JP3487689B2 - Fecal occult blood determination device - Google Patents

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 based on an immune reaction for determining the presence or absence of fecal occult blood.

[0002]

2. Description of the Related Art Fecal occult blood is produced by digestive system diseases resulting from the contamination of blood in the stool in the digestive tract. It has been confirmed that fecal occult blood is associated with diseases such as colorectal cancer in particular, and the fecal occult blood test 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 the dissolution is 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 buffer container containing a buffer solution, a stool collection device having a lid attached to the buffer container so as to close the buffer container and a stool collecting rod connected to the lid, and a lid. A fecal occult blood determination device comprising a determination unit provided,
A sealing material is provided in the lower part of the lid so as to close the inside of the determination part in order to prevent the buffer solution from flowing into the determination part, and penetrates the lid into the lid and faces the sealing material. A through hole is provided, and a sealing material opening member is provided in the through hole so that the sealing material can be opened by an operation from the outside. Further, each of the through holes has the following configuration. And

According to the first aspect of the invention, a separation filter for removing the solid content in the buffer solution in which the fecal sample is dissolved is provided between the determination unit and the sealing material, and the determination unit is provided in the determination unit. , Is arranged so that the stool sample solution from which solids have been removed by the separation filter is supplied, and an anti-hemoglobin antibody-sensitized antibody is installed and an anti-hemoglobin antibody-immobilized development layer is The lid is provided with
In order to observe the presence or absence of fecal occult blood and the end of the determination from the outer surface of the lid, a determination window and an end window are formed so that the spreading layer can be partially observed.

In the invention according to claim 2, the judging section comprises a separation filter for removing solids in the buffer solution in which the stool sample is dissolved, and a stool sample from which solids have been removed by the separation filter. So as to be supplied with a solution, it is arranged on the opposite side of the encapsulating material across the separation filter,
And the anti-hemoglobin antibody-sensitized carrier is movably installed by a buffer, the anti-hemoglobin antibody-sensitized carrier can pass, the product of the antigen-antibody reaction of the anti-hemoglobin antibody-sensitized carrier and hemoglobin is And an immune filter having pores that do not pass through. Further, at least one of the immune filter of the determination unit provided in the lid and the space above it is configured to be visible from the outer surface.

In the third aspect of the present invention, the determination unit includes a separation filter for removing solids in the buffer solution in which the stool sample is dissolved, and a stool sample from which solids have been removed by the separation filter. So as to be supplied with a solution, it is arranged on the opposite side of the encapsulating material across the separation filter,
And the anti-hemoglobin antibody sensitized carrier is movably installed by a buffer, and further the anti-human hemoglobin antibody having a different recognition epitope from the anti-human hemoglobin antibody is immobilized immovably, and the sensitized carrier passes through. And an immune filter with possible pores. Further, at least one of the immune filter of the determination unit provided in the lid and the space above it is configured to be visible from the outer surface.

Further, in the invention according to any one of claims 1 to 3, the sealing material is capable of sealing between the buffer solution container main body and the through hole or the judging portion, and the sealing material opening member. If it can be released by the force applied from
It is not particularly limited, and examples thereof include a synthetic resin film having an appropriate film thickness as shown in the below-described first embodiment, and a plug body as shown in the third embodiment.

[0016] Further, in the invention according to claims 2 and 3, at least one of the immune filter of the determination portion provided in the lid and the space above it is visible from the outer surface. Comprises, for example, the immunofilter of the lid and a portion facing at least one of the spaces above the transparent material; the entire lid is composed of a transparent material; the immunofilter of the lid and the upper portion thereof. It is possible to provide a transparent window in a portion facing at least one of the spaces.

(Operation) In determining fecal occult blood using the fecal occult blood determining apparatus according to the first to third aspects of the present invention, first, the stool sampling rod portion of the stool collection instrument is pierced into the stool sample,
A stool sample is collected by withdrawing. 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 solution container, and a buffer solution in which the stool sample is dissolved is created.

Next, the sealant release member in the through hole provided in the lid is pressed from the outside to release the sealant. Thereafter, the feces occult blood determination apparatus is turned upside down to remove the solid content of the buffer solution in which the stool sample is dissolved by the separation filter, and then the invention according to claim 1,
It is supplied to the spreading layer, and in the invention described in claims 2 and 3, it is supplied to the immune filter.

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

Further, in the invention according to the first aspect, since the judgment window and the end window are formed in the judgment portion, the judgment can be easily performed from the outside and the end of the judgment can be confirmed. . According to the second and third aspects of the present invention, at least one of the immune filter and the space above it of the determination section provided in the lid is configured to be visible from the outer surface, so that it can be easily performed from the outside. Can be determined.

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 solution container, the sealing material release member is opened. The presence or absence of fecal occult blood can be easily determined simply by opening the sealing material and turning the entire fecal occult blood determination device upside down. That is, since the fecal occult blood determination apparatus of the present invention has a relatively simple configuration because the determination unit is provided in the lid, it can reliably and easily perform from the collection of a stool sample to the determination of fecal occult blood. can do.

[0022]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be clarified by describing non-limiting examples of the present invention with reference to the drawings.

First Embodiment FIG. 1 is 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 1 of the first embodiment is composed of a fecal sampling device 2, a buffer solution container 3 and a determination unit 4. Hereinafter, each member will be described in detail.

The stool collection device 2 has a lid 5 attached to the container 3 so as to close the buffer solution container 3, and a stool collection rod 6 connected to the lid 5. In this embodiment, the lid 5 and the stool collecting rod 6 are integrally formed of synthetic resin.
The lid 5 and the stool collecting rod 6 may be formed of separate members and then fixed to each other with an adhesive or the like.

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

The stool collection part 6a 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 6a 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 cylindrical 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 5 a of the lid body 5.

The surplus feces removing member 9 may be 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. Further, the excess feces removing member 9 is formed so as to cover almost the entire opening of the buffer solution container 3, except that a portion where a sealing material described later is located is opened. However, the area where the excess feces removing member 9 covers the opening of the buffer solution container 3 is not limited to this, and may be smaller. 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, the stool stick 6
When the stool is inserted through the through hole 9a, the peripheral edge of the surplus stool removing member 9 facing the through hole 9a comes into contact with the outer surface of the stool collection portion 6a. Therefore, the surplus stool sample among the stool samples attached to the stool collection unit 6a 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.
However, even if the excess stool removal member 9 is not provided, it is not necessary to provide the excess stool removal member 9 when a predetermined amount of stool sample is inserted into the buffer solution container 3. In addition, the second, which will be described later,
Although the illustration of the surplus feces removing member 9 is omitted in Examples 4, 6, 8 and 10 to 12, it is needless to say that it may be attached if necessary.

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.

At the lower part of the lid 5, a determination unit 4 for the buffer solution 10 is provided.
A sealing material 11 is provided so as to close the inside of the determination unit 4 in order to prevent inflow into the inside, and the sealing material 11 is provided in a through hole 12 that vertically penetrates the lid body 5. The lower end is also attached so that it closes. A sealing material opening member 13 is arranged inside the through hole 12. Sealing material release member 1
3 has an elongated cylindrical shape, the upper portion of which is located in a hole 14 provided in the lid body 5, and the upper end portion has a diameter larger than that of the cylindrical portion to form an operating portion 13a. 13a
A spring 13b is attached to the outer peripheral portion of the lower cylindrical portion. The lower end portion 13c of the sealing material opening member 13 is sharp, and can be pierced through the sealing material 11 when pressed against the sealing material 11 by an external force. Further, a lid 15 is openably attached to the hole 14 provided in the lid body 5 in order to protect the operation portion 13a.

The sealing material opening member 13 is used so that the lower end portion 13c pierces the sealing material 11 when the operating portion 13a is pressed downward by an external force, and after the piercing, the spring is released. 13b restores the original state. A packing 16 such as an O-ring is attached to an outer peripheral portion of a part of the columnar part of the sealing material release member 13, and the fecal occult blood determination device 1 is tilted after the sealing material 11 is pierced. As a result, the buffer solution in which the feces are dissolved is prevented from leaking into the hole 14 when the buffer solution flows into the through hole 12. Further, as the material of the sealing material releasing member 13, a synthetic resin or metal having a relatively high rigidity is used, and as the material of the sealing material 11, a synthetic resin film having an appropriate film thickness, a hot melt heat seal, or the like is used. Used. Further, the sealing material 11 may be a film-shaped one as described above, or a plug body for sealing the lower end portion of the through hole 12 as shown in a third embodiment described later. Further, also in the fifth and seventh embodiments described later, a plug body is preferably used as the sealing material 11. Further, in other embodiments, a plug body can be used as the sealing material 11 by devising the shape of the device.

Between the lower end of the through hole 12 and the lower end of the determination unit 4, that is, between the determination unit 4 and the sealing material 11, the solid content in the buffer solution in which the stool sample is dissolved is removed. In order to do so, a separation filter 17 is provided. The separation filter 17 can be made of, for example, a porous material made of a cellulosic material or a synthetic polymer material. Further, it may be composed of a known filter paper, a separation membrane such as nitrocellulose, or a filter material.

The judging section 4 is provided vertically in the lid 5 and has a circular cross section. The determination unit 4 is a lid 5
May be located relatively close to the surface or may be fairly internal. Further, the shape of the determination unit 4 does not need to be circular in cross section as described above, and may be a free shape such as a square cross section.

A development layer 18 for determining the presence or absence of fecal occult blood in the stool sample based on the principle of immunochromatography is provided in the determination section 4. The shape of the spreading layer 18 is such that the lower end of the spreading layer 18 is the separation filter 17
It is not particularly limited as long as it can be brought into contact with a stool sample solution supplied through
Examples include strips. Further, the development layer 18 is the determination unit 4
It does not necessarily have to be fixed to the inner side wall.

Further, in order to observe the presence or absence of fecal occult blood and the end of the determination from the outer surface of the lid body 5,
The judgment window 1 so that the spreading layer 18 can be partially observed.
9 and an end window 20 are formed. The end window 20 is formed above the determination window 19. Also, the judgment window 19
The end window 20 and the end window 20 are each closed by a transparent film. As such a film, an appropriate synthetic polymer film such as polyethylene, polyvinyl chloride or polypropylene can be used.

As described above, the spreading layer 18 is provided to carry out the 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 18a. In addition, a portion 18b which is separated from the installation site of the first antibody-sensitized carrier by a certain distance.
A second antibody that recognizes an epitope different from that of the first antibody is immobilized.

When the liquid sample containing the test substance is spread from the starting end 18a of the spread layer, the sample moves on the spread layer 18,
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 18b. as a result,
Due to the antigen-antibody reaction between the test substance and the second antibody, the test substance-first antibody-sensitized carrier complex becomes the second antibody immobilization site 1 described above.
8b is trapped. Therefore, if the carrier has a color tone having a clear contrast with the spreading layer 18, the presence of the carrier, that is, the presence of the test substance can be easily confirmed by visual observation.

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. As the material for forming the spreading layer 18, for example, a cellulosic material such as nitrocellulose or cellulose acetate, a synthetic polymer material such as nylon, fluorinated ethylene resin, polypropylene, polyvinyl chloride resin, or the like is used as a conventional membrane. Various materials used as the filter material can be used.

Further, the spreading layer 18 is required to have a function of exhibiting a clear contrast with the color tone of the carrier. Therefore, it is preferable to use a white carrier when the developing layer 18 is black and a colored carrier when the developing layer 18 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 use a monoclonal antibody obtained 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 physical adsorption method or a chemical binding 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 installation of the obtained first antibody-sensitized carrier on the spreading layer 18 is carried out, for example, by adding a 0.01-10% by weight dispersion of the antibody-sensitized carrier in an 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 18, the judgment window 19
The second antibody is immobilized on the site 18b observed by. As the second antibody, an anti-human hemoglobin antibody having a different epitope recognized by the first antibody can be used.
As the immobilization method, 1 to 5 mg / ml of the second antibody 0.1
An example is a method of dropping -10 μl to a predetermined position, allowing it to stand at room temperature for 12 hours or more, and further dropping BSA or the like to the same position if necessary and drying.

In the fecal occult blood determining apparatus of this 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.

In the fecal occult blood determining apparatus of the present embodiment, the method for manufacturing the above-mentioned fecal sampling device 2 is not particularly limited, but, for example, the lid 5 is manufactured in the form of being divided into two parts. There is a method in which the penetrating portion 12 and the determining portion 4 are provided on one side, the sealing material release member 13, the spreading layer 18, etc. are provided on each of them, and then the other one is integrated by a method such as bonding.

The buffer 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. 1 and 2. Note that FIG.
FIG. 4 is a diagram schematically showing only a main part of the fecal occult blood determination device to explain how to use the device.For example, a part that is originally invisible from the outside is also shown by a solid line and drawn according to a drawing method. It is not a drawing.

First, in the fecal occult blood determining apparatus 1 assembled as shown in FIG. 1, the fecal sampling device 2 is removed from the buffer solution container 3. Next, the stool sample 6 of the stool collection tool 2 is pierced into the stool sample A and pulled out to collect the stool sample in the stool collection part 6a (see FIG. 2 (a)).

Next, after collecting the stool sample, the stool collection device 2 is attached to the buffer solution container 3 again. At the time of this attachment, an excess amount of the stool sample is removed by the excess stool removal member 9 shown in FIG. 1, and a predetermined amount of the stool sample is immersed in the buffer solution 10 while being held in the stool collection part 6a. To be done. Also, the lid 5
By engaging the female screw 5a 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, the entire fecal occult blood determination device 1 is turned. By the turning, the stool sample held in the stool collection part 6a is dissolved in the buffer solution 10. Then, as shown in FIG. 2B, the lid 15 is opened, and the operating portion 13a of the sealing portion opening member 13 is pressed downward with a finger or the like to break through the sealing portion 11. Next, when the finger is released, the sealing portion opening member 13 is closed by the spring 1.
It returns to the original position by the recoil of 3b (see FIG. 2 (c)).

Next, as shown in FIG. 2D, the fecal occult blood determining apparatus 1 is turned upside down. Then, the buffer solution 10 in which the stool sample is dissolved enters the penetrating portion 12 from the opened sealing portion 11, the solid content is further removed by the separation filter 17, and the stool sample solution in which the solid content is removed is determined. It enters the section 4 and reaches the starting end 18a of the spreading layer 18 in the determination section 4.
In addition, the buffer solution 1 in which the stool sample that has entered the penetration portion 12 is dissolved
Although not shown in FIG. 2, 0 is prevented from leaking into the hole 14 by the packing 16 shown in FIG.

Next, after the stool sample solution sufficiently wets the starting end 18a of the spreading layer 18, as shown in FIG. 2 (e), the stool occult blood determining apparatus 1 is turned upside down again to dissolve the stool sample. Breaks new contact between the liquid and the spreading layer 18.

After that, immunochromatography is started in the developing layer 18 in the judging section 4, and the buffer solution in which the stool sample is dissolved diffuses on the developing layer 18 by the capillary phenomenon, so that the first antibody-sensitized carrier is obtained. Along with this, it moves toward the end window 20 side. At this time, when human hemoglobin is contained in the buffer solution in which the stool sample is dissolved, the first sample on the carrier is used.
An antigen-antibody reaction occurs with the antibody. At the site where the second antibody is located, when reacting with human hemoglobin, only the first antibody-sensitized carrier causes an antigen-antibody reaction with the second antibody and is trapped (sandwich method). On the other hand, when human hemoglobin is not contained in the buffer solution in which the stool sample is dissolved, the first antibody-sensitized carrier is not trapped by the second antibody. That is, the buffer solution entirely moves to the portion of the spreading layer 18 that faces the end window 20. Therefore, the judgment window 1
When observed from No. 9, the color tone of the carrier is observed (positive) when the human hemoglobin is contained in the stool sample solution, and when it is not contained, the development layer 1
Only 8 colors are observed (negative). Therefore, the presence or absence of fecal occult blood can be easily determined by visually observing through the determination window 19.

The determination window 19 need only 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 18 through the end window 20.

In the above operation, the stool occult blood determination apparatus 1 was turned upside down in order to supply the stool sample solution to the spreading layer 18, but the stool sample solution could only be supplied to the spreading layer 18. For example, the fecal occult blood determining apparatus 1 may be overturned. In this case, it is preferable that at least one of the lid body 5 and the buffer solution container body 7 of the stool collection device 2 has a rectangular shape in order to improve the stability of the fecal occult blood determination device 1 when it is turned over. .

Second Embodiment FIG. 3 is a vertical sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the present invention. The fecal occult blood determination device 31 of the second embodiment is different from the fecal occult blood determination device 1 of the first embodiment in that the through hole 12 in the stool collection device 2 is horizontally configured, and the sealing material 11 is used. Are configured vertically, the shape of the stool collection part 6a of the stool collection rod 6 is different, the position of the separation filter 17 is below the determination part 4, and the like of the first embodiment. It is constructed similarly to the one. Since each member of the second embodiment has the same function as that of the first embodiment, the same reference numerals are given and the description thereof is omitted.

The method of using the fecal occult blood determining apparatus 31 of the second embodiment is the same as that of the fecal occult blood determining apparatus 1 of the first embodiment.

Third Embodiment FIG. 4 is a vertical sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the present invention. In the fecal occult blood determination device 41 of the third embodiment, compared with the fecal occult blood determination device 1 of the first embodiment, the determination part 4 is configured horizontally, and the sealing material 11 is the lower end of the through hole 12. The lower end portion 13 of the sealing material opening member 13 is a plug body that seals the portion.
The shape of a is rounder than that of the first embodiment, the shape of the stool collection part 6a of the stool collection stick 6 is different, and the position of the separation filter 17 is located on the side of the determination part 4. Other than that, the configuration is similar to that of the first embodiment. Since each member of the third embodiment has the same function as that of the first embodiment, the same reference numerals are given and the description thereof is omitted.

The method of using the fecal occult blood determining apparatus 41 of the third embodiment is the same as that of the fecal occult blood determining apparatus 1 of the first embodiment.

Fourth Embodiment FIG. 5 is a vertical sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the invention described in claim 1. Compared with the fecal occult blood determination device 1 of the first embodiment, the fecal occult blood determination device 51 of the fourth embodiment has a configuration in which the through hole 12 in the stool collection device 2 is horizontally configured, It is configured horizontally, that the sealing material 11 is configured vertically, that the shape of the stool collection part 6a of the stool collection rod 6 is different, and that the position of the separation filter 17 is on the side of the determination part 4. Other than that, the configuration is similar to that of the first embodiment. Since each member of the fourth embodiment has the same function as that of the first embodiment, the description thereof is omitted by giving the same reference numeral.

The method of using the fecal occult blood determining apparatus 51 of the fourth embodiment is the same as that of the fecal occult blood determining apparatus 1 of the first embodiment.

Fifth Embodiment FIG. 6 is 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 61 of the fifth embodiment is composed of a fecal sampling device 62, a buffer solution container 63, and a determination unit 64. Hereinafter, each member will be described in detail.

The stool collection tool 62 is the first one except that the shape of the stool collection part 6a of the stool collection tool 2 shown in the first embodiment is slightly different.
The configuration is similar to that of the above-mentioned embodiment. Therefore, the stool collection device 6
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 63 is also constructed in the same manner as the buffer solution container 3 of the first embodiment. Therefore, the same parts are designated by the same reference numerals, and the description thereof will be omitted.

In this embodiment, a sealing material 11 is provided in the lower portion of the lid 5 so as to close the inside of the judging portion 64 in order to prevent the buffer solution from flowing into the judging portion 64. Sealing material 1
The lid 5 is provided with a sealing material opening member 13 so that the sealing material opening member 1 can be opened by an external operation.
3 is provided to vertically penetrate the lid body 5 and is provided with a sealing material 11
Is disposed in the through hole 12 facing the. These encapsulant, encapsulant opening member, and through-hole are configured similarly to those in the first embodiment. Therefore, the same parts as described above are designated by the same reference numerals, and the description thereof will be omitted.

The determination unit 64 is provided vertically in the lid body 5, is closed by the sealing material 11, and after the sealing material 11 is opened by the sealing material opening member 13, fecal occult blood. By turning over the determination device 61, the stool sample solution can flow in. In the determination unit 64, a separation filter 65 and an immune filter 66 are arranged above the space B where the sealing material 11 and the sealing material opening member 13 are arranged. The separation filter 65 is attached so as to close the space B above. The separation filter 65 is composed of an appropriate material capable of removing solids 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 65 may be made of a cellulose-based polymer material or a porous material.

Further, an immune filter 66 is arranged above the separation filter 65 with a predetermined distance. The immune filter 66 is arranged at a position where it can receive the stool sample solution that has been filtered by the separation filter 65. The lid 5 is transparent so that the determination unit 64 can visually observe it from the outside. If the lid 5 is opaque,
It is necessary to provide, for example, a transparent window so that at least one of the immune filter and the space C on the immune filter can be observed. The window may be formed of a transparent synthetic resin film by forming an opening on the outer wall of the lid body 5. Further, a through hole 67 is provided so as to face the space C, and a cap 68 is attached to the through hole 67. The through-hole 67 and the cap 68 are provided for communicating the space C with the outside air, and are provided for facilitating the movement of air and thereby facilitating the later-described operation.

Next, the structure of the immune filter 66 will be described. The immune filter 66 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 an aggregate.

By carrying out the above-mentioned immunoreaction on a filter having a large number of pores through which the above-mentioned aggregates cannot pass but 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 judgment, preferably, the color tone of the carrier has a clear contrast with the immune filter or the space above the filter, whereby the judgment 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.

As described above, in the immune filter 66, the product of the antigen-antibody reaction, that is, the aggregate does not pass through the immune filter 66, but the antibody-sensitized carrier originally installed is dispersed by the stool sample solution. After being processed, it passes through the immune filter 66. As described above, if the color tone of the carrier and the color tone of the immune filter 66 or the space on the immune filter 66 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 latex particles obtained by a method of contacting with and coloring.

The anti-human hemoglobin antibody used in the present invention is the same as that described in the description of the fecal occult blood determining apparatus of the first embodiment.

Next, the fecal occult blood determining device 61 of the fifth embodiment.
The method of using will be described with reference to FIGS. 6 and 7. In addition,
FIG. 7 is a diagram schematically showing only a main part of the fecal occult blood determination device 61 to explain how to use it.
Originally, the part that cannot be seen from the outside is also shown by a solid line, and is not a drawing drawn according to the drawing method.

Also in the fecal occult blood determining apparatus of the fifth embodiment, the fecal sampling device 62 is initially attached to the buffer solution container 63 as shown in FIG. Therefore, the stool collection device 62
Is removed from the buffer solution container 63, and the stool collection part 6a of the stool collection stick 6 is removed.
A stool sample is collected by (see FIG. 7 (a)).

Thereafter, the stool collection rod 6 is inserted into the buffer solution container 63, and the lid 5 of the stool collection device 63 is attached to the buffer solution container 63. That is, the lid 5 is attached to the buffer solution container 63 by the meshing of the internal thread 8a and the external thread 5a, 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 6a of the stool collection rod 6 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 part 6a.

Next, the entire stool occult blood determination device 61 is swirled and dissolved in the stool sample buffer solution 10. Then, Figure 7
As shown in (b), similar to the first embodiment, the opening member 1
The operation portion 13a of No. 3 is pressed downward with a finger or the like to move the sealing portion 1
Break through 1. Next, when the finger is released, the sealing portion opening member 13
Is returned to its original position by the reaction of the spring 13b (see FIG. 7C).

Next, as shown in FIG. 7D, the fecal occult blood determining device 61 is turned upside down and the cap 6
Remove 8. As a result, the buffer solution in which the stool sample is dissolved flows into the space B of the determination unit 64. The buffer solution in which the inflowing stool sample is dissolved enters into the space B.
As a result, the stool sample solution is supplied to the separation filter 65, 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 provided to the immune filter 66. In the immunofilter 66, 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 66 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 C 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 C (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 antibody-sensitized carrier is accompanied by the buffer solution in which the stool sample is dissolved, and the immune filter 66
To reach the space C. Therefore, the color tone of the antibody-sensitized carrier is observed (negative).

The judgment is completed when the fecal sample solution appears in the space C and the color tone of the antibody-sensitized carrier can be judged. After the judgment, the through hole 67 is sealed with the cap 68. As a result, it is possible to prevent the stool solution from leaking.

In the fifth embodiment, the stool sample solution can be guided to the immune filter 66 side by vacuum suction from the through hole 67.

In the fecal occult blood determining apparatus of the present embodiment, the above-mentioned stool collection device 62 is entirely made of synthetic resin material, but it may be made of other material such as metal. However, at least one of the immune filter of the determination unit provided in the lid and the space above it needs to be visible from the outer surface. The content of the buffer solution container 63 is not particularly limited,
About 10 ml is desirable in terms of downsizing of the device.

In the above operation, the fecal occult blood determination device 61 was turned upside down in order to supply the fecal sample dissolution liquid to the determination unit 64. If possible, fecal occult blood determination device 61
It does not matter even if it is overturned. In this case, it is preferable that at least one of the lid body 5 and the buffer solution container body 7 of the stool collection device 62 has a rectangular shape in order to improve the stability of the fecal occult blood determination device 1 when it is overturned. .

Sixth Embodiment FIG. 8 is a vertical cross-sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the present invention. The fecal occult blood determination device 71 of the eighth embodiment is different from the fecal occult blood determination device 61 of the fifth embodiment in that the through hole 12 in the stool collection device 62 is different.
Is configured horizontally, the sealing material 11 is configured vertically, and the shape of the stool sampling portion 6a of the stool sampling rod 6 is different, and the like from the fifth embodiment. It is similarly configured. Since each member of the sixth embodiment has the same function as that of the fifth embodiment, the same reference numerals are given and the description thereof is omitted.

The method of using the fecal occult blood determining device 71 of the sixth embodiment is the same as that of the fecal occult blood determining device 61 of the fifth embodiment.

Seventh Embodiment FIG. 9 is a vertical sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the present invention. The fecal occult blood determination device 81 of the seventh embodiment is different from the fecal occult blood determination device 61 of the fifth embodiment in that the determination unit 64 in the fecal sampling device 62.
Is configured horizontally, and the stool collection part 6 of the stool collection rod 6
The structure is the same as that of the fifth embodiment except that the shape of a is different. Since each member of the seventh embodiment has the same function as that of the fifth embodiment, the same reference numerals are given and the description thereof is omitted.

In the method of using the fecal occult blood determining device 81 of the seventh embodiment, in order to facilitate the flow of the stool sample lysate at the time of determination, the fecal occult blood determining device 81 is not turned upside down but the device 81 is used. It is the same as the fecal occult blood determination device 61 of the fifth embodiment, except that the determination unit 64 is preferably rotated halfway to be used vertically.

Eighth Embodiment FIG. 10 is a vertical cross-sectional view for explaining a fecal occult blood determining apparatus according to another embodiment of the present invention. The fecal occult blood determination device 91 of the eighth embodiment is different from the fecal occult blood determination device 61 of the fifth embodiment in that the through hole 1 in the stool collection device 62 is different.
2 and the determination unit 64 are configured horizontally, the sealing material 11 is configured vertically, and the shape of the stool collection unit 6a of the stool collection rod 6 is different. The configuration is similar to that of the embodiment. Then, since each member of the eighth embodiment has the same function as that of the fifth embodiment, the same reference numerals are attached to them.
The description is omitted.

The method of using the fecal occult blood determining apparatus 91 of the eighth embodiment is such that the fecal occult blood determining apparatus 91 is not turned upside down in order to facilitate the flow of the stool sample lysis solution at the time of determination. It is the same as the fecal occult blood determination device 61 of the fifth embodiment, except that the determination unit 64 is preferably rotated halfway to be used vertically.

Ninth to Twelfth Embodiments The ninth to twelfth embodiments are configured in the same manner as the fifth to eighth embodiments, respectively, except that the immune filter 66 in the fifth to eighth embodiments is different. ing. In the ninth to twelfth examples, the immunofilter has an anti-human hemoglobin antibody-sensitized carrier movably set by a buffer solution, and further comprises an anti-human hemoglobin antibody having a different recognition epitope from the anti-human hemoglobin antibody. It is fixed so that it cannot be moved. Further, the immune filter is made of a material having pores through which the sensitizing carrier can pass.

The method of using the fecal occult blood determining apparatus of the ninth to twelfth embodiments is as follows.
The same as 1, 81, 91.

In the fecal occult blood determination devices of the ninth to twelfth embodiments, when the buffer solution in which the fecal sample from which the solid content is removed is dissolved is applied to the immunofilter, the immunofilter uses the immunofilter method described above. The reaction proceeds. That is, when human hemoglobin is present in the stool sample lysate, on the immunofilter, between the anti-human hemoglobin antibody-sensitized carrier, human hemoglobin, and the anti-human hemoglobin antibody having a different recognition epitope from the anti-human hemoglobin antibody. The immune reaction product (aggregate) is generated in 1. and the anti-human hemoglobin antibodies having different recognition epitopes are immobilized on the immune filter immovably, so that the immune reaction product is immobilized on the immune filter. Therefore, the antibody-sensitized carrier does not flow into the space C. Therefore, the color tone of the antibody-sensitized carrier is not observed in the space C (positive).

On the other hand, when human hemoglobin is not present in the stool sample solution, the antibody-sensitized carrier does not cause the antigen-antibody reaction. Therefore, since the immune filter is configured to have pores through which the sensitizing carrier can pass, the antibody sensitizing carrier is passed through the immunofilter by being accompanied by the buffer solution in which the stool sample is dissolved, It reaches the space C. Therefore, the color tone of the antibody-sensitized carrier is observed (negative).

Further, as described above, the determination can be made not only by the presence or absence of the antibody-sensitized carrier in the space C, but also by the change in the color tone of the immune filter itself. That is, in this case, when human hemoglobin is present in the stool sample solution, the color tone of the antibody-sensitized carrier is observed in the immune filter, and when human hemoglobin is not present, the antibody-sensitized carrier is detected in the immune filter. No color tone is observed.

[0094]

As described above, according to the first aspect of the invention, after collecting a stool sample from the stool collection stick of the stool collection tool, the stool collection stick is inserted into the buffer container to remove the stool collection tool. After attaching to the buffer solution container, swirl or vibrate the whole stool occult blood determination device to dissolve the stool sample in the buffer solution, and then operate the sealing material release member from the outside to open the sealing material. In addition, the stool occult blood determination apparatus is turned upside down to introduce a buffer solution containing no solids into the development layer of the determination unit by inverting the stool sample and performing a series of operations based on the immunochromatographic method. The presence or absence of occult blood is determined, and the presence or absence of fecal occult blood can be easily observed from the outside through the determination window.

As described above, in the inventions described in claims 2 and 3, after the stool sample is collected on the stool collection stick of the stool collection tool, the stool collection stick is inserted into the buffer solution container to remove the stool collection tool. After attaching to the buffer solution container, swirl or vibrate the entire fecal occult blood determination device,
Dissolve the stool sample in a buffer solution, and then operate the sealing material release member from the outside to open the sealing material, and then turn the entire stool occult blood determination device upside down to buffer the stool sample. By supplying the liquid to the separation filter of the determination unit to remove the solid content, and then performing a series of operations to introduce into the immune filter,
The presence or absence of fecal occult blood is determined based on the immune filter method, and the presence or absence of fecal occult blood can be easily observed from the outside.

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.

[Brief description of drawings]

FIG. 1 is a vertical cross-sectional view of a fecal occult blood determining apparatus according to a first embodiment.

2 (a) to 2 (e) are explanatory views schematically showing only a main part for explaining a method of using the fecal occult blood determining apparatus of the first embodiment.

FIG. 3 is a vertical cross-sectional view of a fecal occult blood determining device according to a second embodiment.

FIG. 4 is a vertical cross-sectional view of a fecal occult blood determining device according to a third embodiment.

FIG. 5 is a vertical cross-sectional view of a fecal occult blood determining device according to a fourth embodiment.

FIG. 6 is a vertical cross-sectional view of a fecal occult blood determining apparatus according to a fifth embodiment.

7 (a) to 7 (d) are explanatory views schematically showing only a main part for explaining a method of using the fecal occult blood determination apparatus of the fifth embodiment.

FIG. 8 is a vertical cross-sectional view of the fecal occult blood determining apparatus according to the sixth embodiment.

FIG. 9 is a vertical cross-sectional view of a fecal occult blood determining device according to a seventh embodiment.

FIG. 10 is a vertical cross-sectional view of the fecal occult blood determination device of the eighth embodiment.

[Explanation of symbols]

1, 31, 41, 51, 61, 71, 81, 91 Fecal occult blood determination device 2, 62 Stool collection device 3, 63 Buffer solution container 4, 64 Determination part 5 Lid body 6 Stool collection stick 7 Container body 10 Buffer solution 11 Sealing material 12 Through hole 13 Sealing material opening member 17 Separation filter 18 Development layer 19 Judgment window 20 End window 65 Separation filter 66 Immune filter

─────────────────────────────────────────────────── ─── Continuation of the front page (58) Fields surveyed (Int.Cl. ⁷ , DB name) G01N 33/50 G01N 1/04 G01N 33/48

Claims (3)

(57) [Claims] 1. A stool collection tool having a buffer container containing a buffer solution, a lid attached to the buffer container so as to close the buffer container, and a stool collecting rod connected to the lid. A fecal occult blood determination device comprising a determination unit provided inside the lid, and a sealing material is provided below the lid to close the inside of the determination unit to prevent the buffer solution from flowing into the determination unit. A through-hole is provided in the lid, the through-hole penetrating the lid and facing the sealing material. A stopper release member is provided, a separation filter for removing solids in the buffer solution in which the fecal sample is dissolved is provided between the determination unit and the sealing material, and the separation filter is provided in the determination unit. It is arranged so that the stool sample solution from which solids have been removed is supplied by
And an anti-hemoglobin antibody-sensitized antibody is installed and an anti-hemoglobin antibody-immobilized development layer is provided, and the lid body is observed from the outer surface of the lid body for the presence or absence of fecal occult blood and the end of the determination. In order to achieve this, a determination window and a termination window are formed so that the developed layer can be partially observed, and a fecal occult blood determination apparatus. 2. A stool collection tool having a buffer container containing a buffer solution, a lid attached to the buffer container so as to close the buffer container, and a stool collecting rod connected to the lid. A fecal occult blood determination device comprising a determination unit provided inside the lid, and a sealing material is provided below the lid to close the inside of the determination unit to prevent the buffer solution from flowing into the determination unit. A through-hole is provided in the lid body, the through-hole penetrating the lid body and facing the sealing material, and the sealing material is sealed in the through-hole so as to be opened by an external operation. A stopper release member is provided, and the determination unit supplies a separation filter for removing solids in the buffer solution in which the stool sample has been dissolved, and a stool sample solution in which solids have been removed by the separation filter. So that it is placed on the side opposite to the encapsulant across the separation filter. It is, and anti-hemoglobin antibody-sensitized carrier are movably installed at the buffer,
The anti-hemoglobin antibody-sensitized carrier is allowed to pass, but the anti-hemoglobin antibody-sensitized carrier and an immune filter having pores through which a product of an antigen-antibody reaction of hemoglobin is not passed are provided in the lid body. At least one of the immune filter of the determination unit and the space above it is configured to be visible from the outer surface. 3. A stool collection device 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. A fecal occult blood determination device comprising a determination unit provided inside the lid, and a sealing material is provided below the lid to close the inside of the determination unit to prevent the buffer solution from flowing into the determination unit. A through-hole is provided in the lid body, the through-hole penetrating the lid body and facing the sealing material, and the sealing material is sealed in the through-hole so as to be opened by an external operation. A stopper release member is provided, and the determination unit supplies a separation filter for removing solids in the buffer solution in which the stool sample has been dissolved, and a stool sample solution in which solids have been removed by the separation filter. So that it is placed on the side opposite to the encapsulant across the separation filter. It is, and anti-hemoglobin antibody-sensitized carrier are movably installed at the buffer,
Furthermore, the anti-human hemoglobin antibody and an anti-human hemoglobin antibody having a different recognition epitope are immobilized immovably, and the immunosensitized carrier has an immune filter having a pore through which the sensitized carrier can pass, and is provided in the lid. At least one of the immune filter of the determination unit and the space above it is configured to be visible from the outer surface.