JPH09508525A - エネルギー転移連結の色素によりラベルされたプローブ - Google Patents
エネルギー転移連結の色素によりラベルされたプローブInfo
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- JPH09508525A JPH09508525A JP7520679A JP52067995A JPH09508525A JP H09508525 A JPH09508525 A JP H09508525A JP 7520679 A JP7520679 A JP 7520679A JP 52067995 A JP52067995 A JP 52067995A JP H09508525 A JPH09508525 A JP H09508525A
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer
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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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/81—Packaged device or kit
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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- Inks, Pencil-Leads, Or Crayons (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Steroid Compounds (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
- Thermal Transfer Or Thermal Recording In General (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.注目の少なくとも2種の構成成分を検出するために異なる螢光ラベルを用 いて複数成分の混合物中の構成成分を同定し、そして検出するための方法であっ て、前記ラベルが、 (1)主鎖に結合されるドナー−受容体螢光対を有し、ここで前記ドナーから前 記受容体への効果的なエネルギー転移が生ずるものであり;そして (2)前記個々のラベルが実質的に同じ波長で吸光し、そして異なる波長で発光 すること、 によって特徴づけられており; 前記複数成分の混合物の異なった成分に異なったラベルを結合し; 前記受容体の吸光波長において共通の光源により照射することにより前記ラベ ルされた個々の成分を検出し、そして前記個々のラベルの螢光を検出することを 含んで成る方法。 2.前記ドナーが 350〜 800nmの波長範囲で光を吸収し、そして前記受容体が 450〜1000nmの波長範囲で光を発する請求の範囲第1項記載の方法。 3.前記受容体−ドナー対が9−フェニルキサンテンである請求の範囲第2項 記載の方法。 4.注目の少なくとも2種の構成成分を検出するために異なる螢光ラベルを用 いて複数成分の混合物中の構成成分を同定し、そして検出するための方法であっ て、前記ラベルが、 (1)オリゴヌクレオチド鎖に結合されたドナー−受容体螢光対を有し、ここで 前記ドナーから前記受容体への効果的なエネルギー転移が生ずるものであり;そ して (2)前記個々のラベルが実質的に同じ波長で吸光し、そして異なる波長で発光 する、 ことによって特徴づけられており; 前記複数成分の混合物の異なる成分に異なるラベルを結合し; 前記受容体の吸光波長において共通の光源により照射することにより前記ラベ ルされた個々の成分を検出し、そして前記個々のラベルの螢光を検出することを 含んで成る方法。 5.前記ドナーが 350〜 800nmの波長範囲で光を吸収し、そして前記受容体が 450〜1000nmの波長範囲で光を発する請求の範囲第4項記載の方法。 6.前記受容体−ドナー対が9−フェニルキサンテンである請求の範囲第5項 記載の方法。 7.複数成分の混合物の構成成分を分離するための方法であって、注目の個々 の異なる成分が異なるラベルによりラベルされ、前記ラベルが、 (1)オリゴヌクレオチド鎖に結合されたドナー−受容体螢光対を有し、ここで 前記ドナーから前記受容体への効果的なエネルギー転移が生ずるものであり; (2)前記個々のラベルが実質的に同じ波長で吸光し、そして異なった波長で発 光し;そして (3)前記個々の異なるラベルが、前記オリゴヌクレオチド鎖にそって前記ドナ ー−受容体対のスペーサーを変えることの結果として前記分離において実質的に 同じ移動性を有する、 ことによって特徴づけられており、 前記複数成分の混合物の異なる成分に異なるラベルを結合し; 前記成分を個々の画分に分離し;そして 前記受容体の吸光波長において共通の光源により照することによ り前記ラベルされた個々の成分を検出し、そして前記個々のラベルの螢光を検出 することを含んで成る方法。 8.前記分離が電気泳動によるものである請求の範囲第7項記載の方法。 9.前記ドナーが 350〜 800nmの波長範囲で光を吸収し、そして前記受容体が 450〜1000nmの波長範囲で光を発する請求の範囲第7項記載の方法。 10.前記受容体−ドナー対が9−フェニルキサンテンである請求の範囲第9項 記載の方法。 11.一本鎖核酸をコピーするためのプライマー及び特定のヌクレオチドで前記 コピーに起因する鎖を終結するためのジデオキシヌクレオシドを用いて核酸配列 を配列決定するための方法であって、 配列決定されるべき核酸フラグメントを、プライマーに結合する前記フラグメ ントの5′側配列を含んで成るベクターでクローンニングし; 異なる反応容器において、前記プライマー、dNTP及び少量の異なる1種のジデ オキシヌクレオチドの存在下で DNAポリメラーゼにより前記フラグメントをコピ ーし;そして 一本鎖 DNAフラグメントの得られる混合物を分離し、そしてゲル上に存在する バンドにより配列を決定することを含んで成り、 ここで、(1)前記プライマー結合配列に対して相補的な核酸鎖に結合される 受容体−ドナー螢光対を有し、ここで前記ドナーが前記受容体の螢光のために前 記受容体にエネルギーを効果的に転移し、(2)前記個々のプライマーが実質的 に同じ波長で吸光し、そして異なった波長で発光し;そして(3)前記個々のプ ライマーが、前記核酸鎖にそって前記ドナー−受容体対の空間及び螢光団を変え ることに起因する、前記分離において実質的に同じ移動性を有する ことにより特徴づけられるプライマーを用いることを特徴とする方法。 12.前記受容体−ドナー螢光対のメンバーの1つが前記プライマーの5′末端 に結合される請求の範囲第11項記載の方法。 13.異なった受容体−ドナー対を有する4種のプライマーが存在する請求の範 囲第11項記載の方法。 14.前記受容体−ドナー螢光対が10よりも多くないヌクレオチドにより分離さ れる請求の範囲第11項記載の方法。 15.少なくとも2つの受容体−ドナー螢光対がキサンテン化合物である請求の 範囲第11項記載の方法。 16.前記キサンテン化合物がフルオレセイン及びローダミンを含んで成る請求 の範囲第15項記載の方法。 17.少なくとも2種の螢光化合物を含んで成るキットであって、前記個々の螢 光化合物が、 (1)主鎖に結合された受容体−ドナー螢光対を有し、ここで前記ドナーが前 記受容体の螢光のために前記受容体にエネルギーを効果的に転移し;(2)個々 のラベルが実質的に同じ波長で吸光し、そして異なった波長で発光し;そして( 3)個々のラベルが、前記主鎖にそって前記ドナー−受容体対の空間を変えるこ とに起因する、前記分離において実質的に同じ移動性を有することを特徴とする キット。 18.前記主鎖が核酸鎖であり、そして前記受容体−ドナー螢光対が約10個より も多くないヌクレオチドにより分離される請求の範囲第17項記載のキット。 19.前記ドナーが 350〜 800の波長範囲で光を吸光し、そして前記受容体が 4 50〜1000の波長範囲で光を発する請求の範囲第17項記載のキット。 20.少なくとも2つの受容体−ドナー螢光対がキサンテン化合物である請求の 範囲第19項記載のキット。 21.前記キサンテン化合物がフルオレセイン又はローダミンである請求の範囲 第20項記載のキット。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/189,924 US5654419A (en) | 1994-02-01 | 1994-02-01 | Fluorescent labels and their use in separations |
US08/189,924 | 1994-02-01 | ||
PCT/US1995/001205 WO1995021266A1 (en) | 1994-02-01 | 1995-01-30 | Probes labelled with energy transfer coupled dyes |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP2006257134A Division JP4602956B2 (ja) | 1994-02-01 | 2006-09-22 | エネルギー転移連結の色素によりラベルされたプローブ |
Publications (2)
Publication Number | Publication Date |
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JPH09508525A true JPH09508525A (ja) | 1997-09-02 |
JP3895369B2 JP3895369B2 (ja) | 2007-03-22 |
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Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
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JP52067995A Expired - Lifetime JP3895369B2 (ja) | 1994-02-01 | 1995-01-30 | エネルギー転移連結の色素によりラベルされたプローブ |
JP2006257134A Expired - Lifetime JP4602956B2 (ja) | 1994-02-01 | 2006-09-22 | エネルギー転移連結の色素によりラベルされたプローブ |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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JP2006257134A Expired - Lifetime JP4602956B2 (ja) | 1994-02-01 | 2006-09-22 | エネルギー転移連結の色素によりラベルされたプローブ |
Country Status (9)
Country | Link |
---|---|
US (3) | US5654419A (ja) |
EP (1) | EP0743987B1 (ja) |
JP (2) | JP3895369B2 (ja) |
AT (1) | ATE236994T1 (ja) |
AU (1) | AU692230B2 (ja) |
CA (1) | CA2182516C (ja) |
DE (3) | DE19581489B4 (ja) |
ES (1) | ES2197193T3 (ja) |
WO (1) | WO1995021266A1 (ja) |
Cited By (5)
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JP2003334097A (ja) * | 2002-03-12 | 2003-11-25 | Enzo Life Sciences Inc | リアルタイム核酸検出法と組成物 |
JP2004043819A (ja) * | 1996-05-03 | 2004-02-12 | Applera Corp | 硬質リンカ―を有するエネルギ―転移色素 |
JP2004510144A (ja) * | 2000-09-18 | 2004-04-02 | メディミューン,インコーポレイテッド | ワクチンの免疫原性を測定する試験管内検定 |
JP2006521092A (ja) * | 2003-04-04 | 2006-09-21 | エフ.ホフマン−ラ ロシュ アーゲー | マルチカラーリアルタイムpcr用の改良されたシステム |
JP2014502496A (ja) * | 2010-12-16 | 2014-02-03 | バイオ−ラッド ラボラトリーズ インコーポレーティッド | 定量的増幅のためのユニバーサルリファレンス色素 |
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JP2004043819A (ja) * | 1996-05-03 | 2004-02-12 | Applera Corp | 硬質リンカ―を有するエネルギ―転移色素 |
JP2004068023A (ja) * | 1996-05-03 | 2004-03-04 | Applera Corp | 硬質リンカ―を有するエネルギ―転移色素 |
JP2004510144A (ja) * | 2000-09-18 | 2004-04-02 | メディミューン,インコーポレイテッド | ワクチンの免疫原性を測定する試験管内検定 |
JP2003334097A (ja) * | 2002-03-12 | 2003-11-25 | Enzo Life Sciences Inc | リアルタイム核酸検出法と組成物 |
JP2010017196A (ja) * | 2002-03-12 | 2010-01-28 | Enzo Life Sciences Inc | リアルタイム核酸検出法と組成物 |
JP4547698B2 (ja) * | 2002-03-12 | 2010-09-22 | エンゾ ライフ サイエンシズ、インコーポレイテッド | リアルタイム核酸検出法と組成物 |
JP2006521092A (ja) * | 2003-04-04 | 2006-09-21 | エフ.ホフマン−ラ ロシュ アーゲー | マルチカラーリアルタイムpcr用の改良されたシステム |
JP2009232871A (ja) * | 2003-04-04 | 2009-10-15 | F Hoffmann La Roche Ag | マルチカラーリアルタイムpcr用の改良されたシステム |
JP2014502496A (ja) * | 2010-12-16 | 2014-02-03 | バイオ−ラッド ラボラトリーズ インコーポレーティッド | 定量的増幅のためのユニバーサルリファレンス色素 |
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DE29521620U1 (de) | 1997-11-13 |
JP3895369B2 (ja) | 2007-03-22 |
CA2182516C (en) | 2003-11-25 |
CA2182516A1 (en) | 1995-08-10 |
DE69530286T2 (de) | 2004-04-01 |
AU1736795A (en) | 1995-08-21 |
DE19581489T1 (de) | 1997-01-02 |
ATE236994T1 (de) | 2003-04-15 |
JP2007000151A (ja) | 2007-01-11 |
WO1995021266A1 (en) | 1995-08-10 |
EP0743987A4 (en) | 1998-04-15 |
US5654419A (en) | 1997-08-05 |
ES2197193T3 (es) | 2004-01-01 |
JP4602956B2 (ja) | 2010-12-22 |
DE69530286D1 (de) | 2003-05-15 |
EP0743987A1 (en) | 1996-11-27 |
DE19581489B4 (de) | 2004-07-15 |
EP0743987B1 (en) | 2003-04-09 |
US5688648A (en) | 1997-11-18 |
AU692230B2 (en) | 1998-06-04 |
US5707804A (en) | 1998-01-13 |
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