CN117043601A - Sensitizer for immunochromatographic assay and assay - Google Patents
Sensitizer for immunochromatographic assay and assay Download PDFInfo
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- CN117043601A CN117043601A CN202180083318.9A CN202180083318A CN117043601A CN 117043601 A CN117043601 A CN 117043601A CN 202180083318 A CN202180083318 A CN 202180083318A CN 117043601 A CN117043601 A CN 117043601A
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Abstract
The present invention provides a sensitizer for immunochromatographic assay capable of detecting novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody with high sensitivity, and an assay using the sensitizer. Specifically, a composition comprising a compound represented by the following general formula [4 ]]The polymer is a sensitizer for immunochromatography assay which uses novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured. Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
Description
Technical Field
The present invention relates to a sensitizer for immunochromatographic assay which uses a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, and an immunochromatographic assay which uses a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, which is carried out in the presence of the sensitizer.
Background
Immunochromatography is a method in which a chromatography method utilizing the capillary phenomenon of a membrane and an immunological method utilizing an antigen-antibody reaction are combined. Immunochromatography has high specificity due to antigen-antibody reaction, is easy to handle, can be checked on site without requiring a site or special equipment, and can visually determine the result, and therefore is widely used as a clinical diagnostic method (patent document 1).
Although immunochromatography has an advantage of easy measurement, it is desired to use immunochromatography with higher sensitivity because it has low accuracy of detection sensitivity and is determined to be false negative even when a positive sample is used.
To establish a high-sensitivity immunochromatographic assay, various methods have been studied, and for example, patent document 2 discloses a method of improving sensitivity by changing the addition position of an immunochromatographic reagent. Patent document 3 discloses a method of improving sensitivity by improving a material of a film.
In addition, a method of improving sensitivity by adding an additive to a sample diluent used in a conventional immunochromatography method is known. For example, patent documents 4, 5 and 6 disclose that Bovine Serum Albumin (BSA), a polymer having phosphorylcholine groups, and hyaluronic acid are contained in a sample diluent. These disclosures have the advantage that the addition site of the reagent does not need to be changed and the conventional immunochromatographic membrane can be used.
However, the above patent documents do not disclose or suggest a sensitizer for a method for measuring novel coronavirus (SARS-CoV-2) IgM antibodies and IgG antibodies with high sensitivity.
In particular, since the novel coronavirus (SARS-CoV-2) IgM antibody is an antibody produced at the early stage of infection with the novel coronavirus (SARS-CoV-2), if the IgM antibody can be detected with high sensitivity, the infected person can be isolated at the early stage of infection, and thus spread of infection can be suppressed. Further, since the treatment can be started early, the effect of the treatment can be improved, and therefore, it is considered that a sensitizer capable of detecting an IgM antibody with high sensitivity is highly useful.
In addition, the novel coronavirus (SARS-CoV-2) IgG antibody is an antibody that is produced in vivo for a long period of time, and if the IgG antibody can be detected with high sensitivity, an infected person at the later stage of infection can be grasped. In order to grasp the infected person of the novel coronavirus, which has many infected persons without subjective symptoms, a sensitizer capable of detecting IgG antibodies produced for a long period of time with high sensitivity is considered to be highly useful.
Prior art literature
Patent literature
Patent document 1: japanese patent laid-open No. 01-063865,
patent document 2: japanese patent application laid-open No. 2014-66674,
patent document 3: japanese patent application laid-open No. 2014-62820,
patent document 4: japanese patent application laid-open No. 2008-292326,
patent document 5: japanese patent laid-open No. 2008-058334,
Patent document 6: japanese patent laid-open No. 2003-344413.
Disclosure of Invention
Problems to be solved by the invention
The present invention provides a sensitizer for immunochromatographic assay capable of detecting novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody with high sensitivity, and an assay using the sensitizer.
Means for solving the problems
As a result of intensive studies to find a compound capable of detecting novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody with high sensitivity, the present inventors have found that a polymer or copolymer having phosphorylcholine-like groups on the side chains represented by the following general formulae [1], [3] or [4] has target properties, and have completed the present invention.
Namely, the present invention is constituted as follows.
[1] An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [4 ]:
[ chemical formula 1]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
[2] An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [3 ]:
[ chemical formula 2]
Wherein m and n each represent the number of structural units, and m: n is 100 to 30:0 to 70.
[3] A sample dilution for immunochromatography assay comprising the sensitizer for immunochromatography assay according to [1], wherein a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody is used as a substance to be assayed.
[4] An immunochromatographic assay comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as an analyte, characterized in that the antigen-antibody reaction is carried out in the presence of the sensitizer for immunochromatographic assay according to [1 ].
[5] An apparatus for immunochromatography measurement comprising the sensitizer for immunochromatography measurement according to [1], wherein the specimen is a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody.
[6] An apparatus for immunochromatography measurement comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, wherein the sensitizer for immunochromatography measurement according to [1] is carried on a developed membrane, a sample pad or a conjugate pad.
[7] A kit for detecting novel coronavirus infection (COVID-19) comprising the sensitizer for immunochromatographic assay according to [1], and a substance specifically binding to novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody.
[8] A method for testing a subject for the likelihood of infecting a novel coronavirus infection (covd-19), comprising assaying a biological sample derived from the subject for novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies by performing an antigen-antibody reaction in the presence of a sensitizer according to the immunochromatographic assay of [1 ].
[9] A method for treating a novel coronavirus infection (covd-19), comprising the steps of (1) and (2):
(1) A novel coronavirus (SARS-CoV-2) measurement step in which an antigen-antibody reaction is carried out in the presence of a sensitizer for immunochromatography measurement comprising a polymer represented by the following general formula [4], and
(2) A step of treating a patient with a novel coronavirus infection (COVID-19) based on the measurement result of (1),
[ chemical formula 3]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
[10] A kit for treating novel coronavirus infection (covd-19) comprising a sensitizer for immunochromatographic assay and a therapeutic agent for novel coronavirus infection (covd-19), wherein the sensitizer is formed of a polymer represented by the following general formula [4 ]:
[ chemical formula 4]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
[11] An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [4] and a copolymer represented by the following general formula [5 ]:
[ chemical formula 5]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50;
[ chemical formula 6]
Wherein q R's each independently represent a hydrogen atom or a cation, m and q each represent the number of structural units, and m: q is 99 to 20:1 to 80.
ADVANTAGEOUS EFFECTS OF INVENTION
The method for measuring an immunochromatography assay using a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody as a substance to be measured according to the present invention can detect the novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody with higher sensitivity than the conventional immunochromatography assay for detecting the novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody.
Detailed Description
The sensitizer for immunochromatography assay using a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, a sample dilution for immunochromatography assay using the antibody as a substance to be measured comprising the sensitizer, an immunochromatography assay using the antibody as a substance to be measured by performing an antigen-antibody reaction in the presence of the sensitizer, an instrument for immunochromatography assay using the antibody as a substance to be measured comprising the sensitizer, and a method for treating novel coronavirus infection (COVID-19) comprising a step of performing an antigen-antibody reaction in the presence of the sensitizer are described below.
In the measurement method using the sensitizer for immunochromatographic measurement of the present invention, novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody are used as the measurement target substance, but other measurement target substances may be detected. However, the sensitizer for immunochromatographic assay of the present invention is characterized in that the antibody can be detected with high sensitivity.
In this specification, novel coronavirus refers to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which refers to a strain of coronavirus that causes novel coronavirus infection (covd-19) (also known as coronavirus disease 2019), which is a respiratory disease that is responsible for the pandemic of covd-19. Each SARS-CoV-2 virion is 50-200 nanometers in diameter and, like other coronaviruses, SARS-CoV-2 has 4 structural proteins known as S (spike), E (envelope), M (membrane) and N (nucleocapsid) proteins. The N protein maintains the RNA genome and the S, E and M proteins together form the viral envelope. Spike proteins are proteins that enable the virus to attach and fuse to the membrane of a host cell. SARS-CoV-2 infects humans and humans, assuming that infection is primarily caused initially by respiratory droplets from a range of coughs or sneezes. In addition, since aerosols may infect viruses, it is also suggested that viruses may also be suspended in air.
The polymer for use as a sensitizer for immunochromatographic assay of the present invention, which uses a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, is represented by the following general formula [3 ]:
[ chemical formula 7]
Wherein m and n each represent the number of structural units, and m: n is 100 to 30:0 to 70.
As 1 embodiment, the polymer represented by the above general formula [3] is a polymer having only a monomer { MPC) represented by the following general formula [2 ]: polymers of structural units of 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate.
[ chemical formula 8]
Namely, the polymer can be represented by the following general formula [1 ]:
[ chemical formula 9]
Wherein m represents the number of structural units.
The polymer represented by the above general formula [1] is preferably a water-soluble polymer having a weight average molecular weight of 100,000 ～ 2,000,000, more preferably 500,000 ～ 1,500,000.
In another embodiment, the polymer represented by the general formula [3] is a copolymer having a structural unit based on 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate (MPC) represented by the general formula [2] and a structural unit based on butyl methacrylate.
The type of the copolymer is not particularly limited, and may be a random copolymer or a block copolymer, but a random copolymer is preferable.
In the copolymer, the ratio (m: n) of the number of structural units based on 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate (MPC) to the number of structural units based on butyl methacrylate is preferably 90 to 30:10 to 70.
The copolymer is preferably a water-soluble copolymer having a weight average molecular weight of 10,000 ～ 2,000,000, more preferably 500,000 ～ 1,500,000.
The polymer represented by the above general formula [3] may further contain other structural units within a range not impairing the effects of the present invention. Examples of the other structural unit include those based on
Structural units of monomers such as alkyl (meth) acrylates, cyclic alkyl (meth) acrylates, aromatic group-containing (meth) acrylates, and hydroxyl group-containing (meth) acrylates. Among these monomers, (meth) acrylic esters containing hydroxyl groups are preferable. As the hydroxyl group-containing (meth) acrylate, 2-hydroxy (meth) acrylate, glycerol (meth) acrylate, and glycerol methacrylate are preferable.
The polymer represented by the above general formula [3] which may further contain a structural unit based on a glycerol methacrylate may be represented by the following general formula [ 4):
[ chemical formula 10]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
In the polymer represented by the above general formula [4], the ratio (m: n: p) of the number of structural units based on butyl methacrylate to the number of structural units based on glycerol methacrylate based on the number of structural units of 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate (MPC) represented by the above general formula [2] is preferably 30 to 50:20 to 50.
The polymer represented by the above general formula [4] is preferably a water-soluble copolymer having a weight average molecular weight of 10,000 ～ 100,000, more preferably 20,000 ～ 50,000.
In the case where the polymer represented by the above general formula [4] is a copolymer, the kind of the copolymer is not particularly limited, and may be a random copolymer or a block copolymer, but a random copolymer is preferable.
The polymer represented by the above general formula [3] or [4] used as a sensitizer for immunochromatographic assay is preferably exemplified by the polymers described in the following examples, but is not particularly limited.
The sensitizer for immunochromatographic assay of the present invention may further comprise a copolymer represented by the following general formula [5] in addition to the polymer represented by the above general formula [3] or [4 ]:
[ chemical formula 11]
Wherein q R's each independently represent a hydrogen atom or a cation, m and q each represent the number of structural units, and m: q is 99 to 20:1 to 80.
In the above formula, q R's each independently represent a hydrogen atom or a cation. Examples of the cations include sodium ion, potassium ion, and ammonium ion, and q R may be the same or different. R is preferably a hydrogen atom.
In the above formula, m and q each represent the number of structural units, and m: q is 99 to 20:1 to 80, preferably 90 to 30:10 to 70.
The copolymer represented by the above general formula [5] is a water-soluble copolymer having a weight average molecular weight of 10,000 ～ 5,000,000, preferably 50,000 ～ 2,000,000.
The copolymer represented by the above general formula [5] is a copolymer having a molecular weight based on the monomer { MPC represented by the above general formula [2 ]: copolymers of structural units of 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate and structural units based on methacrylic acid or its salts.
The type of the copolymer is not particularly limited, and may be a random copolymer or a block copolymer, but a random copolymer is preferable.
As examples of methacrylic acid or a salt thereof, methacrylic acid, sodium methacrylate, potassium methacrylate, ammonium methacrylate, and the like can be exemplified. Among them, methacrylic acid is preferable.
The copolymer represented by the above general formula [5] is preferably exemplified by the copolymers described in the following examples, but is not particularly limited.
The content of the copolymer represented by the above general formula [5] in the sensitizer for immunochromatographic assay is not particularly limited, but is 0.1 to 5.0w/v%, preferably 0.5 to 1.0w/v%.
The polymer or copolymer used as the sensitizer for immunochromatographic assay of the present invention can be used by being dissolved in various reagents or samples used for immunochromatographic assay, or can be used by being carried on a kit for immunochromatographic assay, but is preferably used as a sample diluent.
The immunochromatographic assay of the present invention is usually composed of a sample diluent and a test strip as a kit for immunochromatographic assay to be used.
The sample diluent is a suitable diluent for measurement by immunochromatography, and may contain buffer components such as Tris buffer, phosphate buffer, barbital sodium (Veronal) buffer, boric acid buffer, good's buffer, stabilizing components such as albumin, globulin, casein, serum, water-soluble gelatin, surfactant, saccharide, chelating agent, and preservative components such as salicylic acid, benzoic acid, sodium azide.
When the sensitizer of the present invention is used by dissolving it in a sample diluent, the concentration thereof is not particularly limited and is usually used in the range of 0.1 to 5.0w/v%, preferably 0.5 to 1.0w/v% in terms of the concentration meter at the time of the antigen-antibody reaction. In one embodiment, the concentration of the sensitizer of the present invention in the sample diluent is generally 0.1 to 5.0 wt%, preferably 0.5 to 1.0 wt%.
The test strip generally includes at least a first substance capable of undergoing an antigen-antibody reaction at a first epitope of a sample, a second substance capable of undergoing an antigen-antibody reaction at a second epitope of the sample and labeled, and a membrane carrier, wherein the first substance is immobilized in advance on a predetermined position of the membrane carrier to form a capture site, and the second substance is disposed at a position separated from the capture site so as to be capable of being developed chromatographically on the carrier. In one embodiment, in the case where the sample is an antibody (including an IgM antibody and/or an IgG antibody), the first substance and/or the second substance may be an antigen against the antibody.
Typical structures of the test strip for immunochromatography include: the device comprises a sample supply unit (hereinafter, also referred to as a sample pad) for supplying a sample, a conjugate pad for disposing a conjugate, a detection unit for fixing a capture reagent such as an antibody in a linear form, a development membrane for developing and capturing the immunocomplex by a mobile phase, and an absorption pad for absorbing the sample developed on the development membrane downstream of the detection unit.
As specific examples of materials suitable for the sample pad, glass fiber (glass fiber), acrylic fiber, hydrophilic polyethylene material, polyester, dry paper, pulp, cellulose fiber, fabric, and the like are included, but not limited thereto. The sample pad may contain a blocking reagent or a buffer component which is usually used, if necessary, in a range not departing from the object of the present invention and not affecting the reaction system, and may contain a blood coagulant or the like when the sample is blood. In this case, the sample may be contained in at least a part of the sample pad or the whole sample pad.
In one embodiment, the conjugate pad is comprised of a pad-like porous material capable of spreading a sample through the sample pad and capable of retaining the conjugate, with the conjugate retained in a portion or all of it. Examples of the porous material constituting the conjugate pad include pads made of nonwoven fibers such as paper, cellulose mixtures, cellulose fibers, nitrocellulose, polyesters, acrylonitrile copolymers, glass (glass fibers), and rayon.
As the test strip for immunochromatography measurement, commercially available ones can be used, and those produced by a known method can be used. In the case of producing the film by a known method, a substrate generally used in this field may be used as the substrate for the stretched film, and examples of the substrate include cellulose, nitrocellulose, nylon, and the like.
Examples of the immunochromatography measurement instrument containing the sensitizer of the present invention, which uses a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a measurement target substance, include: (1) an appliance comprised of an unrolled film; (2) An instrument which is composed of a sample marking portion and an unfolding membrane, and in which the sample marking portion and the unfolding membrane are formed in a manner capable of moving by capillary phenomenon; (3) An instrument which is composed of a sample marking portion, an unfolding membrane, and a liquid absorbing portion, and in which the sample marking portion, the unfolding membrane, and the liquid absorbing portion are formed in such a manner as to be movable in this order by capillary phenomenon; (4) An instrument or the like which is constituted by a sample dropping section, a sample marking section, a development film, and a liquid absorbing section, and in which the sample dropping section, the sample marking section, the development film, and the liquid absorbing section are formed so as to be movable in this order by capillary phenomenon; but is not particularly limited.
When the sensitizer of the present invention is used by being carried on the immunochromatographic measurement instrument, the amount thereof varies depending on the location to be carried, the type of sensitizer to be used, the type of measurement target substance to be used, the labeling substance to be used, and the like, and may be carried on, for example, a development film of the immunochromatographic measurement instrument, a sample labeling portion, a sample dripping portion such as a sample pad or conjugate pad, and the like. When the sensitizer of the present invention is carried in the development membrane or the sample labeling portion of the immunochromatographic assay device, the sensitizer is expressed as a unit area (cm 2 ) The amount of the sensitizer contained therein is usually 0.01. Mu.g to 10mg, preferably 0.1. Mu.g to 4mg, more preferably 1 to 800. Mu.g. In the case where the sensitizer of the present invention is carried in a sample addition portion, in the case where the sample addition portion is a sample pad, the carried sensitizer of the present invention is usually 0.01. Mu.g to 50mg, preferably 100. Mu.g to 20mg, more preferably 1 to 10mg. When the sample drop section is a conjugate pad, the sensitizer of the present invention is usually supported in an amount of 0.01. Mu.g to 10mg, preferably 0.1. Mu.g to 4mg, more preferably 1 to 800. Mu.g. When 2 or more polymers or copolymers having different ratios of the weight average molecular weight and the number of structural units are supported, the amounts of the sensitizer are each amount. The area to be supported varies depending on the type and size of the immunochromatographic measurement instrument to be used and the amount of the measurement sample, but is usually 5 to 60%, preferably 5 to 15%, of the total area in the case of an developed membrane of the immunochromatographic measurement instrument, and is usually 5 to 40%, preferably 10 to 20%, of the total area in the case of a sample drop-in portion.
The sensitizer of the present invention may be used by being carried on a developed membrane, a sample labeling portion, a sample dripping portion such as a sample pad or a conjugate pad, or the like of an immunochromatographic assay device which is a constituent element of a commercially available kit. Examples of the immunochromatographic measurement instrument that is a component of such a commercially available kit include a novel coronavirus (SARS-CoV-2) IgM test kit (colloidal gold) (manufactured by Ray Biotec. Co.), a novel coronavirus (SARS-CoV-2) IgG test kit (colloidal gold) (manufactured by Ray Biotec. Co.), a novel coronavirus IgM/IgG antibody test kit (COVID-19 IgM/IgG Plus (manufactured by MALDOMINc.), a novel coronavirus (SARS-CoV-2) antibody test kit (RF-NC 001 (manufactured by cone) and a novel coronavirus (SARS-CoV-2) antibody test kit (IgG) RF-NC002 (manufactured by cone) and SARS-CoV-2 antibody test (colloidal gold immunochromatography) (manufactured by Lepu Medical Technology).
The present invention provides a kit for detecting novel coronavirus infection (covd-19) comprising the sensitizer for immunochromatographic assay of the present invention and a substance specifically binding to novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody.
"specific" means that the affinity for the IgM antibody and/or IgG antibody of the novel coronavirus (SARS-CoV-2) is higher than that for other substances (in one embodiment, igM antibody and/or IgG antibody directed against an antigen other than the protein of the novel coronavirus (SARS-CoV-2) is not included in the "other substances"). The specifically bound substance is for example present at about 10 -6 、10 -7 、10 -8 、10 -9 、10 -10 The following KD binds to novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies. In addition, the specifically bound substance is 10 -14 、10 -13 、10 -12 The KD above binds to novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies.
Preferably, the material is isolated or purified. "isolation or purification" means an operation of removing components other than the target component from a naturally occurring state. The purity of the isolated or purified substance (the ratio of the weight of the substance to the weight of the total protein) is usually 50% or more, preferably 70% or more, more preferably 90% or more, and most preferably 95% or more (e.g., substantially 100%).
The substance may be directly or indirectly labeled with a labeling substance. Examples of the labeling substance include fluorescent substances (e.g., FITC and rhodamine) and radioactive substances (e.g., FITC and rhodamine) 14 C、 3 H、 125 I) Enzymes (e.g., alkaline phosphatase, peroxidase), colored particles (e.g., metal colloid particles, colored latex), biotin, and the like.
In the present invention, examples of "substances specifically binding to novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody" include substances specific to novel coronaviruses
Antigens of (SARS-CoV-2) IgM antibodies and/or IgG antibodies, antibodies that specifically bind to novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies, nucleic acids (e.g., aptamers), etc., preferably antigens directed against novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies, antigens directed against novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies.
The antigen is not limited as long as it is an antigen against the novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody to be measured according to the present invention, and examples thereof include novel coronavirus S (spike) protein, E (envelope) protein, M (membrane) protein and N (nucleocapsid) protein.
When a recombinant novel coronavirus (SARS-CoV-2) protein is used as an antigen, the recombinant novel coronavirus (SARS-CoV-2) protein can be produced, for example, by the following method. The polynucleotide encoding the amino acid sequence of the novel coronavirus (SARS-CoV-2) protein is integrated into an appropriate expression vector, and the recombinant novel coronavirus (SARS-CoV-2) protein can be obtained from the disrupted product of the transformed cell by inserting the polynucleotide into an appropriate host for transformation. The host cell is not particularly limited, and various host cells conventionally used in genetic engineering methods, such as E.coli, bacillus subtilis, yeast, plant or animal cells, and the like, can be used.
The novel coronavirus (SARS-CoV-2) protein may be a recombinant protein produced by the transformant, or may be a protein isolated or purified from a natural novel coronavirus (SARS-CoV-2) producing the protein by a per se known protein isolation and purification technique. Alternatively, it may be a chemically synthesized or biochemically synthesized protein in a cell-free translation system.
In one embodiment, the "anti-novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody" includes an anti-IgM antibody and/or anti-IgG antibody of an animal of the same species as the animal producing the IgM antibody and/or IgG antibody.
The antibody may be of any isotype, e.g., igA, igD, igE, igG (e.g., igG1, igG2, igG3, or IgG 4), igM, and the like. The antibody may be a polyclonal antibody or a monoclonal antibody, or may be a chimeric antibody or a single chain antibody. In addition, the antibody may be a part (antibody fragment) of an antibody having antigen binding property such as a Fab fragment or a fragment generated by a Fab expression library. Examples of antibody fragments include: (i) Fab fragments, which are monovalent fragments consisting of the VL, VH, CL and CH1 domains, (ii) F (ab') 2 A fragment which is a bivalent fragment comprising 2 Fab fragments linked at the hinge region by a disulfide bridge, (iii) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (iv) a Fab 'fragment which is a fragment that disrupts F (ab') using mild reducing conditions 2 A disulfide bridge of the fragment, (v) an Fv fragment stabilized with a disulfide (dsFv), and (vi) a domain antibody (dAb) that is a single variable region domain (VH or VL) polypeptide of an antibody that specifically binds an antigen; but is not limited thereto.
The anti-novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody used in the method of the present invention may be produced by mouse, guinea pig, hamster, goat or rabbit. In addition, anti-novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies can be used in commercially available products.
The test kit of the present invention comprises the sensitizer for immunochromatographic assay of the present invention and a substance specifically binding to a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody in different containers.
The test kit of the present invention may further comprise instructions describing that the sensitizer for immunochromatographic assay of the present invention and a substance specifically binding to a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody can be used or should be used for the test of novel coronavirus infection (COVID-19).
The novel kit for the examination of coronavirus infection (covd-19) may comprise any carrier (e.g., a pharmaceutically acceptable carrier), a stabilizer or buffer component, other therapeutic agents or supplements, and the like. Examples of pharmaceutically acceptable carriers include, but are not limited to, diluents such as water and physiological saline. The novel kit for detecting coronavirus infection (covd-19) of the present invention may comprise a sample diluent, a test strip for immunochromatography assay, and an instrument for immunochromatography assay.
The present invention provides a method for testing a subject for the likelihood of infection with a novel coronavirus infection (covd-19) comprising assaying a biological sample derived from the subject for novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies by performing an antigen-antibody reaction in the presence of a sensitizer for an immunochromatographic assay of the present invention.
The biological sample that can be used in this method is exemplified by blood, but is not particularly limited as long as it can detect novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody. As the "blood", blood derived from any tissue is also conceivable, but peripheral blood is generally used because of easy collection. As a blood collection method, a method known per se can be applied. The collected blood may be used as it is in the present method, or may be used as a liquid component (plasma) obtained by separating cellular components (red blood cells, white blood cells, platelets, etc.) by a method known per se, such as centrifugation, filtration, etc. In addition, the method can be used as a liquid component (serum) obtained by coagulating blood to separate platelets or coagulation factors.
In the examination method of the present invention, for example, a biological sample of a subject such as blood is suspended in an extraction solution, and an antigen is extracted from the biological sample solution. Then, for example, in the case of using the immunochromatographic assay device of the above (4), a biological sample solution treated with the extraction solution is dropped into the sample drop portion. Thus, in the presence of the sample dilution liquid for immunochromatography assay comprising the sensitizer for immunochromatography assay of the present invention, the biological sample liquid is developed from the sample drop portion to the sample label portion by capillary phenomenon, and then developed on the membrane (menbrane).
The target antigen (e.g., novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody) in the biological sample solution forms an immunocomplex with the labeling substance (e.g., anti-novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody) carried in the sample labeling part when passing through the sample labeling part, and is directly developed to one end of the developed membrane. The complex then binds and develops color by immunochemical reaction with a substance (e.g., an antigen against a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody) present on (coated on) the test line of the developed membrane, thereby appearing a line that can be seen on the developed membrane. In the case of such a color development, the sample was positive, and when the target antigen in the biological sample solution was a novel coronavirus (SARS-CoV-2) IgM antibody, it was determined that the subject was infected with a novel coronavirus infection (COVID-19). When the target antigen in the biological sample solution is a novel coronavirus (SARS-CoV-2) IgG antibody, it can be determined that the subject is being infected or has been infected with a novel coronavirus infection (COVID-19). In addition, the intensity of the positive can be determined by measuring its color development with a visual or appropriate analytical device (e.g., immunochromatographic reader (immunochromato reader), spectrophotometer). In the case of using a fluorescent dye as a detection method with a labeled antibody, for example, it is possible to determine whether or not the test line is positive by irradiation with ultraviolet rays or the like based on the presence or absence or the degree of color development.
On the other hand, after the test line, there is a control line, and the antibodies present therein (coated thereon) and the labeling substance (e.g., anti-novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody) that did not form an immunocomplex are combined by an immunochemical reaction and developed, so that it can be seen that immunochromatography proceeds without problems.
After the sample solution is spread on the spreading film, unreacted labeled antibody or the like is absorbed by the absorbent pad. In the case where the sample does not contain a target antigen (for example, a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody), the test line is not developed, and only the control line is developed. In this case, the sample was negative, and when the target antigen in the biological sample solution was a novel coronavirus (SARS-CoV-2) IgM antibody, it could not be determined that the subject was infected with the novel coronavirus infection (COVID-19). When the target antigen in the biological sample solution is a novel coronavirus (SARS-CoV-2) IgG antibody, it cannot be determined that the subject is suffering from a novel coronavirus infection (COVID-19) or has been suffering from a past infection.
In one embodiment, if the target antigen in the biological sample solution is negative in the case of a novel coronavirus (SARS-CoV-2) IgM antibody and negative in the case of a novel coronavirus (SARS-CoV-2) IgG antibody, it cannot be determined that the subject is suffering from a novel coronavirus infection (COVID-19) or has been suffering from a past infection. If the target antigen in the biological sample solution is positive in the case of the novel coronavirus (SARS-CoV-2) IgM antibody and negative in the case of the novel coronavirus (SARS-CoV-2) IgG antibody, it can be determined that the subject is suffering from novel coronavirus infection (COVID-19), and that the initial stage of infection can be determined. If the target antigen in the biological sample solution is positive in the case of the novel coronavirus (SARS-CoV-2) IgM antibody and the target antigen in the biological sample solution is positive in the case of the novel coronavirus (SARS-CoV-2) IgG antibody, it can be determined that the subject is suffering from novel coronavirus infection (COVID-19), and that a certain period of time has elapsed after the infection. When the target antigen in the biological sample solution is negative in the case of the novel coronavirus (SARS-CoV-2) IgM antibody and positive in the case of the novel coronavirus (SARS-CoV-2) IgG antibody, it can be determined that the subject has suffered from the novel coronavirus infection (COVID-19).
The biological sample provided in the method for conducting an assay 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 having a possibility of infection with a novel coronavirus infection (covd-19), and among them, rodents such as mice, rats, hamsters and guinea pigs, experimental animals such as rabbits, livestock such as pigs, cattle, goats, horses, sheep and minks, pets such as dogs and cats, primates such as humans, apes, cynomolgus monkeys, rhesus monkeys, marmoset, gorillas and chimpanzees are preferable, and humans are particularly preferable.
The novel method for treating coronavirus infection (COVID-19) of the present invention comprises at least the steps (1) and (2) below.
(1) And a step of measuring a novel coronavirus (SARS-CoV-2) by performing an antigen-antibody reaction in the presence of a sensitizer for immunochromatography measurement comprising the polymer or copolymer.
(2) And (3) a step of treating the patient with a novel coronavirus infection (covd-19) based on the measurement result of (1).
It should be noted that patients also include those who may be infected with the novel coronavirus (SARS-CoV-2). By assaying is meant detecting the presence or absence of novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies in a sample derived from a patient.
In addition, the step of treating a patient with a novel coronavirus infection includes not only administration of a known therapeutic agent, a therapeutic agent to be marketed in the future, and a candidate therapeutic agent in clinical stages to the patient, but also symptomatic therapy (aspirin administration, fluid infusion administration, etc.).
Since the novel coronavirus infection (COVID-19) treatment method of the present invention can detect the novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody with high sensitivity, the novel coronavirus infection (COVID-19) can be rapidly treated for patients suffering from novel coronavirus infection (COVID-19).
Symptoms of the novel coronavirus infection (covd-19) are involved in many aspects from asymptomatic to severe pneumonia and death, and specifically, fever, dry cough, burnout sensation/tiredness, phlegm, olfactory disorder/gustatory disorder, shortness of breath, muscle pain/joint pain, sore throat, headache, aversion to cold, nausea/vomiting, nasal obstruction, diarrhea, hemoptysis, conjunctival congestion, and the like can be cited.
In another embodiment, the method for treating coronavirus infection (COVID-19) of the present invention comprises at least the steps (1) and (2) below.
(1) And a step of measuring a novel coronavirus (SARS-CoV-2) by performing an antigen-antibody reaction in the presence of a sensitizer for immunochromatography measurement comprising the polymer or copolymer.
(2) And (3) a step of treating a subject other than the human with a novel coronavirus infection (covd-19) based on the measurement result of (1).
The subject other than human provided in the treatment method of the present invention is not limited, but preferably means a mammal. Further preferred examples of the mammal include mammals that are likely to be infected with novel coronavirus infection (covd-19), and among them, mammals such as mice, rats, hamsters, guinea pigs, and experimental animals such as rabbits, livestock such as pigs, cattle, goats, horses, sheep, and minks, pets such as dogs and cats, and primates such as apes, cynomolgus monkeys, rhesus monkeys, marmosets, gorillas, and chimpanzees are preferred.
The present invention provides a novel therapeutic kit for coronavirus infection (COVID-19) comprising a sensitizer for immunochromatography assay and a novel therapeutic agent for coronavirus infection (COVID-19), wherein the sensitizer comprises a polymer or copolymer represented by the general formula [1], [3], [4] or [5 ].
Examples of the therapeutic agent include therapeutic agents containing a novel therapeutic agent for coronavirus infection (covd-19), and examples of the therapeutic agent include, but are not limited to, adefovir, dexamethasone, fampicvir (Avigan), ciclesonide, nafamostat, camostat, ivermectin, nelfinavir, and the like. The therapeutic agent may also be tolizumab, baratinib, acartinib, lei Fuli zulizumab, iritolan (ericoran), ibudilast, LY3127804, octreotide Li Shan antigen, HLCM051, ADR-001, antipobate, apremilast, cenicriviroc, as therapeutic agents for severe pneumonia or acute respiratory distress syndrome. Casirivimab, imdevimab and/or a mixture thereof, which are virus-neutralizing antibodies that exhibit neutralizing activity against SARS-CoV-2 by non-competitive binding to the receptor binding site of the viral spike protein, are also possible. Can also be a therapeutic agent to be marketed in the future and a candidate therapeutic agent in clinical stage. The therapeutic agent may also comprise aspirin, fluid infusion/infusion solutions, etc. for symptomatic therapy.
The therapeutic kit of the present invention comprises the sensitizer for immunochromatographic assay of the present invention and the therapeutic agent for novel coronavirus infection (covd-19) in different containers.
The therapeutic kit of the present invention may further comprise instructions describing that the sensitizer for immunochromatographic assay of the present invention and the therapeutic agent for novel coronavirus infection (covd-19) may be used or should be used for the treatment of novel coronavirus infection (covd-19).
The novel therapeutic kit for coronavirus infection (covd-19) may comprise any carrier (e.g., a pharmaceutically acceptable carrier), a stabilizer or buffer component, other therapeutic agents or supplements, and the like. Examples of pharmaceutically acceptable carriers include, but are not limited to, diluents such as water and physiological saline. The novel kit for treating coronavirus infection (covd-19) of the present invention may comprise a sample diluent, a test strip for immunochromatography assay, and an instrument for immunochromatography assay.
The therapeutic kit of the present invention may further comprise a substance that specifically binds to a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody.
Since the novel coronavirus infection (COVID-19) therapeutic kit of the present invention can detect the novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody with high sensitivity, the novel coronavirus infection (COVID-19) can be rapidly treated for a subject (patient) suffering from the novel coronavirus infection (COVID-19).
Hereinafter, the present invention will be described in more detail with reference to the examples, but the present invention is not limited thereto.
Examples
Production example
(Synthesis of copolymer of the invention and comparative example)
Copolymers used as sensitizers for immunochromatographic assays of the present invention and copolymers of comparative examples were synthesized. Details are as follows.
The polymer used in the present invention and comparative example was a polymer of 2- (methacryloyloxy) ethyl-2' - (trimethylammonium) ethyl phosphate (MPC) and a random copolymer synthesized from MPC and the following monomers.
(Polymer 1)
Weight average molecular weight: 1030×10 3 MPC polymers
(Polymer 2)
Weight average molecular weight: 600X 10 3 Ratio of the number of structural units based on MPC monomer to the number of structural units based on butyl methacrylate: 80:20
(Polymer 3)
Weight average molecular weight: 1210 x 10 3 Ratio of the number of structural units based on MPC monomer to the number of structural units based on butyl methacrylate: 80:20
(Polymer 4)
Weight average molecular weight: 22X 10 3 Ratio of the number of structural units based on MPC monomer, the number of structural units based on butyl methacrylate and the number of structural units based on glycerol methacrylate: 40:40:20
(Polymer 5)
Weight average molecular weight: 26×10 3 Ratio of the number of structural units based on MPC monomer, the number of structural units based on butyl methacrylate and the number of structural units based on glycerol methacrylate: 40:20:40
(Polymer 6)
Weight average molecular weight: 680×10 3 Ratio of the number of structural units based on MPC monomer to the number of structural units based on methacrylic acid: 30:70
Examples
(confirmation of detection sensitivity Using a known immunochromatographic assay device)
In this example, it was confirmed whether the above-mentioned polymer can improve the detection sensitivity (whether or not there is a sensitization effect) of a known novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody detection kit. Details are as follows.
Comparative example
The evaluation was performed using a commercially available novel coronavirus (SARS-CoV-2) IgM antibody detection kit, a novel coronavirus (SARS-CoV-2) IgM test kit (colloidal gold) (manufactured by Ray Biotec. Co.) and a novel coronavirus (SARS-CoV-2) IgG antibody detection kit (colloidal gold) (manufactured by Ray Biotec. Co.). More specifically, 25. Mu.L of a novel coronavirus-positive patient serum (CoV-PosM-S-100, manufactured by RayBiotec. Co.) was added to 245. Mu.L of a SARS-CoV-2IgM antibody detection kit or a dilution of a SARS-CoV-2IgG antibody detection kit, and the dilution was developed in an immunochromatography kit.
Example 1
Polymer 1, 2, 3, 4 or 5 was added to the dilution of the kit so that the final concentration became 0.5 wt%, and the solution prepared so that the serum of the novel coronavirus-positive patient was diluted 100-fold with respect to the comparative example was developed, and the detection sensitivity was compared.
Example 2
After adding polymer 1, 2 or 4 to the dilution of the kit to a final concentration of 0.5 wt%, polymer 6 was further added to a final concentration of 0.5 wt%. Except for this, the detection sensitivity was compared in the same manner as in example 1.
Example 3
The sample pad of the kit was loaded with the polymer 1, 2, 3, 4 or 5 so that the loading amount was 5mg, and the solution prepared so that the serum of the novel coronavirus-positive patient was 100-fold diluted with respect to the comparative example was developed, and the detection sensitivity was compared.
Example 4
Polymer 1, 2 or 4 was carried in the sample pad of the kit so that the carrying amount became 5mg, and polymer 6 was carried so that the carrying amount became 5mg. Except for this, the detection sensitivity was compared in the same manner as in example 3.
Example 5
The conjugate pad of the kit carries the polymer 1, 2, 3, 4 or 5 so that the carrying amount becomes 500. Mu.g, and further, a solution prepared so that the serum of a novel coronavirus-positive patient is diluted 100 times with respect to the comparative example is developed, and the detection sensitivity is compared.
Example 6
Polymer 1, 2 or 4 was supported in the conjugate pad of the kit so that the loading amount reached 500. Mu.g, and polymer 6 was further supported so that the loading amount reached 500. Mu.g. Except for this, the detection sensitivity was compared in the same manner as in example 5.
(confirmation result of IgM antibody detection sensitivity Using an apparatus for immunochromatography measurement)
The results of the improvement in detection sensitivity when a polymer was added to the diluent using a known novel coronavirus (SARS-CoV-2) IgM antibody detection kit-novel coronavirus (SARS-CoV-2) IgM test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 1 according to examples 1 and 2.
TABLE 1
When 0.5 wt% of polymer 1, 2, 3, 4 or 5 was added, a test line having a depth equal to or more than or slightly less than that of the control as a comparative example was detected for IgM, and thus it was confirmed that the sensitivity was improved by adding polymer 1, 2, 3, 4 or 5. Preferably, it was confirmed that the sensitivity was further improved by using the polymer 6 in combination.
(confirmation result of IgG antibody detection sensitivity Using immunochromatographic assay device)
The results of the improvement in detection sensitivity when a polymer was added to the diluent using a known novel coronavirus (SARS-CoV-2) IgG antibody detection kit, a novel coronavirus (SARS-CoV-2) IgG test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 2, according to examples 1 and 2.
TABLE 2
When 0.5 wt% of polymer 1, 2, 3, 4 or 5 was added, a test line having a depth equal to or slightly smaller than that of the control as a comparative example was detected for IgG, and thus it was confirmed that the sensitivity was improved by adding polymer 1, 2, 3, 4 or 5. Preferably, it was confirmed that the sensitivity was further improved by using the polymer 6 in combination.
The results of the improvement in detection sensitivity when the polymer was supported in the sample pad using a known novel coronavirus (SARS-CoV-2) IgM antibody detection kit-novel coronavirus (SARS-CoV-2) IgM test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 3 according to examples 3 and 4.
TABLE 3
When the polymer 1, 2, 3, 4 or 5 was supported on the sample pad, the test line having a depth equal to or slightly smaller than that of the control sample as a comparative example was detected for IgM, and thus it was confirmed that the sensitivity was improved by supporting the polymer 1, 2, 3, 4 or 5 on the sample pad. Preferably, it was confirmed that the sensitivity was further improved by using the polymer 6 in combination.
The results of the improvement in detection sensitivity when the polymer was supported on the sample pad using a known novel coronavirus (SARS-CoV-2) IgG antibody detection kit-novel coronavirus (SARS-CoV-2) IgG test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 4, according to examples 3 and 4.
TABLE 4
When the polymer 1, 2, 3, 4 or 5 was supported on the sample pad, the test line having a depth equal to or slightly smaller than that of the control sample, which was the comparative example, was detected for IgG, and thus it was confirmed that the sensitivity was improved by supporting the polymer 1, 2, 3, 4 or 5 on the sample pad. Preferably, it was confirmed that the sensitivity was further improved by using the polymer 6 in combination.
The results of the improvement in detection sensitivity when the polymer was supported in the conjugate pad using a known novel coronavirus (SARS-CoV-2) IgM antibody detection kit-novel coronavirus (SARS-CoV-2) IgM test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 5 according to examples 5 and 6.
TABLE 5
When the polymer 1, 2, 3, 4 or 5 was supported on the conjugate pad, the test line having a depth equal to or slightly smaller than that of the control sample as a comparative example was detected for IgM, and thus it was confirmed that the sensitivity was improved by supporting the polymer 1, 2, 3, 4 or 5 on the conjugate pad. Preferably, it can be confirmed that the sensitivity is further improved by using the polymer 6 in combination.
The results of the improvement in detection sensitivity when the polymer was supported on the conjugate pad using a known novel coronavirus (SARS-CoV-2) IgG antibody detection kit-novel coronavirus (SARS-CoV-2) IgG test kit (colloidal gold) (manufactured by Ray Biotec. Co.) are shown in Table 6 according to examples 5 and 6.
TABLE 6
When the polymer 1, 2, 3, 4 or 5 was supported on the conjugate pad, the test line having a depth equal to or slightly smaller than that of the control sample as a comparative example was detected for IgG, and thus it was confirmed that the sensitivity was improved by supporting the polymer 1, 2, 3, 4 or 5 on the conjugate pad. Preferably, it can be confirmed that the sensitivity is further improved by using the polymer 6 in combination.
According to the above examples, the present invention can provide an immunochromatographic assay having a significantly superior sensitization effect compared with a known immunochromatographic assay using a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody as a measurement target substance.
As described above, the present invention can provide an immunochromatographic assay having an excellent sensitization effect as compared with a known immunochromatographic assay using a novel coronavirus (SARS-CoV-2) IgM antibody and/or an IgG antibody as a substance to be measured.
In addition, since the method for treating novel coronavirus infection (covd-19) of the present invention can detect novel coronavirus (SARS-CoV-2) with high sensitivity, it is possible to rapidly treat novel coronavirus infection in patients suffering from novel coronavirus infection or patients who are likely to be infected with novel coronavirus (SARS-CoV-2), and thus the therapeutic effect is high.
Industrial applicability
The present application provides a sensitizer for immunochromatography assay which uses novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured.
The present application is based on Japanese patent application 2020-206166 (application date: 12/11/2020) and Japanese patent application 2021-006659 (application date: 19/2021/1), the contents of which are all incorporated herein.
Claims (11)
1. An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [4 ]:
[ chemical formula 1]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
2. An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [3 ]:
[ chemical formula 2]
Wherein m and n each represent the number of structural units, and m: n is 100 to 30:0 to 70.
3. A sample dilution for immunochromatography assay comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be assayed, which comprises the sensitizer for immunochromatography assay according to claim 1.
4. An immunochromatographic assay comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as an analyte, characterized in that an antigen-antibody reaction is carried out in the presence of the sensitizer for immunochromatographic assay according to claim 1.
5. An apparatus for immunochromatography measurement comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a measurement target substance, which comprises the sensitizer for immunochromatography measurement according to claim 1.
6. An apparatus for immunochromatographic assay comprising a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as an analyte, wherein the sensitizer for immunochromatographic assay according to claim 1 is supported on a developed membrane, a sample pad or a conjugate pad.
7. A kit for the examination of novel coronavirus infection (COVID-19) comprising the sensitizer for immunochromatographic assay according to claim 1 and a substance specifically binding to novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody.
8. A method for testing a subject for the likelihood of infecting a novel coronavirus infection (covd-19) comprising assaying a biological sample derived from the subject for novel coronavirus (SARS-CoV-2) IgM antibodies and/or IgG antibodies by performing an antigen-antibody reaction in the presence of a sensitizer according to claim 1.
9. A method for treating a novel coronavirus infection (covd-19), comprising the steps of (1) and (2):
(1) A novel coronavirus (SARS-CoV-2) measurement step in which an antigen-antibody reaction is carried out in the presence of a sensitizer for immunochromatography measurement comprising a polymer represented by the following general formula [4], and
(2) A step of treating a patient with a novel coronavirus infection (COVID-19) based on the measurement result of (1),
[ chemical formula 3]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
10. A kit for treating novel coronavirus infection (covd-19) comprising a sensitizer for immunochromatographic assay and a therapeutic agent for novel coronavirus infection (covd-19), wherein the sensitizer is formed of a polymer represented by the following general formula [4 ]:
[ chemical formula 4]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50.
11. An sensitizer for immunochromatography measurement which comprises a novel coronavirus (SARS-CoV-2) IgM antibody and/or IgG antibody as a substance to be measured, said sensitizer comprising a polymer represented by the following general formula [4] and a copolymer represented by the following general formula [5 ]:
[ chemical formula 5]
Wherein m, n and p respectively represent the number of structural units, and m: n: p is 100-30:0-70:0-50;
[ chemical formula 6]
Wherein q R's each independently represent a hydrogen atom or a cation, m and q each represent the number of structural units, and m: q is 99 to 20:1 to 80.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2020-206166 | 2020-12-11 | ||
| JP2021006659 | 2021-01-19 | ||
| JP2021-006659 | 2021-01-19 | ||
| PCT/JP2021/044765 WO2022124270A1 (en) | 2020-12-11 | 2021-12-06 | Sensitizer for immunochromatographic assays, and assay |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117043601A true CN117043601A (en) | 2023-11-10 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202180083318.9A Pending CN117043601A (en) | 2020-12-11 | 2021-12-06 | Sensitizer for immunochromatographic assay and assay |
Country Status (1)
| Country | Link |
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| CN (1) | CN117043601A (en) |
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2021
- 2021-12-06 CN CN202180083318.9A patent/CN117043601A/en active Pending
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