JPH11166929A - Sticking agent for inspection and inspecting method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sticking agent for inspection capable of simply and non-invasively sampling an object to be inspected present in or on the surface of a body in the state that impurities are removed and an inspecting method capable of accurate inspection using the above-mentioned sticking agent for inspection. SOLUTION: A sticking agent for inspection is formed by laminating a support layer 1, a reservoir layer 2. and a sticking layer 4. A control film 3 is provided between the reservoir layer 2 and the sticking layer 4 in the sticking agent for inspection. The sticking layer 4 capable of being stuck at least in an oral cavity, the support layer 1, the reservoir layer 2, and the control film 3 are laminated in the same order. A substance to be inspected is, for example. a carcinoembryonic antigen(CEA), Streptococcus mutans, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a test patch and a test patch for the purpose of collecting a substance which is an indicator of a disease on or inside a human body or an indicator of a physiological condition of the human body. It relates to the inspection method used.

[0002]

2. Description of the Related Art Carcinoembryonic antigen (CEA; carcino)
Embronic antigen) is a substance often used as an index for malignant tumors such as breast cancer,
By measuring the concentration of CEA in the serum, it is possible to diagnose whether or not cancer has occurred in the living body.

[0003] However, in early breast cancer, the concentration of CEA in serum is not so high that it is difficult to detect it.
Is not suitable for early detection of breast cancer.

There is also a method of diagnosing breast cancer by palpation of the breast. However, early breast cancer may be overlooked, and it is necessary for a doctor to perform a direct medical examination for each individual. However, the palpation method is not suitable as a large-scale screening method.

On the other hand, the nipple secretion, which is a secretion from the mammary gland, contains CEA that is directly released from the tumor to the ducts. Therefore, the CEA concentration in the nipple secretion is lower than the CEA concentration in serum. By measuring the concentration, early detection of breast cancer is possible.

[0006] However, since the secretion amount of the nipple secretion itself is extremely small except during pregnancy and lactation, it is difficult to collect the nipple secretion easily. Normally, a method of squeezing the nipple directly from the nipple to the hematocrit tube by compressing the nipple is used, but this method has a problem that it takes time and effort to collect a necessary amount.

In order to solve such a problem, Japanese Patent Application Laid-Open No. 9-149890 discloses a support, a pressure-sensitive adhesive layer spread on the surface thereof, A patch for testing having an adsorbent for adsorbing occult blood has been disclosed. According to this test patch, there is an effect that a very small amount of occult blood can be collected simply and non-invasively in a short time.

However, in this patch for inspection, since the adsorbing carrier and the collecting portion are in direct contact with each other, there is a problem that impurities are apt to be mixed in, so that the inspection accuracy is lowered.

[0009] Streptococcus mutans bacteria (hereinafter referred to as "mutans bacteria") are a type of streptococci.
It is known that it is distributed in large numbers in the human oral cavity and promotes dental caries by decomposing sucrose in the oral cavity to produce acids, and secretes substances causing bad breath. By examining the presence or absence of infection with mutans bacteria, the prevention and early treatment of dental caries can be performed. In the early stage of infection, the number of mutans bacteria in the oral cavity is small, and is easily overlooked, for example, in bacteriological examination by collecting saliva and the like. In addition, since the number of mutans bacteria is greatly affected by the state of the oral cavity such as a meal and tooth brushing, it is difficult to adjust the conditions in the examination, which may be overlooked.

[0010]

SUMMARY OF THE INVENTION In view of the above, it is an object of the present invention to provide a simple and non-invasive method of collecting an object to be inspected present inside or on the surface of a body from which contaminants have been removed. An object of the present invention is to provide a test patch that can be used, and a test method that can perform an accurate test using the test patch.

[0011]

The patch for inspection according to claim 1 of the present invention (hereinafter referred to as the present invention 1) comprises a support layer,
An adhesive patch for inspection comprising a reservoir layer and an adhesive layer, wherein a control film is provided between the reservoir layer and the adhesive layer. Hereinafter, the present invention 1
Will be described in detail.

The patch for examination of the present invention 1 is basically constituted by laminating a support layer, a reservoir layer and a sticking layer. The support layer is a layer that serves to hold the reservoir layer and the like. The support layer needs to hold the reservoir layer and the like, and have a property of not permeating the test substance stored in the reservoir layer. The material of the support layer is not particularly limited as long as it has the above properties, and examples thereof include polyethylene terephthalate. The material is used as a single-layer sheet or film, or as a laminate of two or more layers.

[0013] The reservoir layer is a layer that plays a role in storing the test substance. The reservoir layer is not particularly limited as long as it has a function of dissolving and storing the substance to be tested, and examples thereof include buffers such as physiological saline and phosphate buffer; and polysaccharides such as gelatin and cellulose. Gels to be formed are mentioned. The substance to be tested is CE
In the case of A, the reservoir layer is preferably a phosphate buffer.

The adhesive layer serves to adhere to the surface of the human body when the patch for inspection is applied to the surface of the human body, and includes an adhesive. The adhesive constituting the adhesive layer is not particularly limited as long as it can be applied to the human body. For example, an acrylic adhesive, natural rubber,
Rubber-based pressure-sensitive adhesives containing styrene-butadiene rubber, acrylonitrile-butadiene rubber, and the like; silicone-based pressure-sensitive adhesives containing polydimethylsiloxane as a main component, and the like.
Among them, a silicone-based pressure-sensitive adhesive, an acrylic-based pressure-sensitive adhesive composed of vinylpyrrolidone / 2-ethylhexyl acrylate copolymer, and the like are preferable. These adhesives may be used alone or in combination of two or more.

In order to improve the adhesiveness to the human body, or to improve the feeling of sticking,
For example, additives such as rosin, hydrogenated rosin, tackifiers such as polyterpene resins, etc .; vegetable oils such as olive oil; animal oils such as tallow, higher fatty acid esters such as isopropyl myristate, and liquid paraffin may be added.

Preferred combinations of the pressure-sensitive adhesive and the additives include, for example, a silicone-based pressure-sensitive adhesive and liquid paraffin, a pressure-sensitive adhesive composed of a vinylpyrrolidone / 2-ethylhexyl acrylate copolymer, and isopropyl myristate. With the above combination, the plasticity of the adhesive layer is improved, and the substance to be inspected easily permeates.

The thickness of the above-mentioned adhesive layer is preferably a thickness sufficient to adhere to the surface of the human body and a thickness that does not hinder the permeation of the test substance. From such a viewpoint, the thickness of the adhesive layer is preferably from 10 to 400 μm, more preferably from 20 to 100 μm.

In the patch for inspection of the present invention 1, a control film is provided between the reservoir layer and the adhesive layer. The test substance is not particularly limited as long as it is a secretion secreted inside or on the surface of the human body and these secretions reach the surface of the human body.
Examples include CEA in nipple secretions.

The control membrane controls the penetration and permeation of the substance to be tested, thereby preventing foreign substances such as cells and blood cells in the secretion solution from reaching the reservoir layer, and improving the test accuracy due to the interference of the foreign substances. It plays a role in preventing the decline. The material of the control film is not particularly limited as long as it has the above-mentioned functions, and examples thereof include cellulose derivatives such as cellulose and nitrocellulose; and porous polyethylene. Of these, cellulose and nitrocellulose are preferred, and especially for CEA,
Nitrocellulose is preferred. In consideration of the above characteristics, the thickness of the control film is preferably 20 to 500 μm,
0 to 200 μm is more preferable.

FIG. 1 is a cross-sectional view schematically showing the patch for inspection of the present invention 1. This patch for inspection is composed of the adhesive layer 4
It is composed of a control film 3, a reservoir layer 2 and a support layer 1, which are sequentially formed thereon. Further, in order to protect the adhesive layer 4, a release sheet 5 is adhered below the adhesive layer 4.

Further, a concave portion is formed in a part of the support layer 1, and the concave layer is filled with a material constituting the reservoir layer 2 to form the reservoir layer 2. Reservoir layer 2
May be formed on the control film 3 side. The release sheet 5 is peeled off and used when sticking to the body.

Another aspect of the invention according to claim 6 is an inspection method characterized by using the patch for inspection of the invention 1 described above. In the above-mentioned inspection method, first, the above-mentioned patch for inspection is stuck on the surface of the human body, and the substance to be inspected on or on the surface of the human body is transmitted through a control membrane or the like and stored in the reservoir layer.

Subsequently, the substance to be inspected stored in the reservoir layer is selectively taken out by a method such as extraction or the like, and the specimen is inspected, or the reservoir layer containing the substance to be inspected is collected and directly inspected. I do.

Although the inspection method is not particularly limited, in this specification, CEA is used as an example of the inspection method.
An inspection method will be described. When the reservoir layer in which the test substance is stored is liquid, for example,
A part of the reservoir layer is directly collected by a microsyringe and used as a sample for inspection. When the reservoir layer is solid, for example, the reservoir layer is immersed in a buffer to perform an extraction operation, and the extract is used as a sample for inspection.

The obtained sample was subjected to an enzyme immunoassay (E
IA), immunoturbidimetry (TIA), an immunological method such as a latex agglutination method, etc., are examined, and it is determined whether there is a suspicion of breast cancer or the like based on the CEA concentration. In the method according to the present invention, Since a sample containing no impurities can be used, the test can be performed with high test accuracy.

The patch for inspection according to claim 4 of the present invention (hereinafter referred to as the present invention 2) comprises at least an adhesive layer, a support layer, a reservoir layer and a control film which can be applied in the oral cavity in this order. ing.

The adhesive layer serves to adhere to the oral cavity at least when the adhesive patch for inspection is applied to the oral cavity, and comprises an adhesive. The adhesive constituting the adhesive layer is not particularly limited as long as it can be applied at least to the oral cavity. For example, a natural rubber-based adhesive, an adhesive using a rubber-based resin such as SBR or NBR, An acrylic pressure-sensitive adhesive, a silicone-based pressure-sensitive adhesive, and the like can be given. In the oral cavity, a large amount of water and the like also exists on mucous membranes and the like.
Alternatively, an adhesive which generates an adhesive force after water absorption is preferable. Examples of the water-absorbing adhesive include vinylpyrrolidone / 2
-Ethylhexyl acrylate copolymer and the like,
Examples of pressure-sensitive adhesives that generate pressure-sensitive adhesive strength after water absorption include polyvinylpyrrolidone, polyacrylic acid, and the like.

In order to improve at least the adhesiveness to the oral cavity or to improve the feeling of sticking, a tackifier such as rosin, hydrogenated rosin and polyterpene resin; olive oil, etc. Vegetable oils; animal oils such as beef tallow; higher fatty acid esters such as isopropyl myristate; and additives such as liquid paraffin.

Preferred examples of the combination of the above-mentioned pressure-sensitive adhesive and the above-mentioned additives include a pressure-sensitive adhesive comprising a vinylpyrrolidone / 2-ethylhexyl acrylate copolymer and isopropyl myristate. With the above combination, the plasticity of the adhesive layer is improved, and the substance to be inspected easily permeates.

The thickness of the above-mentioned adhesive layer is sufficient if it is at least sufficient to adhere to the inner surface of the oral cavity.
00 μm is preferable, and 50 to 200 μm is more preferable.

The reservoir layer is a layer that plays a role in storing the test substance. The reservoir layer is not particularly limited as long as it has a function of dissolving and storing the test substance, or if the test substance is a bacterium, as long as it can capture it, for example, physiological saline. And buffer solutions such as phosphate buffers; gels formed from polysaccharides such as gelatin and cellulose; The substance to be tested is Streptococcus mutans (Strept
In the case of ococcus mutans bacteria, agarose gel is preferably used as the reservoir layer.

The support layer serves to hold the reservoir layer. The support layer needs to hold the reservoir layer and have a property of not allowing the test substance stored in the reservoir layer to pass through. Further, in the present invention 2, an adhesive layer is provided on the other side of the support layer, not on the reservoir layer side, and the adhesive layer connects the reservoir layer via the support layer to the oral cavity or the like. Serves to fix to. The material of the support layer is not particularly limited as long as it has the above properties, and examples thereof include polyethylene terephthalate. The material is used as a single-layer sheet or film, or as a laminate of two or more layers.

On the other side of the reservoir layer, not on the support layer side, a control film is provided. The control membrane allows the test substance to permeate, controls its permeation and permeation effects, and prevents the contamination of the oral cavity such as food waste from reaching the reservoir layer. It plays a role in preventing the decline.

The material of the control film is not particularly limited as long as it has the above-mentioned functions.
Cellulose derivatives such as nitrocellulose; porous polyethylene; Among these, a porous cellulose membrane and a nitrocellulose membrane are preferable, and particularly, the substance to be tested is Streptococcus mutans (Str.
In the case of E. mutans bacteria, a porous nitrocellulose membrane is preferred. The thickness of the control film is 20 to 500 μm in consideration of the above characteristics.
Is preferable, and 40 to 200 μm is more preferable.

The test substance is mainly secreted or present in the oral cavity. If the substance can be measured to diagnose the physiological condition of the human body or the disease to be affected, the substance may be used. Although not limited to the type, preferably, for example, Streptococcus mutans (Str
lactic acid, acetic acid, and butyric acid secreted by S. eptococcus mutans and the above Streptococcus mutans bacteria.

FIG. 2 is a cross-sectional view schematically showing the patch for inspection of the present invention 2. This patch for inspection is composed of the adhesive layer 2
4, a support layer 21 and a reservoir layer 22 sequentially formed
And the control film 23. Also, the adhesive layer 2
4 to protect the release sheet 2
5 are adhered.

A recess is formed in a part of the control film 23, and the recess is filled with a material constituting the reservoir layer to form a reservoir layer 22. In addition, a concave portion for forming the reservoir layer 22 may be formed on the support layer 21 side. The release sheet 25 is peeled off and used when sticking to the body.

Another aspect of the invention according to claim 6 is an inspection method characterized by using the patch for inspection according to the second aspect of the present invention. In the above-mentioned inspection method, first, the patch for inspection of the present invention 2 is applied to the surface of the human body, particularly to the oral cavity (the invention according to claim 7), whereby the inspection is performed on the surface of the human body, particularly to the oral cavity. The target substance is permeated through the control membrane and stored in the reservoir layer.

Subsequently, the reservoir layer containing the substance to be inspected is collected and directly inspected, or the substance to be inspected stored in the reservoir layer is selectively taken out by a method such as extraction to perform the inspection. I do.

The inspection method is not particularly limited. For example, the following method can be used. When the reservoir layer in which the substance to be inspected is stored is in a liquid state, for example, a part of the reservoir layer is directly collected by a microsyringe or the like, and used as a sample for inspection. When the reservoir layer is solid, for example, the reservoir layer is immersed in a buffer to perform an extraction operation, and the extract is used as a sample for inspection. When the test substance is a cell such as a mutans bacterium, for example, when the reservoir layer is in a liquid state, the bacteria are identified by smearing the reservoir layer, and when the reservoir layer is in a solid state, the bacterium is directly stained. Or after contact smearing and staining,
Perform identification.

In these test samples, chemical substances and the like can be obtained by immunological methods such as enzyme immunoassay (EIA), immunoturbidimetry (TIA), latex agglutination, etc.
Alternatively, it is inspected by a biochemical method such as an enzyme reaction, and bacteria are inspected by microscopy, for example, after staining.

In the method according to the present invention, for example, a test object in the oral cavity is collected by applying the patch of the present invention 2 and a test sample collected from the reservoir layer is inspected, so that the test can be easily performed. Inspection can be performed noninvasively with high inspection accuracy.

[0043]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1 Solid silicone adhesive (manufactured by Dow Corning)
An adhesive solution prepared by mixing 30 parts by weight of liquid paraffin (produced by Maruishi Pharmaceutical Co., Ltd.) with 70 parts by weight of Silascon 355) was coated on a PET (polyethylene terephthalate) film as a release sheet to a thickness of 300 μm. , And dried at 60 ° C for 1 hour to form an adhesive layer having a thickness of 20 µm.

On the other hand, by forming a nitrocellulose membrane having a thickness of 100 μm for the control membrane, a diameter of 40 m was obtained.
A plurality of concave portions having a depth of 2 m and a depth of 2 mm were formed, and the nitrocellulose membrane subjected to the above-mentioned molding treatment was bonded to the above-mentioned adhesive layer.

Next, the pH of the concave portion of the nitrocellulose membrane was
A phosphate layer was formed by injecting 2 ml of the phosphate buffer solution of 7.5 at a time, and then a PET film was adhered to a nitrocellulose membrane as a support, and the reservoir layer was sealed. Further, the laminate subjected to these treatments was punched into a circle having a diameter of 50 mm centering on the reservoir layer to obtain a patch.

Comparative Example 1 A soft polyvinyl chloride film (Esmedica manufactured by Sekisui Chemical Co., Ltd.) was used as a support, and a styrene-isoprene-styrene (SIS) copolymer, a polyisobutylene rubber, an ester gum, and a fluid were used as an adhesive layer. Each of paraffins blended at a weight ratio of 20: 10: 35: 35 was spread on the above-mentioned support to produce an oval adhesive tape of 56 mm × 43 mm.

On the other hand, a PET film was adhered to a polyester nonwoven fabric as a protective layer, and then a copolymer of vinylpyrrolidone and 2-ethylhexyl acrylate (manufactured by Sekisui Chemical Co., Ltd.) was spread on the PET film. Next, a composite having a pore size of 0.45 μm (manufactured by Bio-Rad Co., Ltd.) composed of three layers based on nitrocellulose / polyester / nitrocellulose and functioning as a water-absorbing / adsorbing member was formed on the adhesive of the protective layer. ). Next, the laminate of the protective layer and the adsorption carrier was cut into a circle having a diameter of 20 mm,
The adhesive layer was adhered to the adhesive tape via the protective layer, and the side of the water-absorbing / adsorbing member / adsorbing carrier was further covered with a PET film as a release coating material to obtain a patch for inspection.

FIG. 3 is a cross-sectional view schematically showing the patch for inspection produced in Comparative Example 1.
A pressure-sensitive adhesive layer 12 is formed thereon, and a water-absorbing / adsorbing member / adsorption carrier 14 is laminated on a part of the pressure-sensitive adhesive layer 12 with a protective layer 13 interposed therebetween. Coated.

Evaluation method: 200 microliters of the nipple secretion of a healthy woman were obtained for 20 persons, and CEA was added to the nipple secretion of 10 of them.
4 μg was added. Next, this CEA-added nipple discharge was applied to a phenol plate in a circular shape with a diameter of 10 mm,
The patches prepared in Example 1 and Comparative Example 1 were applied to the portion to which the nipple discharge was applied.

Thereafter, for the patch of Example 1, 1 ml of the buffer solution in the reservoir layer was collected to obtain a sample, and this sample was used to measure the CEA concentration using an EIA kit (manmotech kit manufactured by Mochida Pharmaceutical Co., Ltd.). did. In the patch of Comparative Example 1, the CEA concentration was measured using the same EIA kit as in Example 1 after extracting the adsorbent with a buffer.

Table 1 shows the evaluation results. In Table 1,
+ Indicates positive and-indicates negative.

[0053]

[Table 1] As is clear from the results shown in Table 1, in the case of Example 1, an inspection result with higher accuracy was obtained than in the case of Comparative Example 1.

Example 2 A pressure-sensitive adhesive solution composed of a 10% by weight ethanol solution of polyvinylpyrrolidone was treated with a silicone-treated P as a release sheet.
ET (polyethylene terephthalate) film (thickness 8
(0 μm) was spread to a thickness of 500 μm, and then dried at 60 ° C. for 1 hour to form a 50 μm thick adhesive layer.

This was pressure-bonded to a PET film (thickness: 50 μm), which had not been subjected to a release treatment, as a support layer.

On the other hand, a plurality of concave portions having a diameter of 20 mm and a depth of 2 mm were formed by molding a porous nitrocellulose membrane having a thickness of 100 μm for a control membrane. Next, a reservoir layer was formed by placing an agarose gel having a diameter of 20 mm in the concave portion of the nitrocellulose membrane.

On the nitrocellulose membrane in which the agarose gel is accommodated in the recess, the release sheet-adhesive layer-support layer obtained above is laminated in this order so that the support layer side is in contact. Then, the nitrocellulose membrane and the support layer were adhered to each other using an adhesive, and the recess where the agarose gel was placed was sealed. Further, the laminate subjected to these treatments was punched out into a circular shape having a diameter of 30 mm around the reservoir layer (concave portion) to obtain a patch.

After removing the release sheet from the patch,
The patch was applied to the mouth of 20 healthy subjects for one day. One day later, it was recovered, the support layer was peeled off to expose the reservoir layer (agarose gel), a part of the reservoir layer was removed, smeared on a slide glass, stained, and then examined under a microscope for Streptococcus mutans bacteria. And the results are shown in Table 2. In Table 2, ○ indicates that bacteria were detected, and X indicates that bacteria were not detected.

Comparative Example 2 5 mL of the same subject's saliva as in Example 2 was collected, a part thereof was smeared on a slide glass, stained, and examined for Streptococcus mutans bacteria under a microscope. It was shown to.

[0060]

[Table 2] As is clear from Table 2, the samples collected by the patch of the present invention also detected those samples that could not be detected by the method of Comparative Example 2, indicating that the method of the present invention is excellent.

[0061]

Since the patch according to the first aspect has the above-mentioned constitution, the test substance existing inside or on the surface of the body can be easily and non-invasively collected without impurities. be able to. Since the patch according to claim 2 has the above-described configuration, it exhibits all of the above effects, and since the reservoir layer is in a liquid state, a part of the reservoir layer can be directly collected by a microsyringe or the like, so that it is more convenient. A sample for inspection can be collected. Since the patch according to the third aspect has the above-mentioned constitution, it can be simply and non-invasively collected in a state in which the carcinoembryonic antigen present inside or on the surface of the body is free of impurities.

Since the patch according to the fourth aspect has the above-mentioned constitution, the test substance existing inside or on the surface of the body, particularly the test substance existing in the oral cavity, can be easily and noninvasively contaminated. It can be collected with things removed.

Since the patch according to the fifth aspect has the above-mentioned constitution, it is easy and non-invasive to exist in the oral cavity Streptococcus mutans (Streptococ).
Cus mutans) can be collected in a state where contaminants have been removed.

Since the inspection method according to the sixth aspect has the above-described configuration, it is possible to accurately inspect the substance to be inspected.

Since the inspection method according to the seventh aspect has the above-described configuration, it is possible to accurately inspect the substance to be inspected existing in the oral cavity.

[Brief description of the drawings]

FIG. 1 is a cross-sectional view schematically showing a patch for inspection of the present invention 1.

FIG. 2 is a cross-sectional view schematically showing a patch for inspection of the present invention 2.

FIG. 3 is a cross-sectional view schematically showing the patch prepared in Comparative Example 1.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Support layer 2 Reservoir layer 3 Control film 4 Adhesion layer 5 Release sheet 21 Support layer 22 Reservoir layer 23 Control film 24 Adhesion layer 25 Release sheet

──────────────────────────────────────────────────続 き Continued on front page (51) Int.Cl. ⁶ Identification code FI G01N 33/574 G01N 33/574 E

Claims (7)

[Claims] 1. An inspection patch comprising a support layer, a reservoir layer and an adhesive layer laminated, wherein a control film is provided between the reservoir layer and the adhesive layer. Patch for inspection. 2. The patch according to claim 1, wherein the reservoir layer is in a liquid state. 3. The patch according to claim 1, wherein the test substance is a carcinoembryonic antigen. 4. An adhesive layer that can be applied at least in the oral cavity,
An inspection patch, wherein a support layer, a reservoir layer, and a control film are laminated in this order. 5. The test substance is Streptococcus mutans (Streptococcus mutans).
The patch for inspection according to claim 4, which is s) a bacterium. 6. An inspection method using the patch for inspection according to any one of claims 1 to 5. 7. An inspection method characterized in that a test substance stored in a reservoir layer is measured by affixing the patch for inspection according to claim 4 or 5 in an oral cavity.