WO2022124270A1 - イムノクロマト測定法用増感剤及び測定法 - Google Patents
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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- 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
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- 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
- G01N33/56983—Viruses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/10—Detection of antigens from microorganism in sample from host
Definitions
- Specific means that the affinity for the new coronavirus (SARS-CoV-2) IgM antibody and / or the IgG antibody is higher than the affinity for other substances (in one embodiment, the new coronavirus).
- IgM antibodies and / or IgG antibodies to antigens other than the protein of (SARS-CoV-2) are not included in the "other substances”).
- the specifically bound substance is, for example, about 10-6 , 10-7 , 10-8 , 10-9 , 10-10 or less, and the new coronavirus (SARS-CoV-2) IgM antibody and / or Binds to IgG antibody.
- the specifically binding substance binds to the new coronavirus (SARS-CoV-2) IgM antibody and / or IgG antibody with KD of 10-14 , 10-13 , 10-12 or higher.
- Example 5 Polymers 1, 2, 3, 4 or 5 are supported on the conjugate pad of the kit so as to have a loading amount of 500 ⁇ g, and the serum of a new coronavirus-positive patient is diluted 100-fold with respect to the comparative example.
- the solution prepared in was developed and the detection sensitivities were compared.
- the present invention is remarkably excellent in sensitizing effect as compared with the known new type coronavirus (SARS-CoV-2) IgM antibody and / or an immunochromatographic measurement method using an IgG antibody as a measurement target substance.
- SARS-CoV-2 known new type coronavirus
- IgM antibody IgM antibody
- IgG antibody immunochromatographic measurement method
- the present invention can provide a sensitizer for an immunochromatographic measurement method using a new type coronavirus (SARS-CoV-2) IgM antibody and / or an IgG antibody as a measurement target substance.
- SARS-CoV-2 new type coronavirus
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Abstract
Description
[1] 下記一般式〔4〕で表される重合体を含んでなる、新型コロナウイルス(SARS-CoV-2)IgM抗体および/またはIgG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。
[3] [1]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抗体および/またはIgG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。
[4] [1]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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定法。
[5] [1]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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定用器具。
[6] [1]に記載のイムノクロマト測定法用増感剤を展開膜、サンプルパッドまた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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定用器具。
[8] [1]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抗体および/またはIgG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。
(1)下記一般式〔4〕で表される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。
[10] 下記一般式〔4〕で表される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。
[11] 下記一般式〔4〕で表される重合体および下記一般式〔5〕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抗体および/またはIgG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。
上記一般式〔1〕で表される重合体は、好ましくは重量平均分子量100,000~2,000,000、より好ましくは500,000~1,500,000の水溶性重合体である。
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。
(1)前記重合体または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。
(1)前記重合体または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。
(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。
(重合体1)
重量平均分子量;1030×103、MPC重合体
(重合体2)
重量平均分子量;600×103、MPC単量体に基づく構成単位数とブチルメタクリレートに基づく構成単位数の比;80:20
(重合体3)
重量平均分子量;1210×103、MPC単量体に基づく構成単位数とブチルメタクリレートに基づく構成単位数の比;80:20
(重合体4)
重量平均分子量;22×103、MPC単量体に基づく構成単位数とブチルメタクリレートに基づく構成単位数とグリセロールメタクリレートに基づく構成単位数の比;40:40:20
(重合体5)
重量平均分子量;26×103、MPC単量体に基づく構成単位数とブチルメタクリレートに基づく構成単位数とグリセロールメタクリレートに基づく構成単位数の比;40:20:40
(重合体6)
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
(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、上記の重合体が、公知の新型コロナウイルス(SARS-CoV-2)IgM抗体および/またはIgG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。
市販の新型コロナウイルス(SARS-CoV-2)IgM抗体検出キットNovel Coronavirus (SARS-CoV-2) IgM Test Kit (Colloidal Gold) (Ray Biotec.社製)およびSARS-CoV-2 IgG抗体検出キットNovel Coronavirus (SARS-CoV-2) IgG Test Kit (Colloidal Gold) (Ray Biotec.社製)(で評価した。より詳しくは、SARS-CoV-2 IgM抗体検出キットまたはSARS-CoV-2 IgG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。
当該キットの希釈液に終濃度0.5重量%となるように重合体1、2、3、4または5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。
当該キットの希釈液に終濃度0.5重量%となるように重合体1、2または4を添加したのち、さらに終濃度0.5重量%となるように重合体6を添加した。それ以外は実施例1と同様に検出感度を比較した。
当該キットのサンプルパッドに、重合体1、2、3、4または5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。
当該キットのサンプルパッドに、重合体1、2または4を担持量5mgとなるように担持し、さらに、重合体6を担持量5mgとなるように担持した。それ以外は実施例3と同様に検出感度を比較した。
当該キットのコンジュゲートパッドに、重合体1、2、3、4または5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。
当該キットのコンジュゲートパッドに、重合体1、2または4を担持量500μgとなるように担持し、さらに、重合体6を担持量500μgとなるように担持した。それ以外は実施例5と同様に検出感度を比較した。
実施例1および2に従い、公知の新型コロナウイルス(SARS-CoV-2)IgM抗体検出キットNovel Coronavirus (SARS-CoV-2) IgM Test Kit (Colloidal Gold) (Ray Biotec.社製)を使用しての、希釈液に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。
実施例1および2に従い、公知の新型コロナウイルス(SARS-CoV-2)IgG抗体検出キットNovel Coronavirus (SARS-CoV-2) IgG Test Kit (Colloidal Gold) (Ray Biotec.社製)を使用しての、希釈液に重合体を添加した際の検出感度向上の結果を表2に示す。
Claims (11)
- 請求項1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抗体および/またはIgG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。
- 請求項1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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定法。
- 請求項1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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定用器具。
- 請求項1に記載のイムノクロマト測定法用増感剤を展開膜、サンプルパッドまた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抗体および/またはIgG抗体を測定対象物質とするイムノクロマト測定用器具。
- 請求項1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抗体および/またはIgG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。
- 請求項1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抗体および/またはIgG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。
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KR20230118916A (ko) | 2023-08-14 |
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