Hemoglobin detection method, solution, fecal storage container, and fecal-containing liquid evaluation method

Abstract

This method for detecting hemoglobin includes: preparing a feces-containing liquid that contains feces and at least one selected from the group consisting of sulfurous acid, sulfite, disulfurous acid, disulfites, dithionous acid, and dithionites, and that has been stored for an arbitrary time; and detecting the hemoglobin contained in the feces-containing liquid by immunochromatography.

Description

[Technical Field]

[0001] The present disclosure relates to methods for detecting hemoglobin, solutions, fecal storage containers, and methods for evaluating fecal-containing fluids. This application claims priority based on Japanese Patent Application No. 2020-063212, filed on March 31, 2020, the contents of which are incorporated herein by reference. [Background technology]

[0002] A fecal occult blood test is known as a useful test method for diagnosing tumors in the lower gastrointestinal tract, such as the large intestine. In a fecal occult blood test, hemoglobin in stool is generally detected. Meanwhile, various methods for detecting hemoglobin in stool are known, such as a chemical assay, an immunoassay using a latex agglutination method, and an immunoassay using an immunochromatography method.

[0003] In any of the above detection methods, stool that has been stored for a certain period after collection may be used for testing. Patent Document 1 discloses a method for measuring components in a stool sample, which comprises contacting a collected stool sample with a desiccant to dry the stool sample, storing the dried stool sample in the desiccant, adding an aqueous medium to the desiccant in which the dried stool sample has been stored, dissolving the components in the stool sample in the aqueous medium, and measuring the components in the stool sample dissolved in the aqueous medium. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2017 / 104132 Summary of the Invention [Problem to be solved by the invention]

[0005] From the perspective of utilizing fecal occult blood tests for diagnosing diseases, there is a need in clinical settings for a method that can detect hemoglobin contained in stool with high accuracy through simple procedures and without requiring a long time.

[0006] Therefore, the present disclosure provides a hemoglobin detection method that is simple, stable, and rapid, allowing for hemoglobin detection. The present disclosure also provides a solution and a feces storage container suitable for the simple, stable, and rapid hemoglobin detection method. Furthermore, the present disclosure provides a feces-containing liquid evaluation method that is simple, stable, and rapid, allowing for evaluation of feces-containing liquid. [Means for solving the problem]

[0007] Examples of embodiments are given below: The present invention is not limited to the following embodiments.

[0008] One embodiment relates to a method for detecting hemoglobin, which includes preparing a feces-containing liquid that contains at least one selected from the group consisting of sulfurous acid, sulfite salts, disulfite, disulfite salts, dithionous acid, and dithionite salts and feces, and that has been stored for a given period of time, and detecting hemoglobin contained in the feces-containing liquid by immunochromatography.

[0009] Another embodiment relates to a solution containing at least one selected from the group consisting of sulfurous acid, sulfite salts, disulfite, disulfite salts, dithionous acid, and dithionite salts, and used as a preservative solution for stool and as a developing solution for immunochromatography capable of detecting hemoglobin.

[0010] Another embodiment relates to a stool storage container comprising a stool collection container and the solution of the above embodiment contained within the stool collection container, the stool collection container having a stool collection stick and a container body, the stool collection stick having a handle portion on one side and a rod portion on the other side, and the rod portion having a stool collection portion at or near the tip of the other side.

[0011] Another embodiment relates to a method for evaluating a feces-containing liquid, which comprises evaluating whether or not the feces-containing liquid contains hemoglobin using the hemoglobin detection method of the above embodiment. [Effects of the Invention]

[0012] According to the present disclosure, a hemoglobin detection method that is simple, stable, and rapid is provided, and a solution and feces storage container suitable for the simple, stable, and rapid hemoglobin detection method are also provided. Furthermore, according to the present disclosure, a feces-containing liquid evaluation method that is simple, stable, and rapid is provided, and a feces-containing liquid evaluation method is also provided. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 is a perspective schematic diagram illustrating an example of an immunochromatography device. [Figure 2] FIG. 2 is a schematic front view showing an example of a waste storage container. DETAILED DESCRIPTION OF THE INVENTION

[0014] The following describes embodiments of the present invention. The present invention is not limited to the following embodiments. The following embodiments can be implemented alone or in combination.

[0015] <Hemoglobin detection method> A hemoglobin detection method according to one embodiment of the present invention includes at least preparing a feces-containing liquid containing feces and at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite, the feces-containing liquid having been stored for an arbitrary period of time, and detecting hemoglobin contained in the feces-containing liquid by immunochromatography. The hemoglobin detection method may further include an optional step. In one embodiment of the detection method, immunochromatography is used to detect hemoglobin. Immunochromatography is a simple method capable of detecting hemoglobin.

[0016] Generally, in a fecal occult blood test, a certain amount of time elapses between the collection of stool and the start of the operation to detect hemoglobin contained in the stool. The stool is stored between collection and detection. According to a detection method that is one embodiment of the present invention, stable and rapid detection of hemoglobin can be achieved. In the detection method that is one embodiment of the present invention, a stool-containing liquid containing the above-mentioned acid and / or salt is used. It is presumed that the acid and / or salt acts as a stabilizer for hemoglobin in the stool-containing liquid, thereby preventing decomposition and denaturation of hemoglobin even after storage. Therefore, it is considered possible to stably detect hemoglobin even when using a stool-containing liquid that has been prepared for some time.

[0017] Furthermore, the acid and / or salt can prevent the viscosity of the feces-containing liquid from increasing compared to when sugars or the like, which are commonly known as stabilizers, are used. As a result, it is believed that the development time of the feces-containing liquid in the immunochromatography method, i.e., the time required to detect hemoglobin by the immunochromatography method, can be shortened, making it possible to perform a rapid hemoglobin test. However, the present invention is not limited by these assumptions.

[0018] In the hemoglobin detection method, the stool-containing liquid may be a liquid containing a solution (developer liquid) described below and stool. The hemoglobin detection method may further include providing the developer liquid and receiving the stool-containing liquid containing the developer liquid and stool. Examples of providing the developer liquid include a medical professional such as a doctor or nurse handing the developer liquid to a patient, and a testing professional who performs hemoglobin detection handing the developer liquid to a subject. The patient or test subject who receives the developer liquid can collect stool and mix the collected stool with the developer liquid to obtain the stool-containing liquid. Examples of receiving the stool-containing liquid include a medical professional receiving the stool-containing liquid from a patient, and a testing professional receiving the stool-containing liquid from a subject.

[0019] [Stool-containing fluid] The feces-containing liquid contains at least one selected from the group consisting of sulfurous acid, sulfite salts, disulfite, disulfite salts, dithionous acid, and dithionite salts, and feces. The feces-containing liquid preferably contains water as a liquid capable of dissolving and / or dispersing the acid and / or salt (sulfur-containing acid) and feces. The feces-containing liquid may further contain optional components other than water. Examples of optional components include at least one selected from the group consisting of buffers, proteins, antibiotics, preservatives, sugars, ferrocyanide compounds, chelating agents, protease inhibitors, surfactants, inorganic acids and salts thereof, and organic acids and salts thereof.

[0020] Each component that can be contained in the feces-containing liquid will be described later. The feces-containing liquid can be obtained, for example, by mixing at least a solution (developing liquid) described later with feces. Examples of the content of each component contained in the feces-containing liquid include the contents exemplified for each component in the developing liquid described later. In this case, the volume of the feces-containing liquid can be used as the basis for the content, instead of the volume of the developing liquid.

[0021] The feces are not particularly limited, and examples thereof include feces excreted or collected from mammals, such as humans, monkeys, gorillas, orangutans, pandas, dogs, cats, horses, pigs, sheep, wild boars, rabbits, mice, squirrels, hamsters, tigers, and lions. The hemoglobin detection method is suitable for humans.

[0022] The total content of components derived from feces contained in the feces-containing liquid may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more, based on the volume of the feces-containing liquid. The total content of components derived from feces contained in the feces-containing liquid may be, for example, 5% (w / v) or less, 3% (w / v) or less, 2% (w / v) or less, 1% (w / v) or less, or 0.5% (w / v) or less, based on the volume of the feces-containing liquid.

[0023] In the present disclosure, "% (w / v)" refers to the percentage of mass (g) based on the volume (100 mL). For example, "the content of component X is x% (w / v) based on the volume of the feces-containing liquid" means that component X is contained in an amount of x (g) per 100 mL of the feces-containing liquid. In addition, in the present disclosure, for a certain content, any numerical value selected from the example upper and lower limit values ​​and a numerical range obtained by combining these values ​​are also considered to be exemplified in the present disclosure. The same applies to other numerical values ​​such as pH, viscosity, temperature, and time.

[0024] The hemoglobin detection method uses a feces-containing liquid that has been stored for a desired period of time. The feces-containing liquid that has been stored may be a feces-containing liquid that has been stored for a desired period of time after preparation of a feces-containing liquid containing at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite. The state in which the feces-containing liquid is stored is not particularly limited, and examples include leaving it still, transporting it, shaking it, etc. The feces-containing liquid may be a feces-containing liquid that has been stored in a temperature-controlled atmosphere. Alternatively, the feces-containing liquid may be a feces-containing liquid that has been stored without any particular temperature control, for example, in a normal living environment.

[0025] The lower limit of the arbitrary time is not particularly limited, but may be, for example, 30 minutes or more, 1 hour or more, 5 hours or more, 10 hours or more, 1 day or more, 3 days or more, or 5 days or more, taking into account the time required from obtaining the fecal-containing liquid to performing the immunochromatographic test. The arbitrary time may be, for example, 21 days or less, 14 days or less, 10 days or less, 8 days or less, 7 days or less, 5 days or less, or 3 days or less. A time of 21 days or less tends to more easily prevent denaturation or degradation of hemoglobin. The starting point of the arbitrary time may be when the feces are contacted with at least one selected from the group consisting of sulfite, sulfite salt, disulfite, disulfite salt, dithionous acid, and dithionite salt, i.e., when the fecal-containing liquid containing at least both is obtained. The ending point of the arbitrary time may be when the fecal-containing liquid is supplied to an apparatus used for immunochromatography.

[0026] The storage temperature may be, for example, 0°C or higher, 2°C or higher, 4°C or higher, 8°C or higher, 10°C or higher, or 15°C or higher. The storage temperature may be, for example, 50°C or lower, 48°C or lower, 45°C or lower, 40°C or lower, or 38°C. When the temperature is 50°C or lower, denaturation or decomposition of hemoglobin tends to be easily prevented. For example, it is preferable that the temperature of the atmosphere during storage falls within the above range. For example, it is preferable that the temperature of the feces-containing liquid during storage falls within the above range.

[0027] The pH of the feces-containing liquid may be, for example, 6.0 to 7.5. Examples of the pH of the feces-containing liquid include the pH values ​​exemplified for the developing liquid described below. The viscosity of the feces-containing liquid may be, for example, 1.00 to 1.80 cP. Examples of the viscosity of the feces-containing liquid include the viscosity values ​​exemplified for the developing liquid described below. The pH and viscosity of the feces-containing liquid can be measured by the same method as the pH and viscosity of the developing liquid.

[0028] [Immunochromatography] In the hemoglobin detection method, hemoglobin is detected by immunochromatography. The immunochromatography method is not particularly limited, and a general method utilizing immune reaction and capillary action can be used. Unlike latex agglutination, immunochromatography does not require large equipment or complicated processes, and therefore hemoglobin can be easily detected. The hemoglobin detection method using immunochromatography is a method suitable for so-called POCT (Point of Care Test).

[0029] The immunochromatography method includes, for example, providing a membrane having an area where a first capture substance capable of capturing hemoglobin is immobilized, and a second capture substance labeled with a label and capable of capturing hemoglobin; forming a complex containing the first capture substance, hemoglobin, and the second capture substance in the area; and detecting the complex. The immunochromatography method may further include any optional steps.

[0030] In the immunochromatography method, the provision may be, for example, provision of an immunochromatography device having at least a sample pad, a conjugation pad, a membrane having an area where a first capture substance capable of capturing hemoglobin is immobilized, and a second capture substance. The immunochromatography device may further include an optional portion such as an absorbent pad.

[0031] The immunochromatography method may further include supplying a feces-containing liquid to a sample pad. In the immunochromatography method, detecting the complex may be detecting the complex after a given time has elapsed since supplying the feces-containing liquid to the sample pad. The given time is, for example, 10 seconds to 30 minutes. The longer the given time, the greater the effect of speeding up a single detection. Even if the given time is short, in places such as hospitals and testing institutions where tests are performed on many feces-containing liquids (e.g., specimens), the greater the effect of shortening the overall time by speeding up a single detection.

[0032] Examples of capture substances include antibodies, antibody fragments, antigens, and antigen fragments. One or both of the first capture substance and the second capture substance may comprise at least one selected from the group consisting of antibodies and antibody fragments. Examples of labels include metal colloid particles such as gold colloid particles, silver colloid particles, and platinum colloid particles; colored latex particles such as poly(meth)acrylic polymer particles and polystyrene particles containing a colorant; and fluorescent particles. The label may comprise at least one selected from the group consisting of metal colloid particles and colored latex particles. The membrane, sample pad, conjugation pad, and absorption pad may be, for example, filter paper or nonwoven fabric. Examples of materials for the filter paper or nonwoven fabric include cellulose, polyester, polyolefin, cotton, and the like.

[0033] FIG. 1 is a perspective schematic diagram illustrating an example of an immunochromatography device. The immunochromatography device 10 has a sample pad 11, a conjugation pad 12, a membrane 14 having an area 13 where a first capture substance A1 is immobilized, and a second capture substance A2 contained in the conjugation pad 12. The second capture substance A2 is labeled with a label M and is a substance capable of capturing hemoglobin. The immunochromatography device 10 further has an area 15 where a third capture substance A3 is immobilized, and an absorption pad 16. The third capture substance A3 is a substance capable of capturing the second capture substance A2.

[0034] In a hemoglobin detection method using an immunochromatography device 10, a feces-containing liquid 17 can be supplied to a sample pad 11. A complex of hemoglobin Hb contained in the supplied feces-containing liquid 17, a second capture substance A2, and a first capture substance A1 is formed in region 13.

[0035] In the present disclosure, the hemoglobin detection method can be used to diagnose or predict diseases accompanied by bleeding from the lower gastrointestinal tract, such as the large intestine. Examples of diseases accompanied by bleeding from the lower gastrointestinal tract include colon cancer, ischemic enteritis, drug-induced enteritis, infectious enteritis, ulcerative colitis, and Crohn's disease.

[0036] <Solution> One embodiment of the present invention relates to a solution for use as a stool preservation solution and as a developing solution in an immunochromatography method capable of detecting hemoglobin. The solution contains at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite. In the present disclosure, this solution may be referred to as a "developing solution." In the present disclosure, at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite may be referred to as a "sulfur-containing acid." In the present disclosure, sulfur-containing acids other than sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite (e.g., sulfuric acid) are not included in the term "sulfur-containing acid." It is believed that the inclusion of a sulfur-containing acid in the feces-containing solution can prevent the feces-containing solution from becoming too viscous, shorten the development time, and suppress variations in detection results that may occur due to the passage of time between obtaining the feces-containing solution and the start of testing. According to the developing solution of one embodiment of the present invention, rapid and stable hemoglobin testing can be achieved.

[0037] The developing solution preferably contains water as a liquid capable of dissolving the sulfur-containing acid. The developing solution may further contain optional components other than water. Examples of optional components include at least one selected from the group consisting of buffers, proteins, antibiotics, preservatives, sugars, ferrocyanide compounds, chelating agents, protease inhibitors, surfactants, inorganic acids and their salts, and organic acids and their salts. Specific examples of the developing solution include a developing solution containing a sulfur-containing acid, a buffer, a preservative, and water; a developing solution containing a sulfur-containing acid, a buffer, a protein, an antibiotic, a preservative, and water; and the like.

[0038] The stool, stool preservation, and immunochromatography method are as described above. The developing solution can be preferably used to obtain a stool-containing solution in the hemoglobin detection method of the above embodiment. The developing solution can function as a mobile phase in the immunochromatography method.

[0039] (sulfur-containing acid) The developing solution contains at least one selected from the group consisting of sulfurous acid, sulfites, disulfites, disulfites, dithionous acid, and dithionites. The developing solution may contain two or more selected from the group consisting of sulfurous acid, sulfites, disulfites, disulfites, dithionous acid, and dithionites.

[0040] From the viewpoint of stably performing detection, the developing solution may contain, for example, at least one selected from the group consisting of sulfite, disulfite, and dithionite. From the viewpoint of stably performing detection, the developing solution may contain, for example, at least one selected from the group consisting of disulfite and disulfite. From the viewpoint of stably performing detection, the developing solution may contain, for example, disulfite.

[0041] Examples of cations contained in salts include alkali metal ions such as lithium ion, sodium ion, and potassium ion; alkaline earth metal ions such as magnesium ion and calcium ion; inorganic cations such as ammonium ion; and organic cations such as organic ammonium ions such as trimethylammonium and triethylammonium. The cation may include, for example, an inorganic cation, an alkali metal ion, or a sodium ion.

[0042] Specifically, the developing solution may contain at least one selected from the group consisting of sulfurous acid, lithium sulfite, sodium sulfite, potassium sulfite, ammonium sulfite, disulfite, lithium disulfite, sodium disulfite, potassium disulfite, ammonium disulfite, dithionous acid, lithium dithionite, sodium dithionite, potassium dithionite, and ammonium dithionite.

[0043] The content of the sulfur-containing acid, based on the volume of the developing solution, may be, for example, 0.0001% (w / v) or more, 0.0005% (w / v) or more, 0.001% (w / v) or more, 0.002% (w / v) or more, 0.003% (w / v) or more, 0.004% (w / v) or more, 0.005% (w / v) or more, 0.006% (w / v) or more, 0.007% (w / v) or more, 0.008% (w / v) or more, 0.009% (w / v) or more, 0.01% (w / v) or more, 0.02% (w / v) or more, 0.03% (w / v) or more, 0.04% (w / v) or more, or 0.05% (w / v) or more. The content of the sulfur-containing acid, based on the volume of the developing solution, may be, for example, 5% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, 0.1% (w / v) or less, 0.09% (w / v) or less, 0.08% (w / v) or less, 0.07% (w / v) or less, 0.06% (w / v) or less, 0.05% (w / v) or less, 0.04% (w / v) or less, 0.03% (w / v) or less, 0.02% (w / v) or less, 0.01% (w / v) or less, 0.009% (w / v) or less, 0.008% (w / v) or less, 0.007% (w / v) or less, 0.006% (w / v) or less, or 0.005% (w / v) or less. When the amount is within the above range, the effect of stabilizing hemoglobin sufficiently tends to be easily obtained.

[0044] (water) The water contained in the developing solution is not particularly limited, and examples thereof include deionized water, distilled water, etc. The content of water may be the balance of the other components.

[0045] (buffering agent) The developing solution may contain a buffer. The buffer may be one that, when dissolved in water, yields a buffer solution such as a phosphate buffer, a carbonate buffer, an ammonia buffer, an acetate buffer, a lactate buffer, a citrate buffer, a tartrate buffer, a borate buffer, a glycine buffer, a Tris buffer, or a Good's buffer. Examples of buffers that yield a Good's buffer include 2-morpholinoethanesulfonic acid (MES), piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), N-(2-acetamido)-2-aminoethanesulfonic acid (ACES), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris), 3-[N,N-bis(2-hydroxyethyl)]-2-aminoethanesulfonic acid (BES), and the like. (2-hydroxyethyl)amino]-2-hydroxypropanesulfonic acid (DIPSO), 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid [(H)EPPS], 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES), 3-[4-(2-hydroxyethyl)-1-piperazinyl]-2-hydroxypropanesulfonic acid (HEPPSO), 3-(morpholino)propanesulfonic acid (M OPS), 3-(morpholino)-2-hydroxypropanesulfonic acid (MOPSO), piperazine-N,N'-bis(2-hydroxypropanesulfonic acid) (POPSO), N-[tris(hydroxymethyl)methyl]-2-hydroxy-3-aminopropanesulfonic acid (TAPSO), N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid (TES), N-(2-acetamido)iminodiacetic acid (ADA), N,N-bis(2-hydroxypropanesulfonic acid) Examples of suitable buffers include N-(hydroxyethyl)glycine (Bicine), N-[tris(hydroxymethyl)methyl]glycine (Tricine), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), and N-cyclohexyl-3-amino-2-hydroxypropanesulfonic acid (CAPSO). From the viewpoint of stable detection in particular, the buffer may contain N-(2-acetamido)iminodiacetic acid (ADA).The developing solution may contain one buffer agent alone or two or more buffer agents in combination.

[0046] When the developing solution contains a buffer, the content of the buffer, based on the volume of the developing solution, may be, for example, 0.01% (w / v) or more, 0.05% (w / v) or more, 0.1% (w / v) or more, 0.3% (w / v) or more, or 0.5% (w / v) or more. The content of the buffer, based on the volume of the developing solution, may be, for example, 10% (w / v) or less, 5% (w / v) or less, 3% (w / v) or less, 2% (w / v) or less, or 1% (w / v) or less. When the content is within the above range, fluctuations in the pH of the feces-containing solution tend to be suppressed, and degradation and denaturation of hemoglobin tend to be easily prevented.

[0047] (protein) The developing solution may contain a protein. Examples of proteins include albumin and iron protein. Examples of albumin include serum albumin, ovalbumin, lactalbumin, etc., with serum albumin being preferred. Examples of serum albumin that can be used include serum albumin prepared in accordance with standard methods from the serum of mammals such as humans, cows, and horses; commercially available serum albumin, etc. Suitable examples of serum albumin include bovine serum albumin (BSA) and human serum albumin. The developing solution may contain one type of protein alone or two or more types in combination.

[0048] When the developing solution contains a protein, the protein content, based on the volume of the developing solution, may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.05% (w / v) or more, or 0.1% (w / v) or more. The protein content, based on the volume of the developing solution, may be, for example, 10% (w / v) or less, 5% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.3% (w / v) or less. When the protein content is within the above range, hemoglobin detection tends to be more stable and rapid.

[0049] (antibiotics) The developer may contain an antibiotic. Examples of antibiotics include β-lactam antibiotics, tetracycline antibiotics, macrolide antibiotics, aminoglycoside antibiotics, nucleoside antibiotics, ansamycin antibiotics, polypeptide antibiotics, and other antibiotics. The antibiotic may include an aminoglycoside antibiotic, such as streptomycin, streptomycin B, dehydrostreptomycin, oxystreptomycin, kanamycin, kasugamycin, gentamicin A, gentamicin C, lincomycin, bleomycin, and mannoside hydroxystreptomycin.

[0050] The antibiotic may be a salt. Examples of the salt include acid addition salts (hydrochloride, sulfate, phosphate, acetate, fumarate, oxalate, tartrate, etc.), ammonium salt, organic amine addition salts (triethylamine salt, etc.), and metal salts (lithium salt, sodium salt, potassium salt, magnesium salt, calcium salt, etc.). The developing solution may contain one antibiotic alone or two or more antibiotics in combination.

[0051] When the developer solution contains an antibiotic, the content of the antibiotic, based on the volume of the developer solution, may be, for example, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.05% (w / v) or more, 0.1% (w / v) or more, or 0.2% (w / v) or more. The content of the antibiotic, based on the volume of the developer solution, may be, for example, 5% (w / v) or less, 3% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.4% (w / v) or less. When the content is within the above range, the inclusion of an antibiotic in the developer solution tends to be sufficiently effective.

[0052] (preservatives) The developing solution may contain a preservative. Examples of the preservative include azides and chelating agents. The preservative may include azides, and examples of the azides include sodium azide. The preservative may include a chelating agent, and examples of the chelating agent include the chelating agents described below. The developing solution may contain one type of preservative alone or two or more types in combination.

[0053] When the developing solution contains a preservative, the content of the preservative, based on the volume of the developing solution, may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more. The content of the preservative, based on the volume of the developing solution, may be, for example, 2% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, 0.3% (w / v) or less, or 0.1% (w / v) or less. When the content is within the above range, the inclusion of a preservative in the developing solution tends to be sufficiently effective.

[0054] (Sugars) The developing solution may contain a sugar. Examples of sugars include glucose, sucrose, maltose, cyclodextrins, fructose, sorbose, saccharose, lactose, trehalose, galacturonic acid, mannitol, D-glucosamine, mannose, cellobiose, glycidol, inositol, etc. The developing solution may contain one type of sugar alone or two or more types in combination.

[0055] When the developing solution contains a sugar, the sugar content, based on the volume of the developing solution, may be, for example, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, 0.05% (w / v) or more, or 0.1% (w / v) or more. The sugar content, based on the volume of the developing solution, may be, for example, 5% (w / v) or less, 3% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.3% (w / v) or less. When the sugar content is 0.005% (w / v) or more, stable detection of hemoglobin tends to be facilitated. When the sugar content is 5% (w / v) or less, high viscosity of the developing solution tends to be prevented. From the viewpoint of performing a more rapid test, it is preferable that the developing solution does not contain a sugar, or if it does contain one, the content is small. The sugar content may be, for example, 0.1% (w / v) or less, 0.01% (w / v) or less, 0.001% (w / v) or less, or 0.000% (w / v).

[0056] (ferrocyanide compounds) The developing solution may contain a ferrocyanide compound. Examples of ferrocyanide compounds include sodium ferrocyanide, potassium ferrocyanide, 11-ferrocenyl-1-undecanethiol, 8-ferrocenyl-1-octanethiol, 6-ferrocenyl-1-hexanethiol, 11-ferrocenylundecyl-polyoxyethylene ether, and 11-ferrocenyltrimethylundecylammonium bromide. The developing solution may contain one ferrocyanide compound alone or two or more ferrocyanide compounds in combination.

[0057] (chelating agent) The developing solution may contain a chelating agent. Examples of the chelating agent include ethylenediaminetetraacetic acid, O,O'-bis(2-aminophenyl)ethyleneglycol-N,N,N',N'-tetraacetic acid, diethylenetriamine-N,N,N',N'',N''-pentaacetic acid, ethylenediamine-N,N'-diacetic acid, and ethylenediamine-N,N'-bis(methylenephosphonic acid). The chelating agent may be a salt. Examples of the salt include ammonium salt, sodium salt, potassium salt, magnesium salt, and calcium salt. The developing solution may contain one chelating agent alone or two or more chelating agents in combination.

[0058] (Protease inhibitors) The developing solution may contain a protease inhibitor. Examples of protease inhibitors include cOmplete (trademark, manufactured by Roche), antipapain dihydrochloride, aprotinin, chymostatin, E-64, leupeptin, pefabloc (trademark, manufactured by Roche), bepstatin, phosphoramidon, antithrombin III, (4-amidinophenyl)methanesulfonyl fluoride, calpain inhibitor 1,3,4-dichloroisocoumarin, α-macroglobulin, and bestatin. The developing solution may contain one protease inhibitor alone or two or more protease inhibitors in combination.

[0059] (surfactant) The developing liquid may contain a surfactant. Examples of surfactants include nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants. The developing liquid may contain one surfactant alone or two or more surfactants in combination.

[0060] (Inorganic acids and their salts) The developing solution may contain an inorganic acid and / or a salt of an inorganic acid. Examples of inorganic acids and salts of inorganic acids include inorganic acids and salts of inorganic acids other than the optional components described above (hereinafter, sometimes referred to as "other inorganic acids and their salts"). Examples of inorganic acids include hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, etc. Examples of salts include ammonium salts, sodium salts, potassium salts, calcium salts, magnesium salts, etc. The developing solution may contain one or more of the other inorganic acids and salts thereof, either singly or in combination.

[0061] The content of the other inorganic acid and its salt, based on the volume of the developing solution, may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more. The content of the other inorganic acid and its salt, based on the volume of the developing solution, may be, for example, 10% (w / v) or less, 5% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.1% (w / v) or less. When the content is within the above range, hemoglobin detection tends to be more stable and rapid. The content of the other inorganic acid and its salt may be, for example, 0.05% (w / v) or less, 0.01% (w / v) or less, 0.001% (w / v) or less, or 0.000% (w / v).

[0062] In particular, when the developing solution contains an alkali metal halide salt, the content of the alkali metal halide salt may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more, based on the volume of the developing solution. The content of the alkali metal halide salt may be, for example, 10% (w / v) or less, 5% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.1% (w / v) or less, based on the volume of the developing solution. When the content is within the above range, hemoglobin detection tends to be more stable and rapid. The alkali metal halide salt is, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more, based on the volume of the developing solution. - M +(X represents a halogen ion, and M represents an alkali metal ion.) Examples of alkali metal halide salts include sodium chloride and potassium chloride. For example, it is preferable that sodium chloride satisfies the above content range. From the viewpoint of performing more stable tests, it is preferable that the developing solution does not contain an alkali metal halide salt, or if it does contain one, the content is small, for example, 1% (w / v) or less. The content of the alkali metal halide salt may be, for example, 0.05% (w / v) or less, 0.01% (w / v) or less, 0.001% (w / v) or less, or 0.000% (w / v).

[0063] The total content of all inorganic acids and inorganic acid salts contained in the developing solution may be, for example, 0.001% (w / v) or more, 0.005% (w / v) or more, 0.01% (w / v) or more, 0.03% (w / v) or more, or 0.05% (w / v) or more, based on the volume of the developing solution. The total content of all inorganic acids and inorganic acid salts contained in the developing solution may be, for example, 15% (w / v) or less, 10% (w / v) or less, 5% (w / v) or less, 1% (w / v) or less, 0.5% (w / v) or less, or 0.1% (w / v) or less, based on the volume of the developing solution. When the content is within the above range, degradation and denaturation of hemoglobin tend to be easily prevented.

[0064] The content of the sulfur-containing acid may be, for example, 5% by mass or more, 10% by mass or more, 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, or 90% by mass or more, based on the total mass of all inorganic acids and inorganic acid salts contained in the developing solution. The content of the sulfur-containing acid may be, for example, 100% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, 25% by mass or less, or 20% by mass or less, based on the total mass of all inorganic acids and inorganic acid salts contained in the developing solution. When the content is within the above range, hemoglobin decomposition and denaturation tend to be easily prevented.

[0065] (organic acids and their salts) The developing solution may contain an organic acid and / or a salt of an organic acid. Examples of organic acids and salts of organic acids include organic acids and salts of organic acids other than the optional components described above. Examples of organic acids include malic acid, succinic acid, fumaric acid, glycolic acid, 2-ketoglutaric acid, isocitric acid, lactic acid, pyruvic acid, uric acid, and oxalacetic acid. Examples of salts include ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. The developing solution may contain one organic acid and / or a salt of an organic acid alone or two or more organic acids in combination.

[0066] The developing solution may be a solution containing a sulfur-containing acid, a buffer, a protein, an antibiotic, a preservative, and water. In this case, the total content of the sulfur-containing acid, buffer, protein, antibiotic, preservative, and water in the developing solution may be, for example, 90% (w / v) or more, 92% (w / v) or more, 95% (w / v) or more, 98% (w / v) or more, 99% (w / v) or more, 99.3% (w / v) or more, 99.5% (w / v) or more, 99.8% (w / v) or more, or 100.0% (w / v).

[0067] [pH] The pH of the developing solution may be, for example, 6.0 or higher, 6.3 or higher, 6.5 or higher, 6.6 or higher, 6.7 or higher, or 6.8 or higher. The pH of the developing solution may be, for example, 7.5 or lower, 7.2 or lower, 7.0 or lower, 6.9 or lower, 6.8 or lower, or 6.7 or lower. When the pH is within the above range, degradation and denaturation of hemoglobin tend to be easily prevented. The pH of the developing solution can be measured using a general pH meter. An example of a pH meter is the "pH / ion meter HM-42X" manufactured by DKK-TOA Corporation. The temperature of the developing solution during measurement is 25°C.

[0068] [viscosity] The viscosity of the developing solution may be, for example, 1.00 cP or more, 1.05 cP or more, 1.10 cP or more, 1.15 cP or more, 1.20 cP or more, 1.25 cP or more, 1.30 cP or more, 1.35 cP or more, 1.40 cP or more, 1.45 cP or more, or 1.50 cP or more. The viscosity of the developing solution may be, for example, 1.80 cP or less, 1.75 cP or less, 1.70 cP or less, 1.65 cP or less, 1.60 cP or less, 1.55 cP or less, 1.50 cP or less, 1.45 cP or less, 1.40 cP or less, 1.35 cP or less, or 1.30 cP or less. When the viscosity is within the above range, hemoglobin tends to be detected more quickly. From the viewpoint of rapid detection, a low viscosity is preferable. The viscosity of the developing solution can be measured using a vibration viscometer. An example of a vibration viscometer is "Viscomate VM-1G" manufactured by Sekonic Co., Ltd. The temperature of the developing solution during measurement is 25°C.

[0069] [Application] One embodiment of the present invention, a solution (developing solution), is used as a preservative solution for stool and as a developing solution for immunochromatography that can detect hemoglobin. A feces-containing solution is obtained by mixing the solution with stool. The stool can be stored for any length of time in a dissolved or dispersed state in the preservative solution. Furthermore, the feces-containing solution can be used in immunochromatography as a specimen sample containing the developing solution and stool.

[0070] For mixing the solution with the stool and / or storing the stool, a stool collection container as described below can be used, but is not particularly limited, and any known container that can be used for fecal occult blood tests may be used. The immunochromatography method in which the solution is used may be the method in the detection method of the above-mentioned embodiment, but is not particularly limited, and may be any known method.

[0071] <Fecal storage device> One embodiment of the present invention relates to a stool storage container, which includes a stool collection container and a solution (developing liquid) contained within the stool collection container. The stool collection container has a stool collection stick and a container body. The stool collection stick has a grip portion on one side and a rod portion on the other side, and the rod portion has a stool collection portion at or near the tip of the other side. "Near the tip" refers, for example, to the portion extending from the tip of the other side of the rod portion to the center of the rod portion. The stool storage container may further include optional portions.

[0072] Figure 2 is a front view schematic diagram showing an example of a stool storage container. Stool storage container 20 includes a container body 21, a solution (developing liquid) 22, and a stool collection stick 23. Using the stool storage container shown in Figure 2, stool is collected using the stool collection stick, and the stool collection stick with the stool attached is inserted into a stool collection container, thereby obtaining a stool-containing liquid in the stool collection container. The stool-containing liquid can be stored in the stool storage container for any desired period of time.

[0073] <Method for evaluating fecal fluid> A method for evaluating a feces-containing liquid according to one embodiment of the present invention includes evaluating whether or not the feces-containing liquid contains hemoglobin using the hemoglobin detection method according to the above-described embodiment. The method for evaluating a feces-containing liquid may further include an optional step.

[0074] When hemoglobin is detected by immunochromatography according to the hemoglobin detection method, it can be determined that the feces-containing liquid contains hemoglobin. On the other hand, when hemoglobin is not detected by immunochromatography, it can be determined that the feces-containing liquid does not contain hemoglobin.

[0075] <Example of embodiment> Preferred examples of the present invention are listed below: The present invention is not limited to the following examples. [1] Preparing a feces-containing liquid containing at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite, and feces, and having been stored for a given period of time; and Detecting hemoglobin contained in the feces-containing liquid by immunochromatography. A method for detecting hemoglobin, comprising: [2] The immunochromatography method preparing a membrane having a region where a first capture substance capable of capturing the hemoglobin is immobilized, and a second capture substance that is labeled with a label and is capable of capturing the hemoglobin; forming a complex in the region, the complex including a first capture agent, hemoglobin, and a second capture agent; and detecting the complex. The detection method according to [1] above, comprising: [3] The detection method according to [2] above, wherein the first capture substance comprises at least one substance selected from the group consisting of an antibody and an antibody fragment. [4] The detection method according to [2] or [3] above, wherein the second capture substance comprises at least one substance selected from the group consisting of an antibody and an antibody fragment. [5] The detection method according to any one of the above [2] to [4], wherein the label comprises at least one kind selected from the group consisting of metal colloid particles and colored latex particles. [6] The detection method according to any one of [1] to [5] above, wherein the given period of time is 1 hour to 10 days. [7] The detection method according to any one of [1] to [6] above, wherein the storage temperature is 4 to 45°C. [8] The detection method described in any one of [2] to [7] above, wherein the preparing step includes preparing an immunochromatography device having a sample pad, a conjugation pad, the membrane, and the second capture substance. [9] The immunochromatography method further comprises supplying the stool-containing liquid to the sample pad; The detection method described in [8] above, wherein detecting the complex includes detecting the complex after a given time has elapsed since the feces-containing liquid was supplied to the sample pad.

[10] A solution containing at least one selected from the group consisting of sulfurous acid, sulfites, disulfites, disulfites, dithionous acid, and dithionites, and used as a stool preservation solution and as a developing solution for immunochromatography capable of detecting hemoglobin. Or, [10'] Use of a solution containing at least one selected from the group consisting of sulfurous acid, sulfites, disulfites, disulfites, dithionous acid, and dithionites as a stool preservation solution and as a developing solution for immunochromatography capable of detecting hemoglobin.

[11] The solution according to

[10] above, wherein the pH is 6.5 to 6.9. Or, [11'] The use according to [10'] above, wherein the pH of the solution is 6.5 to 6.9.

[12] The solution according to

[10] or

[11] above, having a viscosity of 1.65 cP or less at 25°C. Or, [12'] The use according to [10'] or [11'] above, wherein the viscosity of the solution at 25°C is 1.65 cP or less.

[13] The solution according to any one of the above

[10] to

[12] , further comprising a buffering agent, a preservative, and water. Or, [13'] The use according to any one of the above [10'] to [12'], wherein the solution further comprises a buffering agent, a preservative, and water.

[14] The solution according to

[13] above, wherein the buffer comprises N-(2-acetamido)iminodiacetic acid. Or, [14'] The use according to [13'] above, wherein the buffer comprises N-(2-acetamido)iminodiacetic acid.

[15] The solution according to any one of the above

[10] to

[14] , which does not contain an alkali metal halide salt, or if it contains an alkali metal halide salt, the content of the alkali metal halide salt is 1% (w / v) or less. Or, [15'] The use according to any one of the above [10'] to [14'], which does not contain an alkali metal halide salt, or if it contains an alkali metal halide salt, the content of the alkali metal halide salt is 1% (w / v) or less.

[16] The solution according to any one of the above

[10] to

[15] , wherein the content of the inorganic acid and the salt of the inorganic acid is 0.1 to 1% (w / v). Or, [16'] The use according to any one of the above [10'] to [15'], wherein the content of the inorganic acid and the salt of the inorganic acid in the solution is 0.1 to 1% (w / v).

[17] The solution according to any one of

[10] to

[16] above, which is used in the detection method according to any one of [1] to [9] above. Or, [17'] Use according to any one of [10'] to [16'] above in the detection method according to any one of [1] to [9] above.

[18] The detection method according to any one of [1] to [9] above, wherein the feces-containing liquid contains the solution according to any one of

[10] to

[16] above and the feces.

[19] Providing the solution according to any one of

[10] to

[16] above; and The detection method described in

[18] above, further comprising receiving the feces-containing liquid.

[20] A stool collection container and the solution according to any one of

[10] to

[17] above, which is contained in the stool collection container; The stool collection container has a stool collection stick and a container body, The fecal collection stick has a grip portion on one side and a stick portion on the other side, The rod portion has a stool collection portion at or near the tip of the other side, in this stool storage container.

[21] A method for evaluating a feces-containing liquid, comprising evaluating whether or not the feces-containing liquid contains hemoglobin using the hemoglobin detection method described in any one of [1] to [9],

[18] and

[19] above. [Example]

[0076] The embodiments of the present invention will be described in more detail with reference to examples, but the embodiments of the present invention are not limited to the following examples.

[0077] Example A: Preparation and Evaluation of Solution (Developer) A developing solution containing a sulfur-containing acid and a developing solution not containing a sulfur-containing acid were prepared, and the stability of hemoglobin in each developing solution was evaluated.

[0078] [Preparation of developing solution] (Preparation of Developer A1) Developer A1 was prepared containing 30 mmol / L N-(2-acetamido)iminodiacetic acid (ADA), 0.2% (w / v) bovine serum albumin (BSA), 0.38% (w / v) streptomycin sulfate (3,000 kU / L), 0.1% (w / v) sodium azide, 1 mM sodium sulfite (0.013% (w / v)), sodium hydroxide for pH adjustment, and water.

[0079] (Preparation of developing solutions A2 to A7) Sodium sulfite and its concentration in the developing solution are 1 mM ("mM" stands for 10 -3 Developing solutions A2 to A7 were obtained in the same manner as developing solution A1, except that the additives and concentrations (mol / L) were changed to those shown in Table 1.

[0080] The reagents used to prepare the developing solutions A1 to A7 are as follows. N-(2-acetamido)iminodiacetic acid (ADA, Dojindo Laboratories) Bovine serum albumin (BSA, manufactured by Boval) Streptomycin sulfate (Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium azide (Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium sulfite (manufactured by Junsei Chemical Co., Ltd.) Sodium hydroxide (Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium disulfite (Junsei Chemical Co., Ltd.) Sodium dithionite (Tokyo Chemical Industry Co., Ltd.) Sodium lactate 70% solution (Fujifilm Wako Pure Chemical Industries, Ltd.) Trehalose (Hayashibara Co., Ltd.) α-Ketoglutaric acid (2-oxoglutaric acid, Fujifilm Wako Pure Chemical Industries, Ltd.) Iron(II) sulfate (heptahydrate, Fujifilm Wako Pure Chemical Industries, Ltd.)

[0081] [pH measurement] The pH of each of the developing solutions A1 to A7 prepared above was measured at a liquid temperature of 25° C. The pH was measured using a pH / ion meter HM-42X (manufactured by DKK-TOA Corporation). The results are shown in Table 1.

[0082] [Table 1]

[0083] [Evaluation of the developer] The stability of hemoglobin in the developing solution was evaluated by calculating the residual rate of hemoglobin after storage in the developing solution.

[0084] (Evaluation of Developer A1) (1) Preparation of sample solution To 100 mL of the developing solution A1 prepared above, 3 mg of hemoglobin was added to prepare sample solution A1. (2) Measurement of hemoglobin in sample solution immediately after preparation The hemoglobin concentration in sample solution A1 prepared in (1) above was measured using Extell "Hemo Auto" HS antibody-sensitized latex suspension and Extell "Hemo Auto" HS hemoglobin buffer solution with the fully automated fecal human hemoglobin analyzer HM-JACKarc.

[0085] (2-1) Preparation of calibration curve A calibration curve showing the relationship between hemoglobin concentration and turbidity was prepared using Extell Hemoglobin Standard HS according to the method described in the package insert for Extell Hemo Auto HS. (2-2) Measurement of hemoglobin concentration in sample solution immediately after preparation Extell "Hemo-Auto" HS antibody-sensitized latex suspension (90 μL) and Extell "Hemo-Auto" HS hemoglobin buffer solution (190 μL) were added to an HM-JACKarc cup, followed by the addition of sample solution A1 (20 μL) prepared in (1) above, and the reaction was carried out at 25°C. The turbidity was measured 72 seconds and 288 seconds after the addition of sample solution A1, and the turbidity after 72 seconds was subtracted from the turbidity after 288 seconds to obtain the measured value. The measured value was compared with the calibration curve prepared in (2-1) to determine the hemoglobin concentration in sample solution A1.

[0086] (3) Preparation of sample solutions for storage stability evaluation The sample solution A1 prepared in (1) above was stored at 30°C for 7 days to prepare sample solution A1 (after storage).

[0087] (4) Measurement of hemoglobin concentration in the sample solution (after storage) The hemoglobin concentration in sample solution A1 (after storage) was determined in the same manner as in (2) above, except that sample solution A1 (after storage) prepared in (3) above was used instead of sample solution A1 immediately after preparation.

[0088] (5) Calculation of the residual hemoglobin rate in the sample solution (after storage) The hemoglobin concentration in sample solution A1 immediately after preparation, determined in (2) above, was taken as 100%, and the percentage of the hemoglobin concentration in sample solution A1 (after storage), determined in (4) above, relative to this concentration (hemoglobin residual rate (%)) was calculated. The results are shown in Table 1.

[0089] (Evaluation of Developers A2 to A7) Sample solutions A2 to A7 were prepared in the same manner as sample solution A1, except that developing solutions A2 to A7 prepared above were used instead of developing solution A1.

[0090] The residual hemoglobin rate (%) for sample solutions A2 to A7 was calculated in the same manner as in (2) to (5), except that sample solutions A2 to A7 were used instead of sample solution A1. The results are shown in Table 1.

[0091] The reagents, instruments, etc. used in the evaluation of the developing solutions A1 to A7 are as follows. Hemoglobin (lyophilized human hemoglobin powder, manufactured by Sigma) Fully automated fecal human hemoglobin analyzer HM-JACKarc (Hitachi Chemical Diagnostics Systems) Extell "Hemo-Auto" HS antibody-sensitized latex suspension, Extell "Hemo-Auto" HS hemoglobin buffer solution, Extell hemoglobin standard HS (all manufactured by Hitachi Chemical Diagnostics Systems Co., Ltd.) pH / ion meter HM-42X (manufactured by DKK-TOA Corporation)

[0092] As shown in Table 1, hemoglobin exhibited good stability in a developing solution containing a sulfur-containing acid. By using a developing solution containing a sulfur-containing acid, hemoglobin can be stably detected.

[0093] Example B: Detection of hemoglobin by immunochromatography As the developer for immunochromatography, a developer containing a sulfur-containing acid or a developer not containing a sulfur-containing acid was used, and the rapidity of hemoglobin detection was evaluated.

[0094] [Preparation of developing solution] (Developing solution B1) Developing solution B1 (pH 6.8) was prepared in the same manner as developing solution A1.

[0095] (Developing solution B2) A 10 mM phosphate buffer solution (PBS) (pH 7.4) containing 140 mM sodium chloride was prepared. The resulting PBS was used as developing solution B2.

[0096] (Developing solution B3) A buffer solution containing 30 mmol / L N-(2-acetamido)iminodiacetic acid (ADA), 0.2% (w / v) bovine serum albumin (BSA), 0.38% (w / v) streptomycin sulfate (3,000 kU / L), 0.1% (w / v) sodium azide, 3.5% (w / v) sodium lactate, sodium hydroxide for pH adjustment, and water was prepared. The resulting buffer solution was designated Developer B3 (pH 7.2).

[0097] (Developing solution B4) Developing solution B4 (pH 7.2) was obtained in the same manner as developing solution B3, except that it further contained 1% (w / v) trehalose.

[0098] (Developing solution B5~B8) Developing solutions B5 to B8 (all pH 7.2) were obtained in the same manner as developing solution B4, except that the concentration of trehalose in the developing solution was changed to 3% (w / v) to 10% (w / v).

[0099] The reagents used to prepare developing solutions B1 and B3 to B8 are as described above. The reagents used to prepare developing solution B2 (PBS) are as follows: Sodium chloride (Fujifilm Wako Pure Chemical Industries, Ltd.) Disodium hydrogen phosphate (Kanto Chemical Co., Ltd.) Sodium dihydrogen phosphate (Kanto Chemical Co., Ltd.)

[0100] [Viscosity measurement] The viscosity of the developing solutions B1 to B8 and purified water prepared above was measured at a liquid temperature of 25° C. A viscometer Viscomate VM-1G (manufactured by Sekonic Corporation) was used to measure the viscosity. The results are shown in Table 2.

[0101] [Table 2]

[0102] [Hemoglobin detection] (Preparation of evaluation half strips and antibody solution) The following anti-human hemoglobin antibody-immobilized half strips and gold colloid-labeled anti-human hemoglobin antibody solutions were prepared.

[0103] (1) Preparation of anti-human hemoglobin antibody-immobilized half-strip A Hi-Flow Plus HFC070504 membrane measuring 50 mm vertically and 250 mm horizontally was prepared. Next, a line of anti-human hemoglobin antibody solution (1 mg / mL anti-human hemoglobin antibody in 5 mM phosphate buffer, pH 6.0) was applied at a rate of 1.0 μL / cm using the Biojet Quanti dispensing module at a position 21 mm from the bottom of the membrane to form a detection area (the coated surface is referred to as the membrane surface). After application, the membrane was dried at 25°C for 2 hours. After drying, the membrane was immersed in blocking buffer (50 mM borate buffer, pH 8.5, containing 0.5% (w / v) casein) for 30 minutes, followed by a wash solution (50 mM Tris-HCl buffer, pH 7.5, containing 0.5% (w / v) sucrose and 0.05% (w / v) sodium cholate) for 30 minutes. After washing, the membrane was dried overnight in a desiccator. The backside of the membrane was then attached to the adhesive surface of a precut backing sheet. Cellulose fiber sample pads were then attached to the membrane surface as absorbent pads, overlapping the membrane's upper surface 20 mm from the top. The membrane was then cut lengthwise into 50 mm x 5 mm strips to prepare anti-human hemoglobin antibody-immobilized half strips.

[0104] (2) Preparation of colloidal gold-labeled anti-human hemoglobin antibody solution Gold Colloid: 40 nm (BBI Solution) was prepared as a gold colloid solution. Next, 900 μL of the gold colloid solution, 100 μL of 20 mM borate buffer (pH 8.5), and 100 μL of 30 μg / mL anti-human hemoglobin antibody solution were mixed and allowed to stand at 25°C for 30 minutes. After that, 55 μL of 1% (w / v) polyethylene glycol 20,000 solution and 110 μL of BSA solution (BSA solution adjusted to pH 8.0 with sodium hydroxide) were added to the resulting mixture and mixed. The resulting mixture was then centrifuged at 8,000 g for 10 minutes at 10°C. After removing the supernatant, 2 mL of colloidal gold storage buffer (20 mM Tris-HCl buffer, pH 8.2, containing 0.05% (w / v) polyethylene glycol 20,000, 150 mM sodium chloride, 1% (w / v) BSA, and 0.1% (w / v) sodium azide) was added to the precipitate, and the precipitate was dispersed in the buffer using a benchtop ultrasonic cleaner W-113Mk II. The process from centrifugation to dispersion of the precipitate was then repeated two more times under the same conditions, after which colloidal gold storage buffer was added to adjust the concentration, yielding a colloidal gold-labeled anti-human hemoglobin antibody solution. The concentration of the colloidal gold-labeled anti-human hemoglobin antibody solution was adjusted so that the turbidity at 525 nm when diluted to 5% (v / v) with colloidal gold storage buffer was 0.3.

[0105] (Hemoglobin detection time measurement) The developer solutions B1 to B8 were used as developer solutions for immunochromatography, and the time required for hemoglobin detection was measured. The immunochromatography used the anti-human hemoglobin antibody-immobilized half-strip prepared above and a gold colloid-labeled anti-human hemoglobin antibody solution.

[0106] (Evaluation of Developer B1) (1) Preparation of the mixed solution To one well of a 96-well microplate, 90 μL of developing solution B1, 5 μL of a 50 μg / mL aqueous hemoglobin solution, and 5 μL of a gold colloid-labeled anti-human hemoglobin antibody solution were added to prepare 100 μL of a mixed solution.

[0107] (2) Penetration of the mixed liquid into the half strip The lower end of the anti-human hemoglobin antibody-immobilized half strip (the end not having the CELLULOSE FIBER SAMPLE PADS attached) was immersed in 100 μL of the mixture prepared in (1) above in the well, and the mixture was allowed to penetrate the half strip.

[0108] (3) Hemoglobin detection Hemoglobin was detected by confirming the pink coloration resulting from the gold colloid in the detection area on the anti-human hemoglobin antibody-immobilized half-strip. The time from immersion of the lower end of the anti-human hemoglobin antibody-immobilized half-strip in the mixed solution until hemoglobin was detected was measured. The results are shown in Table 2.

[0109] (Evaluation of Developers B2 to B8) The time required for hemoglobin detection was measured in the same manner as for developing solution B1, except that developing solutions B2 to B8 were used instead of developing solution B1. The results are shown in Table 2.

[0110] The reagents, instruments, etc. used in the evaluation of the developing solutions B1 to B8 are as follows. Anti-human hemoglobin antibody (anti-human hemoglobin sheep polyclonal antibody, 4870-3979G, Bio-Rad Laboratories) Casein (Megmilk Snow Brand) Boric acid (Fujifilm Wako Pure Chemical Industries, Ltd.) Sucrose (Fujifilm Wako Pure Chemical Industries, Ltd.) Sodium cholate (Fujifilm Wako Pure Chemical Industries, Ltd.) Trishydroxymethylaminomethane (Tris, manufactured by Tokyo Chemical Industry Co., Ltd.) Polyethylene glycol 20,000 (Fujifilm Wako Pure Chemical Industries, Ltd.) Hi-Flow Plus HFC070504 (membrane, manufactured by Merck Millipore) Pre-cut backing sheet (manufactured by Nippon Engineering Co., Ltd.) CELLULOSE FIBER SAMPLE PADS (Merck Millipore) Gold Colloid: 40 nm (colloidal gold solution, BBI Solution) Biojet Quanti dispensing module (dispenser, manufactured by BioDot) Tabletop ultrasonic cleaner W-113Mk II (manufactured by Honda Electronics Co., Ltd.) Viscosity measuring instrument Viscomate VM-1G (manufactured by Sekonic)

[0111] As shown in Table 2, hemoglobin could be detected in a short time when a developing solution containing a sulfur-containing acid was used. Table 2 also shows that hemoglobin could be detected in a short time when a developing solution with low viscosity was used. [Explanation of symbols]

[0112] 10 Immunochromatography device 11 Sample Pad 12 Conjugation Pad 13 Region where the first capture substance is immobilized 14 Membrane 15 Region where the third capture substance is immobilized 16 absorbent pads 17 Fecal fluid A1 First capture substance A2 Second capture agent A3 Third capture substance M-labeled substance Hb hemoglobin 20 stool storage container 21 Container body 22 Solution (developer) 23 Stool collection stick 23a Gripping part 23b Rod part 23c Fecal collection section 24 Stool collection container

Claims

1. A feces-containing liquid containing at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite and feces, the feces-containing liquid having been stored for a given period of time, and detecting hemoglobin contained in the feces-containing liquid by immunochromatography; The immunochromatography method comprises: providing an immunochromatography device having a sample pad, a conjugation pad, a membrane having an area where a first capture substance capable of capturing the hemoglobin is immobilized, and a second capture substance that is labeled with a label and is capable of capturing the hemoglobin; supplying the feces-containing liquid directly to the sample pad; forming a complex in the region, the complex including a first capture substance, hemoglobin, and a second capture substance; and and detecting the complex after a given time has elapsed since the feces-containing liquid was supplied to the sample pad. How to detect hemoglobin.

2. The detection method according to claim 1, wherein the feces-containing liquid contains at least one selected from the group consisting of dithionous acid and dithionites.

3. The detection method according to claim 1 or 2, wherein the first capture substance comprises at least one substance selected from the group consisting of an antibody and an antibody fragment.

4. The detection method according to any one of claims 1 to 3, wherein the second capture substance comprises at least one selected from the group consisting of an antibody and an antibody fragment.

5. 5. The detection method according to claim 1, wherein the label comprises at least one kind selected from the group consisting of metal colloid particles and colored latex particles.

6. The detection method according to any one of claims 1 to 5, wherein the feces-containing liquid after storage for an arbitrary period of time is feces-containing liquid after storage for 1 hour to 10 days.

7. The detection method according to any one of claims 1 to 6, wherein the storage temperature is 4 to 45°C.

8. A solution containing at least one selected from the group consisting of sulfurous acid, sulfite, disulfite, disulfite, dithionous acid, and dithionite, which is used as a preservative solution for stool and as a developing solution for immunochromatography in a method capable of detecting hemoglobin, The method comprises: Preparing a feces-containing liquid containing the solution and feces, and storing the solution for a desired period of time; using the stool-containing liquid as a specimen sample in an immunochromatography method; The immunochromatography method comprises: providing an immunochromatography device having a sample pad, a conjugation pad, a membrane having an area where a first capture substance capable of capturing the hemoglobin is immobilized, and a second capture substance labeled with a label and capable of capturing the hemoglobin; supplying the feces-containing liquid directly to the sample pad; When the feces-containing liquid contains hemoglobin, it further comprises: forming a complex in the region, the complex including a first capture substance, hemoglobin, and a second capture substance; and and detecting the complex after a given time has elapsed since the feces-containing liquid was supplied to the sample pad. solution.

9. The solution according to claim 8, which contains at least one selected from the group consisting of dithionous acid and dithionites.

10. 10. The solution according to claim 8, wherein the pH is from 6.5 to 6.

9.

11. The solution according to any one of claims 8 to 10, having a viscosity at 25°C of 1.65 cP or less.

12. The solution of any one of claims 8 to 11, further comprising a buffer, a preservative, and water.

13. 13. The solution of claim 12, wherein the buffer comprises N-(2-acetamido)iminodiacetic acid.

14. The solution according to any one of claims 8 to 13, which does not contain an alkali metal halide salt, or if it contains an alkali metal halide salt, the content of the alkali metal halide salt is 1% (w / v) or less.

15. The solution according to any one of claims 8 to 14, wherein the content of the inorganic acid and the salt of the inorganic acid is 0.1 to 1% (w / v).

16. The solution according to any one of claims 8 to 15, which is used in the detection method according to any one of claims 1 to 7.

17. The detection method according to any one of claims 1 to 7, wherein the feces-containing liquid contains the solution according to any one of claims 8 to 15 and the feces.

18. Providing a solution according to any one of claims 8 to 15, and 18. The method of claim 17, further comprising receiving the feces-containing fluid.

19. A method for evaluating a feces-containing liquid, comprising evaluating whether or not the feces-containing liquid contains hemoglobin using the hemoglobin detection method according to any one of claims 1 to 7, 17 and 18.

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