JP3302097B2 - 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 test apparatus for determining the presence or absence of fecal occult blood, and more particularly to a fecal occult blood determining apparatus utilizing immunochromatography.

[0002]

2. Description of the Related Art Fecal occult blood develops due to digestive diseases caused by blood contamination in the stool in the digestive tract. In particular, a 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 Japan.

[0003] Currently used fecal occult blood test methods include:
There are roughly two methods. The first method is a conventional chemical occult blood reaction (hereinafter, referred to as “chemical method”), which is a non-specific reaction for detecting a peroxidase-like activity of hemoglobin (hereinafter, referred to as Hb). It is. This method is based on a mechanism in which a chromogen (eg, guaiac or orthotolidine) is oxidized in the presence of a peroxidase-like activity-retaining substance and aqueous hydrogen peroxide to form a color. Therefore, in the present method, the drug and the food containing Hb, which are substances having a peroxidase-like activity, show a positive result, so that dosing and dietary restriction are required.

As an inspection apparatus based on the present chemical method, simple measurement systems have been disclosed, and some of them are widely used. For example, Japanese Patent Publication No. 2-45826,
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, etc. There is disclosed a system in which the state of coloring is checked by dropping hydrogen water. In these systems,
By applying an appropriate amount of a stool sample to a part of the guaiac oil attached to the card type main body and further dropping hydrogen peroxide water on the part on the back side of the application part, whether or not the guaiac oil at the dripping part develops color Is determined. It is advantageous that it can be performed without opening the stool sample application section, can be handled hygienically, and has a simple configuration, but the drawbacks derived from the above measurement principle have not been solved.

The second method, which has been recently developed to solve the above-mentioned disadvantages of the chemical method, 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 as in a chemical method. The determination is performed by a general immunoserological test such as a latex agglutination method or an enzyme-labeled immunoassay, and 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 and mixed on a determination plate, and the determination is made based on the presence or absence of an aggregation image. Although the specificity is high in principle compared to the chemical method, skill is required to judge the agglutination method. In addition, the test system based on the immunization method is provided as a kit including a determination plate, a latex reagent, a stir bar, a stool, a lysing solution, and the like. The system has not been developed.

[0007] The simplification of these immunization methods is a step unique to the immunization method not included in the chemical method, that is, a step of dissolving a stool sample in a buffer solution and the like, and removing undissolved solids in the lysate after dissolution by filtration. The process is performed. That is, as for these operations, a feces collection device capable of relatively quantitative and hygienic handling is disclosed, for example, Japanese Utility Model Laid-Open No. 3-44663, Japanese Utility Model Publication No. 2-3977.
JP, JP-A-1-66071, JP-A-1-8571
In Japanese Patent Application Publication No. 668 and Japanese Utility Model Application Laid-Open No. 64-42454, a collection rod for collecting stool is attached to the lid of a container containing a buffer, and the lid is set in the buffer container after stool collection. Thereby, the stool at the tip of the collection rod is dispersed in the buffer solution. In addition, a filter mechanism is installed in the dropping portion when the dropping portion is dropped onto the determination plate after shaking well, so that insoluble solids are removed. After being dropped on the judgment plate, the operation is carried out according to the above-mentioned conventional latex agglutination method.

[0008] The simplification of the fecal occult blood measurement system by the immunization method seems to be achieved by integrating the above-mentioned steps and the antigen-antibody reaction, but the technology 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 immunochromatography has been established. The biggest problem when using the immunochromatography method for fecal occult blood testing is that the stool sample lysis and filtration steps are difficult to incorporate into the system.

It is an object of the present invention to provide a fecal occult blood test apparatus using an immunochromatography method, which has a relatively simple structure and simplifies the operation throughout the fecal occult blood determination operation. It is an object of the present invention to provide a fecal occult blood determination device that enables even an unskilled person to easily and accurately determine the presence or absence of fecal occult blood.

[0011]

Means for Solving the Problems The inventions according to the first and second aspects of the present invention have been made to achieve the above-mentioned objects, and have the following configurations.

That is, the invention according to claim 1 is an apparatus for determining fecal occult blood, which comprises a feces collection device for collecting feces, an opening for inserting the feces collection device into an outer surface, and A main body having a judgment window for externally judging the presence or absence of fecal occult blood, a buffer solution container provided in the main body, and arranged so that the feces collection device is inserted through the opening, A filter disposed below the liquid container, for filtering a buffer in which the stool sample is dissolved,
A developing layer in which a starting end is located below the filter and a determination unit faces the determination window, and a developing layer on which an anti-hemoglobin antibody sensitized carrier and an anti-human hemoglobin antibody are fixed; By moving the feces collection device inserted in the liquid container or the buffer solution container up and down,
The filter from at least a part of the lower surface of the buffer solution container
The buffer in which the stool sample is dissolved flows and then the filter
In such a manner that the buffer solution filtered in the developing layer flows to the developing layer, it comprises a flow path forming means for forming a flow path between at least a part of the buffer solution container lower surface and the developing layer, Fecal occult blood determination device.

According to a second aspect of the present invention, there is provided an apparatus for determining fecal occult blood, comprising: a feces collection device for collecting feces; and an opening for inserting the feces collection device into an outer surface. A main body having a judgment window for externally judging the presence or absence of fecal occult blood, a buffer solution container provided in the main body, and arranged so that a feces collection device is inserted through the opening, and the buffer solution A filter for filtering a buffer solution in which the stool sample is dissolved, which is disposed below the container, and a determination unit is disposed below the filter so as to face the determination window , and anti-human hemoglobin A developing layer on which the antibody-sensitized carrier and the anti-human hemoglobin antibody are fixed,
The buffer in which the stool sample is dissolved flows from at least a part of the lower surface of the buffer solution container to the filter, and then the filter
Flow path forming means for forming a flow path between at least a part of the lower surface of the buffer solution container and the developing layer so that the buffer solution filtered in the developing layer flows through the developing layer; By pressing the one end, the flow path forming means is operated, whereby the buffer solution in which the stool sample is dissolved can be dropped from the buffer solution container toward the developing layer. And a switch configured as follows.

As described above, the invention according to claims 1 and 2 is a fecal occult blood determination device using an immune reaction,
This is a device that makes the fecal occult blood determination kit using the conventional immunization method simpler and enables even an unskilled person to easily determine the presence or absence of fecal occult blood.

A specific example of the structure of the fecal occult blood determination device according to the first and second aspects of the present invention will be described below with reference to the drawings.
FIG. 1 is a schematic perspective view for explaining a first structural example of the fecal occult blood determination device of the present invention.

The fecal occult blood judging device of this embodiment has a main body 1 and a feces collection device 2. On the upper surface of the main body 1, a feces collection device insertion hole 1a into which the above-mentioned feces collection device 2 is inserted, and a determination window 1b for enabling a determination result by an immunochromatography method to be described later to be known by looking up from above. Is formed.

In the present embodiment, the feces collection device 2 is provided with
Having. The recess 2a is provided for quantitatively or semi-quantitatively collecting a stool sample. That is, the feces sampling device 2 is pierced into the stool, and thereafter, the excess stool is wiped off, whereby a fixed amount of stool sample corresponding to the size of the recess 2a is collected in the recess 2a. In order to eliminate excess stool, the stool collection device 2 may be wiped off after stool is collected. However, in this example, since an excess stool elimination mechanism is provided inside the main body 1 as described later, It is also possible to eliminate an excess stool by this surplus stool elimination mechanism.

Since the stool collection device 2 is provided for collecting stool, its material is not particularly limited.
What is necessary is just to be configured with a rigidity sufficient to maintain the shape as shown in the figure. Although FIG. 1 shows the round bar-shaped stool device 2, it may have a square bar shape or a plate shape. In addition, since the feces collection device 2 is to be inserted into the feces collection device insertion hole 1a of the main body 1 described above, it is necessary to be configured to have a size that allows the insertion.

In this embodiment, the stool device 2 having the recess 2a formed near the lower end is used.
(A)-(c) and FIGS. 3 (a)-(c),
The shape of the stool device 2 can be variously changed.

In the stool device 2 shown in FIG.
b and 2b are sharpened, respectively, so that they can easily penetrate an upper partition of a buffer solution container described later. In addition, as shown in FIG.
a may be constituted by a V-shaped groove, or as shown in FIG. 2 (b), a plurality of recesses 2a are formed along both side edges of the faecal collection device 2 in a lower portion of the faecal collection device 2. You may. Further, as shown in FIG. 2 (c), a toilet device 2 having a semicircular concave portion 2a when viewed from the front may be used. Further, as shown in FIG. 3 (a), a toilet device 2 having a concave portion 2a having a lower wall extending in a horizontal direction may be used.

3 (b) and 3 (c) show outer stools 2c, 2c and insertion rods 2d, 2d, respectively.
2d. On the outer surfaces of the outer cylinders 2c, 2c, there are concave portions 2a, 2a as in the case of the toilet device 2 shown in FIG.
The inner surface of the recess 2a is formed so as not to reach the inside of the outer cylinders 2c, 2c, that is, the hollow portion. Also,
Each of the insertion rods 2d, 2d has a pointed tip, and is configured to easily penetrate an upper partition wall of a buffer solution container described later. Furthermore, the feces collection device 2 shown in FIG.
Then, a pair of stirring blades 2e and 2
e is formed, and by rotating the insertion rod 2d thereby, a buffer solution described later can be stirred.

FIG. 4 shows a state in which stool is collected using the stool collection device 2 of FIG. 1 and inserted through the stool collection device insertion hole 1 a of the main body 1. In FIG. 4, the feces collection device 2 is inserted through the feces collection device insertion hole 1a.
Is inserted inside. In the main body 1, a buffer solution container 3 is disposed at a portion where the toilet device 2 is inserted, and a buffer solution 4 is stored in the buffer solution container 3. The buffer solution container 3 is normally configured so as not to leak the buffer solution 4 to the outside, and is made of, for example, a film-like resin such as polyethylene, polypropylene, and vinyl chloride, or a rubber such as styrene-butadiene rubber.

An upper partition 5 is attached to an upper portion of the buffer solution container 3. The upper partition 5 may be attached to the upper surface of the buffer solution container 3 or may be formed so as to form an opening in the upper surface of the buffer solution container 3 and close the opening. In any case, the upper partition wall 5 needs to be configured so that the internal buffer solution 4 does not leak from the upper surface side.

The upper partition 5 can be easily penetrated by the above-mentioned feces-collecting device 2 when the above-mentioned feces-collecting device 2 is inserted. It is made of a material that does not leak the buffer solution 4 from the upper portion of the container 3, various synthetic rubbers or polymer resins that can achieve a liquid-tight seal.

A step 3 a is provided on the bottom surface of the buffer solution container 3, and the bottom surface above the step 3 a and the bottom 1
The pressing spring 7 is disposed between the pressing spring 7 and the pressing spring 7. Buffer solution container 3
Is normally urged upward by the elastic force of the pressing spring 7, as shown in FIG. In addition, a lower partition 8 is formed on the bottom surface below the step 3a. The lower partition 8 is easily penetrated by a drip needle 9 described later, and after the drip needle 9 is pulled out, the buffer solution container 3
, That is, various rubbers or soft polymer resins capable of achieving a liquid-tight seal.

Below the lower partition 8, a drip needle 9 is fixed so that its tip is directed upward. Below the dropping needle 9, a filter 10 is arranged. The positions of the drip needle 9 and the filter 10 are fixed. Dripping needle 9
Is provided to allow the internal buffer solution 4 to be dropped on the filter 10 side through the lower partition 8 described above. The filter 10 is provided for filtering the dropped stool solution and removing solid components.

The filter 10 is made of known filter paper or filter material. Below the filter 10, a developing layer 11 for constituting a chromatography section
Is arranged. The spreading layer 11 has a starting end side 11a
Are disposed below the filter 10. Therefore,
The stool lysate that has been filtered through the filter 10 is
a. Further, the determination unit 11b of the development layer 11
Are arranged so as to face the determination window 1b as shown in the figure.

In the present embodiment, an O-ring 14 is attached to the above-mentioned sample from the feces taking instrument insertion hole 1a on the upper partition 5 of the buffer solution container 3. The inner diameter of the O-ring 14 is substantially equal to the outer diameter of the feces collection instrument 2 or slightly smaller than the outer diameter of the feces collection instrument 2. The O-ring 14
It is fixed to constitute the above-described stool rejection mechanism, and since the inner diameter is selected as described above, the stool rejection mechanism is attached around the stool collection device 2 by passing the stool collection device 2 therethrough. Excess stool is removed. Note that 12
Indicates a space for storing the excess stool discharged as described above.

The O-ring 14 constituting the above-mentioned excess stool discharging device is not limited to the position shown in the figure, and may be fixed directly to the upper part of the buffer solution container, or may be provided on the side of the fecal sampling instrument insertion hole 1a, ie, the buffer solution. You may arrange | position separately from the container 3.

Next, the chromatography section including the developing layer 11 will be described. The chromatography unit is provided for performing so-called immunochromatography, and an antibody to a test substance (hereinafter, abbreviated as a first antibody) is sensed at one end of a developing layer 11 made of a porous material such as a filter. The carrier made is fixed. In addition, the first
At a certain distance from the fixed part of the antibody sensitized carrier,
A second antibody that recognizes an epitope different from the first antibody is immobilized. When a liquid sample containing a test substance is developed from the first antibody immobilization section side, that is, from the start end 11a side, the sample moves on the developing layer 11 and the test substance
Causes an antigen-antibody reaction with the antibody. The test substance-first antibody-sensitized carrier complex further moves and reaches the second antibody-immobilized site, where an antigen-antibody reaction between the test substance and the second antibody causes
The test substance-first antibody-sensitized carrier complex is trapped in the second antibody-immobilized site. Therefore, if the carrier has a color tone having a clear contrast with the developing layer 11, 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, and the test substance-containing liquid sample is the stool lysate described above.

Examples of the material constituting the spreading layer 11 include cellulose-based materials such as nitrocellulose and cellulose acetate; and synthetic polymer-based materials such as nylon, ethylene fluoride resin, polypropylene, and polyvinyl vinyl salt. A known material used as a membrane filter can be used.

Further, since the developing layer 11 needs to exhibit a clear contrast with the color tone of the carrier 16, when the developing layer 11 is black, the carrier is a white carrier, a developing layer. When 11 is white, it is necessary to use a colored carrier. Latex particles are most preferred as the carrier, but since the latex particles are usually white, when applied to a white developing layer, for example, styrene, latex obtained by emulsion polymerization of vinyl toluene, etc. Particles, in a reaction medium containing water and an organic solvent compatible with water, a method of coloring by contacting with a poorly water-soluble agent dissolved in the reaction medium,
Latex particles obtained by emulsion polymerization and having anionic functional groups, for example, latex particles comprising a copolymer containing a monomer having anionic functional groups such as (meth) acrylic acid, in an aqueous reaction medium, It is preferable to use colored latex particles obtained by a method of contacting with a coloring agent, which is an amphoteric electrolyte dissolved in the reaction medium, for coloring.

The first antibody used in the present invention is a rabbit,
It can be obtained by purifying an antiserum obtained by immunizing sheep, goats, horses, guinea pigs or the like with human Hb by affinity chromatography or the like. Monoclonal antibodies can also be produced by a known cell fusion method using immunized mouse spleen cells. In any case, it is desirable that the first antibody used be sufficiently purified.

The sensitization of the first antibody to the carrier can be performed by a known method such as a separation adsorption method or a chemical bonding method. For example, by adding a buffer solution of an anti-human Hb antibody to latex and heating the mixture to 37 ° C. with stirring for 30 to 120 minutes, the first antibody can be physically adsorbed on the latex surface. When a latex having a carboxyl group is used, the first 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,
Write "BSA". ) May be added.

Developed layer 1 of the obtained first antibody-sensitized carrier 16
The immobilization on 1 is carried out, for example, by dropping 0.05 to 1 ml of a 0.01 to 1% by weight dispersion at a predetermined position, and
It can be carried out by drying for more than an hour.

Further, on the development layer 11, the judgment window 1b
Is added to the site observed by
Antibody 17 is immobilized. As the second antibody 17,
Anti-human Hb antibodies with different epitopes that recognize the first antibody can be used. As an immobilization method, 1 to 5 mg / ml
0.1 to 10 μl of the second antibody is dropped at a predetermined position, left at room temperature for 12 hours or more, and if necessary, BSA or the like is dropped at the same position and dried.

The buffer 4 is preferably a neutral salt solution such as an ammonia buffer, a glycine buffer, a phosphate buffer, and a Tris buffer, and particularly preferably, a pH of 6.5 to 8.0. 5 ranges of the above buffers. The amount of the buffer depends on the amount of the stool sample to be collected, but is preferably about 0.5 to 10 ml.

Next, an operation method of the fecal occult blood determination apparatus of this embodiment will be described. First, the feces sampling device 2 shown in FIG. 1 is pierced into the stool, then pulled out, and inserted into the main body 1 from the feces sampling device insertion hole 1a as shown in FIG. In this case, the excess stool is removed when passing through the O-ring 14, and the space 12
Is stored in. Therefore, a substantially constant amount of sample stool is inserted into the buffer solution 4 by the stool collection device 2. In this state, the stool is dissolved by the buffer solution 4. In this case, the main body 1 may be slightly shaken in order to speed up the dissolution of the stool in the buffer solution 4 or to ensure the dissolution.

Next, the stool is dissolved, and a stool solution is obtained. In the state shown in FIG. 4, the stool collection device 2 is further pressed downward. As a result, the lower end of the toilet device 2 comes into contact with the bottom of the buffer solution container 3 above the step 3a, and when further pressed, the pressing spring 7 is compressed and the buffer solution container 3 is moved downward. As a result, the lower partition wall 8 of the buffer solution container 3 is pierced by the dropping needle 9, and the stool solution flows downward through the portion pierced by the dropping needle 9, whereby the buffer solution container 3
A flow path is formed between the lower partition 8 and the development layer 11.
The stool solution that has flowed out is filtered by the filter 10, thereby removing solids. The stool solution from which the solids have been removed is dropped on the start end portion 11 a of the developing layer 11.

The stool solution dropped from the dropping needle 9 diffuses on the developing layer 11 and moves toward the terminal end 11c together with the first antibody-sensitized carrier 16. At this time, when human stool is contained in the stool lysate, an antigen-antibody reaction occurs with the first antibody on the carrier. At the site where the second antibody 17 is located, only the human Hb-adsorbed carrier causes an antigen-antibody reaction with the second antibody 17 and is trapped (sandwich method). When human stool lysate contained no human Hb,
-The first antibody-sensitized carrier complex is not trapped by the second antibody 17, but moves to the terminal end 11 c of the spreading layer 11. Therefore, when observed from the judgment window 1b, it is found that human Hb
, The color tone of the carrier is observed (positive), and when it is not included, it is not observed (negative: the color of the developing layer 11 remains).

The shape and size of the judgment window 1b are not limited as long as it is set at a position where the above judgment is clearly observed. As described above, in the fecal occult blood determination device of this example, after the fecal sampling device 2 was pierced into the feces, it was inserted into the main body 1 through the fecal collection device insertion hole 1a of the main body 1, and the feces was dissolved in the buffer solution. By simply pressing the stool device 2 downward at an appropriate time, solid components are removed from the stool solution and the stool solution is dropped onto the developing layer 11 of the chromatography unit.
Therefore, the stool solution containing the first antibody-sensitized carrier 16 that has been developed in the developing layer 11 penetrates into the developing layer 11 by capillary action, and the first antibody-sensitized carrier 16 moves in the developing layer 13 and the determination window It is immobilized by the second antibody 17 immobilized at an appropriate position of the determination section 11b observable from 1b. Therefore, by observing the presence or absence of the immobilized carrier 16 through the determination window 1b, the presence or absence of fecal occult blood can be determined. That is, when the color tone of the carrier 16 shows a clear contrast with the color tone of the developing layer 11, it is understood that fecal occult blood exists.

FIGS. 5 (a) and 5 (b) are perspective views showing modified examples of the fecal sampling device and the buffer solution container in the fecal occult blood determination device of the first structural example. In the first structural example, the feces collection instrument 2 of a round bar, had been inserted from the feces collection instrument insertion hole 1a in the buffer vessel 3 in the body, as shown in FIG. 5 (a),
A cap 2f may be fixed to the upper end of the stool device 2 so that the cap 2f is fixed to the buffer solution container 3. That is, a female screw is formed on the inner peripheral surface of the cap 2f, and a screw thread meshing with the female screw is formed in the buffer solution container 3f.
Is formed on the outer peripheral edge of the upper end. Therefore, as shown in FIG. 5 (b), the stool is pierced into the stool, the stool is collected in the recess 2a, and then inserted into the buffer solution container 3, and the cap 2f is inserted.
May be engaged with the cap mounting portion 3b and fixed, and the stool may be dissolved in the buffer solution 4 by shaking the buffer solution container 3 by hand as shown in the figure.

As described above, the cap 2 is placed in the buffer solution container 3.
In the case of employing a structure in which the feces collection device 2 is fixed using f, the buffer solution container 3 can be inserted into the main body 1 from the upper surface of the main body 1 instead of the feces collection device insertion hole 1a of FIG. A relatively large opening may be formed as described above. And cap 2f
, The buffer solution container 3 may be moved downward, and the pressing spring 7 shown in FIG. 4 may be compressed.

[0044] In FIG. 5 (b), the although the bottom of the buffer vessel 3 is uniformly shown, when it is applied to the first structure example shown in FIG. 5 (b) buffer A step 3a may be formed at the bottom of the liquid container 3 in the same manner as the buffer solution container 3 shown in FIG.

FIGS. 6A and 6B are schematic cross-sectional views for explaining another modification of the first structural example. In this modified example, a tubular member 21 is arranged to form a flow path between the lower partition 8 on the bottom surface of the buffer solution container 3 and the developing layer 11. The upper end of the tubular member 21 is fixed to the lower partition 8 in a liquid-tight manner. In this modification, the lower partition 8
Is configured to be separable from the bottom surface of the buffer solution container 3.
On the other hand, the sample inflow hole 21 is located at an intermediate height of the cylindrical member 21.
a is formed, and the filter 10 is arranged at the lower end.

In the modification shown in FIG. 6, the stool collection instrument 2 is inserted into the buffer solution container 3, and the stool collected is dissolved in the buffer solution 4 as shown in FIG. The stool device 2 is pressed. As a result, the buffer solution container 3 compresses the pressing spring 7 and moves downward. At this time, the tubular member 21 and the lower partition 8 enter the buffer solution container 3, and as a result, the stool lysate flows into the tubular member 21 from the sample inflow hole 21 a of the tubular member 21. Then, the stool solution that has flowed in is filtered by the filter 10 and dropped onto the developing layer 11 below.

FIG. 7 is a cross-sectional view for explaining a fecal occult blood determination apparatus according to a second structural example of the present invention, and is a view corresponding to FIG. 4 showing the first structural example. Judgment window 1b
A portion subsequent to the filter 10, that is, a chromatographic unit, is configured in the same manner as in the example of the first structure. Therefore, the corresponding parts are given the same reference numerals, and the description thereof will be omitted.

In the second structural example, a rod-shaped switch 22 is inserted into a switch insertion hole 1e on the upper surface of the main body 1. The switch 22 is inserted into the main body 1 through the switch insertion hole 1 e and is configured to reach the inside of the buffer solution container 3. In the space 23 below the switch insertion hole 1e, a flange 2 is attached to the outer peripheral surface of the switch 22.
2a is formed integrally with the switch 22. A pressing spring 24 is disposed below the flange 22a, and the switch 22 is normally urged upward by the pressing spring 24.

The lower end 22b of the switch 22 is pointed as shown, and is configured to penetrate the lower partition 8 provided at a part of the bottom of the buffer solution container 3. In this example, after the stool is collected by the stool collection device 2, FIG.
As shown in (1), the fecal dispenser 2 is inserted into the buffer solution container 3 in the main body 1 through the fecal respirator insertion hole 1a to obtain a feces solution. Also in the second example, since the O-ring 14 is provided,
Excess flights are discharged to the space 12.

Next, the switch 22 is pressed downward in anticipation of the time when the stool is dissolved in the buffer solution 4. As a result, the pressing spring 24 is compressed, the lower partition 8 is pierced by the tip 22b of the switch 22, and the stool solution is dropped on the filter 10 below. Subsequent steps are the same as in the first structure example.

That is, the second structural example is characterized in that the structure for obtaining the stool solution by the switch 2 is separated from the structure for dropping the stool solution on the filter 10 side.
Other configurations are the same as those of the first structural example. Therefore, as in the first example, the presence or absence of fecal occult blood can be determined in the developing layer 11 by looking through the determination window 1b.

FIGS. 8A and 8B show the state of the switch 2 shown in FIG.
It is each sectional drawing which shows the modification of Example 2. In the second example, a rod-shaped member having a sharpened tip 22b is shown as the switch 22. However, as shown in FIG. 8A, a groove 22c communicating with the lower end on the lower end side of the switch 22 is formed. Alternatively, as shown in FIG. 8B, the internal flow path 22 leading to the lower end
d may be formed, whereby the stool dissolving solution can be dripped on the filter 10 more smoothly. FIG.
In (b), the internal flow path 22d is connected to the switch 2
2 having an inflow hole 22e opened at an intermediate height position,
In this example, when the switch 22 is pressed, the buffer solution 4 flows from the inflow hole 22e and drops downward as shown in a schematic partial cutaway sectional view in FIG. In this case, the height position of the inflow hole 22e needs to be selected so as to be located in the buffer solution container 3 with the switch 22 pushed down as shown in FIG.

FIG. 10 is a partially cutaway sectional view for explaining still another modification of the second structural example. here,
The switch 22 has an outer cylinder 22f and an inner cylinder 22g. The outer cylinder 22f is inserted into the main body 1 through the switch insertion hole 1e, and the inner cylinder 22g is fixed to the upper surface of the filter 10. In the outer cylinder 22f and the inner cylinder 22g, through holes 22h and 22i are formed on the outer peripheral surface so as to be arranged vertically. Therefore, in this example, the stool collection device 2
Is inserted into the buffer solution container 3 to obtain a stool solution, and then the outer tube 22f of the switch 22 is pushed down to overlap the through hole 22h and the through hole 22i, so that the stool solution is contained in the inner tube 22g. It flows and is dropped on the filter 10 below. On the other hand, after dropping the stool solution, the outer tube 22f is pulled up and the through-hole 22h and the through-hole 22i are arranged so as not to overlap with each other, whereby the inflow of the stool solution can be stopped.

In the modification shown in FIG. 10, preferably, a flange is provided on the outer peripheral surface at a portion located in the space 23 of the outer cylinder 22f, and a pressing spring is disposed on the lower surface of the flange, so that the outer cylinder 22f can be moved outwardly. The cylinder 22f may be urged upward, in which case the outer cylinder 22f can be returned to the initial state shown in FIG. 10 simply by releasing the hand after pressing the outer cylinder 22f.

As is apparent from FIG. 10, the switch mechanism in the second structural example has a flow path formed by an appropriate structure for allowing the stool solution to flow between the bottom surface of the buffer solution container 3 and the developing layer 11. Various modifications can be made so that they can be formed.

[0056]

According to the first aspect of the present invention, the stool sample is dissolved in the buffer solution container in the main body, and the stool sample is dissolved in the buffer solution (hereinafter referred to as stool) by the filter disposed in the main body. Lysate) is filtered and also provided in the main body, and the presence or absence of fecal occult blood is determined by immunochromatography using a developing layer on which the anti-human hemoglobin antibody-sensitized carrier and the anti-human hemoglobin antibody are fixed. Is determined.

That is, in each of the first and second aspects of the present invention, the stool sample dissolving step, the stool lysate filtration step, and the immune reaction, which are essential steps in the fecal occult blood test by the immunoassay, are performed. Are performed in the same apparatus, that is, in the main body. Therefore, the configuration of the fecal occult blood determination device using the immunization method is simplified.

Further, according to the first aspect of the present invention, by moving the buffer solution container or the stool collection device up and down, a flow path is formed between the lower surface of the buffer solution container and the developing layer, whereby The stool solution is dropped on the spreading layer.

According to the second aspect of the present invention, by pressing the switch, a flow path is similarly formed between the lower surface of the buffer solution container and the developing layer, and the stool solution is dropped on the developing layer. You.

Therefore, after stool is collected by the stool collection device,
The fecal sampling device is inserted from the opening of the main body, and thereafter, the above operations are performed, and the determination of fecal occult blood can be easily performed by observing from the determination window. That is, claim 1,
In the invention described in 2, the fecal occult blood can be reliably determined by a relatively simple operation.

[0061]

The present invention will be clarified below by describing non-limiting examples of the present invention. Example 1 (1) Preparation of a carrier sensitized with a first antibody: a phosphate buffer solution (pH 7.0) of an anti-human Hb monoclonal antibody (mouse MU-110: manufactured by Japan Biotest Laboratory) as a first antibody.
2: 200 μg / ml). Further, a blue polystyrene-based latex (average particle size of 0.12 μm) prepared by the method of coloring latex particles described above.
m: 5% by weight of solid content).
Stirred for minutes. The obtained dispersion is treated at 4 ° C. for 18,000 r.
Centrifuged at pm for 50 minutes. After removing the supernatant, a 1% by weight BSA (manufactured by Sigma) phosphate buffer solution (p
H7.2) was added to adjust the latex concentration to 1% by weight. (2) Preparation of first antibody-sensitized carrier and second antibody binding spreading layer: 1 m of anti-human Hb monoclonal antibody (mouse MU-107: Nippon Biotest Laboratories) as second antibody
5 μl of a g / ml phosphate buffer solution (pH 7.2) was added dropwise to the developing layer (nitrocellulose membrane: 1 × 5 cm, manufactured by Bio-Rad) and dried at room temperature for 12 hours. After further drying, it was immersed in a 1% by weight BSA phosphate buffer solution (pH 7.2) at 37 ° C. for 30 minutes.
It was dried again at room temperature for 12 hours.

Further, 0.5 μl of the first antibody-sensitized carrier obtained in the above (1) was immobilized on the developing layer in the same manner as described above. (3) Creation of system: A system similar to the structural example shown in FIG. 4 was created. As the stool collection device 21, a polyethylene container having the shape shown in FIG. 2 (b) is used, a buffer container is made of polyethylene (capacity: 2.5 ml), and 2.0 ml of a phosphate buffer (pH 7.2) is used. Injected. 1 drop
Set to 00 μl. As the filtration device, a filter for liquid chromatography (manufactured by Sekisui Chemical Co., Ltd.) having an average pore diameter of 2 μm was used.

Test Example 1 In this example, a sensitivity test was performed using a human Hb standard solution. Human Hb (manufactured by Sigma) in phosphate buffer (pH 7.2)
And the solution was added to the buffer container 3 of the system prepared in Example 1 so as to have a concentration of 0 to 1000 μg / ml. After gently shaking, the stool collection device 2 was pushed in and dropped onto the developing layer 11 from the dropping portion. After standing for 5 minutes,
The second antibody-immobilized portion 17 of the developing layer 11 was observed from the judgment window 1b of the main body, and whether the colored latex was blue or not was judged as positive or negative. Table 1 shows the results. From the results in Table 1, it can be seen that human Hb by the system of the present invention
Was determined to be 0.25 μg / ml.

Test Example 2 In this example, a fixed amount of a human Hb preparation was added to a human stool sample, and the sample was applied to the same system as in Test Example 1.

In the same manner as in Test Example 1, human H
b Standard was added. Further, a fixed amount of a sample was collected from the stool of a healthy person using the stool collection instrument 2, set in the buffer solution container 3, and dissolved. Table 1 shows the results of the same operation and judgment as in Test Example 1 above. As is clear from Table 1, the same results as in Test Example 1 were obtained. Therefore, it is understood that the system of the present invention can be applied to stool sample inspection.

[0066]

[Table 1]

Example 2 (1) Preparation of the first antibody-sensitized carrier and (2) preparation of the first antibody-sensitized carrier and the second antibody-binding spreading layer were carried out in the same manner as in Example 1. (3) Production of System A system similar to the structural example shown in FIG. 7 was produced. The stool collection device 2 is made of polyethylene having the shape shown in FIG. 2B, and the buffer solution container 3 is made of polyethylene (with a capacity of 2.5
2.0 ml of a phosphate buffer (pH 7.2)
Was injected. The drop amount was set to 100 μl. The filter used was a liquid chromatography filter (manufactured by Sekisui Chemical Co., Ltd.) having an average pore diameter of 2 μm.

Test Example 3 In this example, a sensitivity test was performed using a human Hb standard solution. Human Hb (manufactured by Sigma) in phosphate buffer (pH 7.2)
And the solution was added to the buffer container 5 of the system prepared in Example 2 so as to have a concentration of 0 to 1000 μg / ml. After gentle shaking, the drop switch 22 was pressed, and the stool solution was dropped onto the developing layer 11. After standing for 5 minutes,
The second antibody-immobilized portion 17 of the developing layer 11 was observed from the judgment window 1b of the main body, and whether the colored latex was blue or not was judged as positive or negative. Table 2 shows the results. From the results in Table 2, it can be seen that human Hb by the system of the present invention
It can be seen that the measurement sensitivity is 0.25 μg / ml.

Test Example 4 In this example, a fixed amount of human Hb standard was added to a human stool sample, and the sample was applied to the same system as in Test Example 3.

In the same manner as in Test Example 3, human H
b Standard was added. Further, a fixed amount of a sample was collected from the stool of a healthy person using the stool collection device 2 shown in FIG. The stool collection instrument 2 was set on the main body and shaken lightly. The operation was performed in the same manner as in Test Example 3 above, and the results of the determination are shown in Table 2. From the obtained results, it was confirmed that the system of the present invention can be applied to stool sample inspection.

[0071]

[Table 2]

[0072]

As described above, according to the first and second aspects of the present invention, the buffer solution container for dissolving the stool and the solid matter in the stool sample in the main body combined with the stool collection device are used. A filter for filtration and a developing layer constituting a chromatography section are integrally formed. Therefore,
According to the first aspect of the present invention, after a stool sample is collected from a stool collection instrument, the stool solution is inserted into the main body and the buffer solution container or the stool collection apparatus is moved up and down, and the stool lysate flows through the channel forming means. Thus, the presence or absence of fecal occult blood can be determined by the developed layer. Similarly, in the invention described in claim 2,
By pressing the switch after inserting the stool collection instrument into the main body, the stool lysate is dropped on the developing layer, and the presence or absence of fecal occult blood can be similarly determined.

Therefore, according to the first and second aspects of the present invention,
Since all the steps of fecal occult blood determination using the immunochromatography method are performed in the main body, the configuration of the system required for fecal occult blood determination is simplified, and extremely easily, even if it is unskilled. It is possible to easily determine the presence or absence of fecal occult blood.

[Brief description of the drawings]

FIG. 1 is a schematic perspective view for explaining a fecal occult blood determination device of a first structural example of the present invention.

FIGS. 2A to 2C are side views for explaining a modified example of the stool collection device.

FIGS. 3A to 3C are side views for explaining other modified examples of the stool collection device.

FIG. 4 is a schematic cross-sectional view of the fecal occult blood determination device of the first structural example.

FIGS. 5 (a) and (b) are perspective views respectively illustrating a modified example of a stool collection device and a buffer solution container in which the stool collection device is set.

FIGS. 6A and 6B show a modification of the first structural example, FIG. 6A is a schematic cross-sectional view showing a state in which the stool is inserted into a buffer solution container, and FIG. FIG. 4 is a schematic cross-sectional view showing a state in which a stool is moved downward to form a flow path between a buffer solution container and a developing layer.

FIG. 7 is a schematic cross-sectional view for explaining a fecal occult blood determination device of a second structural example.

FIGS. 8A and 8B are cross-sectional views each showing a modification of the switch.

9 is a partially cutaway sectional view showing a state where the switch shown in FIG. 8B is pressed.

FIG. 10 is a partially cutaway schematic cross-sectional view for explaining a modification of the second structural example.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Main body 1a ... Feces collection device insertion hole 1b ... Judgment window 1e ... Switch insertion hole 2 ... Feces collection device 3 ... Buffer solution container 4 ... Buffer solution 7 ... Press spring 8 ... Lower partition wall 9 ... For forming a flow path Dripping needle 10 ... Filter 11 ... Development layer 22 ... Switch

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-6-201701 (JP, A) JP-A-6-194363 (JP, A) JP-A-4-351959 (JP, A) 28735 (JP, U) Hikaru 2-140468 (JP, U) (58) Fields investigated (Int. Cl. ⁷ , DB name) G01N 33/48-33/98

Claims (2)

(57) [Claims] 1. An apparatus for judging fecal occult blood, comprising: a fecal sampling device for collecting feces; an opening for inserting the fecal collecting device on an outer surface; and presence / absence of fecal occult blood. A main body having a determination window for; a buffer solution container provided in the main body, the buffer solution container being arranged through the opening so that the feces collection device is inserted therein; and a buffer solution container being disposed below the buffer solution container. A filter for filtering a buffer solution in which the stool sample is dissolved, and a start portion located below the filter, and a judging portion arranged so as to face the judging window, the anti-hemoglobin antibody and a spreading layer created carriers and anti-human hemoglobin antibody is fixed, by moving the inserted feces collection instrument to the buffer vessel or the buffer vessel vertically, the at least a portion of said buffer container bottom surface Fi The stool sample is dissolved in Ruta
Buffer flowed and then filtered through the filter
A fecal occult blood determination device, comprising: flow path forming means for forming a flow path between at least a part of the lower surface of the buffer solution container and the spread layer so that flows into the spread layer . 2. An apparatus for judging fecal occult blood, comprising: a fecal sampling device for collecting stool, an opening for inserting the fecal sampling device on an outer surface, and the presence or absence of fecal occult blood from outside. A main body having a determination window for, a buffer solution container provided in the main body, and arranged so that a feces collection device is inserted through the opening, and disposed below the buffer solution container, A filter for filtering a buffer solution in which the stool sample is dissolved, a determination unit is provided below the filter so as to face the determination window , and an anti-human hemoglobin antibody-sensitized carrier and an anti-human hemoglobin antibody-sensitized carrier are provided. and a spreading layer human hemoglobin antibody is fixed, the flight buffer the sample is dissolved to flow from at least a portion of the buffer container bottom surface to the filter, then the filter
Flow path forming means for configuring a flow path between at least a part of the lower surface of the buffer solution container and the developing layer so that the buffer solution filtered in the developing layer flows into the developing layer; By pressing the one end, the flow path forming means is operated, whereby the buffer solution in which the stool sample is dissolved can be dropped from the buffer solution container toward the developing layer. A fecal occult blood determination device, comprising: a switch configured to: