JP5706315B2 - 検査試薬及びそれを用いた被検試料中の被測定物の測定方法 - Google Patents
検査試薬及びそれを用いた被検試料中の被測定物の測定方法 Download PDFInfo
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Images
Classifications
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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
- G01N33/531—Production of immunochemical test materials
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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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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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
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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
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
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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
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
- G01N33/545—Synthetic resin
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Description
緩衝液であるA液に含まれる緩衝剤は、検査試薬に通常用いられている周知の緩衝剤であってよく、例えばアラニン、アルギニン、アスパラギン、アスパラギン酸、システイン、グルタミン、グルタミン酸、グリシン、ヒスチジン、イソロイシン、ロイシン、リシン、メチオニン、フェニルアラニン、プロリン、セリン、スレオニン、トリプトファン、チロシン、バリン等のアミノ酸;クエン酸、マレイン酸、グルタル酸等のカルボン酸塩;リン酸ナトリウム、リン酸水素ナトリウム等のリン酸塩;炭酸カルシウム、炭酸マグネシウム等の炭酸塩;グッドの緩衝剤等が好ましく使用可能であり、トリス緩衝剤、グッド緩衝剤、リン酸系緩衝剤が好適に用いられるが、緩衝剤はこれらに限定されるものではない。
上記の通り、B液中には、被測定物捕捉物質が不溶性担体粒子上に担持された粒子が浮遊状態で含まれる。この浮遊粒子は、何ら限定されるものではなく、従来から検査試薬において用いられている周知の粒子であってよい。
上記本発明の検査試薬を用いた本発明の測定方法は、A液と、B液と、被検試料とを混合する工程と、得られた混合液中の前記粒子の凝集の程度を測定する工程とを含む。
上記本発明の検査試薬を用いた本発明の検査試薬キットは、A液、B液及び容器等、検査に必要な薬品及び部品からなる。
以下の通り、浮遊されたタンパク質を担持した不溶性担体粒子を含む粒子浮遊液(B液)の自己凝集の違いを測定した。また、生理食塩水をブランク検体とし、緩衝液(A液)の電気伝導度を変えた反応条件下での非特異的凝集の違いを評価した。また、CRPの測定により感度の違いを評価した。
A液:100mMトリス含有の緩衝液に、電気伝導度をモニターしながら塩化ナトリウムを添加し、所定の電気伝導度とした。電気伝導度8ms/cm(塩化ナトリウム濃度50mM、比較例1、5)、16ms/cm(塩化ナトリウム濃度150mM、比較例2)、35ms/cm(塩化ナトリウム濃度400mM、実施例1)、75ms/cm(塩化ナトリウム濃度1000mM、実施例2、比較例3,4)、200ms/cm(塩化ナトリウム濃度5000mM、実施例3)。
上記のB液の初期の吸光度と1ヵ月後の吸光度を測定して、自己凝集の有無を確認した。
自己凝集は、0.1%ポリスチレンラテックス粒子分散液の吸光度と、B液の吸光度の差を保存の前後でそれぞれ算出し、以下の判定基準に従って各電気伝導度における自己凝集を評価した。
0.1%ポリスチレンラテックス粒子分散液の吸光度を基準として、
○:差が15%以下
×:差が15%超
○:吸光度変化量ΔAbsの10000倍が20以下
×:吸光度変化量ΔAbsの10000倍が20超
CRPの測定は、ブランクである生理食塩液とCRP0.05mg/dL溶液測定時の吸変化量の差を算出し、感度を比較した。
A液及びB液の電気伝導度の測定は、東亜電波工業(株)社製電気伝導率計CM−60G型を用いて行った。
(電極:CT−57101B、温度:25℃)
結果を下記表1に示す。
検量線作成
上記実施例2の試薬を用い、種々の濃度のCRPを含む標準溶液を用いて上記と同様に測定を行い、キャリブレーションを行った。得られた吸光度変化量とCRP濃度の関係を図4に示す。
血清検体の測定
上記実施例2の試薬を用い、血清検体について上記と同様にCRPの測定を行った。測定を20回繰返し行い、その再現性を調べた。結果を表2に示す。
Claims (6)
- 30ms/cm以上の電気伝導度を有する緩衝液であるA液と、被測定物捕捉物質が不溶性担体粒子上に担持された粒子が浮遊している粒子浮遊液であって6.5 ms/cm以下の電気伝導度を有するB液とを少なくとも含む、前記被測定物を測定するための検査試薬。
- 前記A液の電気伝導度が200ms/cm以下である請求項1に記載の検査試薬。
- 前記被測定物捕捉物質が抗体若しくはその抗原結合性断片、又は抗原である請求項1又は2に記載の検査試薬。
- 前記不溶性担体粒子がラテックス粒子である請求項1ないし3のいずれか1項に記載の検査試薬。
- 前記ラテックス粒子がポリスチレンラテックス粒子である請求項4に記載の検査試薬。
- 請求項1ないし5のいずれか1項に記載の検査試薬を少なくとも含む、検査試薬キット。
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AU2017321642B2 (en) * | 2016-08-31 | 2024-06-13 | Eiken Kagaku Kabusiki Kaisha | Antibody measurement method using antigen-carrying insoluble carrier particles on which antigen is immobilized by different methods, and reagent for antibody measurement |
JP6224217B1 (ja) * | 2016-12-27 | 2017-11-01 | Jsr株式会社 | ラテックス粒子分散液の保管方法 |
JP6919499B2 (ja) * | 2017-08-01 | 2021-08-18 | 藤倉化成株式会社 | 不溶性担体粒子を含有する免疫測定試薬の劣化防止手段 |
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Citations (7)
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JPS57182168A (en) * | 1981-05-02 | 1982-11-09 | Mitsubishi Chem Ind Ltd | Immunochemical reagent |
JPH0688821A (ja) * | 1992-09-08 | 1994-03-29 | Tokuyama Soda Co Ltd | 免疫学的凝集反応試薬の製造方法 |
JPH06167495A (ja) * | 1992-07-14 | 1994-06-14 | S R L:Kk | 凝集イムノアッセイ法 |
JPH0720129A (ja) * | 1993-07-02 | 1995-01-24 | Tokuyama Corp | 免疫学的凝集反応試薬 |
JPH0875746A (ja) * | 1994-09-02 | 1996-03-22 | Tokuyama Corp | 免疫学的凝集反応用担体粒子の凝集特性評価方法 |
JP2005351643A (ja) * | 2004-06-08 | 2005-12-22 | Dai Ichi Pure Chem Co Ltd | 免疫学的測定試薬 |
WO2007063616A1 (ja) * | 2005-11-30 | 2007-06-07 | Nihon University | 超高感度c-反応性タンパク質測定試薬及び測定方法 |
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JP2525486B2 (ja) | 1989-09-07 | 1996-08-21 | 富士写真フイルム株式会社 | 写真焼付方法 |
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS57182168A (en) * | 1981-05-02 | 1982-11-09 | Mitsubishi Chem Ind Ltd | Immunochemical reagent |
JPH06167495A (ja) * | 1992-07-14 | 1994-06-14 | S R L:Kk | 凝集イムノアッセイ法 |
JPH0688821A (ja) * | 1992-09-08 | 1994-03-29 | Tokuyama Soda Co Ltd | 免疫学的凝集反応試薬の製造方法 |
JPH0720129A (ja) * | 1993-07-02 | 1995-01-24 | Tokuyama Corp | 免疫学的凝集反応試薬 |
JPH0875746A (ja) * | 1994-09-02 | 1996-03-22 | Tokuyama Corp | 免疫学的凝集反応用担体粒子の凝集特性評価方法 |
JP2005351643A (ja) * | 2004-06-08 | 2005-12-22 | Dai Ichi Pure Chem Co Ltd | 免疫学的測定試薬 |
WO2007063616A1 (ja) * | 2005-11-30 | 2007-06-07 | Nihon University | 超高感度c-反応性タンパク質測定試薬及び測定方法 |
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ES2440326T3 (es) | 2014-01-28 |
EP2418486B1 (en) | 2013-11-06 |
US9250235B2 (en) | 2016-02-02 |
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