at the tip of the stool collection rod 5.

The stool collection section 6 is constructed by giving unevenness to the surface. The unevenness is provided for semi-quantitatively collecting the stool sample when the stool sample is stuck and pulled out. The stool collection unit 6 can be configured in any shape as long as it can semi-quantitatively collect a stool sample, and can be provided with depressions of various shapes or projections.
Alternatively, it can be configured by forming a hole. The buffer solution container 3 has a rectangular tube-shaped container body 7 and a cylindrical mounting portion 8 provided at the upper end of the container body 7 and having an opening at the top. A male screw 8a is formed on the outer peripheral surface of the cylindrical mounting portion 8, and the male screw 8a is
It is formed so as to mesh with the female screw 4 a of the lid body 4.

A surplus feces removing member 9 is attached from the upper surface of the cylindrical attaching portion 8 toward the center. The surplus stool removing member 9 has a substantially conical shape provided so that the central side thereof extends downward, and a through hole 9a is formed at the lower end. The surplus feces removing member 9 is made of synthetic resin or metal having an appropriate thickness so as to have elasticity, and a slit is formed around the through hole 9a. Therefore, when the stool collecting rod 5 is inserted through the through hole 9a,
The peripheral part of the excess stool removing member 9 facing the through hole 9a is the stool collection part 6
Will be abutted against the outer surface of the. Therefore, the surplus stool sample among the stool samples attached to the stool collection unit 6 is removed at the peripheral portion, and a predetermined amount of the stool sample collected by the stool collection unit 6 is removed. Only the stool sample will be inserted into the buffer solution container 3.

A buffer solution 10 is stored inside the buffer solution container 3. As the buffer solution 10, for example, a near neutral salt solution such as an ammonia buffer solution, a glycine buffer solution, a phosphate buffer solution, a Tris buffer solution is preferable, and particularly preferably,
The said buffer solution of pH = 6.5-8.5 is mentioned. The amount of buffer solution is preferably about 0.5 to 10 ml, though it depends on the amount of the stool sample to be collected.

In the side wall of the container body 7 of the buffer solution container 3,
The determination unit 11 is configured. In the determination unit 11,
A closure member 12 is provided to prevent the buffer solution in which the fecal sample is dissolved from flowing into the determination unit 11. The closing member 12 is fixed to the inner surface of the side wall so as to close the through hole facing the inner peripheral surface of the side wall of the container body 7. Closure material 12
Is made of a material that can be pierced by the closed state opening member 13 described later. As such materials,
A sealing material such as a synthetic resin film having an appropriate film thickness or hot melt heat sealing can be used.

Further, in the judging section 11, a closed state opening member 13 is provided as a member for piercing the closing member 12 by operating from the outside. In this embodiment, the closed state opening member 13 is used which has a needle-shaped portion 13a protruding inward and a planar operation portion 13b connected to the needle-shaped portion 13a. Such a closed state opening member 13 can be made of metal or relatively hard synthetic resin. The outer surface of the operating portion 13b of the closed state opening member 13 is covered with the elastic film 14. The elastic film 14 is provided to surround the closed state opening member 13 and prevent the buffer solution from leaking to the outside of the container body 7. Therefore, the elastic film 14 is provided on the outer wall of the container body 7 at its peripheral portion. Is stuck. The elastic film 14 can be made of, for example, synthetic rubber, natural rubber, or a synthetic resin film having elasticity.

In the judgment section 11, a separation filter 15 is provided on the side of the closing member 12. The separation filter 15 is provided to remove solids in the buffer solution in which the stool sample is dissolved, and can be composed of, for example, a known filter paper, a separation membrane such as nitrocellulose, or a filter material. The separation filter 15 may be made of a porous material such as a cellulosic material or a synthetic polymer material.

Above the separation filter 15, a spreading layer 16 is provided.
Are integrally formed with the separation filter 15. Also,
A judgment window 17 and an end window 18 are formed on the side wall of the container body 7 in order to allow the development layer 16 to be partially observed. The end window 18 is formed above the determination window 17. Further, the judgment window 17 and the end window 18 are
Each is closed by a transparent film. As such a film, an appropriate synthetic polymer film such as polyethylene, polyvinyl chloride or polypropylene can be used.

The spreading layer 16 is provided to carry out a known immunochromatographic method, and is made of a porous material. That is, a carrier sensitized with an antibody against a test substance (hereinafter, abbreviated as a first antibody) is installed at the starting end 16a. In addition, a second antibody that recognizes an epitope different from that of the first antibody is immobilized at a position separated from the installation site of the first antibody-sensitized carrier by a certain distance.

When the liquid sample containing the test substance is spread from the starting end 16a of the spread layer, the sample moves on the spread layer 16,
In addition, the test substance causes an antigen-antibody reaction with the first antibody. The test substance-first antibody-sensitized carrier complex further moves in the spreading layer and reaches the second antibody immobilization site. As a result, due to the antigen-antibody reaction between the test substance and the second antibody, the test substance-first
The antibody-sensitized carrier complex is trapped at the second antibody immobilization site. Therefore, if the carrier has a color tone having a clear contrast with the spreading layer 16, the presence of the carrier, that is, the presence of the test substance can be easily confirmed visually.

In this example, the test substance is human hemoglobin, which is an index for determining fecal occult blood, and the test substance-containing liquid sample is the above-mentioned fecal lysate. Examples of the material forming the spreading layer 16 include cellulosic materials such as nitrocellulose and cellulose acetate, synthetic polymer materials such as nylon, fluorinated ethylene resin, polypropylene, polyvinyl chloride resin, etc. Various materials used as the filter material can be used.

Further, the spreading layer 16 is required to have a function of exerting a clear contrast with the color tone of the carrier. Therefore, it is preferable to use a white carrier when the spreading layer 16 is black and a colored carrier when the spreading layer 16 is white. The most preferred carrier is latex particles, but since latex particles are usually white, when the background is white,
For example, styrene, latex particles obtained by emulsion polymerization of vinyltoluene, etc., in a reaction medium containing water and an organic solvent having compatibility with water, with a poorly water-soluble colorant dissolved in the reaction medium. Method of contacting and coloring, latex particles obtained by emulsion polymerization and having an anionic functional group, for example, (meth) latex particles made of a copolymer containing a monomer having an anionic functional group such as acrylic acid, It is preferable to use colored latex particles obtained by, for example, a method of contacting with a colorant which is an amphoteric electrolyte dissolved in the reaction medium to color it in an aqueous reaction medium.

The anti-human Hb antibody used in the present invention (first
Antibodies) can be obtained by purifying antisera obtained by immunizing rabbits, sheep, goats, horses, guinea pigs, etc. with human Hb by affinity chromatography or the like. It is also possible to produce a monoclonal antibody by a known cell fusion method using immunized mouse spleen cells. In any case, it is desirable that the anti-human Hb antibody used be sufficiently purified.

The anti-human Hb antibody can be sensitized to the carrier by a known method such as a separation adsorption method or a chemical bonding method. For example, a buffer solution of anti-human Hb antibody is added to latex, and the mixture is stirred at 37 ° C. for 30 to 120 minutes.
The antibody can be physically adsorbed on the latex surface by heating to. If a latex having a carboxyl group is used, the antibody can be bound to the latex by a covalent bond via a water-soluble carbodiimide or the like. If necessary, bovine serum albumin (hereinafter referred to as "B
SA ”. ) And the like may be added.

The first antibody-sensitized carrier thus obtained is placed on the spreading layer 16, for example, a 0.01-10% by weight dispersion of the antibody-sensitized carrier is applied in a predetermined amount of about 0.05-1 ml. It can be carried out by dropping it in a position and drying it at room temperature for 12 hours or more.

Further, on the spreading layer 16, the judgment window 17
The second antibody is immobilized on the site observed by. As the second antibody, an anti-human hemoglobin antibody having a different epitope recognized by the first antibody can be used. The immobilization method is as follows: 1-5 mg / ml of the second antibody 0.1-1
An example is a method in which 0 μl is dropped at a predetermined position, left at room temperature for 12 hours or more, and then BSA or the like is dropped at the same position if necessary and dried.

In the fecal occult blood determining apparatus of the present embodiment, the above-mentioned stool collection device 2 is entirely made of synthetic resin material, but it may be made of other material such as metal. The inner volume of the buffer solution container 3 is not particularly limited, but it is preferably about 1 to 10 ml in terms of downsizing of the device.

The buffer solution container 3 can be made of synthetic resin such as polyethylene, polypropylene or vinyl chloride resin, but may be made of other material such as metal.

Next, a method of using the fecal occult blood determining apparatus shown in FIG. 1 will be described with reference to FIGS. 2 and 3. First, FIG.
In the fecal occult blood determination device 1 assembled as shown in (4), the fecal sampling device 2 is removed from the buffer solution container 3. Next, a stool sample 5 is collected in the stool collection part 6 by piercing the stool sample A with the stool collection rod 5 of the stool collection device 2 and pulling it out (FIG.
(See (a)).

Next, after collecting a stool sample, the stool collection device 2 is attached to the buffer solution container 3 again (see FIG. 2 (b)). At the time of this attachment, an excess amount of the stool sample is removed by the excess stool removing member 9 shown in FIG. 1B, and the buffer solution 10 is held in a state where a predetermined amount of the stool sample is held in the stool collection unit 6. Is immersed in. Further, by engaging the female screw 4a of the lid body 4 with the male screw 8a of the buffer solution container 3, the buffer solution 10 is prevented from leaking to the outside. In this state,
As shown by the arrow in (b), the entire fecal occult blood determination apparatus 1 is turned. By the turning, the stool sample held in the stool collection unit 6 is dissolved in the buffer solution 10. Thereafter, as shown by an arrow B in FIG. 3A, the operating portion 13b of the closed-state opening member 13 is pressed with a finger from the outside of the elastic film 14 to form the needle-shaped portion 13a.
The closing member 12 is pierced by the tip of. As a result, the buffer solution 10 in which the fecal sample is dissolved will flow into the determination unit 11.

Then, the solid content in the stool sample solution is removed by the separation filter 15 and the stool sample solution from which the solid content has been removed is supplied to the starting end 16a of the spreading layer 16.
In the spreading layer 16, the buffer solution in which the stool sample is dissolved is guided to the spreading layer 16 by the capillary phenomenon.

Next, as shown in FIG. 3 (b), the fecal occult blood determining apparatus 1 is placed sideways so that the side opposite to the side on which the determining unit 11 is provided faces downward. Immunochromatography is started in the developing layer 16, the buffer solution in which the stool sample is dissolved diffuses on the developing layer 16, and moves toward the end window 18 side together with the first antibody-sensitized carrier. At this time, when human hemoglobin is contained in the buffer solution in which the stool sample is dissolved, an antigen-antibody reaction occurs with the first antibody on the carrier. Then, at the site where the second antibody is located, only the human hemoglobin adsorption carrier causes an antigen-antibody reaction with the second antibody and is trapped (sandwich method). On the other hand, when human hemoglobin was not contained in the buffer solution in which the stool sample was dissolved, the human hemoglobin-first antibody-sensitized carrier complex was not trapped by the second antibody. That is, the buffer solution entirely moves to the portion of the spreading layer 16 that faces the end window 18. Therefore, when observed from the judgment window 17, the color tone of the carrier is observed when the hemoglobin in the stool sample solution contains human hemoglobin (positive), and when it does not contain, the development layer. Only 16 colors are observed (negative). Therefore, the presence or absence of fecal occult blood can be easily determined by visually observing through the determination window 17.

The determination window 17 is only required to be installed at a position where the above determination can be clearly observed, and its shape and size are not particularly limited. The end of the determination is performed by observing the development layer 16 through the end window 18.

Second Embodiment FIGS. 4 (a) and 4 (b) are a plan view and a vertical sectional view for explaining a fecal occult blood determining apparatus according to an embodiment of the present invention. .

The fecal occult blood determination device 21 has a fecal collection device 22 and a buffer solution container 23. Before use, the stool collection device 22 is attached to the buffer solution container 23 as shown in FIG. The details of each member will be described below.

The stool collection device 22 is constructed in the same manner as the stool collection device 2 shown in the above embodiment. Therefore, the stool collection device 2
Regarding 2, the description is omitted by giving each part the same reference numeral as each part of the stool collection device 2 of the first embodiment.

The buffer solution container 23 is also configured in the same manner as the buffer solution container 3 of the first embodiment except for the determination section 31. Therefore, the same parts are designated by the same reference numerals, and the description thereof will be omitted.

In this embodiment, the judging section 31 is composed of the closing material 32.
It is separated from the buffer solution 10 by. The closure member 32 is fixed to the inner wall of the buffer solution container 3 so as to close the opening facing the inner wall of the container body 7. The closing member 32 is a material that can be pierced by a closed state opening member 33 described later,
For example, it is composed of a synthetic resin film having an appropriate thickness or a hot melt heat closing material.

The judging section 31 is provided with a closed state opening member 33 for penetrating the closing member 32. The closed state opening member 33 is made of metal or synthetic resin and has a needle-shaped portion 33a having a sharp tip inside. The outer end of the needle-shaped portion 33a is connected to the plate-shaped operation portion 33b. Further, the elastic film 34 is attached so as to surround the outer portion of the closed state opening member 33. The elastic film 34 is made of natural rubber, synthetic rubber, or an appropriate synthetic resin film having elasticity, and its peripheral portion is fixed to the outer wall of the container body 7. Therefore, the buffer solution that has flowed into the determination unit 31 is prevented from leaking to the outside by the elastic film 34.

Preferably, the outer surface of the operating portion 33b of the closed state opening member 33 is fixed to the inner surface of the elastic film 34. Thereby, the elastic film 34 of the closed state opening member 33 is
It is possible to prevent rattling and movement inside.

The needle-shaped portion 33a is constructed so as to penetrate a through hole provided in the outer wall of the container body 7 in the determination portion 31. The feature of this embodiment is that the above-mentioned closing member 32
The separation filter 35 and the immune filter 36 are arranged above the space D in which the closed state opening member 33 is arranged. The separation filter 35 is attached so as to close the space D above. Separation filter 3
5 is composed of an appropriate material capable of removing the solid content in the buffer solution in which the stool sample is dissolved, for example, a known filter paper or a filter material such as nitrocellulose. The separation filter 35 may be made of a cellulose-based polymer material or a porous material.

An immune filter 36 is arranged above the separation filter 35 with a predetermined distance. The immune filter 36 is arranged at a position where it can receive the stool sample solution that has been filtered by the separation filter 35. Further, the container body 7 is preferably transparent so that the determination unit 31 can visually observe it from the outside. Further, a through hole 38 is provided so as to face the space E, and a cap 39 is attached to the through hole 38. The through hole 38 and the cap 39 are provided to communicate the space E with the outside air, and are provided to facilitate the movement of air and thereby facilitate the later-described operation.

Next, the structure of the immune filter 36 will be described. The immune filter 36 is provided to carry out a known immune filter method, and uses a latex agglutination reaction. In the latex agglutination reaction method,
For example, an antigen-antibody reaction between an antibody-sensitized latex and an antigen causes an immune reaction product to form a structure and produce an aggregate.

By carrying out the above-mentioned immunoreaction on a filter having a large number of pores through which the agglomerates cannot pass but the non-aggregates can pass, the presence or absence of latex on the filter after the reaction, or on the filter. Positive or negative is judged depending on the presence or absence of colored latex accumulated in the space.

At the time of the determination, preferably, the color tone of the carrier is constituted so as to have a clear contrast with the immune filter or the space below the filter, whereby the determination can be made more reliable and easy.

In the present invention, the above-mentioned antigen is human hemoglobin in the stool lysate, and the antibody is anti-human hemoglobin antibody. The anti-human hemoglobin antibody is immobilized on the latex particles, but is immobilized by the buffer solution so that it can be dissolved.

As described above, in the immune filter 36, the product of the antigen-antibody reaction, that is, the aggregate does not pass through the immune filter 36, but the antibody-sensitized carrier originally installed is dispersed by the stool sample solution. After being processed, it passes through the immune filter 36. As described above, if the color tone of the carrier and the color tone of the immune filter 36 or the space on the immune filter 36 have a clear contrast, positive and negative determinations can be made easily and reliably.

In order to make the color tone of the carrier different from the color tone of the background where the carrier is present and the agglomerate, it is preferable to use a colored carrier. For example, it is preferable to use a white carrier when the background is black and a colored carrier when the background is white. Most preferred as the carrier is latex particles, but since latex particles are usually white, when the background is white, for example, styrene, latex particles obtained by emulsion polymerization of vinyltoluene, etc. And in a reaction medium containing an organic solvent compatible with water, a method of coloring by contacting with a poorly water-soluble colorant dissolved in the reaction medium, or obtained by emulsion polymerization and having an anionic functional group In an aqueous reaction medium, latex particles, for example, latex particles composed of a copolymer containing a monomer having an anionic functional group such as (meth) acrylic acid, are dissolved in the reaction medium, and the coloring agent is an amphoteric electrolyte. It is preferable to use colored latex particles obtained by a method of contacting with and coloring.

The anti-human hemoglobin antibody used in the present invention can be obtained by purifying the antiserum obtained by immunizing rabbits, sheep, goats, horses, guinea pigs, etc. with human hemoglobin by affinity chromatography or the like. It is also possible to produce a monoclonal antibody by a known cell fusion method using immunized mouse spleen cells. In any case, it is desirable that the anti-human hemoglobin antibody used be sufficiently purified.

As a determination method, as described above,
In addition to the method of observing the latex reacted on the immunofilter, a method of observing the liquid remaining in the space below the immunofilter may be adopted. In the method of observing the space below the immunofilter, when it is negative (when hemoglobin is not detected), unreacted latex is observed in the space below the immunofilter, and when it is positive, the unreacted latex is observed. No latex will be observed.

Next, a method of using the fecal occult blood determining apparatus of the second embodiment will be described. Also in the fecal occult blood determination apparatus of the second embodiment, initially, the stool collection device 22 is attached to the buffer container 23 as shown in FIGS. 4 (a) and 4 (b).
Therefore, remove the stool collection device 22 from the buffer container 23,
A stool sample is collected by the stool collection unit 6 of the stool collection stick 5 (Fig. 5).
(See (a)).

Thereafter, the stool collection rod 5 is inserted into the buffer solution container 3, and the lid 4 of the stool collection device 22 is attached to the buffer solution container 3. That is, the lid 4 is attached to the buffer solution container 3 by the engagement of the internal thread 4a and the external thread 8a, and the leakage of the internal buffer solution 10 is prevented. Further, as in the case of the first embodiment, the excess stool sample attached to the stool collection part 6 of the stool collection rod 5 is removed by the excess stool removal part 9. Therefore,
A desired amount of stool sample is inserted into the buffer solution 10 while being held by the stool collection unit 6.

Next, as shown by the arrow in FIG. 5B, the entire fecal occult blood determination device 21 is swirled and dissolved in the fecal sample buffer solution 10. After that, the elastic film 34 is pressed from the outside with a finger, the closing member 32 is pierced by the needle-like portion 33a of the closed state opening member 33, and the cap 39 is removed. As a result, the buffer solution in which the stool sample is dissolved is determined by the determination unit 3.
It will flow into the space D of 1. The buffer solution in which the inflowing stool sample is dissolved enters into the space D (see FIG. 4). Next, the whole is turned upside down (FIG. 6 (b)). As a result, the stool sample solution is supplied to the separation filter 35, the solid content therein is removed, and the buffer solution in which the stool sample from which the solid content is removed is dissolved is supplied to the immune filter 36. In the immunofilter 36, when the reaction by the above-mentioned immunofilter method progresses and human hemoglobin is present in the stool sample solution, an antigen-antibody reaction occurs on the immunofilter 36, and the antibody-sensitized carrier and human hemoglobin The reaction of (1) produces an immune reaction product (aggregate). Therefore, the antibody-sensitized carrier is dispersed in the buffer solution, but does not flow into the space E because it forms an aggregate due to the antigen-antibody reaction. Therefore, the color tone of the antibody-sensitized carrier is not observed in the space E, and therefore the color tone of the antibody-sensitized carrier is observed on the immune filter 36 (positive).

On the other hand, when human hemoglobin is not present in the stool sample solution, the antibody-sensitized carrier does not cause the above antigen-antibody reaction. Therefore, the buffer solution in which the stool sample is dissolved passes through the immune filter 36 and reaches the space E. Therefore, the color tone of the antibody-sensitized carrier is observed (negative).

Then, after the judgment is completed, the through hole 38 is sealed with the cap 39, so that the liquid leakage of the fecal solution can be prevented and the measurement is completed. In addition,
In the second embodiment, the stool sample solution can be guided to the immune filter 36 side by vacuum suction from the through hole 38.

Further, in the first and second embodiments, the portions for operating the closed state opening members 13, 33 from the outside, that is, the elastic films 14, 34 are exposed to the outside. Therefore, if the elastic films 14 and 34 are erroneously pressed before use, the buffer solution 10 may flow into the determination units 11 and 31. In order to prevent such an accident, preferably, as shown in FIG. 7, a protective cover 20 made of a relatively hard synthetic resin may be provided so as to cover the portion where the elastic film 14 is provided. In this case, the protective cover 20 needs to be attached so that it can be easily removed when pressing the elastic film 14.

Next, a fecal occult blood determining apparatus according to a third embodiment of the present invention will be described. Mostly, the fecal occult blood determination apparatus of the third embodiment is different only in the configuration of the immune filter and the determination method, and other specific configurations are as follows.
Since it is the same as the second embodiment, the description of the fecal occult blood determining apparatus of the second embodiment will be omitted by referring to it.

In the fecal occult blood determining apparatus of the third embodiment, the second
Unlike the embodiment described above, the anti-hemoglobin antibody-sensitized carrier is arranged below the separation filter 35, and the anti-human hemoglobin antibody-sensitized carrier is movably set by the buffer solution. Furthermore, a second anti-human hemoglobin antibody having a recognition site different from that of the anti-human hemoglobin antibody carried on the anti-human hemoglobin antibody-sensitized carrier is immobilized immovably in the immune filter 36, and The immune filter 36 is configured to have pores through which the human hemoglobin antibody-sensitized carrier can pass.

The operation of the fecal occult blood determining apparatus of the third embodiment is performed in the same manner as the fecal occult blood determining apparatus of the second embodiment. In the determination, the buffer solution in which the stool sample is dissolved is supplied to the immune filter 36, and when hemoglobin is present in the stool sample, human Hb reacts with the anti-human hemoglobin antibody-sensitized carrier and the antigen-antibody reaction. The Hb-anti-hemoglobin antibody-sensitized carrier complex is moved by the buffer solution, and the second anti-human hemoglobin antibody immobilized on the immune filter 36 and human Hb cause an antigen-antibody reaction to cause the second anti-hemoglobin antibody. The above-mentioned immune complex will be immobilized on the portion where the human hemoglobin antibody is immobilized.

Therefore, when positive, that is, when human Hb was present in the stool sample, the color tone of the colored latex reacted as described above was observed on the immunofilter, and No latex can be observed in the space below the filter. On the other hand, when it is negative, that is, when human Hb is not present in the stool sample, the color tone of unreacted colored latex is observed in the space below the immune filter. In this way, also in the third embodiment, as in the case of the second embodiment, even an unskilled person can easily visually determine the presence or absence of fecal occult blood.

In the first embodiment shown in FIG. 1B, the closing member 12 that closes the inside of the judging portion is made of a sealing material such as synthetic resin, and the closed state opening member 13 has a needle shape. By being pierced by the portion 13a, the closed state in the determination part by the closing material 12 was opened.

However, the closing material in the present invention is not limited to the sealing material made of the synthetic resin film as described above. That is, as shown in FIG. 8, an inside plug type closing material 41 may be used. The inner plug type closing member 41 is made of a material having an appropriate shape-retaining property such as synthetic resin or metal, and in the initial state, the determination unit 11 is used.
In order to close the container, it is press-fitted into the through hole facing the inner peripheral surface of the side wall of the container body 7. In opening the closed state, the operating portion 42b of the closed state opening member 42 is pressed from the outside to the inside, and the closing member 41 is pushed inward by the tip portion 42a to remove the closing member 41 from the through hole (Fig. 9). In this way, the closed state by the inside plug type closing member 41 is opened by operating the closed state opening member 42 from the outside.

Similarly, in the second and third embodiments as well,
The closing member for closing the inside of the determination unit 11 is not limited to the closing member 12 made of the sealing material shown in FIG. 7, and as shown in FIG.
Can be used. The inner plug type closing member 43 can also be made of a material having an appropriate shape-retaining property such as a synthetic resin or a metal, similarly to the closing member 41 shown in FIG. As shown in FIG. 10, in the initial state, the closing member 43 is press-fitted into the through hole facing the inner peripheral surface of the side wall of the container body 7, and is closed by the operation from the outside.
The closed state of the closing member 43 can be released by pressing the closing member 43 inward by the tip portion 44a of the above and removing the closing member 43 from the through hole.

As described above, the closed state may be released by the closing member according to the present invention by piercing the closing member, or when the inside plug type closing member is used, the closing member is penetrated. It may be done by removing it from the hole.

[0080]

As described above, according to the inventions described in claims 1 to 3, after collecting a stool sample on the stool sampling rod of the stool collection tool, the stool collection tool is inserted into the buffer solution container to collect stool. After attaching the device to the buffer solution container, swirl or vibrate the entire stool occult blood determination device to dissolve the stool sample in the buffer solution, and then operate the closed state opening member from the outside to dissolve the stool sample. An immune reaction can be caused only by performing a series of operations for introducing the prepared buffer solution into the determination unit. That is, in the invention according to claim 1, the presence or absence of fecal occult blood is determined in the spreading layer based on immunochromatography, and in the inventions according to claims 2 and 3, the presence or absence of fecal occult blood based on the immunofilter method. Is determined.

Therefore, according to the inventions described in claims 1 to 3, it is possible to reliably determine the presence or absence of fecal occult blood with an extremely simple operation and without requiring a great deal of skill. In addition, in the invention according to claims 1 to 3, a determination unit that configures immunochromatography or an immune filter is provided in the side wall of the buffer solution container, and is physically a fecal sampling device and a buffer solution container. The system consists of only one. Therefore, the structure of the fecal occult blood determination device can be simplified,
Moreover, miniaturization can be promoted.

[Brief description of drawings]

1A and 1B are a plan view and a vertical cross-sectional view of a fecal occult blood determining apparatus according to a first embodiment.

2A and 2B are schematic cross-sectional views for explaining a step of collecting a stool sample and a step of preparing a stool sample solution in the fecal occult blood determination apparatus of the first embodiment. .

3 (a) and 3 (b) are schematic cross-sectional views for explaining the step of breaking the closing material and the step of making a judgment in the fecal occult blood determining apparatus of the first embodiment.

4 (a) and 4 (b) are respectively a plan view and a vertical cross-sectional view of the fecal occult blood determining devices of the second and third embodiments.

5 (a) and 5 (b) are schematic views for explaining a step of collecting a stool sample and a step of preparing a stool sample solution in the fecal occult blood determining apparatuses of the second and third examples. Sectional view.

6 (a) and 6 (b) are schematic cross-sectional views for explaining a step of piercing a closing material and a step of making a determination using the fecal occult blood determining devices of the second and third embodiments.

FIG. 7 is a partially cutaway sectional view showing a structure in which a cover for protecting the closed state opening member from the outside is attached.

FIG. 8 is a vertical cross-sectional view for explaining a modified example of the fecal occult blood determining apparatus according to the first embodiment, which uses an inside plug type closing material.

9 is a schematic side view showing a state in which the inside plug type closing material is removed and the closed state is opened in the fecal occult blood determining apparatus shown in FIG.

FIG. 10 is a second example in which the closing material is of the inside plug type.
FIG. 8 is a vertical cross-sectional view for explaining a modified example of the fecal occult blood determining apparatus of the embodiment.

[Explanation of symbols]

 1, 21 ... Fecal occult blood determination device 2, 22 ... Stool collection device 3, 23 ... Buffer container 4 ... Lid body 5 ... Stool collection rod 6 ... Stool collection part 7 ... Container body 11 ... Judgment part 12 ... Closing material 13 ... Closed state open member 14 ... Elastic membrane 15 ... Separation filter 16 ... Deployment layer 17 ... Judgment window 18 ... End window 31 ... Judgment part 32 ... Closing material 33 ... Closed state open member 34 ... Elastic film 35 ... Separation filter 36 ... Immune filter 41 ... Closed material 42 ... Closed state opening member 43 ... Closed material 44 ... Closed state opening member

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Yuji Kaneko 2-1 Hyakuyama, Shimamoto-cho, Mishima-gun, Osaka Prefecture Sekisui Chemical Co., Ltd. (72) Takuya Wada 2-1 Hyakuyama, Shimamoto-cho, Mishima-gun, Osaka 2-1 Sekisui Chemical Industry Co., Ltd.

Claims (3)

[Claims] 1. A stool collection container having a buffer solution container containing a buffer solution, a lid attached to the buffer solution container so as to close the buffer solution container, and a stool collection rod connected to the lid body. An instrument and a determination section provided on the side wall of the buffer solution container, and a closure provided on the inner wall of the buffer solution container to close the determination section to prevent the buffer solution from flowing into the determination section. A material, a closed state opening member attached to the buffer solution container so that the closed state by the closed material can be opened by an operation from the outside, and a stool that has flowed in when the closed state is opened. In order to remove the solid content in the buffer solution in which the sample is dissolved, a separation filter provided in the determination unit and a fecal sample solution in which the solid content is removed by the separation filter are arranged to be supplied. And anti-hemoglobin antibody sensitized carrier And has a spreading layer on which an anti-hemoglobin antibody is immobilized, and in order to observe the presence or absence of fecal occult blood from the outer surface of the buffer solution container and the end of the determination, the side wall of the buffer solution container is An apparatus for determining fecal occult blood, wherein a determination window and an end window are formed so that the developed layer can be partially observed. 2. A stool collection container having a buffer solution container containing a buffer solution, a lid body attached to the buffer solution container so as to close the buffer solution vessel, and a stool collection rod connected to the lid body. An instrument and a determination section provided on the side wall of the buffer solution container, and a closure provided on the inner wall of the buffer solution container to close the determination section to prevent the buffer solution from flowing into the determination section. A closed material, a closed state opening member attached to the buffer solution container so that the closed state of the closed material can be opened by an operation from the outside, and an inflow when the closed state of the closed material is released In order to remove the solid content in the buffer solution in which the stool sample has been dissolved, the separation filter provided in the determination unit and the stool sample solution in which the solid content is removed by the separation filter are supplied. The closing filter with the separation filter in between. Is arranged on the opposite side of the material, and the anti-hemoglobin antibody-sensitized carrier is movably installed by a buffer solution,
Fecal occult blood determination characterized by having an immunofilter having pores through which the anti-hemoglobin antibody-sensitized carrier can pass but the product of the antigen-antibody reaction between the anti-hemoglobin antibody-sensitized carrier and hemoglobin does not pass apparatus. 3. A stool collection container having a buffer solution container containing a buffer solution, a lid attached to the buffer solution container so as to close the buffer solution container, and a stool collection rod connected to the lid body. An instrument and a determination section provided on the side wall of the buffer solution container, and a closure provided on the inner wall of the buffer solution container to close the determination section to prevent the buffer solution from flowing into the determination section. A closed material, a closed state opening member attached to the buffer solution container so that the closed state of the closed material can be opened by an operation from the outside, and an inflow when the closed state of the closed material is released In order to remove the solid content in the buffer solution in which the stool sample has been dissolved, the separation filter provided in the determination unit and the stool sample solution in which the solid content is removed by the separation filter are supplied. And, with the separation filter sandwiched between the closing material and Is arranged on the opposite side, and the anti-hemoglobin antibody sensitized carrier is movably installed by the buffer solution, and the anti-hemoglobin antibody sensitized by the anti-hemoglobin antibody sensitized carrier is a recognition site. The second that is different
Anti-hemoglobin antibody of is immobilized immovably,
An apparatus for determining fecal occult blood, comprising: an immune filter having pores through which the anti-hemoglobin antibody-sensitized carrier can pass.