WO2022091923A1 - Sensitizer for immunochromatographic assays, and assay - Google Patents
Sensitizer for immunochromatographic assays, and assay Download PDFInfo
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- WO2022091923A1 WO2022091923A1 PCT/JP2021/038885 JP2021038885W WO2022091923A1 WO 2022091923 A1 WO2022091923 A1 WO 2022091923A1 JP 2021038885 W JP2021038885 W JP 2021038885W WO 2022091923 A1 WO2022091923 A1 WO 2022091923A1
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- sensitizer
- cov
- sars
- coronavirus
- copolymer
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-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
Definitions
- the present invention is a sensitizer for an immunochromatographic measurement method using a new coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, and a new coronavirus (SARS-CoV-2) performed in the coexistence of the sensitizer.
- the present invention relates to an immunochromatographic measurement method using an IgM antibody as a measurement target substance.
- the immunochromatography method is a method that combines a chromatography method that utilizes the capillary phenomenon of the membrane and an immunological method that utilizes an antigen-antibody reaction.
- the immunochromatographic method has high specificity due to the antigen-antibody reaction, is easy to operate, can be inspected on the spot without the need for a place or special equipment, and the result can be visually judged, so that it is a clinical diagnostic method. It is widely used (Patent Document 1).
- the immunochromatographic method has the advantage that it can be easily measured, but the accuracy of detection sensitivity is low, and even if a positive sample is used, it may be judged as false negative. Therefore, a higher-sensitivity immunochromatographic measurement method is desired.
- Patent Document 2 discloses a method for increasing the sensitivity by changing the addition position of a reagent for an immunochromatographic test.
- Patent Document 3 discloses a method for increasing the sensitivity by improving the material of the membrane.
- Patent Documents 4, 5 and 6 disclose that bovine serum albumin (BSA), a polymer having a phosphorylcholine group, and hyaluronic acid are contained in a sample diluent.
- BSA bovine serum albumin
- a polymer having a phosphorylcholine group a polymer having a phosphorylcholine group
- hyaluronic acid are contained in a sample diluent.
- the new coronavirus (SARS-CoV-2) IgM antibody is an antibody produced in the early stage of infection with the new coronavirus (SARS-CoV-2), and if the IgM antibody can be detected with high sensitivity, Since the infected person can be isolated at the initial stage of infection, it is possible to suppress the spread of infection. In addition, since the treatment can be started at an early stage, the effect of the treatment can be enhanced, so that a sensitizer that can detect the IgM antibody with high sensitivity is considered to be highly useful.
- Japanese Unexamined Patent Publication No. 01-063865 Japanese Unexamined Patent Publication No. 2014-66674 Japanese Unexamined Patent Publication No. 2014-62820 Japanese Unexamined Patent Publication No. 2008-292326 Japanese Unexamined Patent Publication No. 2008-058334 Japanese Patent Application Laid-Open No. 2003-344413
- the present invention provides a sensitizer for an immunochromatographic measurement method capable of detecting a new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, and a measurement method using the sensitizer.
- SARS-CoV-2 new type coronavirus
- a sensitizer for an immunochromatographic measurement method which comprises a copolymer represented by the following general formula [1] and contains a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
- SARS-CoV-2 new type coronavirus
- n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.
- a sensitizer for immunochromatography which uses a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
- the ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 99 to 20: 1.
- the ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 90 to 30:10.
- SARS-CoV- new coronavirus
- the new coronavirus (SARS-CoV-) according to any one of [1] to [4], wherein the copolymer is a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000.
- a sensitizer for an immunochromatographic measurement method using an IgM antibody as a measurement target substance [6] Dilution of a sample for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, which contains the sensitizer for the immunochromatographic measurement method according to any one of [1] to [5]. liquid. [7] A new type coronavirus (SARS-CoV-2) IgM characterized in that an antigen-antibody reaction is carried out in the presence of the sensitizer for the later lateral flow test according to any one of [1] to [5]. An immunochromatographic measurement method using an antibody as a measurement target substance.
- SARS-CoV-2 new type coronavirus
- a new type coronavirus in which the sensitizer for the later lateral flow test according to any one of [1] to [5] is carried on a developing membrane, a sample pad, or a conjugate pad.
- An instrument for measuring immunochromatography that uses IgM antibody as the measurement target substance.
- a new type coronavirus (SARS) in a biological sample derived from a subject is performed.
- SARS coronavirus
- -CoV-2) A method for testing the possibility that a subject is infected with a new coronavirus infection (COVID-19), which comprises measuring IgM antibody.
- COVID-19 A method for treating a new type of coronavirus infection (COVID-19), which comprises the following steps (1) and (2).
- n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.
- the new coronavirus (SARS-CoV-2) IgM antibody of the present invention uses the new coronavirus (SARS-CoV-2) IgM antibody of the present invention as the measurement target substance
- the conventional new coronavirus (SARS-CoV-2) IgM antibody is used. It is possible to detect the new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity as compared with the immunochromatographic measurement method to be detected.
- a method for treating a new type of coronavirus infection (COVID-19) including a step of causing an antibody reaction will be described below.
- the measurement target substance is an IgM antibody, but other measurement target substances can also be detected.
- the sensitizer for the immunochromatographic measurement method of the present invention is characterized in that the above antibody can be detected with high sensitivity.
- the new coronavirus refers to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and is a new coronavirus infection (COVID) which is a respiratory disease that caused the COVID-19 pandemic. -19) Refers to a coronavirus strain that causes (also called coronavirus disease 2019).
- SARS-CoV-2 virion has a diameter of 50-200 nanometers, and like other coronaviruses, SARS-CoV-2 has S (spikes), E (envelopes), M (membranes), and There are four structural proteins known as N (nucleocapsid) proteins.
- the N protein carries the RNA genome and the S, E, and M proteins together form the viral envelope.
- Peplomers are proteins that allow a virus to attach to and fuse with the membrane of a host cell.
- SARS-CoV-2 is transmitted from person to person, and the infection was initially assumed to occur primarily through respiratory droplets from a range of coughs and sneezes. It has also been suggested that the virus may be levitating because the aerosol can infect the virus.
- the copolymer used as a sensitizer for an immunochromatographic measurement method using the new coronavirus IgM antibody of the present invention as a measurement target substance is represented by the following general formula [1].
- each of n Rs independently represents a hydrogen atom or a cation.
- examples of the cation include sodium ion, potassium ion, ammonium ion and the like, and n R's may be the same or different.
- R is preferably a hydrogen atom.
- n 99 to 20: 1 to 80, preferably 90 to 30:10 to 70.
- the copolymer represented by the above formula is preferably a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000, more preferably 50,000 to 2,000,000.
- the copolymer represented by the general formula [1] of the present invention is a monomer represented by the above general formula [2] ⁇ MPC: 2- (methacrylic acid) ethyl 2'-(trimethylammonio) ethyl phosphate. ⁇ And a copolymer having a structural unit based on methacrylic acid or a salt thereof.
- the type of the copolymer is not particularly limited, and it may be a random copolymer or a block copolymer, but a random copolymer is preferable.
- methacrylic acid or a salt thereof examples include methacrylic acid, sodium methacrylate, potassium methacrylate, ammonium methacrylate and the like. Of these, methacrylic acid is preferable.
- copolymer used as a sensitizer for an immunochromatographic measurement method is preferably described in the following examples, but is not particularly limited.
- the copolymer used as the sensitizer for the immunochromatographic measurement method of the present invention may be used by dissolving it in various reagents and samples used for the immunochromatographic measurement method, or may be used by being carried on a kit for the immunochromatographic measurement method. Although it may be preferable, it is preferably used as a sample diluent.
- the immunochromatographic measurement method of the present invention generally consists of a sample diluent and a test strip as a kit for the lateral chromatographic measurement method to be used.
- the sample diluent is an appropriate diluent for measurement by the immunochromatography method, and is a buffer component such as Tris buffer, phosphate buffer, veronal buffer, boric acid buffer, Good buffer, albumin, etc. It can contain stabilizing components such as globulin, casein, serum, water-soluble gelatin, surfactants, sugars and chelating agents, and antiseptic components such as salicylic acid, benzoic acid and sodium azide.
- a buffer component such as Tris buffer, phosphate buffer, veronal buffer, boric acid buffer, Good buffer, albumin, etc. It can contain stabilizing components such as globulin, casein, serum, water-soluble gelatin, surfactants, sugars and chelating agents, and antiseptic components such as salicylic acid, benzoic acid and sodium azide.
- the concentration thereof is not particularly limited as the concentration at which the antigen-antibody reaction is carried out, but is usually 0.1 to 5.0 w / v%. It is preferably used so as to be 0.5 to 1.0 w / v%. In one embodiment, the concentration of the sensitizer of the present invention in the sample diluent is usually 0.1 to 5.0% by weight, preferably 0.5 to 1.0% by weight.
- a test strip generally refers to a first substance capable of an antigen-antibody reaction with a first antigen-determining group of a sample and a second substance capable of an antigen-antibody reaction with a second antigen-antibody group of the sample.
- the substance and the membrane carrier are provided at least, the first substance is preliminarily fixed at a predetermined position of the membrane carrier to form a capture site, and the second substance is the carrier at a position isolated from the capture site. It is arranged and configured so that it can be chromatographed at.
- the first substance and / or the second substance can be an antigen against the antibody.
- Typical configurations of a test strip for immunochromatography include a sample supply unit for supplying a sample (hereinafter, also referred to as a sample pad), a conjugate pad for arranging a conjugate, and a capture reagent such as an antibody.
- a sample pad for supplying a sample
- a conjugate pad for arranging a conjugate
- a capture reagent such as an antibody.
- the configuration in which the pads are arranged can be mentioned.
- sample pads include, but are not limited to, glass fiber (glass fiber), acrylic fiber, hydrophilic polyethylene material, dried paper, paper pulp, and woven fabric.
- the sample pad includes a blocking reagent, a buffer solution component, and a blood flocculant when the sample is blood, as needed, as long as it does not deviate from the object of the present invention and does not affect the reaction system. Etc. can be included. In that case, it suffices to be included in at least a part of the sample pad, and can be included in all.
- the conjugate pad is made of a pad-like porous material that has passed through the sample pad, is expandable of the sample, and is capable of retaining the conjugate, and retains the conjugate in part or all thereof.
- the porous material constituting the conjugated pad include pads made of non-woven fibers such as paper, cellulose mixture, nitrocellulose, polyester, acrylonitrile copolymer, glass, rayon and the like.
- the test strip for immunochromatography measurement a commercially available product or a product produced by a known method may be used.
- the base material used in the developing film may be one usually used in this field, and for example, cellulose, nitrocellulose, nylon and the like are preferable.
- the immunochromatographic measuring instrument using the new coronavirus IgM antibody containing the sensitizer of the present invention as a measurement target substance is, for example, (1) a developing membrane, (2) a sample labeling portion and a developing membrane, and The sample labeling part and the developing film are formed so as to be movable by capillarity, (3) the sample labeling part, the developing film and the liquid absorbing part, and the sample labeling part, the developing film and the developed film.
- the liquid absorbing portion is formed so as to be movable in this order by capillarity, (4) a sample dropping portion, a sample labeling portion, a developing film, and a liquid absorbing portion, and the sample dropping portion and the sample label. Examples thereof include those in which the portion, the developing film, and the liquid absorbing portion are formed so as to be movable by a capillary phenomenon in this order, but the present invention is not particularly limited.
- the amount thereof depends on the place to be supported, the type of sensitizer used, the type of the substance to be measured, the labeling substance used, and the like. It varies, but can be supported, for example, on a developing membrane, sample pad, or conjugate pad of an immunochromatographic measuring instrument.
- the amount of the sensitizer contained per unit area (cm 2 ) is usually 0.01 ⁇ g to 10 mg, preferably 0. .1 ⁇ g to 4 mg, more preferably 1 to 800 ⁇ g.
- the supported sensitizer of the present invention When the sensitizer of the present invention is supported on the sample dropping portion, when the sample dropping portion is a sample pad, the supported sensitizer of the present invention is usually 0.01 ⁇ g to 50 mg, preferably 100 ⁇ g to 20 mg. , More preferably 1-10 mg. When the sample dropping portion is a conjugate pad, the supported sensitizer of the present invention is usually 0.01 ⁇ g to 10 mg, preferably 0.1 ⁇ g to 4 mg, and more preferably 1 to 800 ⁇ g.
- the area to be supported varies depending on the type and size of the immunochromatographic measuring instrument used and the amount of the measuring sample, but in the case of the developed film of the immunochromatographic measuring instrument, it is usually 10 to 60% of the total area. It is preferably 5 to 15%, and in the case of a sample dropping portion, it is usually 5 to 40% of the total area, preferably 10 to 20%.
- the present invention is used for testing a new type coronavirus infection (COVID-19), which comprises a sensitizer for an immunochromatography method of the present invention and a substance that specifically binds to a new type coronavirus (SARS-CoV-2) IgM antibody.
- COVID-19 a new type coronavirus infection
- SARS-CoV-2 a new type coronavirus
- IgM antibody a new type coronavirus
- “Specific” means that the affinity for the new coronavirus (SARS-CoV-2) IgM antibody is higher than the affinity for other substances (in one embodiment, the new coronavirus (SARS-CoV-). IgM antibodies against antigens other than the protein of 2) are not included in the "other substances”).
- the specifically binding substance binds to the new coronavirus (SARS-CoV-2) IgM antibody, for example, with a KD of about 10-6 , 10-7 , 10-8 , 10-9 , 10-10 or less. ..
- the specifically binding substance binds to the new coronavirus (SARS-CoV-2) IgM antibody with a KD of 10-14 , 10-13 , 10-12 or higher.
- the substance is isolated or purified.
- isolation or purification means that an operation has been performed to remove components other than the desired component from the natural state.
- the purity (ratio of the weight of the substance to the total protein weight) of the isolated or purified substance is usually 50% or more, preferably 70% or more, more preferably 90% or more, and most preferably 95% or more (the ratio of the weight of the substance to the total protein weight). For example, substantially 100%).
- the substance may be directly or indirectly labeled with a labeling substance.
- Labeling substances include fluorescent substances (eg, FITC, rhodamine), radioactive substances (eg, 14 C, 3 H, 125 I), enzymes (eg, alkaline phosphatase, peroxidase), colored particles (eg, metal colloidal particles, coloring). Latex), biotin and the like.
- the "substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody” includes an antigen against the new coronavirus (SARS-CoV-2) IgM antibody and a new coronavirus (SARS-CoV-). 2) Antibodies that specifically bind to IgM antibodies, nucleic acids (eg, aptamers), etc. may be mentioned, but new coronavirus (SARS-CoV-2) antigens against IgM antibodies, anti-new coronavirus (SARS-CoV-2). IgM antibody is preferred.
- the antigen is not limited as long as it is an antigen against the new coronavirus (SARS-CoV-2) IgM antibody to be measured by the present invention, but for example, the new coronavirus S (spike) protein, E (envelope) protein, and the like.
- the new coronavirus S spike
- E envelope
- M membrane
- N nucleocapsid
- the recombinant new coronavirus (SARS-CoV-2) protein can be produced, for example, by the following method.
- a polynucleotide encoding a new coronavirus (SARS-CoV-2) protein amino acid sequence is incorporated into an appropriate expression vector, inserted into an appropriate host for transformation, and targeted from disrupted cells of this transformed cell.
- a new recombinant coronavirus (SARS-CoV-2) protein can be obtained.
- the host cell is not particularly limited, and various host cells conventionally used in genetic engineering techniques such as Escherichia coli, bacillus, yeast, plant or animal cell can be used.
- the new coronavirus (SARS-CoV-2) protein is a protein separation and purification known from the recombinant protein produced from the above transformant and the natural new coronavirus (SARS-CoV-2) that produces the recombinant protein. It may be isolated or purified by technique. Further, the protein may be chemically synthesized or biochemically synthesized by a cell-free translation system.
- the above-mentioned "anti-new coronavirus (SARS-CoV-2) IgM antibody” includes an anti-IgM antibody of an animal of the same species as the animal that produced the IgM antibody.
- the antibody can be any isotype, eg, IgA, IgD, IgE, IgG (eg, IgG1, IgG2, IgG3, or IgG4), IgM, and the like.
- the antibody may be a monoclonal antibody, a monoclonal antibody, a chimeric antibody, or a single chain antibody. Further, it may be a part of an antibody having antigen-binding property (antibody fragment) such as a Fab fragment or a fragment produced by a Fab expression library.
- antibody fragments are (i) Fab fragments, which are monovalent fragments consisting of VL, VH, CL and CH1 domains, and (ii) divalent fragments containing two Fab fragments linked by disulfide bridges in the hinge region.
- F (ab') 2 fragment (iii) Fv fragment consisting of VL and VH domain of single arm of antibody, (iv) disrupt disulfide bridge of F (ab') 2 fragment using mild reduction conditions.
- the anti-new coronavirus (SARS-CoV-2) IgM antibody used in the method of the present invention may be produced in mice, guinea pigs, hamsters, goats or rabbits. Further, as the anti-new coronavirus (SARS-CoV-2) IgM antibody, a commercially available one can be used.
- the test kit of the present invention contains the above-mentioned sensitizer for the immunochromatography method of the present invention and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody in separate containers.
- SARS-CoV-2 new coronavirus
- test kit of the present invention contains a sensitizer for the immunochromatography method of the present invention and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody to a new coronavirus infection (COVID-19). ) May include instructions stating that they can or should be used for inspection.
- SARS-CoV-2 new coronavirus
- COVID-19 new coronavirus infection
- the test kit for the new coronavirus infection may include any carrier, such as a pharmaceutically acceptable carrier, stabilizers and buffer components, other therapeutic agents and supplements.
- a pharmaceutically acceptable carrier include, but are not limited to, diluents such as water and physiological saline.
- the kit for testing a new type coronavirus infection (COVID-19) of the present invention may include a sample diluent, a test strip for measuring an immunochromatography, and an instrument for measuring an immunochromatography.
- the present invention measures a novel coronavirus (SARS-CoV-2) IgM antibody in a biological sample derived from a subject by performing an antigen-antibody reaction in the presence of the sensitizer for the immunochromatography method of the present invention.
- the biological sample examples include blood, but the biological sample is not particularly limited as long as the new coronavirus (SARS-CoV-2) IgM antibody can be detected.
- blood blood derived from any tissue can be assumed, but peripheral blood is usually used because it is easy to collect.
- a method for collecting blood a method known per se can be applied.
- the collected blood may be used as it is in this method, but it is used as a liquid component (plasma) in which cellular components (erythrocytes, leukocytes, platelets, etc.) are separated by a method known per se, for example, centrifugation, filtration, etc. It may be used in the method. It can also be used in this method as a liquid component (serum) in which blood is coagulated and platelets and coagulation factors are separated.
- a biological sample of a subject such as blood is placed in an extract and suspended, and an antigen is extracted from the biological sample solution.
- the biological sample solution treated with the extract is dropped onto the sample dropping portion.
- the biological sample solution in the presence of the sample diluent for the immunochromatography method containing the sensitizer for the immunochromatography method of the present invention moves from the sample dropping part to the sample labeling part and the membrane (membrane) due to the capillary phenomenon.
- the target antigen in the biological sample solution eg, new coronavirus (SARS-CoV-2) IgM antibody
- SARS-CoV-2 new coronavirus
- IgM antibody a labeling substance carried on the sample labeling part as it passes through the sample labeling part (eg, anti-new corona). It forms an immune complex with a virus (SARS-CoV-2) IgM antibody) and is directly expanded to one end of the developing membrane. Then, the complex and a substance (eg, an antigen against the new coronavirus (SARS-CoV-2) IgM antibody) existing on the test line of the developing membrane (coated on the test line) cause an immunochemical reaction. Visible lines appear on the developing membrane by combining and coloring.
- SARS-CoV-2 new coronavirus
- the sample is positive, and it can be determined that the subject is infected with the new coronavirus infection (COVID-19). Further, the intensity of the positive can be determined by visually measuring the degree of color development or by measuring it with an appropriate analytical instrument (for example, an immunochromatographic reader or a spectrophotometer).
- an analytical instrument for example, an immunochromatographic reader or a spectrophotometer.
- a fluorescent dye is used as the detection means of the labeled antibody, it is possible to irradiate the test line with ultraviolet rays or the like and determine whether or not the test line is positive or not depending on the presence or absence of color development and the degree thereof.
- the unreacted labeled antibody or the like is absorbed by the absorption pad. If the sample does not contain a target antigen (eg, new coronavirus (SARS-CoV-2) IgM antibody), the test line will not be colored and only the control line will be colored. .. In this case, the sample is negative and it is not determined that the subject is infected with the new coronavirus infection (COVID-19).
- a target antigen eg, new coronavirus (SARS-CoV-2) IgM antibody
- the biological sample used in the method for testing of the present invention is a sample derived from a mammalian subject.
- the mammal is not particularly limited as long as it is a mammal that may be infected with the new coronavirus infection (COVID-19), but among them, rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like.
- rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like.
- Experimental animals, pigs, cows, goats, horses, sheep, minks and other domestic animals, dogs, cats and other pets, humans, monkeys, crab monkeys, red-tailed monkeys, marmosets, orangutans, chimpanzees and other primates are preferred, and humans are particularly preferred. ..
- the method for treating a new type coronavirus infection (COVID-19) of the present invention includes at least the following steps (1) and (2).
- (1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
- (2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
- Patients include those who may be infected with the new coronavirus (SARS-CoV-2).
- the measurement means detecting the presence or absence of the new coronavirus (SARS-CoV-2) IgM antibody in the sample derived from the patient.
- the process of applying new coronavirus infection treatment to patients is not only to administer known therapeutic agents, therapeutic agents to be launched in the future, and clinical stage therapeutic agent candidates to patients, but also symptomatic treatment (aspirin administration, fluid replacement, etc.). Infusion administration, etc.) is also included.
- the method for treating a new coronavirus infection (COVID-19) of the present invention can detect a new coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, and thus can detect a new coronavirus infection (COVID-19). ) Can be rapidly treated with the new coronavirus infection (COVID-19).
- Symptoms of the new coronavirus infection range from asymptomatic to severe pneumonia and death. Specifically, fever, chills, malaise / dullness, sputum, olfactory / taste disorders, and shortness of breath , Muscle pain / joint pain, sore throat, headache, chills, nausea / vomiting, stuffy nose, diarrhea, hemoptysis, conjunctival hyperemia, etc.
- the method for treating a novel coronavirus infection (COVID-19) of the present invention comprises at least the following steps (1) and (2).
- (1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
- (2) A step of applying a new coronavirus infection (COVID-19) treatment to a subject other than a human based on the measurement result of the above (1).
- Subjects other than humans used in the therapeutic method of the present invention are not limited, but preferably mean mammals. More preferably, the mammals include mammals that may be infected with the new coronavirus infection (COVID-19), and among them, rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like. Experimental animals, pigs, cows, goats, horses, sheep, minks and other domestic animals, dogs, cats and other pets, monkeys, crab monkeys, lizard monkeys, guinea pigs, orchid utans, chimpanzees and other primates are preferred.
- the mammals include mammals that may be infected with the new coronavirus infection (COVID-19), and among them, rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like.
- rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like.
- the present invention includes a new coronavirus infection, which comprises a sensitizer for an immunochromatography method containing a copolymer represented by the above general formula [1] and a therapeutic agent for a new coronavirus infection (COVID-19).
- a therapeutic kit for (COVID-19) is provided.
- the therapeutic agent examples include those containing a therapeutic agent for coronavirus infection (COVID-19), and the therapeutic agent includes, but is not limited to, remdesivir, dexamethasone, favipiravir (Abigan), cyclesonide, and nafamostat. , Camostat, Ibermectin, Favipiravir, etc.
- Therapeutic agents are tocilizumab, baricitinib, acarabrutinib, labrizumab, eritoran, ibzilast, LY3127804, inrimab, HLCM051, ADR-001, squid, apremilast, and cenicriviro, which are therapeutic agents for severe pneumonia and acute respiratory distress syndrome.
- the therapeutic agent may include aspirin, fluid replacement, infusion and the like used for symptomatic treatment.
- the therapeutic kit of the present invention contains the above-mentioned sensitizer for the immunochromatographic measurement method of the present invention and the therapeutic agent for the new coronavirus infection (COVID-19) in separate containers.
- the therapeutic kit of the present invention uses the sensitizer for the immunochromatography method of the present invention and the therapeutic agent for the new coronavirus infection (COVID-19) for the treatment of the new coronavirus infection (COVID-19). It may include instructions stating that it can or should be used.
- the therapeutic kit for the new coronavirus infection may include any carrier, such as a pharmaceutically acceptable carrier, stabilizers and buffer components, other therapeutic agents and supplements.
- a pharmaceutically acceptable carrier include, but are not limited to, diluents such as water and physiological saline.
- the kit for treating a new type of coronavirus infection (COVID-19) of the present invention may include a sample diluent, a test strip for immunochromatography measurement, and an instrument for immunochromatography measurement.
- the therapeutic kit of the present invention may further contain a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody.
- SARS-CoV-2 new coronavirus
- the treatment kit for the new coronavirus infection (COVID-19) of the present invention can detect the new coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, the new coronavirus infection (COVID-) 19)
- the subject (patient) can be rapidly treated with the new coronavirus infection (COVID-19).
- the copolymer used in the present invention and Comparative Examples is a random copolymer synthesized from 2- (methacryloyloxy) ethyl 2'-(trimethylammonio) ethyl phosphate (MPC) and methacrylic acid.
- Example (confirmation of detection sensitivity using a known device for measuring an immunochromatography)
- SARS-CoV-2 known new coronavirus
- IgM antibody detection kit whether it has a sensitizing effect.
- Example 1 Copolymers 1, 2, 3 and 4 are added to the diluted solution of the kit so as to have a final concentration of 0.5% by weight, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example.
- the solutions prepared as described above were developed and the detection sensitivities were compared.
- Example 2 Copolymers 1, 2, 3 and 4 are supported on the sample pad of the kit so as to have a loading amount of 5 mg, and the serum of a new coronavirus-positive patient is prepared to be diluted 100-fold with respect to the comparative example.
- the prepared solutions were developed and the detection sensitivities were compared.
- Example 3 The copolymers 1, 2, 3 and 4 are supported on the conjugate pad of the kit so that the loading amount is 500 ⁇ g, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example.
- the prepared solutions were developed and the detection sensitivities were compared.
- Table 1 shows the results of improving the detection sensitivity when the copolymer was added to the diluted solution using the known novel coronavirus (SARS-CoV-2) IgM antibody detection kit.
- Table 2 shows the results of improving the detection sensitivity when the polymer is supported on the sample pad using the known new coronavirus (SARS-CoV-2) IgM antibody detection kit.
- Table 3 shows the results of improving the detection sensitivity when the polymer is supported on the conjugate pad using the known new coronavirus (SARS-CoV-2) IgM antibody detection kit.
- the present invention provides an immunochromatographic measurement method having a significantly superior sensitizing effect as compared with an immunochromatographic measurement method using a known novel coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance. I was able to provide it.
- SARS-CoV-2 novel coronavirus
- the present invention is remarkably superior in sensitizing effect as compared with the known pharmacochromatographic measurement method using the known novel coronavirus (SARS-CoV-2) IgM antibody as the measurement target substance.
- An immunochromatographic measurement method could be provided.
- the method for treating a new coronavirus infection (COVID-19) of the present invention can detect a new coronavirus (SARS-CoV-2) with high sensitivity, so that a patient with a new coronavirus infection or a new type can be detected. Since it is possible to quickly treat a patient who may be infected with coronavirus (SARS-CoV-2) with a new type of coronavirus infection, the therapeutic effect is high.
- the present invention can provide a sensitizer for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
- SARS-CoV-2 new type coronavirus
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Abstract
The present invention provides: a sensitizer for immunochromatographic assays, which can detect a new coronavirus (SARS-CoV-2) IgM antibody with high sensitivity; and an assay using the sensitizer. More specifically, the present invention provides a sensitizer for an immunochromatographic assay in which a substance to be measured is a new coronavirus (SARS-CoV-2) IgM antibody, the sensitizer comprising a copolymer represented by general formula [1]. (In the formula, n R's independently represent a hydrogen atom or a positive ion; each of m and n represents the number of constituent units; and m:n is 99 to 20:1 to 80.)
Description
本発明は、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤、及び該増感剤の共存下で行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法に関する。
The present invention is a sensitizer for an immunochromatographic measurement method using a new coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, and a new coronavirus (SARS-CoV-2) performed in the coexistence of the sensitizer. The present invention relates to an immunochromatographic measurement method using an IgM antibody as a measurement target substance.
イムノクロマト法は、メンブレンの毛細管現象を利用したクロマトグラフィーの手法と抗原抗体反応を利用した免疫学的手法を組み合わせた方法である。イムノクロマト法は、抗原抗体反応に起因する高い特異性を備え、操作が簡便であり、場所や特別な設備を必要とせずその場で検査でき、かつ結果を目視で判定できることから、臨床診断法として広く普及している（特許文献１）。
The immunochromatography method is a method that combines a chromatography method that utilizes the capillary phenomenon of the membrane and an immunological method that utilizes an antigen-antibody reaction. The immunochromatographic method has high specificity due to the antigen-antibody reaction, is easy to operate, can be inspected on the spot without the need for a place or special equipment, and the result can be visually judged, so that it is a clinical diagnostic method. It is widely used (Patent Document 1).
イムノクロマト法は、簡易に測定できる利点はあるが、検出感度の精度が低く、陽性検体を用いても偽陰性と判定される場合もあるので、より高感度のイムノクロマト測定法が望まれている。
The immunochromatographic method has the advantage that it can be easily measured, but the accuracy of detection sensitivity is low, and even if a positive sample is used, it may be judged as false negative. Therefore, a higher-sensitivity immunochromatographic measurement method is desired.
これまでに、高感度のイムノクロマト測定法を確立する目的で種々の方法が検討されており、例えば、特許文献２では、イムノクロマトの試薬の添加位置を変える事で高感度化する手法が開示されている。また、特許文献３では、メンブレンの素材を改良することで高感度化する手法が開示されている。
So far, various methods have been studied for the purpose of establishing a highly sensitive immunochromatographic measurement method. For example, Patent Document 2 discloses a method for increasing the sensitivity by changing the addition position of a reagent for an immunochromatographic test. There is. Further, Patent Document 3 discloses a method for increasing the sensitivity by improving the material of the membrane.
さらに、従来のイムノクロマト法に用いている検体希釈液に、添加剤を加えることで、高感度化を達成する方法が知られている。例えば、特許文献４、５及び６では、検体希釈液中に、ウシ血清アルブミン（ＢＳＡ）、ホスホリルコリン基を有する重合体、ヒアルロン酸を含有させることが開示されている。これらの開示では、試薬の添加位置を変える必要がなく、また従来のイムノクロマト用メンブレンを使用可能という利点がある。
Further, a method of achieving high sensitivity by adding an additive to the sample diluent used in the conventional immunochromatography method is known. For example, Patent Documents 4, 5 and 6 disclose that bovine serum albumin (BSA), a polymer having a phosphorylcholine group, and hyaluronic acid are contained in a sample diluent. These disclosures have the advantages that it is not necessary to change the reagent addition position and that a conventional membrane for immunochromatography can be used.
しかしながら、上記特許文献では、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体の測定を高感度に行う方法に用いる増感剤については開示又は示唆をされていない。
However, the above patent document does not disclose or suggest a sensitizer used in a method for measuring a new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity.
とくに、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体は、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）への感染初期に産生される抗体であり、ＩｇＭ抗体を高感度に検出可能であれば、感染初期段階で感染者を隔離できることから感染の拡大を抑えることが可能となる。また、早期に治療を開始できることから治療の効果を高めることができるため、ＩｇＭ抗体を高感度に検出可能とする増感剤は有用性が高いと考えられる。
In particular, the new coronavirus (SARS-CoV-2) IgM antibody is an antibody produced in the early stage of infection with the new coronavirus (SARS-CoV-2), and if the IgM antibody can be detected with high sensitivity, Since the infected person can be isolated at the initial stage of infection, it is possible to suppress the spread of infection. In addition, since the treatment can be started at an early stage, the effect of the treatment can be enhanced, so that a sensitizer that can detect the IgM antibody with high sensitivity is considered to be highly useful.
本発明では、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を高感度に検出できるイムノクロマト測定法用増感剤及び該増感剤を使用した測定法を提供することにある。
The present invention provides a sensitizer for an immunochromatographic measurement method capable of detecting a new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, and a measurement method using the sensitizer.
本発明者らは、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を高感度に検出できる化合物を見出すべく鋭意検討した結果、下記一般式〔１〕で示されるホスホリルコリン類似基を有する基を側鎖に有する共重合体が目的の性能を有していることを見出し、本発明を完成するに至った。
As a result of diligent studies to find a compound capable of detecting a new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, the present inventors have side-chained a group having a phosphorylcholine-like group represented by the following general formula [1]. We have found that the copolymer contained in the chain has the desired performance, and have completed the present invention.
すなわち、本発明は以下よりなる。
［１］ 下記一般式〔１〕で表される共重合体を含んでなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 That is, the present invention comprises the following.
[1] A sensitizer for an immunochromatographic measurement method, which comprises a copolymer represented by the following general formula [1] and contains a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
［１］ 下記一般式〔１〕で表される共重合体を含んでなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 That is, the present invention comprises the following.
[1] A sensitizer for an immunochromatographic measurement method, which comprises a copolymer represented by the following general formula [1] and contains a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
（式中、ｎ個のＲはそれぞれ独立して、水素原子又は陽イオンを示し、ｍとｎはそれぞれ構成単位数を示し、ｍ：ｎは９９～２０：１～８０である。）
［２］ 前記共重合体は、下記一般式〔２〕で表される単量体に基づく構成単位と、メタクリル酸またはその塩に基づく構成単位を有する共重合体である［１］に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 (In the equation, n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.)
[2] The copolymer according to [1], wherein the copolymer has a structural unit based on a monomer represented by the following general formula [2] and a structural unit based on methacrylic acid or a salt thereof. A sensitizer for immunochromatography, which uses a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
［２］ 前記共重合体は、下記一般式〔２〕で表される単量体に基づく構成単位と、メタクリル酸またはその塩に基づく構成単位を有する共重合体である［１］に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 (In the equation, n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.)
[2] The copolymer according to [1], wherein the copolymer has a structural unit based on a monomer represented by the following general formula [2] and a structural unit based on methacrylic acid or a salt thereof. A sensitizer for immunochromatography, which uses a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
［３］ 前記共重合体が、一般式〔２〕で表される単量体に基づく構成単位数とメタクリル酸またはその塩に基づく構成単位数の比（ｍ：ｎ）が９９～２０：１～８０の共重合体である［１］又は［２］に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
［４］ 前記共重合体が、一般式〔２〕で表される単量体に基づく構成単位数とメタクリル酸またはその塩に基づく構成単位数の比（ｍ：ｎ）が９０～３０：１０～７０の共重合体である［１］又は［２］に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
［５］ 前記共重合体が、重量平均分子量１０，０００～５，０００，０００の水溶性共重合体である［１］～［４］のいずれかに記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
［６］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤を含む、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用検体希釈液。
［７］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行わせることを特徴とする、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法。
［８］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤を展開膜、サンプルパッド、又はコンジュゲートパッドに担持させてなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定用器具。
［９］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キット。
［１０］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行うことにより、被験体由来の生物学的試料中の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定することを含む、該被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染している可能性を試験するための方法。
［１１］ 以下の（１）、（２）の工程を含む新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法、
（１）下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程、及び
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を患者に施す工程。 [3] The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 99 to 20: 1. A sensitizer for an immunochromatographic measurement method using the new coronavirus (SARS-CoV-2) IgM antibody according to [1] or [2], which is a copolymer of 80 to 80, as a measurement target substance.
[4] The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 90 to 30:10. A sensitizer for an immunochromatographic measurement method using the new coronavirus (SARS-CoV-2) IgM antibody according to [1] or [2], which is a copolymer of ~ 70, as a measurement target substance.
[5] The new coronavirus (SARS-CoV-) according to any one of [1] to [4], wherein the copolymer is a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000. 2) A sensitizer for an immunochromatographic measurement method using an IgM antibody as a measurement target substance.
[6] Dilution of a sample for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, which contains the sensitizer for the immunochromatographic measurement method according to any one of [1] to [5]. liquid.
[7] A new type coronavirus (SARS-CoV-2) IgM characterized in that an antigen-antibody reaction is carried out in the presence of the sensitizer for the later lateral flow test according to any one of [1] to [5]. An immunochromatographic measurement method using an antibody as a measurement target substance.
[8] A new type coronavirus (SARS-CoV-2) in which the sensitizer for the later lateral flow test according to any one of [1] to [5] is carried on a developing membrane, a sample pad, or a conjugate pad. An instrument for measuring immunochromatography that uses IgM antibody as the measurement target substance.
[9] Coronavirus infection containing the sensitizer for the later lateral flow test according to any one of [1] to [5] and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody. Kit for testing for disease (COVID-19).
[10] By performing an antigen-antibody reaction in the presence of the sensitizer for immunochromatography according to any one of [1] to [5], a new type coronavirus (SARS) in a biological sample derived from a subject is performed. -CoV-2) A method for testing the possibility that a subject is infected with a new coronavirus infection (COVID-19), which comprises measuring IgM antibody.
[11] A method for treating a new type of coronavirus infection (COVID-19), which comprises the following steps (1) and (2).
(1) A measurement step of a new type coronavirus (SARS-CoV-2) that undergoes an antigen-antibody reaction in the presence of a sensitizer for an immunochromatographic measurement method containing a copolymer represented by the following general formula [1]. And (2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
［４］ 前記共重合体が、一般式〔２〕で表される単量体に基づく構成単位数とメタクリル酸またはその塩に基づく構成単位数の比（ｍ：ｎ）が９０～３０：１０～７０の共重合体である［１］又は［２］に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
［５］ 前記共重合体が、重量平均分子量１０，０００～５，０００，０００の水溶性共重合体である［１］～［４］のいずれかに記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
［６］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤を含む、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用検体希釈液。
［７］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行わせることを特徴とする、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法。
［８］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤を展開膜、サンプルパッド、又はコンジュゲートパッドに担持させてなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定用器具。
［９］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キット。
［１０］ ［１］～［５］のいずれかに記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行うことにより、被験体由来の生物学的試料中の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定することを含む、該被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染している可能性を試験するための方法。
［１１］ 以下の（１）、（２）の工程を含む新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法、
（１）下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程、及び
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を患者に施す工程。 [3] The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 99 to 20: 1. A sensitizer for an immunochromatographic measurement method using the new coronavirus (SARS-CoV-2) IgM antibody according to [1] or [2], which is a copolymer of 80 to 80, as a measurement target substance.
[4] The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 90 to 30:10. A sensitizer for an immunochromatographic measurement method using the new coronavirus (SARS-CoV-2) IgM antibody according to [1] or [2], which is a copolymer of ~ 70, as a measurement target substance.
[5] The new coronavirus (SARS-CoV-) according to any one of [1] to [4], wherein the copolymer is a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000. 2) A sensitizer for an immunochromatographic measurement method using an IgM antibody as a measurement target substance.
[6] Dilution of a sample for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, which contains the sensitizer for the immunochromatographic measurement method according to any one of [1] to [5]. liquid.
[7] A new type coronavirus (SARS-CoV-2) IgM characterized in that an antigen-antibody reaction is carried out in the presence of the sensitizer for the later lateral flow test according to any one of [1] to [5]. An immunochromatographic measurement method using an antibody as a measurement target substance.
[8] A new type coronavirus (SARS-CoV-2) in which the sensitizer for the later lateral flow test according to any one of [1] to [5] is carried on a developing membrane, a sample pad, or a conjugate pad. An instrument for measuring immunochromatography that uses IgM antibody as the measurement target substance.
[9] Coronavirus infection containing the sensitizer for the later lateral flow test according to any one of [1] to [5] and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody. Kit for testing for disease (COVID-19).
[10] By performing an antigen-antibody reaction in the presence of the sensitizer for immunochromatography according to any one of [1] to [5], a new type coronavirus (SARS) in a biological sample derived from a subject is performed. -CoV-2) A method for testing the possibility that a subject is infected with a new coronavirus infection (COVID-19), which comprises measuring IgM antibody.
[11] A method for treating a new type of coronavirus infection (COVID-19), which comprises the following steps (1) and (2).
(1) A measurement step of a new type coronavirus (SARS-CoV-2) that undergoes an antigen-antibody reaction in the presence of a sensitizer for an immunochromatographic measurement method containing a copolymer represented by the following general formula [1]. And (2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
（式中、ｎ個のＲはそれぞれ独立して、水素原子又は陽イオンを示し、ｍとｎはそれぞれ構成単位数を示し、ｍ：ｎは９９～２０：１～８０である。）
［１２］ 下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キット。 (In the equation, n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.)
[12] A new coronavirus infection (12), which comprises a sensitizer for an immunochromatography method containing a copolymer represented by the following general formula [1] and a therapeutic agent for a new coronavirus infection (COVID-19). COVID-19) treatment kit.
［１２］ 下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キット。 (In the equation, n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.)
[12] A new coronavirus infection (12), which comprises a sensitizer for an immunochromatography method containing a copolymer represented by the following general formula [1] and a therapeutic agent for a new coronavirus infection (COVID-19). COVID-19) treatment kit.
（式中、ｎ個のＲはそれぞれ独立して、水素原子又は陽イオンを示し、ｍとｎはそれぞれ構成単位数を示し、ｍ：ｎは９９～２０：１～８０である。）
(In the equation, n Rs independently represent hydrogen atoms or cations, m and n each indicate the number of constituent units, and m: n is 99 to 20: 1 to 80.)
本発明の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤を使用した測定方法では、従来の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を検出するイムノクロマト測定法と比較して、高感度で新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を検出することができる。
In the measurement method using the sensitizer for immunochromatography, which uses the new coronavirus (SARS-CoV-2) IgM antibody of the present invention as the measurement target substance, the conventional new coronavirus (SARS-CoV-2) IgM antibody is used. It is possible to detect the new type coronavirus (SARS-CoV-2) IgM antibody with high sensitivity as compared with the immunochromatographic measurement method to be detected.
本発明の新型コロナウイルスＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤、該増感剤を含む上記抗体を測定対象物質とするイムノクロマト測定法用検体希釈液、該増感剤の存在下で抗原抗体反応を行わせる上記抗体を測定対象物質とするイムノクロマト測定法、該増感剤を含む上記抗体を測定対象物質とするイムノクロマト測定用器具、及び、該増感剤の存在下で抗原抗体反応を行わせる工程を含む新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法を、以下で説明する。
Presence of a sensitizer for an immunochromatographic measurement method using the new coronavirus IgM antibody of the present invention as a measurement target substance, a sample diluent for an immunochromatographic measurement method using the above antibody containing the sensitizer as a measurement target substance, and the sensitizer. An immunochromatographic measurement method using the above-mentioned antibody as a measurement target substance, an device for measuring an immunochromatography using the above-mentioned antibody as a measurement target substance containing the sensitizer, and an antigen in the presence of the sensitizer. A method for treating a new type of coronavirus infection (COVID-19) including a step of causing an antibody reaction will be described below.
なお、本発明のイムノクロマト測定法用増感剤を使用する測定法では、測定対象物質としてはＩｇＭ抗体であるが、他の測定対象物質も検出できる。しかし、本発明のイムノクロマト測定法用増感剤は、上記抗体を高感度に検出できることを特徴とする。
In the measurement method using the sensitizer for the immunochromatographic measurement method of the present invention, the measurement target substance is an IgM antibody, but other measurement target substances can also be detected. However, the sensitizer for the immunochromatographic measurement method of the present invention is characterized in that the above antibody can be detected with high sensitivity.
本明細書中、新型コロナウイルスとは、重症急性呼吸器症候群コロナウイルス２（ＳＡＲＳ－ＣｏＶ－２）を指し、ＣＯＶＩＤ－１９パンデミックの原因となった呼吸器疾患である新型コロナウイルス感染症（ＣＯＶＩＤ－１９）（コロナウイルス病２０１９ともいう）を引き起こすコロナウイルス株のことを言う。各ＳＡＲＳ－ＣｏＶ－２ビリオンの直径は５０～２００ナノメートルであり、他のコロナウイルスと同様に、ＳＡＲＳ－ＣｏＶ－２には、Ｓ（スパイク）、Ｅ（エンベロープ）、Ｍ（膜）、およびＮ（ヌクレオカプシド）タンパク質として知られる４つの構造タンパク質がある。Ｎタンパク質はＲＮＡゲノムを保持し、Ｓ、Ｅ、およびＭタンパク質が一緒になってウイルスエンベロープを形成する。スパイクタンパク質は、ウイルスが宿主細胞の膜に付着して融合することを可能にするタンパク質である。ＳＡＲＳ－ＣｏＶ－２はヒトからヒトへ感染し、感染は当初、主に一定範囲内の咳やくしゃみからの呼吸器飛沫を介して起こると想定された。また、エアロゾルがウイルスを感染させる可能性があるため、ウイルスも空中浮遊している可能性があることも示唆されている。
In the present specification, the new coronavirus refers to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and is a new coronavirus infection (COVID) which is a respiratory disease that caused the COVID-19 pandemic. -19) Refers to a coronavirus strain that causes (also called coronavirus disease 2019). Each SARS-CoV-2 virion has a diameter of 50-200 nanometers, and like other coronaviruses, SARS-CoV-2 has S (spikes), E (envelopes), M (membranes), and There are four structural proteins known as N (nucleocapsid) proteins. The N protein carries the RNA genome and the S, E, and M proteins together form the viral envelope. Peplomers are proteins that allow a virus to attach to and fuse with the membrane of a host cell. SARS-CoV-2 is transmitted from person to person, and the infection was initially assumed to occur primarily through respiratory droplets from a range of coughs and sneezes. It has also been suggested that the virus may be levitating because the aerosol can infect the virus.
本発明の新型コロナウイルスＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤として用いられる共重合体は、下記一般式〔１〕で表される。
The copolymer used as a sensitizer for an immunochromatographic measurement method using the new coronavirus IgM antibody of the present invention as a measurement target substance is represented by the following general formula [1].
上記式中、ｎ個のＲはそれぞれ独立して、水素原子又は陽イオンを示す。ここで、陽イオンは、ナトリウムイオン、カリウムイオン、アンモニウムイオン等が挙げられ、ｎ個のＲは同一であっても、異なってもよい。Ｒは、好ましくは水素原子である。
In the above formula, each of n Rs independently represents a hydrogen atom or a cation. Here, examples of the cation include sodium ion, potassium ion, ammonium ion and the like, and n R's may be the same or different. R is preferably a hydrogen atom.
上記式中、ｍとｎはそれぞれ構成単位数を示し、ｍ：ｎは、９９～２０：１～８０、好ましくは９０～３０：１０～７０である。
In the above formula, m and n indicate the number of constituent units, respectively, and m: n is 99 to 20: 1 to 80, preferably 90 to 30:10 to 70.
上記式で表される共重合体は、好ましくは重量平均分子量１０，０００～５，０００，０００、より好ましくは５０，０００～２，０００，０００の水溶性共重合体である。
The copolymer represented by the above formula is preferably a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000, more preferably 50,000 to 2,000,000.
本発明の一般式〔１〕で表される共重合体は、上記一般式〔２〕で表される単量体｛ＭＰＣ：２－（メタクリロイルオキシ）エチル ２’－（トリメチルアンモニオ）エチル ホスフェート｝に基づく構成単位と、メタクリル酸またはその塩に基づく構成単位を有する共重合体により成る。
ここで、当該共重合体の種類には特に制限はなく、ランダム共重合体であっても、ブロック共重合体であってもよいが、ランダム共重合体が好ましい。 The copolymer represented by the general formula [1] of the present invention is a monomer represented by the above general formula [2] {MPC: 2- (methacrylic acid) ethyl 2'-(trimethylammonio) ethyl phosphate. } And a copolymer having a structural unit based on methacrylic acid or a salt thereof.
Here, the type of the copolymer is not particularly limited, and it may be a random copolymer or a block copolymer, but a random copolymer is preferable.
ここで、当該共重合体の種類には特に制限はなく、ランダム共重合体であっても、ブロック共重合体であってもよいが、ランダム共重合体が好ましい。 The copolymer represented by the general formula [1] of the present invention is a monomer represented by the above general formula [2] {MPC: 2- (methacrylic acid) ethyl 2'-(trimethylammonio) ethyl phosphate. } And a copolymer having a structural unit based on methacrylic acid or a salt thereof.
Here, the type of the copolymer is not particularly limited, and it may be a random copolymer or a block copolymer, but a random copolymer is preferable.
メタクリル酸またはその塩の例としては、メタクリル酸、メタクリル酸ナトリウム、メタクリル酸カリウム、メタクリル酸アンモニウム等を例示することができる。中でも、メタクリル酸が好ましい。
Examples of methacrylic acid or a salt thereof include methacrylic acid, sodium methacrylate, potassium methacrylate, ammonium methacrylate and the like. Of these, methacrylic acid is preferable.
イムノクロマト測定法用増感剤として用いられる共重合体は、好ましくは、以下の実施例に記載のものを例示することができるが特に限定されない。
The copolymer used as a sensitizer for an immunochromatographic measurement method is preferably described in the following examples, but is not particularly limited.
一般式〔１〕で示される水溶性共重合体の具体的な製造例は、特開平０８－３３３４２１号、特許文献６に記載の方法を例示することができるが特に限定されない。
Specific examples of the production of the water-soluble copolymer represented by the general formula [1] can be exemplified by the methods described in JP-A-08-333421 and Patent Document 6, but are not particularly limited.
本発明のイムノクロマト測定法用増感剤として用いられる共重合体は、イムノクロマト測定法に用いられる各種試薬や試料に溶解させて用いてもよく、また、イムノクロマト測定法用キットに担持させて用いてもよいが、好ましくは、検体希釈液として用いることが好ましい。
The copolymer used as the sensitizer for the immunochromatographic measurement method of the present invention may be used by dissolving it in various reagents and samples used for the immunochromatographic measurement method, or may be used by being carried on a kit for the immunochromatographic measurement method. Although it may be preferable, it is preferably used as a sample diluent.
なお、本発明のイムノクロマト測定法とは、用いられるイムノクロマト測定法用キットとして一般に、検体希釈液とテストストリップからなる。
The immunochromatographic measurement method of the present invention generally consists of a sample diluent and a test strip as a kit for the lateral chromatographic measurement method to be used.
検体希釈液とは、イムノクロマトグラフィー法で測定するに際しての、適当な希釈剤であり、トリス緩衝剤、リン酸緩衝剤、ベロナール緩衝剤、ホウ酸緩衝剤、グッド緩衝剤等の緩衝成分、アルブミン、グロブリン、カゼイン、血清、水溶性ゼラチン、界面活性剤、糖類、キレート剤等の安定化成分、サリチル酸、安息香酸、アジ化ナトリウム等の防腐成分含むことができる。
The sample diluent is an appropriate diluent for measurement by the immunochromatography method, and is a buffer component such as Tris buffer, phosphate buffer, veronal buffer, boric acid buffer, Good buffer, albumin, etc. It can contain stabilizing components such as globulin, casein, serum, water-soluble gelatin, surfactants, sugars and chelating agents, and antiseptic components such as salicylic acid, benzoic acid and sodium azide.
本発明の増感剤を、検体希釈液に溶解させて用いる場合、その濃度は、抗原抗体反応がなされる際の濃度として、とくに限定されないが、通常０．１～５．０ｗ/ｖ％、好ましくは０．５～１．０ｗ/ｖ％となるように用いられる。一態様において、検体希釈液中の本発明の増感剤の濃度は、通常０．１～５．０重量％、好ましくは０．５～１．０重量％である。
When the sensitizer of the present invention is used by dissolving it in a sample diluent, the concentration thereof is not particularly limited as the concentration at which the antigen-antibody reaction is carried out, but is usually 0.1 to 5.0 w / v%. It is preferably used so as to be 0.5 to 1.0 w / v%. In one embodiment, the concentration of the sensitizer of the present invention in the sample diluent is usually 0.1 to 5.0% by weight, preferably 0.5 to 1.0% by weight.
テストストリップとは、一般に、検体の第一の抗原決定基にて抗原抗体反応可能な第一の物質と、前記検体の第二の抗原決定基にて抗原抗体反応可能で且つ標識された第二の物質と、膜担体とを少なくとも備え、前記第一の物質は前記膜担体の所定位置に予め固定されて捕捉部位を形成し、前記第二の物質は前記捕捉部位から隔離した位置で前記担体にてクロマト展開可能なように配置されて構成される。
A test strip generally refers to a first substance capable of an antigen-antibody reaction with a first antigen-determining group of a sample and a second substance capable of an antigen-antibody reaction with a second antigen-antibody group of the sample. The substance and the membrane carrier are provided at least, the first substance is preliminarily fixed at a predetermined position of the membrane carrier to form a capture site, and the second substance is the carrier at a position isolated from the capture site. It is arranged and configured so that it can be chromatographed at.
一態様において、該検体が抗体（ＩｇＭ抗体を含む）の場合、上記第一の物質および／または第二の物質は該抗体に対する抗原であり得る。
In one embodiment, when the sample is an antibody (including an IgM antibody), the first substance and / or the second substance can be an antigen against the antibody.
イムノクロマトグラフィー用テストストリップの典型的な構成としては、サンプルを供給するためのサンプル供給部（以下、サンプルパッドともいうことがある）、コンジュゲートを配置するためのコンジュゲートパッド、抗体などの捕捉試薬がライン状に固定化された検出部を有し、上記免疫複合体を移動相により展開させて捕捉する展開膜、および展開膜を展開してきたサンプルを検出部よりも下流で吸収するための吸収パッドが配置された構成を挙げることができる。
Typical configurations of a test strip for immunochromatography include a sample supply unit for supplying a sample (hereinafter, also referred to as a sample pad), a conjugate pad for arranging a conjugate, and a capture reagent such as an antibody. Has a detection unit immobilized in a line shape, and an expansion membrane that expands and captures the immune complex by a mobile phase, and an absorption for absorbing a sample that has developed the expansion membrane downstream from the detection unit. The configuration in which the pads are arranged can be mentioned.
サンプルパッドに適した材料の具体例として、ガラス繊維（グラスファイバー）、アクリル繊維、親水性ポリエチレン材、乾燥紙、紙パルプ、織物等が含まれるが、これらに限定されない。また、サンプルパッドには、本発明の目的を逸脱せず、反応系に影響のない範囲において、必要に応じ通常使用されるブロッキング試薬、緩衝液成分、サンプルが血液の場合には、血液凝集剤等を含ませておくこともできる。その場合、サンプルパッドの少なくとも一部に含まれていればよく、全部に含ませることもできる。
Specific examples of materials suitable for sample pads include, but are not limited to, glass fiber (glass fiber), acrylic fiber, hydrophilic polyethylene material, dried paper, paper pulp, and woven fabric. Further, the sample pad includes a blocking reagent, a buffer solution component, and a blood flocculant when the sample is blood, as needed, as long as it does not deviate from the object of the present invention and does not affect the reaction system. Etc. can be included. In that case, it suffices to be included in at least a part of the sample pad, and can be included in all.
一態様において、コンジュゲートパッドは、サンプルパッドを通過した、サンプルが展開可能であり、かつ、コンジュゲートを保持可能なパッド状の多孔質材料からなり、その一部あるいは全部にコンジュゲートを保持する。コンジュゲートパッドを構成する多孔質材料としては、紙、セルロース混合物、ニトロセルロース、ポリエステル、アクリロニトリルコポリマー、ガラス、レーヨン等のような不織繊維からなるパッドが挙げられる。
In one aspect, the conjugate pad is made of a pad-like porous material that has passed through the sample pad, is expandable of the sample, and is capable of retaining the conjugate, and retains the conjugate in part or all thereof. .. Examples of the porous material constituting the conjugated pad include pads made of non-woven fibers such as paper, cellulose mixture, nitrocellulose, polyester, acrylonitrile copolymer, glass, rayon and the like.
イムノクロマト測定用テストストリップは、市販品を用いても、公知の方法により作製したものを用いてもよい。公知の方法により作製する場合、上記展開膜で用いられる基材としては、通常この分野で用いられるものを用いればよく、例えば、セルロース、ニトロセルロース、ナイロン等が好ましいものとして挙げられる。
As the test strip for immunochromatography measurement, a commercially available product or a product produced by a known method may be used. When produced by a known method, the base material used in the developing film may be one usually used in this field, and for example, cellulose, nitrocellulose, nylon and the like are preferable.
本発明の増感剤を含む新型コロナウイルスＩｇＭ抗体を測定対象物質とするイムノクロマト測定用器具としては、例えば、（１）展開膜からなるもの、（２）検体標識部並びに展開膜からなり、且つ該検体標識部と該展開膜とが毛細管現象により移動可能なように形成されたもの、（３）検体標識部、展開膜並びに液体吸収部からなり、且つ該検体標識部と該展開膜と該液体吸収部とが、この順に毛細管現象により移動可能なように形成されたもの、（４）検体滴下部、検体標識部、展開膜並びに液体吸収部からなり、且つ該検体滴下部と該検体標識部と該展開膜と該液体吸収部とが、この順に毛細管現象により移動可能なように形成されたもの等が挙げられるが、とくに限定されない。
The immunochromatographic measuring instrument using the new coronavirus IgM antibody containing the sensitizer of the present invention as a measurement target substance is, for example, (1) a developing membrane, (2) a sample labeling portion and a developing membrane, and The sample labeling part and the developing film are formed so as to be movable by capillarity, (3) the sample labeling part, the developing film and the liquid absorbing part, and the sample labeling part, the developing film and the developed film. The liquid absorbing portion is formed so as to be movable in this order by capillarity, (4) a sample dropping portion, a sample labeling portion, a developing film, and a liquid absorbing portion, and the sample dropping portion and the sample label. Examples thereof include those in which the portion, the developing film, and the liquid absorbing portion are formed so as to be movable by a capillary phenomenon in this order, but the present invention is not particularly limited.
本発明の増感剤を、前記イムノクロマト測定用器具に担持させて用いる場合、その量は、担持させる場所、使用する増感剤の種類、使用する測定対象物質の種類、使用する標識物質等により変動するが、例えば、イムノクロマト測定用器具の展開膜、サンプルパッド、又はコンジュゲートパッドに担持させ得る。本発明の増感剤をイムノクロマト測定用器具の展開膜、検体標識部に担持させる場合、単位面積（ｃｍ２）当たりに含まれる増感剤の量として、通常０．０１μｇ～１０ｍｇ、好ましくは０．１μｇ～４ｍｇ、より好ましくは１～８００μｇである。本発明の増感剤を検体滴下部に担持させる場合、該検体滴下部がサンプルパッドである場合は、担持される本発明の増感剤は、通常０．０１μｇ～５０ｍｇ、好ましくは１００μｇ～２０ｍｇ、より好ましくは１～１０ｍｇである。該検体滴下部がコンジュゲートパッドである場合は、担持される本発明の増感剤は、通常０．０１μｇ～１０ｍｇ、好ましくは０．１μｇ～４ｍｇ、より好ましくは１～８００μｇである。また、担持される面積は、用いられるイムノクロマト測定用器具の種類及び大きさ、測定用試料の量により変動するが、イムノクロマト測定用器具の展開膜の場合には、通常総面積の１０～６０％、好ましくは５～１５％、検体滴下部の場合には、通常総面積の５～４０％、好ましくは１０～２０％である。
When the sensitizer of the present invention is supported on the immunochromatographic measuring instrument and used, the amount thereof depends on the place to be supported, the type of sensitizer used, the type of the substance to be measured, the labeling substance used, and the like. It varies, but can be supported, for example, on a developing membrane, sample pad, or conjugate pad of an immunochromatographic measuring instrument. When the sensitizer of the present invention is carried on the developing membrane or the sample labeling portion of the immunochromatographic measurement instrument, the amount of the sensitizer contained per unit area (cm 2 ) is usually 0.01 μg to 10 mg, preferably 0. .1 μg to 4 mg, more preferably 1 to 800 μg. When the sensitizer of the present invention is supported on the sample dropping portion, when the sample dropping portion is a sample pad, the supported sensitizer of the present invention is usually 0.01 μg to 50 mg, preferably 100 μg to 20 mg. , More preferably 1-10 mg. When the sample dropping portion is a conjugate pad, the supported sensitizer of the present invention is usually 0.01 μg to 10 mg, preferably 0.1 μg to 4 mg, and more preferably 1 to 800 μg. The area to be supported varies depending on the type and size of the immunochromatographic measuring instrument used and the amount of the measuring sample, but in the case of the developed film of the immunochromatographic measuring instrument, it is usually 10 to 60% of the total area. It is preferably 5 to 15%, and in the case of a sample dropping portion, it is usually 5 to 40% of the total area, preferably 10 to 20%.
本発明は、本発明のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キットを提供する。
The present invention is used for testing a new type coronavirus infection (COVID-19), which comprises a sensitizer for an immunochromatography method of the present invention and a substance that specifically binds to a new type coronavirus (SARS-CoV-2) IgM antibody. Provide a kit.
「特異的」とは、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に対する親和性が、他の物質に対する親和性よりも高いことを意味する（一態様において、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）のタンパク質以外の抗原に対するＩｇＭ抗体は、該「他の物質」には含まれない）。該特異的に結合する物質は、例えば、約１０－６、１０－７、１０－８、１０－９、１０－１０以下のＫＤで新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に結合する。また、該特異的に結合する物質は、１０－１４、１０－１３、１０－１２以上のＫＤで新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に結合する。
"Specific" means that the affinity for the new coronavirus (SARS-CoV-2) IgM antibody is higher than the affinity for other substances (in one embodiment, the new coronavirus (SARS-CoV-). IgM antibodies against antigens other than the protein of 2) are not included in the "other substances"). The specifically binding substance binds to the new coronavirus (SARS-CoV-2) IgM antibody, for example, with a KD of about 10-6 , 10-7 , 10-8 , 10-9 , 10-10 or less. .. In addition, the specifically binding substance binds to the new coronavirus (SARS-CoV-2) IgM antibody with a KD of 10-14 , 10-13 , 10-12 or higher.
前記物質は単離又は精製されていることが好ましい。「単離又は精製」とは、天然にある状態から目的とする成分以外の成分を除去する操作が施されていることを意味する。単離又は精製された前記物質の純度（全タンパク質重量に対する、前記物質の重量の割合）は、通常50％以上、好ましくは70％以上、より好ましくは90％以上、最も好ましくは95％以上（例えば実質的に100％）である。
It is preferable that the substance is isolated or purified. "Isolation or purification" means that an operation has been performed to remove components other than the desired component from the natural state. The purity (ratio of the weight of the substance to the total protein weight) of the isolated or purified substance is usually 50% or more, preferably 70% or more, more preferably 90% or more, and most preferably 95% or more (the ratio of the weight of the substance to the total protein weight). For example, substantially 100%).
前記物質は、直接的又は間接的に標識物質により標識されていてもよい。標識物質としては、蛍光物質（例、FITC、ローダミン）、放射性物質（例、14C、3H、125Ｉ）、酵素（例、アルカリホスファターゼ、ペルオキシダーゼ）、着色粒子（例、金属コロイド粒子、着色ラテックス）、ビオチン等が挙げられる。
The substance may be directly or indirectly labeled with a labeling substance. Labeling substances include fluorescent substances (eg, FITC, rhodamine), radioactive substances (eg, 14 C, 3 H, 125 I), enzymes (eg, alkaline phosphatase, peroxidase), colored particles (eg, metal colloidal particles, coloring). Latex), biotin and the like.
本発明において「新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質」としては、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に対する抗原、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する抗体、核酸（例、アプタマー）等が挙げられるが、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に対する抗原、抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体が好ましい。
In the present invention, the "substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody" includes an antigen against the new coronavirus (SARS-CoV-2) IgM antibody and a new coronavirus (SARS-CoV-). 2) Antibodies that specifically bind to IgM antibodies, nucleic acids (eg, aptamers), etc. may be mentioned, but new coronavirus (SARS-CoV-2) antigens against IgM antibodies, anti-new coronavirus (SARS-CoV-2). IgM antibody is preferred.
前記抗原としては、本発明が測定対象とする新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に対する抗原であれば限定されないが、例えば、新型コロナウイルスＳ（スパイク）タンパク質、Ｅ（エンベロープ）タンパク質、Ｍ（膜）タンパク質、およびＮ（ヌクレオカプシド）タンパク質が挙げられる。
The antigen is not limited as long as it is an antigen against the new coronavirus (SARS-CoV-2) IgM antibody to be measured by the present invention, but for example, the new coronavirus S (spike) protein, E (envelope) protein, and the like. Examples include M (membrane) protein and N (nucleocapsid) protein.
抗原として組換え新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）タンパク質を用いる場合、組換え新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）タンパク質は、例えば以下の方法で作製することができる。新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）タンパク質アミノ酸配列をコードするポリヌクレオチドを適切な発現ベクターに組み込み、これを適切な宿主に挿入して形質転換し、この形質転換細胞の破砕物から目的とする組換え新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）タンパク質を得ることができる。上記宿主細胞は特に限定されず、従来から遺伝子工学的手法で用いられている各種の宿主細胞、例えば大腸菌、枯草菌、酵母、植物又は動物細胞などを用いることができる。
When a recombinant new coronavirus (SARS-CoV-2) protein is used as an antigen, the recombinant new coronavirus (SARS-CoV-2) protein can be produced, for example, by the following method. A polynucleotide encoding a new coronavirus (SARS-CoV-2) protein amino acid sequence is incorporated into an appropriate expression vector, inserted into an appropriate host for transformation, and targeted from disrupted cells of this transformed cell. A new recombinant coronavirus (SARS-CoV-2) protein can be obtained. The host cell is not particularly limited, and various host cells conventionally used in genetic engineering techniques such as Escherichia coli, bacillus, yeast, plant or animal cell can be used.
新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）タンパク質は、上記形質転換体から産生される組換えタンパク質の他、これを産生する天然の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）から自体公知のタンパク質分離精製技術により単離又は精製されるものであってもよい。また、化学合成若しくは無細胞翻訳系で生化学的に合成されたタンパク質であってもよい。
The new coronavirus (SARS-CoV-2) protein is a protein separation and purification known from the recombinant protein produced from the above transformant and the natural new coronavirus (SARS-CoV-2) that produces the recombinant protein. It may be isolated or purified by technique. Further, the protein may be chemically synthesized or biochemically synthesized by a cell-free translation system.
一態様において、上記「抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体」には、該ＩｇＭ抗体を産生した動物と同種の動物の抗ＩｇＭ抗体が含まれる。
In one embodiment, the above-mentioned "anti-new coronavirus (SARS-CoV-2) IgM antibody" includes an anti-IgM antibody of an animal of the same species as the animal that produced the IgM antibody.
抗体は、任意のアイソタイプ、例、ＩｇＡ、ＩｇＤ、ＩｇＥ、ＩｇＧ（例、ＩｇＧ１、ＩｇＧ２、ＩｇＧ３、又はＩｇＧ４）、ＩｇＭ等であり得る。抗体は、ポリクローナル抗体、モノクローナル抗体の他、キメラ抗体、単鎖抗体であり得る。また、Ｆａｂ断片やＦａｂ発現ライブラリーによって生成される断片等のように抗原結合性を有する抗体の一部（抗体断片）であり得る。抗体断片の例としては、（ｉ）ＶＬ、ＶＨ、ＣＬ及びＣＨ１ドメインからなる１価の断片であるＦａｂ断片、（ｉｉ）ヒンジ領域でジスルフィド架橋によって結びついた２つのＦａｂ断片を含む２価の断片であるＦ（ａｂ’）２断片、（ｉｉｉ）抗体のシングルアームのＶＬ及びＶＨドメインからなるＦｖ断片、（ｉｖ）温和な還元条件を用いてＦ（ａｂ’）２断片のジスルフィド架橋を破壊して得られるＦａｂ’断片、（ｖ）ジスルフィドで安定化したＦｖ断片（ｄｓＦｖ）及び（ｖｉ）抗原に特異的に結合する抗体の単一な可変領域ドメイン（ＶＨ若しくはＶＬ）ポリペプチドであるドメイン抗体（ｄＡｂ）が挙げられるが、これらに限定されない。
The antibody can be any isotype, eg, IgA, IgD, IgE, IgG (eg, IgG1, IgG2, IgG3, or IgG4), IgM, and the like. The antibody may be a monoclonal antibody, a monoclonal antibody, a chimeric antibody, or a single chain antibody. Further, it may be a part of an antibody having antigen-binding property (antibody fragment) such as a Fab fragment or a fragment produced by a Fab expression library. Examples of antibody fragments are (i) Fab fragments, which are monovalent fragments consisting of VL, VH, CL and CH1 domains, and (ii) divalent fragments containing two Fab fragments linked by disulfide bridges in the hinge region. F (ab') 2 fragment, (iii) Fv fragment consisting of VL and VH domain of single arm of antibody, (iv) disrupt disulfide bridge of F (ab') 2 fragment using mild reduction conditions. A domain antibody that is a single variable region domain (VH or VL) polypeptide of an antibody that specifically binds to the Fab'fragment, (v) disulfide-stabilized Fv fragment (dsFv) and (vi) antigen. (DAb), but is not limited to these.
本発明の方法で用いる抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体は、マウス、モルモット、ハムスター、ヤギ又はウサギで作製したものであり得る。また、抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体は市販のものを用いることができる。
The anti-new coronavirus (SARS-CoV-2) IgM antibody used in the method of the present invention may be produced in mice, guinea pigs, hamsters, goats or rabbits. Further, as the anti-new coronavirus (SARS-CoV-2) IgM antibody, a commercially available one can be used.
本発明の検査用キットは、上記本発明のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を別々の容器に含む。
The test kit of the present invention contains the above-mentioned sensitizer for the immunochromatography method of the present invention and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody in separate containers.
更に本発明の検査用キットは、本発明のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査に使用することができる、又は使用すべきであることを記載した説明書を含んでいてもよい。
Further, the test kit of the present invention contains a sensitizer for the immunochromatography method of the present invention and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody to a new coronavirus infection (COVID-19). ) May include instructions stating that they can or should be used for inspection.
上記新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キットは、任意の担体、例えば医薬上許容され得る担体、安定剤や緩衝液成分、他の治療薬やサプリメント等を含み得る。医薬上許容され得る担体としては、例えば、水、生理食塩水等の希釈剤等が挙げられるが、それらに限定されるものではない。また、本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キットは、検体希釈液、イムノクロマト測定用テストストリップ、イムノクロマト測定用器具を含み得る。
The test kit for the new coronavirus infection (COVID-19) may include any carrier, such as a pharmaceutically acceptable carrier, stabilizers and buffer components, other therapeutic agents and supplements. Examples of the pharmaceutically acceptable carrier include, but are not limited to, diluents such as water and physiological saline. Further, the kit for testing a new type coronavirus infection (COVID-19) of the present invention may include a sample diluent, a test strip for measuring an immunochromatography, and an instrument for measuring an immunochromatography.
本発明は、本発明のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行うことにより、被験体由来の生物学的試料中の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定することを含む、該被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染している可能性を試験するための方法を提供する。
The present invention measures a novel coronavirus (SARS-CoV-2) IgM antibody in a biological sample derived from a subject by performing an antigen-antibody reaction in the presence of the sensitizer for the immunochromatography method of the present invention. Provided is a method for testing the possibility that the subject is infected with a new type of coronavirus infection (COVID-19), including the above.
当該方法において用いることのできる生物学的試料としては、血液が挙げられるが、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を検出できる限り、特に限定されない。「血液」としては、いかなる組織由来の血液も想定することができるが、採取の容易から、通常は末梢血が用いられる。血液の採取方法としては、自体公知の方法が適用できる。また採取した血液はそのまま本方法に用いてもよいが、自体公知の方法、例えば遠心分離、濾過などを利用して細胞成分（赤血球、白血球、血小板など）を分離した液体成分（血漿）として本方法に用いてもよい。また血液を凝固させて血小板や凝固因子を分離した液体成分（血清）として本方法に用いることも可能である。
Examples of the biological sample that can be used in the method include blood, but the biological sample is not particularly limited as long as the new coronavirus (SARS-CoV-2) IgM antibody can be detected. As the "blood", blood derived from any tissue can be assumed, but peripheral blood is usually used because it is easy to collect. As a method for collecting blood, a method known per se can be applied. The collected blood may be used as it is in this method, but it is used as a liquid component (plasma) in which cellular components (erythrocytes, leukocytes, platelets, etc.) are separated by a method known per se, for example, centrifugation, filtration, etc. It may be used in the method. It can also be used in this method as a liquid component (serum) in which blood is coagulated and platelets and coagulation factors are separated.
本発明の検査方法を行うに際しては、例えば、血液等の被験体の生物学的試料を抽出液に入れ懸濁し、抗原を生物学的試料液から抽出する。そして、例えば上記（４）のイムノクロマト測定用器具を用いる場合、検体滴下部に抽出液で処理された生物学的試料液を滴下する。そうすることで、本発明のイムノクロマト測定法用増感剤を含む、イムノクロマト測定法用検体希釈液の存在下生物学的試料液は毛細管現象により検体滴下部から検体標識部へ、そしてメンブレン（膜）上に展開される。
When performing the test method of the present invention, for example, a biological sample of a subject such as blood is placed in an extract and suspended, and an antigen is extracted from the biological sample solution. Then, for example, when the device for measuring the immunochromatography of (4) above is used, the biological sample solution treated with the extract is dropped onto the sample dropping portion. By doing so, the biological sample solution in the presence of the sample diluent for the immunochromatography method containing the sensitizer for the immunochromatography method of the present invention moves from the sample dropping part to the sample labeling part and the membrane (membrane) due to the capillary phenomenon. ) Expanded on.
生物学的試料液中の標的抗原（例、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体）は、検体標識部を通過する際に検体標識部に担持された標識物質（例、抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体）と免疫複合体を形成し、そのまま展開膜の一端へと展開される。そして、当該複合体と展開膜のテストライン上に存在する（テストライン上にコーティングされた）物質（例、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に対する抗原）とが免疫化学反応を起こし結合し、呈色することにより視認可能なラインが展開膜上に現れる。このように呈色した場合は、当該検体は陽性であり、被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染していると判定することができる。また、その呈色度合いを目視や適当な分析機器（例えば、イムノクロマトリーダー、分光光度計）で測ることにより当該陽性の強度を判定することができる。なお、標識抗体が有する検出手段として、例えば蛍光色素を用いた場合には、テストラインに紫外線等を照射し、それによる呈色の有無やその程度により陽性か否かを判定することができる。
The target antigen in the biological sample solution (eg, new coronavirus (SARS-CoV-2) IgM antibody) is a labeling substance carried on the sample labeling part as it passes through the sample labeling part (eg, anti-new corona). It forms an immune complex with a virus (SARS-CoV-2) IgM antibody) and is directly expanded to one end of the developing membrane. Then, the complex and a substance (eg, an antigen against the new coronavirus (SARS-CoV-2) IgM antibody) existing on the test line of the developing membrane (coated on the test line) cause an immunochemical reaction. Visible lines appear on the developing membrane by combining and coloring. When the color is developed in this way, the sample is positive, and it can be determined that the subject is infected with the new coronavirus infection (COVID-19). Further, the intensity of the positive can be determined by visually measuring the degree of color development or by measuring it with an appropriate analytical instrument (for example, an immunochromatographic reader or a spectrophotometer). When, for example, a fluorescent dye is used as the detection means of the labeled antibody, it is possible to irradiate the test line with ultraviolet rays or the like and determine whether or not the test line is positive or not depending on the presence or absence of color development and the degree thereof.
一方、テストラインの後にはコントロールラインがあり、そこに存在する（そこにコーティングされた）抗体と免疫複合体未形成の標識物質（例、抗新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体とが免疫化学反応を起こし結合し呈色することにより、イムノクロマトが問題なく行われたことを視認することができる。
On the other hand, there is a control line after the test line, with the antibody present (coated there) and the immunocomplex-unformed label substance (eg, anti-new coronavirus (SARS-CoV-2) IgM antibody). By causing an immunochemical reaction, binding and coloring, it can be visually confirmed that the immunochromatography was performed without any problem.
検体サンプル液が展開膜上を展開通過した後は、吸収パッドにより未反応の標識抗体等が吸収される。なお、検体中に標的抗原（例、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体）が含まれていなかった場合は、テストラインは呈色せず、コントロールラインのみが呈色することになる。この場合、当該検体は陰性であり、被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染しているとは判定されない。
After the sample sample solution has expanded and passed over the developing membrane, the unreacted labeled antibody or the like is absorbed by the absorption pad. If the sample does not contain a target antigen (eg, new coronavirus (SARS-CoV-2) IgM antibody), the test line will not be colored and only the control line will be colored. .. In this case, the sample is negative and it is not determined that the subject is infected with the new coronavirus infection (COVID-19).
本発明の、試験するための方法に供される生物学的試料は、哺乳動物の被験体に由来する試料である。当該哺乳動物としては新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染する可能性がある哺乳動物であれば特に限定されないが、なかでも、マウス、ラット、ハムスター、モルモット等のげっ歯類やウサギ等の実験動物、ブタ、ウシ、ヤギ、ウマ、ヒツジ、ミンク等の家畜、イヌ、ネコ等のペット、ヒト、サル、カニクイザル、アカゲザル、マーモセット、オランウータン、チンパンジーなどの霊長類が好ましく、特にヒトが好ましい。
The biological sample used in the method for testing of the present invention is a sample derived from a mammalian subject. The mammal is not particularly limited as long as it is a mammal that may be infected with the new coronavirus infection (COVID-19), but among them, rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like. Experimental animals, pigs, cows, goats, horses, sheep, minks and other domestic animals, dogs, cats and other pets, humans, monkeys, crab monkeys, red-tailed monkeys, marmosets, orangutans, chimpanzees and other primates are preferred, and humans are particularly preferred. ..
本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法は、少なくとも以下の（１）及び（２）の工程を含む。
（１）前記共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程。
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を患者に施す工程。 The method for treating a new type coronavirus infection (COVID-19) of the present invention includes at least the following steps (1) and (2).
(1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
(2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
（１）前記共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程。
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を患者に施す工程。 The method for treating a new type coronavirus infection (COVID-19) of the present invention includes at least the following steps (1) and (2).
(1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
(2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
なお、患者は、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）に感染しているおそれのある人も含む。なお、測定とは、患者由来の検体中に新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体が存在するかどうかを検出することを意味する。
Patients include those who may be infected with the new coronavirus (SARS-CoV-2). The measurement means detecting the presence or absence of the new coronavirus (SARS-CoV-2) IgM antibody in the sample derived from the patient.
また、新型コロナウイルス感染症治療を患者に施す工程とは、公知治療薬、今後上市される治療薬、臨床段階の治療薬候補を患者に投与するだけではなく、対症療法（アスピリン投与、補液・輸液投与等）も含む。
In addition, the process of applying new coronavirus infection treatment to patients is not only to administer known therapeutic agents, therapeutic agents to be launched in the future, and clinical stage therapeutic agent candidates to patients, but also symptomatic treatment (aspirin administration, fluid replacement, etc.). Infusion administration, etc.) is also included.
本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法は、高感度で新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を検出することができるので、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の患者に迅速に新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を施すことができる。
The method for treating a new coronavirus infection (COVID-19) of the present invention can detect a new coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, and thus can detect a new coronavirus infection (COVID-19). ) Can be rapidly treated with the new coronavirus infection (COVID-19).
新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の症状は、無症状から重症の肺炎、死亡まで多岐にわたるが、具体的には、発熱、空咳、倦怠感・だるさ、痰、嗅覚障害・味覚障害、息切れ、筋肉痛・関節痛、のどの痛み、頭痛、悪寒、吐き気・嘔吐、鼻づまり、下痢、喀血、結膜充血等が挙げられる。
Symptoms of the new coronavirus infection (COVID-19) range from asymptomatic to severe pneumonia and death. Specifically, fever, chills, malaise / dullness, sputum, olfactory / taste disorders, and shortness of breath , Muscle pain / joint pain, sore throat, headache, chills, nausea / vomiting, stuffy nose, diarrhea, hemoptysis, conjunctival hyperemia, etc.
別の態様において、本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法は、少なくとも以下の（１）及び（２）の工程を含む。
（１）前記共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程。
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を、ヒトを除く被験体に施す工程。 In another embodiment, the method for treating a novel coronavirus infection (COVID-19) of the present invention comprises at least the following steps (1) and (2).
(1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
(2) A step of applying a new coronavirus infection (COVID-19) treatment to a subject other than a human based on the measurement result of the above (1).
（１）前記共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程。
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を、ヒトを除く被験体に施す工程。 In another embodiment, the method for treating a novel coronavirus infection (COVID-19) of the present invention comprises at least the following steps (1) and (2).
(1) A step of measuring a new type of coronavirus (SARS-CoV-2) in which an antigen-antibody reaction is carried out in the presence of a sensitizer for an immunochromatographic measurement method containing the copolymer.
(2) A step of applying a new coronavirus infection (COVID-19) treatment to a subject other than a human based on the measurement result of the above (1).
本発明の治療方法に供されるヒトを除く被験体は、限定されないが、好ましくは哺乳動物を意味する。更に好ましくは、当該哺乳動物としては新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染する可能性がある哺乳動物が挙げられ、なかでも、マウス、ラット、ハムスター、モルモット等のげっ歯類やウサギ等の実験動物、ブタ、ウシ、ヤギ、ウマ、ヒツジ、ミンク等の家畜、イヌ、ネコ等のペット、サル、カニクイザル、アカゲザル、マーモセット、オランウータン、チンパンジーなどの霊長類が好ましい。
Subjects other than humans used in the therapeutic method of the present invention are not limited, but preferably mean mammals. More preferably, the mammals include mammals that may be infected with the new coronavirus infection (COVID-19), and among them, rodents such as mice, rats, hamsters and guinea pigs, rabbits and the like. Experimental animals, pigs, cows, goats, horses, sheep, minks and other domestic animals, dogs, cats and other pets, monkeys, crab monkeys, lizard monkeys, guinea pigs, orchid utans, chimpanzees and other primates are preferred.
本発明は、上記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キットを提供する。
The present invention includes a new coronavirus infection, which comprises a sensitizer for an immunochromatography method containing a copolymer represented by the above general formula [1] and a therapeutic agent for a new coronavirus infection (COVID-19). A therapeutic kit for (COVID-19) is provided.
該治療剤としては、例えば新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療薬を含むものが挙げられ、該治療薬としては、限定されないが、レムデシビル、デキサメタゾン、ファビピラビル（アビガン）、シクレソニド、ナファモスタット、カモスタット、イベルメクチン、ネルフィナビル等が挙げられる。治療薬は、重症肺炎や急性呼吸窮迫症候群の治療薬であるトシリズマブ、バリシチニブ、アカラブルチニブ、ラブリズマブ、エリトラン、イブジラスト、ＬＹ３１２７８０４、オチリマブ、HLCM051、ADR-001、イカチバント、アプレミラスト、ｃｅｎｉｃｒｉｖｉｒｏｃであってもよい。ウイルスのスパイクタンパク質の受容体結合部位に非競合的に結合することで、ＳＡＲＳ－ＣｏＶ－２に対して中和活性を示すウイルス中和抗体であるカシリビマブ、イムデビマブ及び／又はそれらの混合物であってもよい。今後上市される治療薬、臨床段階の治療薬候補であってもよい。治療剤は、対症療法に用いられるアスピリン、補液・輸液等を含むものであってもよい。
Examples of the therapeutic agent include those containing a therapeutic agent for coronavirus infection (COVID-19), and the therapeutic agent includes, but is not limited to, remdesivir, dexamethasone, favipiravir (Abigan), cyclesonide, and nafamostat. , Camostat, Ibermectin, Favipiravir, etc. Therapeutic agents are tocilizumab, baricitinib, acarabrutinib, labrizumab, eritoran, ibzilast, LY3127804, ochirimab, HLCM051, ADR-001, squid, apremilast, and cenicriviro, which are therapeutic agents for severe pneumonia and acute respiratory distress syndrome. A virus-neutralizing antibody that exhibits neutralizing activity against SARS-CoV-2 by binding non-competitively to the receptor binding site of the viral spike protein, such as regibimab, imdevimab, and / or a mixture thereof. May be good. It may be a therapeutic drug to be launched in the future or a candidate for a therapeutic drug in the clinical stage. The therapeutic agent may include aspirin, fluid replacement, infusion and the like used for symptomatic treatment.
本発明の治療用キットは、上記本発明のイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を別々の容器に含む。
The therapeutic kit of the present invention contains the above-mentioned sensitizer for the immunochromatographic measurement method of the present invention and the therapeutic agent for the new coronavirus infection (COVID-19) in separate containers.
更に本発明の治療用キットは、本発明のイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療に使用することができる、又は使用すべきであることを記載した説明書を含んでいてもよい。
Further, the therapeutic kit of the present invention uses the sensitizer for the immunochromatography method of the present invention and the therapeutic agent for the new coronavirus infection (COVID-19) for the treatment of the new coronavirus infection (COVID-19). It may include instructions stating that it can or should be used.
上記新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キットは、任意の担体、例えば医薬上許容され得る担体、安定剤や緩衝液成分、他の治療薬やサプリメント等を含み得る。医薬上許容され得る担体としては、例えば、水、生理食塩水等の希釈剤等が挙げられるが、それらに限定されるものではない。また、本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キットは、検体希釈液、イムノクロマト測定用テストストリップ、イムノクロマト測定用器具を含み得る。
The therapeutic kit for the new coronavirus infection (COVID-19) may include any carrier, such as a pharmaceutically acceptable carrier, stabilizers and buffer components, other therapeutic agents and supplements. Examples of the pharmaceutically acceptable carrier include, but are not limited to, diluents such as water and physiological saline. In addition, the kit for treating a new type of coronavirus infection (COVID-19) of the present invention may include a sample diluent, a test strip for immunochromatography measurement, and an instrument for immunochromatography measurement.
本発明の治療用キットは、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を更に含んでもよい。
The therapeutic kit of the present invention may further contain a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody.
本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キットは、高感度で新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を検出することができるので、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の被験体（患者）に迅速に新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を施すことができる。
Since the treatment kit for the new coronavirus infection (COVID-19) of the present invention can detect the new coronavirus (SARS-CoV-2) IgM antibody with high sensitivity, the new coronavirus infection (COVID-) 19) The subject (patient) can be rapidly treated with the new coronavirus infection (COVID-19).
以下、本実施例により本発明をさらに詳細に説明するが、本発明はこれらに限定されない。
Hereinafter, the present invention will be described in more detail with reference to the present examples, but the present invention is not limited thereto.
製造例
（本発明及び比較例の共重合体の合成）
本発明のイムノクロマト測定法用増感剤として用いられる共重合体及び比較例の共重合体を合成した。詳細は、以下の通りである。 Production Example (Synthesis of Copolymers of the Present Invention and Comparative Examples)
A copolymer used as a sensitizer for the immunochromatographic measurement method of the present invention and a copolymer of a comparative example were synthesized. The details are as follows.
（本発明及び比較例の共重合体の合成）
本発明のイムノクロマト測定法用増感剤として用いられる共重合体及び比較例の共重合体を合成した。詳細は、以下の通りである。 Production Example (Synthesis of Copolymers of the Present Invention and Comparative Examples)
A copolymer used as a sensitizer for the immunochromatographic measurement method of the present invention and a copolymer of a comparative example were synthesized. The details are as follows.
本発明及び比較例で使用する共重合体は、２－（メタクリロイルオキシ）エチル ２’－（トリメチルアンモニオ）エチル ホスフェート（ＭＰＣ）とメタクリル酸から合成したランダム共重合体である。
（共重合体１）
重量平均分子量；２２０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；９０：１０
（共重合体２）
重量平均分子量；５５０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；７０：３０
（共重合体３）
重量平均分子量；１１００×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；５０：５０
（共重合体４）
重量平均分子量；６８０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；３０：７０ The copolymer used in the present invention and Comparative Examples is a random copolymer synthesized from 2- (methacryloyloxy) ethyl 2'-(trimethylammonio) ethyl phosphate (MPC) and methacrylic acid.
(Copolymer 1)
Weight average molecular weight; 220 × 103, ratio of number of building blocks based on MPC monomer to number of building blocks based on methacrylic acid; 90:10
(Copolymer 2)
Weight average molecular weight; 550 × 10 3 , ratio of number of constituent units based on MPC monomer to number of constituent units based on methacrylic acid; 70:30
(Copolymer 3)
Weight average molecular weight; 1100 × 103, ratio of number of building blocks based on MPC monomer to number of building blocks based on methacrylic acid; 50:50
(Copolymer 4)
Weight average molecular weight; 680 × 103, ratio of number of constituent units based on MPC monomer to number of constituent units based on methacrylic acid; 30:70
（共重合体１）
重量平均分子量；２２０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；９０：１０
（共重合体２）
重量平均分子量；５５０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；７０：３０
（共重合体３）
重量平均分子量；１１００×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；５０：５０
（共重合体４）
重量平均分子量；６８０×１０３、ＭＰＣ単量体に基づく構成単位数とメタクリル酸に基づく構成単位数の比；３０：７０ The copolymer used in the present invention and Comparative Examples is a random copolymer synthesized from 2- (methacryloyloxy) ethyl 2'-(trimethylammonio) ethyl phosphate (MPC) and methacrylic acid.
(Copolymer 1)
Weight average molecular weight; 220 × 103, ratio of number of building blocks based on MPC monomer to number of building blocks based on methacrylic acid; 90:10
(Copolymer 2)
Weight average molecular weight; 550 × 10 3 , ratio of number of constituent units based on MPC monomer to number of constituent units based on methacrylic acid; 70:30
(Copolymer 3)
Weight average molecular weight; 1100 × 103, ratio of number of building blocks based on MPC monomer to number of building blocks based on methacrylic acid; 50:50
(Copolymer 4)
Weight average molecular weight; 680 × 103, ratio of number of constituent units based on MPC monomer to number of constituent units based on methacrylic acid; 30:70
実施例
（公知のイムノクロマト測定用器具を使用しての検出感度の確認）
本実施例では、上記の共重合体が、公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体の検出キットの検出感度を向上させることができるか（増感効果があるか）どうかを確認した。詳細は、以下の通りである。 Example (confirmation of detection sensitivity using a known device for measuring an immunochromatography)
In this example, it is confirmed whether the above-mentioned copolymer can improve the detection sensitivity of the known new coronavirus (SARS-CoV-2) IgM antibody detection kit (whether it has a sensitizing effect). bottom. The details are as follows.
（公知のイムノクロマト測定用器具を使用しての検出感度の確認）
本実施例では、上記の共重合体が、公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体の検出キットの検出感度を向上させることができるか（増感効果があるか）どうかを確認した。詳細は、以下の通りである。 Example (confirmation of detection sensitivity using a known device for measuring an immunochromatography)
In this example, it is confirmed whether the above-mentioned copolymer can improve the detection sensitivity of the known new coronavirus (SARS-CoV-2) IgM antibody detection kit (whether it has a sensitizing effect). bottom. The details are as follows.
（比較例）
市販の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キット（ＲａｙＢｉｏｔｅｃ．社製）で評価した。より詳しくは、ＳＡＲＳ－ＣｏＶ－２ ＩｇＭ抗体検出キットの希釈液 ２４５μＬに新型コロナウイルス陽性患者血清（ＣｏＶ－ＰｏｓＭ－Ｓ－１００、ＲａｙＢｉｏｔｅｃ．社製） ２５μＬを添加し、希釈液をイムノクロマトキットに展開した。 (Comparative example)
It was evaluated with a commercially available new coronavirus (SARS-CoV-2) IgM antibody detection kit (manufactured by RayBiotec.). More specifically, 25 μL of new coronavirus-positive patient serum (CoV-PosM-S-100, manufactured by RayBiotec.) Was added to 245 μL of the diluted solution of the SARS-CoV-2 IgM antibody detection kit, and the diluted solution was developed into an immunochromatographic kit. bottom.
市販の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キット（ＲａｙＢｉｏｔｅｃ．社製）で評価した。より詳しくは、ＳＡＲＳ－ＣｏＶ－２ ＩｇＭ抗体検出キットの希釈液 ２４５μＬに新型コロナウイルス陽性患者血清（ＣｏＶ－ＰｏｓＭ－Ｓ－１００、ＲａｙＢｉｏｔｅｃ．社製） ２５μＬを添加し、希釈液をイムノクロマトキットに展開した。 (Comparative example)
It was evaluated with a commercially available new coronavirus (SARS-CoV-2) IgM antibody detection kit (manufactured by RayBiotec.). More specifically, 25 μL of new coronavirus-positive patient serum (CoV-PosM-S-100, manufactured by RayBiotec.) Was added to 245 μL of the diluted solution of the SARS-CoV-2 IgM antibody detection kit, and the diluted solution was developed into an immunochromatographic kit. bottom.
（実施例１）
当該キットの希釈液に終濃度０．５重量％となるように共重合体１、２、３、４を添加し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 1)
Copolymers 1, 2, 3 and 4 are added to the diluted solution of the kit so as to have a final concentration of 0.5% by weight, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example. The solutions prepared as described above were developed and the detection sensitivities were compared.
当該キットの希釈液に終濃度０．５重量％となるように共重合体１、２、３、４を添加し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 1)
Copolymers 1, 2, 3 and 4 are added to the diluted solution of the kit so as to have a final concentration of 0.5% by weight, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example. The solutions prepared as described above were developed and the detection sensitivities were compared.
（実施例２）
当該キットのサンプルパッドに、共重合体１、２、３、４を担持量５ｍｇとなるように担持し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 2)
Copolymers 1, 2, 3 and 4 are supported on the sample pad of the kit so as to have a loading amount of 5 mg, and the serum of a new coronavirus-positive patient is prepared to be diluted 100-fold with respect to the comparative example. The prepared solutions were developed and the detection sensitivities were compared.
当該キットのサンプルパッドに、共重合体１、２、３、４を担持量５ｍｇとなるように担持し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 2)
Copolymers 1, 2, 3 and 4 are supported on the sample pad of the kit so as to have a loading amount of 5 mg, and the serum of a new coronavirus-positive patient is prepared to be diluted 100-fold with respect to the comparative example. The prepared solutions were developed and the detection sensitivities were compared.
（実施例３）
当該キットのコンジュゲートパッドに、共重合体１、２、３、４を担持量５００μｇとなるように担持し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 3)
The copolymers 1, 2, 3 and 4 are supported on the conjugate pad of the kit so that the loading amount is 500 μg, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example. The prepared solutions were developed and the detection sensitivities were compared.
当該キットのコンジュゲートパッドに、共重合体１、２、３、４を担持量５００μｇとなるように担持し、さらに、新型コロナウイルス陽性患者血清が比較例に対して１００倍希釈になるように調製した溶液を展開し、検出感度を比較した。 (Example 3)
The copolymers 1, 2, 3 and 4 are supported on the conjugate pad of the kit so that the loading amount is 500 μg, and the serum of the new coronavirus-positive patient is diluted 100-fold with respect to the comparative example. The prepared solutions were developed and the detection sensitivities were compared.
（イムノクロマト測定用器具を使用しての検出感度の確認結果）
公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キットを使用しての、希釈液に共重合体を添加した際の検出感度向上の結果を表１に示す。 (Confirmation result of detection sensitivity using an instrument for immunochromatography measurement)
Table 1 shows the results of improving the detection sensitivity when the copolymer was added to the diluted solution using the known novel coronavirus (SARS-CoV-2) IgM antibody detection kit.
公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キットを使用しての、希釈液に共重合体を添加した際の検出感度向上の結果を表１に示す。 (Confirmation result of detection sensitivity using an instrument for immunochromatography measurement)
Table 1 shows the results of improving the detection sensitivity when the copolymer was added to the diluted solution using the known novel coronavirus (SARS-CoV-2) IgM antibody detection kit.
０．５重量％の共重合体１、２、３、４を添加した場合、ＩｇＭについて、比較例であるコントロールと同等以上か、やや薄い濃さのテストラインが検出できたことから、共重合体１、２、３、４の添加により、感度が向上していることが確認された。
When 0.5% by weight of the copolymers 1, 2, 3, and 4 were added, a test line having a concentration equal to or higher than that of the control as a comparative example or a slightly lighter density could be detected for IgM. It was confirmed that the sensitivity was improved by the addition of the coalesced 1, 2, 3 and 4.
公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キットを使用しての、サンプルパッドに重合体を担持した際の検出感度向上の結果を表２に示す。
Table 2 shows the results of improving the detection sensitivity when the polymer is supported on the sample pad using the known new coronavirus (SARS-CoV-2) IgM antibody detection kit.
共重合体１、２、３、４をサンプルパッドに担持した場合、ＩｇＭについて、比較例であるコントロールと同等以上か、やや薄い濃さのテストラインが検出できたことから、共重合体１、２、３、４のサンプルパッドへの担持により、感度が向上していることが確認された。
When the copolymers 1, 2, 3 and 4 were supported on the sample pad, a test line having a density equal to or higher than that of the control as a comparative example or a slightly lighter density could be detected for IgM. It was confirmed that the sensitivity was improved by supporting the sample pads of 2, 3 and 4.
公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体検出キットを使用しての、コンジュゲートパッドに重合体を担持した際の検出感度向上の結果を表３に示す。
Table 3 shows the results of improving the detection sensitivity when the polymer is supported on the conjugate pad using the known new coronavirus (SARS-CoV-2) IgM antibody detection kit.
共重合体１、２、３、４をコンジュゲートパッドに担持した場合、ＩｇＭについて、比較例であるコントロールと同等以上か、やや薄い濃さのテストラインが検出できたことから、共重合体１、２、３、４のコンジュゲートパッドへの担持により、感度が向上していることが確認された。
When the copolymers 1, 2, 3, and 4 were supported on the conjugate pad, a test line having a density equal to or higher than that of the control as a comparative example or a slightly lighter density could be detected for IgM, so that the copolymer 1 was detected. It was confirmed that the sensitivity was improved by supporting the conjugate pads of 2, 3 and 4.
以上の実施例より、本発明は、公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法と比較して、著しく増感効果が優れているイムノクロマト測定法を提供することができた。
From the above examples, the present invention provides an immunochromatographic measurement method having a significantly superior sensitizing effect as compared with an immunochromatographic measurement method using a known novel coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance. I was able to provide it.
以上述べたことから明らかなように、本発明は、公知の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法と比較して、著しく増感効果が優れているイムノクロマト測定法を提供することができた。
As is clear from the above, the present invention is remarkably superior in sensitizing effect as compared with the known pharmacochromatographic measurement method using the known novel coronavirus (SARS-CoV-2) IgM antibody as the measurement target substance. An immunochromatographic measurement method could be provided.
さらに、本発明の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法は、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）を高感度に検出することができるので、新型コロナウイルス感染症の患者又は新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）に感染しているおそれのある患者に迅速に新型コロナウイルス感染症治療を施すことができるので治療効果が高い。
Furthermore, the method for treating a new coronavirus infection (COVID-19) of the present invention can detect a new coronavirus (SARS-CoV-2) with high sensitivity, so that a patient with a new coronavirus infection or a new type can be detected. Since it is possible to quickly treat a patient who may be infected with coronavirus (SARS-CoV-2) with a new type of coronavirus infection, the therapeutic effect is high.
本発明は、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤を提供することができる。
The present invention can provide a sensitizer for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance.
本出願は日本で出願された特願２０２０－１８１４８４（出願日：２０２０年１０月２９日）を基礎としており、その内容は本明細書に全て包含されるものである。
This application is based on Japanese Patent Application No. 2020-181484 (Filing date: October 29, 2020), the contents of which are all included in the present specification.
Claims (12)
- 下記一般式〔１〕で表される共重合体を含んでなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
- 前記共重合体は、下記一般式〔２〕で表される単量体に基づく構成単位と、メタクリル酸またはその塩に基づく構成単位を有する共重合体である請求項１に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。
- 前記共重合体が、一般式〔２〕で表される単量体に基づく構成単位数とメタクリル酸またはその塩に基づく構成単位数の比（ｍ：ｎ）が９９～２０：１～８０の共重合体である請求項１又は請求項２に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 99 to 20: 1 to 80. A sensitizer for an immunochromatographic measurement method using the novel coronavirus (SARS-CoV-2) IgM antibody according to claim 1 or 2, which is a copolymer, as a measurement target substance.
- 前記共重合体が、一般式〔２〕で表される単量体に基づく構成単位数とメタクリル酸またはその塩に基づく構成単位数の比（ｍ：ｎ）が９０～３０：１０～７０の共重合体である請求項１又は請求項２に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 The ratio (m: n) of the number of structural units based on the monomer represented by the general formula [2] to the number of structural units based on methacrylic acid or a salt thereof (m: n) of the copolymer is 90 to 30:10 to 70. A sensitizer for an immunochromatographic measurement method using the novel coronavirus (SARS-CoV-2) IgM antibody according to claim 1 or 2, which is a copolymer, as a measurement target substance.
- 前記共重合体が、重量平均分子量１０，０００～５，０００，０００の水溶性共重合体である請求項１～４のいずれか１項に記載の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用増感剤。 The new coronavirus (SARS-CoV-2) IgM according to any one of claims 1 to 4, wherein the copolymer is a water-soluble copolymer having a weight average molecular weight of 10,000 to 5,000,000. A sensitizer for immunochromatography, which uses an antibody as a measurement target substance.
- 請求項１～５のいずれか１項に記載のイムノクロマト測定法用増感剤を含む、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法用検体希釈液。 A sample diluted solution for an immunochromatographic measurement method containing a new type coronavirus (SARS-CoV-2) IgM antibody as a measurement target substance, which contains the sensitizer for the immunochromatographic measurement method according to any one of claims 1 to 5.
- 請求項１～５のいずれか１項に記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行わせることを特徴とする、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定法。 Measurement of a novel coronavirus (SARS-CoV-2) IgM antibody, which comprises causing an antigen-antibody reaction in the presence of the sensitizer for the later lateral flow test according to any one of claims 1 to 5. An immunochromatographic measurement method for the target substance.
- 請求項１～５のいずれか１項に記載のイムノクロマト測定法用増感剤を展開膜、サンプルパッド、又はコンジュゲートパッドに担持させてなる、新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定対象物質とするイムノクロマト測定用器具。 A novel coronavirus (SARS-CoV-2) IgM antibody obtained by carrying the sensitizer for the later lateral flow test according to any one of claims 1 to 5 on a developing membrane, a sample pad, or a conjugate pad. An instrument for measuring immunochromatography, which is the substance to be measured.
- 請求項１～５のいずれか１項に記載のイムノクロマト測定法用増感剤及び新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体に特異的に結合する物質を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の検査用キット。 A new coronavirus infection (COVID) comprising the sensitizer for the later lateral flow test according to any one of claims 1 to 5 and a substance that specifically binds to the new coronavirus (SARS-CoV-2) IgM antibody. -19) Inspection kit.
- 請求項１～５のいずれか１項に記載のイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行うことにより、被験体由来の生物学的試料中の新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）ＩｇＭ抗体を測定することを含む、該被験体が新型コロナウイルス感染症（ＣＯＶＩＤ－１９）に感染している可能性を試験するための方法。 By performing an antigen-antibody reaction in the presence of the sensitizer for the immunochromatography method according to any one of claims 1 to 5, a novel coronavirus (SARS-CoV-) in a biological sample derived from a subject is performed. 2) A method for testing the possibility that the subject is infected with Severe Acute Respiratory Syndrome (COVID-19), which comprises measuring IgM antibody.
- 以下の（１）、（２）の工程を含む新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療方法、
（１）下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤の存在下で抗原抗体反応を行う新型コロナウイルス（ＳＡＲＳ－ＣｏＶ－２）の測定工程、及び
（２）上記（１）の測定結果に基づき、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）治療を患者に施す工程。
(1) A measurement step of a new type coronavirus (SARS-CoV-2) that undergoes an antigen-antibody reaction in the presence of a sensitizer for an immunochromatographic measurement method containing a copolymer represented by the following general formula [1]. And (2) A step of treating a patient with a new type of coronavirus infection (COVID-19) based on the measurement result of (1) above.
- 下記一般式〔１〕で表される共重合体を含んでなるイムノクロマト測定法用増感剤及び新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療剤を含む、新型コロナウイルス感染症（ＣＯＶＩＤ－１９）の治療用キット。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0783923A (en) * | 1993-09-14 | 1995-03-31 | Nippon Oil & Fats Co Ltd | Protein adsorption inhibitor |
| JP2003344406A (en) * | 2002-05-24 | 2003-12-03 | Towns:Kk | Developing solvent for immuno-chromatography, measuring method and kit |
| JP2003344413A (en) * | 2002-05-24 | 2003-12-03 | Wako Pure Chem Ind Ltd | Sensitivity enhancing agent for immuno-chromatographic measuring method, measuring method and apparatus for measuring method |
| WO2005031316A1 (en) * | 2003-09-29 | 2005-04-07 | National Institute Of Advanced Industrial Science And Technology | Automatic analyzing method and analyzer |
| CN111398597A (en) * | 2020-03-12 | 2020-07-10 | 中科欧蒙未一(北京)医学技术有限公司 | Kit for detecting IgM antibody against novel coronavirus SARS-CoV-2 in sample |
-
2021
- 2021-10-21 WO PCT/JP2021/038885 patent/WO2022091923A1/en active Application Filing
- 2021-10-21 JP JP2022559065A patent/JPWO2022091923A1/ja active Pending
- 2021-10-21 TW TW110139098A patent/TW202227816A/en unknown
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0783923A (en) * | 1993-09-14 | 1995-03-31 | Nippon Oil & Fats Co Ltd | Protein adsorption inhibitor |
| JP2003344406A (en) * | 2002-05-24 | 2003-12-03 | Towns:Kk | Developing solvent for immuno-chromatography, measuring method and kit |
| JP2003344413A (en) * | 2002-05-24 | 2003-12-03 | Wako Pure Chem Ind Ltd | Sensitivity enhancing agent for immuno-chromatographic measuring method, measuring method and apparatus for measuring method |
| WO2005031316A1 (en) * | 2003-09-29 | 2005-04-07 | National Institute Of Advanced Industrial Science And Technology | Automatic analyzing method and analyzer |
| CN111398597A (en) * | 2020-03-12 | 2020-07-10 | 中科欧蒙未一(北京)医学技术有限公司 | Kit for detecting IgM antibody against novel coronavirus SARS-CoV-2 in sample |
Non-Patent Citations (2)
| Title |
|---|
| KIM JEONG-HUI, RYOSUKE KIKUCHI, ATSUO SUZUKI, RIKA WATARAI, SATORU YOKOYAMA, MASAHIRO MORISE, TETSUYA YAGI, TADASHI MATSUSHITA: "Comparative Study of anti-SARS-CoV-2 antibody testing: Relationship between target antigen and immunoglobulin isotype", IGAKU KENSA - JAPANESE JOURNAL OF MEDICAL TECHNOLOGY, NIHON RINSHO EISEI KENSA GISHIKAI, TOKYO,, JP, vol. 69, no. 4, 25 October 2020 (2020-10-25), JP , pages 554 - 561, XP055926574, ISSN: 0915-8669, DOI: 10.14932/jamt.20-71 * |
| SILVEIRA MARIâNGELA F.; BARROS ALUíSIO J. D.; HORTA BERNARDO L.; PELLANDA LúCIA C.; VICTORA GABRIEL D.; DELLAGOSTIN: "Population-based surveys of antibodies against SARS-CoV-2 in Southern Brazil", NATURE MEDICINE, NATURE PUBLISHING GROUP US, NEW YORK, vol. 26, no. 8, 8 July 2020 (2020-07-08), New York, pages 1196 - 1199, XP037215683, ISSN: 1078-8956, DOI: 10.1038/s41591-020-0992-3 * |
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| Publication number | Publication date |
|---|---|
| TW202227816A (en) | 2022-07-16 |
| JPWO2022091923A1 (en) | 2022-05-05 |
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