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WO2016013599A1 - Coton-tige pour la collecte de matières fécales et kit de détection immunochromatographique contenant celui-ci - Google Patents

Coton-tige pour la collecte de matières fécales et kit de détection immunochromatographique contenant celui-ci Download PDF

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Publication number
WO2016013599A1
WO2016013599A1 PCT/JP2015/070912 JP2015070912W WO2016013599A1 WO 2016013599 A1 WO2016013599 A1 WO 2016013599A1 JP 2015070912 W JP2015070912 W JP 2015070912W WO 2016013599 A1 WO2016013599 A1 WO 2016013599A1
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WIPO (PCT)
Prior art keywords
cotton
stool
swab
cotton body
sample
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PCT/JP2015/070912
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English (en)
Japanese (ja)
Inventor
公良 西谷
結城 久美子
Original Assignee
積水メディカル株式会社
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Application filed by 積水メディカル株式会社 filed Critical 積水メディカル株式会社
Priority to JP2015556884A priority Critical patent/JP5936795B1/ja
Priority to CN201590000636.4U priority patent/CN206671337U/zh
Publication of WO2016013599A1 publication Critical patent/WO2016013599A1/fr

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/02Devices for withdrawing samples
    • G01N1/04Devices for withdrawing samples in the solid state, e.g. by cutting
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals

Definitions

  • the present invention relates to a swab for stool collection and a detection kit using immunochromatography including the swab.
  • Detection methods using immunochromatography are widely used because detection with high specificity and sensitivity is possible.
  • Samples applied to detection using immunochromatography include blood, urine, feces and the like, which are suitable as clinical test samples and widely used as diagnostically useful samples.
  • a substance to be detected contained in stool using immunochromatography it is necessary to collect a specific amount of stool, and various contrivances have been made for the stool collection rod.
  • Patent Document 1 discloses a stool collection container in which a characteristic stool collection rod is accommodated in the center of the container.
  • the stool collection rod is composed of a rod-like portion and a stool collection portion at the tip, and the stool collection portion is provided with a plurality of annular convex portions that are larger in diameter than the outer diameter of the rod-like portion.
  • the action of collecting (scraping) stool becomes stronger, and even when the stool to be collected is hard stool, a necessary amount can be easily collected.
  • the following are known as cotton swabs that are not particularly specialized for collecting stool, but are devised in shape.
  • Patent Document 2 discloses a cleaning swab for a nostril. A cotton ball is attached to the tip of the shaft of the cotton swab, and a ring-shaped depression like a groove is formed around the cotton ball.
  • Patent Document 3 discloses an ear cleaning cotton swab.
  • a cotton body part is provided at the end of the shaft body of the cotton swab, a stopper part larger than the outer ear hole is provided in the shaft body, and the tip of the cotton body part is prevented from entering deeper than the cartilage ear canal, This ensures the safety of ear cleaning.
  • Patent Document 4 discloses a cotton swab having a shaft portion and a contact portion at the tip of the shaft portion, and a mark indicating a depth that can be inserted according to each insertion site of the human body is formed on the shaft portion. ing.
  • feces collected to detect infant rotavirus and the like include rectal stool inserted into the infant's anus in addition to excretion excreted in the infant's diaper and the like. Since rectal stool is generally harder than excretory stool, when the same kind of stool collection rod is used in the same way, the amount of collected stool is large, and the amount of stool collected is small. There is a problem that the amount varies. Furthermore, since the collected stool is suspended in an appropriate liquid and detected using immunochromatography, the construction of the swab for stool collection has little effect on the detection system using the immunochromatography. desirable.
  • JP 2004-317481 A JP 2003-339767 A Japanese Patent Laid-Open No. 7-194650 JP 2005-13423 A
  • the conventional stool collection rod is not configured to collect both rectal stool and excretory stool. Moreover, even if the cotton swab inserted into the nose or ear of the prior art is used as it is for collecting feces, the feces cannot be collected properly. That is, in Patent Document 1, a convex portion is formed around the stool collection rod in order to enhance the scraping action so that even a hard stool can be sufficiently collected. It is formed. Since the stool collection rod having such a convex portion damages the rectum, it cannot be directly inserted into the rectum for collecting feces, and is not suitable for collecting rectal stool.
  • Patent Document 4 is also provided with a stopper on the shaft part to prevent it from being inserted into the ear for insertion into the ear, and the cotton body part is not devised to collect feces. Cannot be used.
  • the present invention provides a necessary amount for both collecting feces as a sample, collecting rectal feces by inserting into the rectum, and collecting excreted excreted feces. It is an object to provide a swab for stool collection that can be collected appropriately and a detection kit using immunochromatography including the swab.
  • a ring-shaped recess or projection is formed on the outer peripheral surface of the cotton body part from the tip in the surface area of the cotton body part. It was found that a necessary amount of feces can be collected by forming at a position corresponding to 15 to 40%.
  • feces can be attached to the tip part from the groove in the cotton body part, and when collecting hard stool represented by rectal stool, This is because the entire cotton body part can be inserted into the rectum so that feces can adhere to the entire cotton body part.
  • a detection kit using immunochromatography comprising the following (1) and (2).
  • An immunochromatographic test strip for detecting a complex of a detected substance and a conjugate in a sample by developing in a carrier, wherein the conjugate is an antibody that reacts immunologically with the detected substance Alternatively, the antigen is immobilized on a label, and the insoluble carrier is A sample supply section for supplying a sample; A detection unit on which an antibody or antigen that immunologically reacts with a target substance is immobilized; The detection kit using the immunochromatography according to the above ⁇ 1>, wherein the shape of the cotton body part of the test strip ⁇ 2> (1) has a cylindrical shape whose diameter is larger than the outer periphery of the shaft body.
  • ⁇ 3> The diameter of the cylindrical cotton body part of (1) is 4.0 to 5.5 mm, the length is 13.0 to 15.0 mm, and the ring-shaped concave or convex part is the tip of the cotton body part.
  • ⁇ 4> The detection kit according to any one of ⁇ 1> to ⁇ 3>, wherein the recess is a groove.
  • ⁇ 5> The detection kit according to any one of ⁇ 1> to ⁇ 4>, wherein the conjugate part having a conjugate is disposed downstream of the sample supply part of the test strip.
  • a cotton swab provided with a cotton body portion at the end of the shaft body, and on the outer peripheral surface of the cotton body portion, The cotton swab in which the ring-shaped groove is formed at a position corresponding to 15 to 40% of the surface area of the cotton body part from the tip.
  • the diameter of the cylindrical cotton body part is 4.0 to 5.5 mm, the length is 13.0 to 15.0 mm, and the ring-shaped groove is 2.0 to 6.5 mm from the tip of the cotton body part.
  • the swab for stool collection according to the above ⁇ 7> which is formed on an outer peripheral surface of 0 mm.
  • the detection method using immunochromatography when collecting stool as a sample, when collecting hard stool such as rectal stool, the whole cotton body part is used, and feces are excreted.
  • the tip part from the concave part or convex part of the cotton body part by using the tip part from the concave part or convex part of the cotton body part, the necessary amount of feces with different properties and types can be appropriately collected with one kind of cotton swab can do. Therefore, according to such a swab for stool collection, a sample collected can be applied to immunochromatography to enable accurate detection.
  • the concave or convex portion of the swab for stool collection of the present invention is formed of the same material as the cotton body without adding components that affect the immune reaction, the cotton body is immersed in a liquid such as a specimen dilution seat.
  • a liquid such as a specimen dilution seat.
  • stool is used as a sample.
  • the stool may be used as a sample as it is, or may be diluted with a diluent as appropriate. Moreover, you may use as a sample what was diluted and filtered suitably.
  • the substance to be detected of the present invention may be any substance as long as it is contained in feces as a sample and can be detected using an antigen-antibody reaction, and examples thereof include viruses, parasites, and proteins.
  • infectious gastroenteritis is mainly caused by viruses and parasites.
  • viruses that can be detected include norovirus, adenovirus, rotavirus, sapovirus, enterovirus, etc.
  • insects include cryptospiridium, amoeba dysentery, giardia and the like.
  • influenza virus as a virus to be detected, and proteins to be detected include human hemoglobin, hepatitis B virus antibody, hepatitis C virus antibody, human immunodeficiency virus antibody, and the like.
  • a swab for stool collection according to the present invention is a cotton swab in which a cotton body portion is provided at an end of a shaft body, and a ring-shaped recess or projection is formed on the outer peripheral surface of the cotton body portion. Is formed at a position corresponding to 15 to 40% from the tip. It is only necessary that one concave portion or convex portion is formed.
  • the cotton body portion is made of a fibrous body such as absorbent cotton or chemical fiber, and is provided at the end of the shaft body.
  • the shape of the cotton body part needs to be a shape that can be inserted into the anus and collected for rectal feces in addition to collecting excretory feces.
  • the cross-sectional shape includes a circle, an ellipse, a polygon, and the like. Among these, a cylindrical shape whose diameter is larger than the outer periphery of the shaft body is desirable.
  • the overall shape is preferably a columnar shape (so-called saddle shape), a water drop shape (saddle shape), or an elliptical cross section, and a cylindrical shape (so-called saddle shape) is most desirable.
  • the cross-sectional diameter is 4.0 to 5.5 mm, preferably 4.0 to 4.5 mm.
  • the length of the cylinder is 13.0 to 15.0 mm, preferably 13.5 to 14.5 mm.
  • the position of the ring-shaped concave portion or convex portion is formed on the outer peripheral surface of 2.0 to 6.0 mm from the tip of the cotton body portion, preferably 2.6 to 6.0 mm, and more preferably 2 .6 to 3.5 mm.
  • the cotton body part is formed, for example, by winding a fiber body around the tip of the shaft body via an adhesive or the like.
  • the surface of the cotton body part can be subjected to various types of processing as long as it does not interfere with stool collection, insertion into the anus, and immune reaction. Examples include processing. Variations on the shape of the cotton body are shown in FIG. In FIG. 4a, the overall shape of the cotton body part is a so-called water droplet type (saddle type).
  • FIG. 4a the overall shape of the cotton body part is a so-called water droplet type (saddle type).
  • FIG. 4B shows an elliptical shape whose cross-sectional shape in the major axis direction is an elongated ellipse.
  • FIG. 4 c shows a so-called saddle shape (columnar shape)
  • FIG. 4 d shows that the surface of the saddle shape is embossed.
  • a ring-shaped groove is formed on the outer peripheral surface of the cotton body part at a position corresponding to 15 to 40% of the surface area of the cotton body part from the tip.
  • FIG. 4e shows a saddle shape, and a smooth convex portion is formed in a ring shape on the outer peripheral surface instead of a groove.
  • the tip of the cotton body part has a rounded shape. However, when it is molded with a mold, it becomes such a shape for convenience of manufacture.
  • (C) to (e), including those having a round shape are referred to as saddle type (cylindrical type).
  • the shaft body is made of a material such as paper or plastic, and is formed so as to have a rigidity capable of collecting feces by contacting the cotton body portion provided at the end portion with the surface of the excretory stool or inserting it into the rectum. ing.
  • the shaft body is preferably in a shape that does not slip when the stool collector grips the shaft body, and the cross-sectional shape includes a circular shape and a polygonal shape such as a triangle and a quadrangle.
  • the length of the shaft should be long enough to allow the cotton body to contact the stool without touching the stool, and to be able to insert the cotton body without touching the anus.
  • the shaft body has a diameter of 1.5 to 3.5 mm, more preferably 2.0 to 3.0 mm when the cross section is circular.
  • the concave portion or the convex portion is substantially formed in a ring shape on the peripheral surface of the cotton body portion, and may be in a position to attach an appropriate amount for collecting feces from excrement feces. It is formed at a position corresponding to 15 to 40% from the tip.
  • the position corresponding to 15 to 40% of the surface area of the cotton body part is for collecting a predetermined amount of stool even if it is a stool having a different property. It is for collecting hard stool such as rectal stool as a whole, desirably 20 to 40%, and more desirably 25 to 35%.
  • the shape of the cotton body portion is a cylindrical shape whose diameter is larger than the outer periphery of the shaft body, and the diameter of the cylindrical cotton body portion is 4.0 to 5.5 mm and the length is 13.0.
  • the ring-shaped recess or projection is formed on the outer peripheral surface of 2.0 to 6.0 mm from the tip of the cotton body. The fact that one concave portion or convex portion is formed is sufficient if it can be recognized that it is visually divided into two parts when the cotton body part is observed.
  • the depth and width of the concave portion or the convex portion may be formed so that the boundary between the two portions of the cotton body portion can be visually recognized.
  • the depth of the concave portion or the height of the convex portion is not particularly limited as long as it can be visually recognized as described above, and is 0.5 to 4.0 mm, more preferably 2.0 to 3.0 mm.
  • the height and shape of the convex portion do not hinder the insertion, and there is little discomfort after insertion
  • the shape is more desirable.
  • the height of the convex portion is desirably about 0.5 to 1.5 mm, and the shape is desirably smooth.
  • a typical example of the concave portion is a groove, and the method for forming the groove in the cotton body portion may be any method as long as the groove is engraved.
  • a part of the inner wall of the mold for molding the cotton body portion A method of forming a groove by forming a groove by providing a protrusion on the protrusion and press-contacting the protrusion when the cotton body part passes through the mold.
  • a striated body can be mentioned, and the method of forming the striated body on the cotton body portion may be any method, for example, on the outer peripheral surface of the molded cotton body portion, and the same material. Examples of the method include winding a cotton striated body with an adhesive and molding the wound portion so that the wound portion is lighter than the other portions.
  • the cotton swab for stool collection of the present invention can appropriately change the site of the cotton body part to which the stool is attached depending on the nature and type of stool and the collection site. For example, when excreted stool is collected, feces are attached to the tip from the concave or convex portion of the cotton body, and when the rectal stool is collected by insertion into the anus, the entire cotton body is anal. Insert feces and attach feces. Moreover, what is necessary is just to make it adhere to the front-end
  • the immunochromatographic test strip of the present invention has an insoluble carrier that detects a complex of a detected substance and a conjugate in a sample by developing a stool-derived sample, and the insoluble carrier supplies the sample.
  • a sample supply unit, and a detection unit on which an antibody or antigen that immunologically reacts with the substance to be detected is immobilized.
  • the conjugate is an antibody or antigen that immunologically reacts with a substance to be detected, immobilized on a label, and may be arranged as a conjugate part on a part of the test strip. Alternatively, it may exist as a separate conjugate reagent to be mixed with the analyte.
  • the sample supply part is a part for supplying a sample containing a substance to be detected, and is formed on a part of the insoluble carrier, and the sample supply part is on the upstream side of the conjugate part.
  • the conjugate part is a part containing a conjugate, and is formed on an insoluble carrier on the downstream side of the sample supply part.
  • the detection part is a part where an antibody or antigen that immunologically reacts with a substance to be detected is immobilized, and is formed on an insoluble carrier on the downstream side of the conjugate part.
  • the sample supply unit, the conjugate unit, and the detection unit are arranged in this order from upstream to downstream, and these may be formed in the same insoluble carrier, or laminated so that separate insoluble carriers partially overlap each other. It may be what was done.
  • a typical example is a test strip that is composed of separate carriers, each of which includes a sample pad that serves as a sample supply unit, a conjugate pad that serves as a conjugate unit, and a detection pad that serves as a detection unit. Each carrier is laminated so that a part thereof contacts the upstream and downstream carriers. When a sample containing a substance to be detected is supplied to the sample pad, the sample flows through the sample pad to the conjugate pad on the downstream side.
  • the substance to be detected in the sample and the conjugate form a complex (aggregate).
  • the sample is then developed into a detection pad that is placed in contact with the bottom surface of the conjugate pad. Since the antibody or antigen that immunologically reacts with the substance to be detected is immobilized on the detection pad, the complex is bound and immobilized here by an immune reaction.
  • the immobilized complex is detected by a means for detecting absorbance or reflected light derived from the conjugate.
  • an individual conjugate reagent for example, a filter chip in which a conjugate is incorporated in a filter can be mentioned.
  • the conjugate and the substance to be detected are bonded to each other by passing the specimen diluent, thereby forming a complex (aggregate).
  • the substance to be detected can be detected.
  • the immunochromatographic test strip is preferably disposed on a solid support such as a plastic adhesive sheet.
  • the solid support is composed of a material that does not interfere with the capillary flow of the sample and conjugate.
  • the immunochromatographic test strip may be fixed on a solid support with an adhesive or the like.
  • the adhesive component and the like are also composed of a substance that does not interfere with the capillary flow of the sample and the conjugate.
  • the immunochromatographic test strip is stored in an appropriate container (housing) in consideration of the size of the immunochromatographic test strip, the sample addition method and addition position, the formation position of the insoluble carrier detection part, the signal detection method, and the like. -It can be mounted and used, and the state stored and mounted in this way is called "device".
  • the antibody or antigen that immunologically reacts with the substance to be detected used in the present invention is an antibody or antigen that can bind to the substance to be detected.
  • the substance to be detected is a virus or an antigen
  • the antibody or the substance to be detected When is an antibody, an antigen is preferred.
  • An antibody or antigen that reacts immunologically with a substance to be detected is immobilized on a label and a detection part described later.
  • the antibody or antigen immobilized on the label and the detection unit may be the same, but it is preferable that the label and the detection unit are different.
  • Marker As the label used in the present invention, a known label conventionally used for a test strip for immunochromatography can be used.
  • colloidal metal particles such as gold colloid particles and platinum colloid particles, color latex particles, magnetic particles, and fluorescent particles are preferable, and gold colloid particles and color latex particles are particularly preferable.
  • the conjugate used in the present invention is obtained by immobilizing an antibody or antigen that immunologically reacts with a substance to be detected on the label as described above.
  • the conjugate is preferably one in which an anti-rotavirus monoclonal antibody and an anti-adenovirus monoclonal antibody are immobilized on colloidal gold particles.
  • Examples of a method for immobilizing an antibody or antigen that immunologically reacts with a substance to be detected to a label include physical adsorption and chemical bonding, and are generally immobilized by physical adsorption.
  • the immunochromatographic test strip of the present invention may further contain other reagents and components depending on measurement conditions and samples.
  • examples of other reagents include blocking agents that prevent non-specific reactions.
  • Other configurations include, for example, a 3rd pad for removing components unnecessary for measurement in the sample, and an absorption pad for controlling the development of the sample by absorbing the sample that has moved and passed through the insoluble carrier. .
  • the production of the immunochromatographic test strip of the present invention can be carried out by appropriately modifying or altering the method described in the examples.
  • the detection kit using the immunochromatography of the present invention includes the stool swab and the immunochromatographic test strip.
  • the detection kit may further include a reagent necessary for detection, a sample diluent, a test tube, an instruction manual, a housing for storing a test strip, and the like.
  • Test Example 1 Test for collecting stool as a sample using the swab for stool collection of the present invention.
  • the swab of this invention The swab used for this test example is shown in FIGS.
  • the cotton swab of the present invention is provided with a cotton body portion 2 at the end of the shaft body 1 and one ring-shaped groove 3 formed on the outer surface near the tip of the cotton body portion.
  • the length of the shaft is 145 mm
  • the cotton end is 14.5 mm from the tip of the shaft
  • the groove is formed at a position 3 mm from the tip.
  • the cotton body part is formed by winding fibrous cotton around the shaft body, and the groove is molded by being compressed by a convex state formed on the inner surface of the mold when the cotton body part passes through the mold. It was.
  • the shaft body has a circular cross-sectional shape and a diameter of 2.5 mm.
  • the cotton body has a cylindrical shape (a saddle shape) and a cross-sectional diameter of 4.5 to 5 mm.
  • the depth of the groove is about 1 mm, and therefore the cross-sectional diameter of the constricted portion of the groove is 2.5 to 3 mm. 2.
  • Fecal collection Samples were prepared from 7 samples of volunteer stool. The breakdown of the properties of the samples were 3 soft stool samples and 4 hard stool samples. The sample (feces) is collected from the specimen as described in 1. above.
  • the stool of each specimen was wiped to the position of 3 mm, 5 mm, and 10 mm from the tip of the cotton body using a cotton swab, and the stool was collected by attaching the stool to the cotton body.
  • the weight of the collected stool was measured.
  • the results are shown in Table 1. 3. Results and Discussion
  • the average value of the amount of soft stool collected from the tip of the cotton body part to 3 mm was 54 mg, which almost coincided with the average value of 58 mg of the hard stool collected from the tip of 10 mm. Accordingly, it was confirmed that the amount collected could be averaged by changing the position where the cotton body part was collected according to the properties of the sample. For example, in the case of hard stool and soft stool, in the case of soft stool, it may be collected up to 30% of the surface area of the entire cotton body part, and in the case of hard stool, it may be collected almost entirely.
  • Test Example 2 Example of production of test strip for immunochromatography of the present invention (3-pad type) A membrane (b) on which an antibody (c1) against an antigen to be detected and a control reagent (c2) are immobilized is attached to a plastic adhesive sheet (a), and then a gold colloid is impregnated with a conjugate sensitized with the antibody.
  • the conjugate pad (d) was placed and mounted. Further, the sample pad (e) was disposed and mounted so as to overlap the conjugate pad, and the absorbent pad (f) was disposed and mounted on the opposite end.
  • the structure which piled up each component was cut
  • Detection can be performed by adding a solution obtained by suspending the stool collected in Test Example 1 in a diluent to the sample pad of the immunochromatographic test strip of the present invention and developing the conjugate pad and the membrane. .
  • the detection method using immunochromatography when collecting stool as a sample, when collecting hard stool such as rectal stool, the whole cotton body part is used, and feces are excreted.
  • the necessary amount of feces with different properties and types can be appropriately collected with one kind of swab for collecting stool can do. Therefore, according to such a swab for collection of stool, since it is possible to collect a certain amount just by changing the collection site in the cotton body part regardless of the nature of the feces, the collection method, it is applied to immunochromatography, Accurate detection is possible.

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Abstract

La présente invention concerne le problème de la fourniture d'un coton-tige pour la collecte de matières fécales, ledit coton-tige permettant la collecte de matières fécales en une quantité appropriée pour détection indépendamment de la propriété ou du type des matières fécales lorsqu'il est utilisé pour collecter des matières fécales en tant qu'échantillon dans un procédé de détection immunochromatographique, et un kit de détection immunochromatographique contenant le coton-tige. Afin de résoudre le problème ci-dessus, l'invention concerne un kit comprenant une combinaison d'un coton-tige pour collecte de matières fécales avec une bandelette de test immunochromatographique, ledit coton-tige comprenant une partie de boule de coton formée à une extrémité d'un axe et, sur la surface périphérique externe de la partie de boule de coton, des parties évidées en forme d'anneau ou des parties faisant saillie étant formées au niveau d'une position correspondant à 15 à 40 %, en termes de surface, depuis l'extrémité supérieure.
PCT/JP2015/070912 2014-07-25 2015-07-23 Coton-tige pour la collecte de matières fécales et kit de détection immunochromatographique contenant celui-ci WO2016013599A1 (fr)

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JP2015556884A JP5936795B1 (ja) 2014-07-25 2015-07-23 採便用の綿棒およびこれを含むイムノクロマトグラフィーを利用した検出キット
CN201590000636.4U CN206671337U (zh) 2014-07-25 2015-07-23 采便用的棉棒及包含了该棉棒的利用了免疫层析法的检测试剂盒

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US20220011306A1 (en) * 2018-11-19 2022-01-13 Sekisui Medical Co., Ltd. Immunochromatographic test strip and immunochromatographic detection kit

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