JP5218423B2 - アッセイシステムの正規化方法 - Google Patents
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- G01N27/416—Systems
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
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- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
- G01N21/278—Constitution of standards
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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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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Description
本出願は、2006年12月28日に提出された米国特許出願No.60/882、514の優先権を主張し、参照によって本明細書に組み入れられる。
本発明は、アッセイの内部標準化(standardization)方法及びそのシステムに関する。
あらかじめ決められた情報は、塩濃度又はイオン濃度の情報を内部標準溶液の濃度と関連付けるためのデータとしてもよい。
αフェトプロテイン(AFP)アッセイを、国際公開公報WO2007/027495号の実施例1の記載に準じて、図5で示すマイクロ流体デバイスを使用することにより実施した。以下の実施例では、表1に示した組成の各アッセイ用緩衝液組成物を100%緩衝濃度として用いた。蛍光末端標識2kb DNAを内部標準(IS)として用いた。
電気泳動中にキャピラリーチャネルで抗原−抗体反応と分離が起こる。キャピラリーチャネル(分離ゾーン)の特定ポイントで、分離されたISシグナル及び抗原−抗体複合体シグナルが検出された。電気泳動中は、TBウェル−LBウェル間の電流シグナル及びHOウェル−LBウェル間の電流シグナルをモニタリングし、それらをISピークを正規化(normalize)するのに用いた。
ISピークを以下の式によって正規化(normalized)した。
Claims (25)
- 以下の構成を有するアッセイシステムの正規化方法、
アッセイシステム内に内部標準溶液を含有する緩衝液を具備し;
アッセイに先立って前記緩衝液の電気伝導率測定値を求め;
前記電気伝導率測定値に基づいて前記緩衝液の濃度変化を求め;
アッセイ結果を正規化するために前記緩衝液の前記濃度変化に基づいて前記内部標準溶液の濃度を決定する。 - 前記緩衝液が内部標準のために用意される、請求項1に記載の方法。
- 前記緩衝液は複数の成分を含み、その成分の内の1つが内部標準溶液である、請求項1に記載の方法。
- 前記緩衝液は複数の成分を含み、その成分の内の1つが前記内部標準溶液であり;そして前記緩衝液の電気伝導率測定値は前記複数の成分の総電気伝導率である、請求項1に記載の方法。
- 前記内部標準溶液の濃度は前記緩衝液の前記濃度変化を外挿することによって求められる、請求項4に記載の方法。
- 前記緩衝液の前記電気伝導率は前記緩衝液の塩又はイオンの濃度とほぼ比例する、請求項1に記載の方法。
- さらに、アッセイに先立って前記電気伝導率測定値を求める前に、前記緩衝液の初期濃度及び対応する前記緩衝液の電気伝導率測定値を求めることを含む、請求項1に記載の方法。
- 前記システムがマイクロ流体電気泳動アッセイシステムである、請求項1に記載の方法。
- 前記アッセイ結果は、蛍光内部標準シグナル及び前記内部標準溶液の濃度に基づいて正規化される、請求項1に記載の方法。
- 以下の構成を有するアッセイシステムの正規化方法、
アッセイシステム内に複数の緩衝液成分を具備し、その成分の内の1つが内部標準溶液を含有し;
アッセイに先立って前記複数の緩衝液成分の電気伝導率測定値を求め;
前記電気伝導率測定値に基づいて前記複数の緩衝液成分内の塩又はイオンの濃度変化を求め;
アッセイ結果を正規化するために塩又はイオンの前記濃度変化に基づいて前記内部標準溶液の濃度を決定する。 - 前記緩衝液成分の前記電気伝導率測定値は、前記緩衝液成分の総電気伝導率測定値である、請求項10に記載の方法。
- 前記内部標準溶液の濃度は、前記複数の緩衝液成分内の塩又はイオンの前記濃度変化を外挿することによって求められる、請求項11に記載の方法。
- 前記緩衝液の電気伝導率は前記緩衝液の塩又はイオンの濃度とほぼ比例する、請求項10記載の方法。
- さらに、アッセイに先立って前記電気伝導率測定値を求める前に、前記緩衝液成分の初期濃度及び前記緩衝液成分の対応する電気伝導率測定値をあらかじめ決めることを含む、請求項10に記載の方法。
- 前記システムがマイクロ流体電気泳動アッセイシステムである、請求項10に記載の方法。
- 前記アッセイ結果は蛍光内部標準シグナル及び内部標準溶液の濃度に基づいて正規化される、請求項10に記載の方法。
- 以下の構成を有するアッセイシステム、
緩衝液試薬を収容するウェルを有し、そのウェルの内の1つが内部標準溶液を収容するウェルを具備する基板;
前記緩衝液試薬を収容する前記ウェル間で電気伝導率を測定するための回路部材;
前記ウェル間で電気伝導率測定値から前記緩衝液試薬の塩濃度又はイオン濃度を測定するための制御部材;
あらかじめ決められた情報を記憶する記憶部材を含み;
前記制御部材が前記記憶部材中のあらかじめ決められた情報に基づいて前記塩濃度又はイオン濃度から前記内部標準溶液の濃度を外挿する。 - 前記システムがマイクロ流体電気泳動アッセイシステムである、請求項17に記載のアッセイシステム。
- 前記制御部材は前記内部標準溶液の濃度に基づいてアッセイ結果を正規化する、請求項17に記載のアッセイシステム。
- 前記制御部材は前記内部標準溶液の濃度及び前記記憶部材に記憶された蛍光内部標準シグナルに基づいてアッセイ結果を正規化する、請求項17に記載のアッセイシステム。
- あらかじめ決められた情報は、塩濃度情報又はイオン濃度情報を前記内部標準溶液の濃度と関連付けるデータである、請求項17に記載のアッセイシステム。
- 前記制御部材は、電気伝導率測定値を前記記憶部材に記憶された標準情報と比較することによって前記システムの形状寸法の変動に対してアッセイ結果を正規化する、請求項17に記載のアッセイシステム。
- 以下の構成を有するアッセイシステムの正規化方法、
前記アッセイシステム内に内部標準溶液を含有する緩衝液を具備し;
アッセイに先立って前記緩衝液の電気伝導率測定値を求め;そして、
前記電気伝導率測定値に基づいてアッセイ結果を正規化する。 - さらに、前記電気伝導率測定値に基づいて前記緩衝液の濃度変化を求め;
前記アッセイ結果を正規化するために前記緩衝液の濃度変化に基づいて前記内部標準溶液の濃度を決定すること、を含む請求項23に記載の正規化方法。 - さらに、前記電気伝導率測定値をあらかじめ決められたデータと比較し;そして、
その比較に基づいて前記アッセイ結果を正規化することを、含む請求項23に記載の正規化方法。
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US88251406P | 2006-12-28 | 2006-12-28 | |
US60/882,514 | 2006-12-28 | ||
PCT/US2007/026523 WO2008082670A2 (en) | 2006-12-28 | 2007-12-27 | Method and system for internal standardization of assays |
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JP2010515053A JP2010515053A (ja) | 2010-05-06 |
JP2010515053A5 JP2010515053A5 (ja) | 2013-02-14 |
JP5218423B2 true JP5218423B2 (ja) | 2013-06-26 |
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US (1) | US20100117666A1 (ja) |
EP (1) | EP2100138B1 (ja) |
JP (1) | JP5218423B2 (ja) |
ES (1) | ES2388005T3 (ja) |
WO (1) | WO2008082670A2 (ja) |
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CN102414559B (zh) * | 2009-04-28 | 2014-03-19 | 和光纯药工业株式会社 | 使用内标物质的测定方法 |
US9075042B2 (en) | 2012-05-15 | 2015-07-07 | Wellstat Diagnostics, Llc | Diagnostic systems and cartridges |
US9625465B2 (en) | 2012-05-15 | 2017-04-18 | Defined Diagnostics, Llc | Clinical diagnostic systems |
US9213043B2 (en) | 2012-05-15 | 2015-12-15 | Wellstat Diagnostics, Llc | Clinical diagnostic system including instrument and cartridge |
WO2014205230A1 (en) * | 2013-06-19 | 2014-12-24 | Step Ahead Innovations Inc. | Aquatic environment water parameter testing systems and methods |
KR101729685B1 (ko) * | 2015-05-21 | 2017-05-11 | 한국기계연구원 | 이온 농도 검출 방법 및 장치 |
CN104897634A (zh) * | 2015-06-16 | 2015-09-09 | 任志海 | 一种原子荧光测定易化学蒸气发生反应元素时的内标法 |
DE102016009442A1 (de) * | 2016-08-03 | 2018-02-08 | Fresenius Medical Care Deutschland Gmbh | Verfahren zur Überwachung des Bicarbonat-Gehalts und des Natrium-Gehalts einer Dialyselösung |
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US4753888A (en) * | 1986-04-09 | 1988-06-28 | Bionostics, Inc. | Multiple control standard for blood analysis |
US5605662A (en) * | 1993-11-01 | 1997-02-25 | Nanogen, Inc. | Active programmable electronic devices for molecular biological analysis and diagnostics |
WO1995010041A1 (en) * | 1993-10-07 | 1995-04-13 | Beckman Instruments, Inc. | Use of capillary electrophoresis for quantitating the concentration of protein components and of the total protein in fluids |
US5948684A (en) * | 1997-03-31 | 1999-09-07 | University Of Washington | Simultaneous analyte determination and reference balancing in reference T-sensor devices |
GB9612264D0 (en) * | 1996-06-12 | 1996-08-14 | Samsoondar James | Quality control material for monitoring calibration of instruments designed to measure serum and plasma specimen integrity |
US6632676B1 (en) * | 1999-09-24 | 2003-10-14 | Clinical Diagnostic Solutions | Multi-purpose reagent system and method for enumeration of red blood cells, white blood cells and thrombocytes and differential determination of white blood cells |
US6265170B1 (en) * | 2000-01-24 | 2001-07-24 | Ingeneus Corporation | Homogenous assay of duplex of triplex hybridization by means of multiple measurements under varied conditions |
US6358387B1 (en) * | 2000-03-27 | 2002-03-19 | Caliper Technologies Corporation | Ultra high throughput microfluidic analytical systems and methods |
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JP3643011B2 (ja) * | 2000-05-18 | 2005-04-27 | アークレイ株式会社 | 定量分析法 |
CA2327527A1 (en) | 2000-12-27 | 2002-06-27 | Geneka Biotechnologie Inc. | Method for the normalization of the relative fluorescence intensities of two rna samples in hybridization arrays |
EP1404878A4 (en) * | 2001-07-11 | 2006-12-06 | Applera Corp | INTERNAL CALIBRATION STANDARDS FOR ELECTROPHORETIC ANALYZES |
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US20100117666A1 (en) | 2010-05-13 |
WO2008082670A3 (en) | 2008-11-27 |
EP2100138A4 (en) | 2010-08-25 |
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WO2008082670A2 (en) | 2008-07-10 |
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