JP2016533746A - 多孔質基材における核酸増幅の検出 - Google Patents
多孔質基材における核酸増幅の検出 Download PDFInfo
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Abstract
Description
Claims (27)
- 核酸の増幅を評価する方法であって、
複数の増幅試薬が多孔質基材全体に分布した多孔質基材を用意する工程と、
核酸を含む試料を多孔質基材の所定の初期位置に塗布する工程と、
試料を多孔質基材上の増幅試薬と所定時間反応させて関心核酸配列の増幅産物を生成する工程であって、幅時に増幅産物が生成したときにシグナルを与えるように構成された1以上の検出可能な部分を増幅産物が含んでいる、工程と、
基材でのシグナルの経時的進展に基づいて、試料の初期濃度を決定する工程と
を含む、方法。 - 初期濃度を決定する工程が、所定時間の終わりの増幅産物から検出されたシグナルの最も遠い位置を決定することを含んでいて、最も遠い位置が所定の初期位置に対して検討され、検出されたシグナルが閾値を超える、請求項1に記載の方法。
- 初期濃度を決定する工程が、増幅産物から検出されたシグナルが、初期位置に対する所定の距離で閾値を超える時間を求めることを含む、請求項1に記載の方法。
- 多孔質基材が、初期位置に対応する印刷マーカー及び初期位置からの異なる距離に対応する複数の印刷マーカーを含む、請求項1に記載の方法。
- 増幅試薬が等温核酸増幅試薬を含む、請求項1に記載の方法。
- 増幅試薬がDNAポリメラーゼ及び1以上の核酸プライマーを含む、請求項1に記載の方法。
- 増幅試薬がRNAポリメラーゼを含む、請求項1に記載の方法。
- 検出されたシグナルが初期位置からの最小距離に達しない場合に、試料が閾値量の関心配列を含んでいないと決定することを含む、請求項1に記載の方法。
- 試料中の目標物の初期濃度を求める工程が、増幅試薬中の1以上のプライマーと相補的な関心配列の核酸配列の出発コピー数を求めることを含む、請求項1に記載の方法。
- 試料の初期濃度に基づいて試料を採取した患者の臨床的特徴を決定することを含み、臨床的特徴が関心核酸配列の有無に関連し、増幅試薬が関心核酸配列と相補的な1以上のプライマーを含む、請求項1に記載の方法。
- 多孔質基材が液体で飽和されている、請求項1に記載の方法。
- 増幅時に基材中を液体が移動する、請求項1に記載の方法。
- 多孔質基材を用意する工程が、増幅試薬を単一溶液中の多孔質基材に塗布することを含む、請求項1に記載の方法。
- 多孔質基材を加熱して1以上の増幅試薬を活性化させることを含む、請求項1に記載の方法。
- 加熱が等温温度までである、請求項14に記載の方法。
- シグナルが、視認できるシグナル及び/又は蛍光シグナルを含む、請求項1に記載の方法。
- シグナルが、電気化学的シグナル又はpH変化を含む、請求項1に記載の方法。
- 多孔質基材が不均一なポロシティを有する、請求項1に記載の方法。
- 多孔質基材が、ポロシティが1の開孔チャネルを含む、請求項18に記載の方法。
- 初期位置からの設定距離でのシグナル強度が、試料中の目標アナライトの出発量と相関する、請求項1に記載の方法。
- 初期位置からの設定距離でのシグナルの有無を用いて、試料中の目標アナライトの出発量を計算する、請求項1に記載の方法。
- シグナルが、色素の放出によって生成する、請求項1に記載の方法。
- シグナルが、アンプリコン、H+又はピロリン酸塩の1以上によって生成する、請求項1に記載の方法。
- 増幅システムであって、
多孔質基材と、
増幅時に検出可能な部分を与えるように構成され、基材全体に分布したシグナル分子、及び目標核酸配列と相補的な1以上のプライマーを含む複数の増幅試薬と、
多孔質基材の複数の位置で部分を検出し、複数の位置の各々で検出されたシグナルに関する出力を与えるように構成された検出器と。
出力を受信して、シグナルが閾値を超える多孔質基材の試料塗布点から最も遠い距離又は多孔質基材の試料塗布点から所定の距離でシグナルが閾値を超える時間に基づいて、多孔質基材に塗布された目標核酸配列の初期濃度を決定するように構成されたプロセッサと
を備えるシステム。 - プロセッサが、経時的に、反応速度の曲線フィッティングに基づいて初期濃度の品質基準を決定するように構成された、請求項24に記載のシステム。
- 複数の増幅試薬が多孔質基材全体に分布した多孔質基材を用意する工程であって、増幅試薬が、増幅時にシグナルを与えるように構成された1以上のシグナル分子、及び目標核酸配列と相補的な1以上のプライマー、及び目標核酸配列の既知出発コピー数を含む試料を多孔質基材の初期位置に含んでいる、工程と、
シグナルが複数の時点で閾値を超える、初期位置から最も遠い距離を決定する工程と
を含む方法。 - 第2の試料が既知出発コピー数の試料を有するか否かを決定する場合に、複数の時点での最も遠い距離を用いる、請求項26に記載の方法。
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5958698A (en) * | 1992-10-26 | 1999-09-28 | Institut Belka | Method for amplification and expression of nucleic acids in solid media and its application for nucleic acid cloning and diagnostics |
JP2002532059A (ja) * | 1998-08-27 | 2002-10-02 | エックストラナ、インコーポレイテッド | 核酸抽出、増幅および検出を統合する自給自足型デバイス |
WO2006025264A1 (ja) * | 2004-08-31 | 2006-03-09 | Eiken Kagaku Kabushiki Kaisha | 核酸解析方法 |
JP2006524993A (ja) * | 2003-05-07 | 2006-11-09 | コリス バイオコンセプト エスピーアールエル | 一ステップオリゴクロマトグラフィー装置及びその使用方法 |
JP2008500820A (ja) * | 2004-04-07 | 2008-01-17 | アクセス バイオ, アイエヌシー. | 核酸検出システム |
US20080280285A1 (en) * | 2005-05-11 | 2008-11-13 | Chen Zongyuan G | Systems and Methods For Testing using Microfluidic Chips |
JP2009528037A (ja) * | 2006-03-01 | 2009-08-06 | エフ.ホフマン−ラ ロシュ アーゲー | 核酸増幅用基材 |
US20100221718A1 (en) * | 2007-06-22 | 2010-09-02 | Aj Innuscreen Gmbh | Method and rapid test for detection of specific nucleic acid sequences |
US20110039261A1 (en) * | 2007-12-20 | 2011-02-17 | Aj Innuscreen Gmbh | Mobile rapid test system for nucleic acid analysis |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5853990A (en) | 1996-07-26 | 1998-12-29 | Edward E. Winger | Real time homogeneous nucleotide assay |
US6387621B1 (en) | 1999-04-27 | 2002-05-14 | University Of Utah Research Foundation | Automated analysis of real-time nucleic acid amplification |
AUPQ495700A0 (en) | 2000-01-05 | 2000-02-03 | Johnson & Johnson Research Pty. Limited | Method for concurrent amplification and real time detection of polymorphic nucleic acid sequences |
US7364848B2 (en) | 2002-09-02 | 2008-04-29 | Pamgene B.V. | Integrated microarray analysis |
US20100173290A1 (en) | 2006-08-14 | 2010-07-08 | Koninklijke Philips Electronics N.V. | Monitoring of enzymatic processes by using magnetizable or magnetic objects as labels |
ITMI20112177A1 (it) | 2011-11-29 | 2013-05-30 | Genefast S R L | Metodo per rilevare la sintesi e/o l'amplificazione di un acido nucleico |
-
2013
- 2013-08-19 US US13/970,315 patent/US9714447B2/en active Active
-
2014
- 2014-08-19 WO PCT/EP2014/067685 patent/WO2015024948A1/en active Application Filing
- 2014-08-19 JP JP2016535465A patent/JP6374967B2/ja active Active
- 2014-08-19 EP EP14753074.5A patent/EP3036341B1/en active Active
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5958698A (en) * | 1992-10-26 | 1999-09-28 | Institut Belka | Method for amplification and expression of nucleic acids in solid media and its application for nucleic acid cloning and diagnostics |
JP2002532059A (ja) * | 1998-08-27 | 2002-10-02 | エックストラナ、インコーポレイテッド | 核酸抽出、増幅および検出を統合する自給自足型デバイス |
JP2006524993A (ja) * | 2003-05-07 | 2006-11-09 | コリス バイオコンセプト エスピーアールエル | 一ステップオリゴクロマトグラフィー装置及びその使用方法 |
JP2008500820A (ja) * | 2004-04-07 | 2008-01-17 | アクセス バイオ, アイエヌシー. | 核酸検出システム |
WO2006025264A1 (ja) * | 2004-08-31 | 2006-03-09 | Eiken Kagaku Kabushiki Kaisha | 核酸解析方法 |
US20080280285A1 (en) * | 2005-05-11 | 2008-11-13 | Chen Zongyuan G | Systems and Methods For Testing using Microfluidic Chips |
JP2009528037A (ja) * | 2006-03-01 | 2009-08-06 | エフ.ホフマン−ラ ロシュ アーゲー | 核酸増幅用基材 |
US20100221718A1 (en) * | 2007-06-22 | 2010-09-02 | Aj Innuscreen Gmbh | Method and rapid test for detection of specific nucleic acid sequences |
US20110039261A1 (en) * | 2007-12-20 | 2011-02-17 | Aj Innuscreen Gmbh | Mobile rapid test system for nucleic acid analysis |
Non-Patent Citations (3)
Title |
---|
CHEN, D., ET AL.: "An integrated, self-contained microfluidic cassette for isolation, amplification, and detection of n", BIOMEDICAL MICRODEVICES, vol. 12, no. 4, JPN6018023642, 2010, pages 705 - 719, XP019814145, ISSN: 0003822934 * |
DUDEK, M., ET AL.: "Development of a Point of Care Lateral Flow Device for Measuring Human Plasma Fibrinogen", ANALYTICAL CHEMISTRY, vol. 82, no. 5, JPN6018023640, 2010, pages 2029 - 2035, XP055146036, ISSN: 0003822933, DOI: 10.1021/ac902763a * |
SOOD, A., ET AL.: "Terminal Phosphate-Labeled Nucleotides with Improved Substrate Properties for Homogeneous Nucleic Ac", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 127, no. 8, JPN6018023643, 2005, pages 2394 - 2395, ISSN: 0003822935 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20190121649A (ko) * | 2018-04-18 | 2019-10-28 | 가천대학교 산학협력단 | 병원균 검출을 위한 비색 검출 장치 및 그의 제조방법 |
KR102082898B1 (ko) | 2018-04-18 | 2020-02-28 | 가천대학교 산학협력단 | 병원균 검출을 위한 비색 검출 장치 및 그의 제조방법 |
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