JP2004159657A - 電気的信号を用いてpcr産物を検出する方法 - Google Patents
電気的信号を用いてpcr産物を検出する方法 Download PDFInfo
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- JP2004159657A JP2004159657A JP2003380668A JP2003380668A JP2004159657A JP 2004159657 A JP2004159657 A JP 2004159657A JP 2003380668 A JP2003380668 A JP 2003380668A JP 2003380668 A JP2003380668 A JP 2003380668A JP 2004159657 A JP2004159657 A JP 2004159657A
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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
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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
- C12Q1/6825—Nucleic acid detection involving sensors
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- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
- G01N27/22—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
- G01N27/221—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance by investigating the dielectric properties
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
【解決手段】 (a)PCR反応容器内に一対以上の電極を提供する段階と、(b)PCRを行う段階と、(c)前記電極間に電場を発生させる段階と、(d)前記PCR反応溶液の誘電特性の変化を測定する段階と、を含むPCR増幅産物を検出する方法。本発明によれば、PCR産物をリアルタイムで検出できる。
【選択図】 図1
Description
公知のPCRチューブに孔を開け、この孔を介してPCR溶液の中間部分に電極を挿し込んだ。次いで、この電極をPCR機器と連結した後、PCRを行いつつ、各PCRサイクル直後に電気的信号を検出した。本実施例では、増幅したDNAをインピーダンスを測定した。
図6ないし7に示したような過程を経て、チャンバーの上部及び下部に電極が具現されたPCRチップを製作した。
前記実施例2において、上部及び下部電極を形成したシリコンウェーハを、シリコン直接接合法を用いて、直接結合した以外は、実施例2と同様にして、2層構造のPCRチップを製造した。
本実施例では、実施例1と同様にして製造された電極を備えたPCRチューブ及び前記電極に作動的に連結されたインピーダンス測定装置(インピーダンスセンサ)を含むPCR機器を利用して、PCR産物をリアルタイムで検出した。
一定の周波数でのPCRサイクルに対するインピーダンスサイズの変化を測定することを除いては、実施例4と同じ方法で、PCR産物のリアルタイム検出を行なった。実施例4では、約1,000Hzの周波数でPCR理論に最適の結果を示すことが示されたので、この近くの周波数である1,077HzでのPCRサイクルによるDNA増幅時及び増幅が起らない時のインピーダンスサイズ(電気的抵抗)を測定した。その結果を図12及び13に示した。
Claims (6)
- (a)PCR溶液を含む容器内に一対以上の電極を提供する段階と、
(b)PCRを行う段階と、
(c)前記電極間に電場を発生させる段階と、
(d)前記PCR溶液の誘電特性の変化を測定する段階と、
を含むPCR産物を検出する方法。 - 前記(b)段階において、前記PCRは、イオンで標識されたプライマーを使用せずに行う、請求項1に記載の方法。
- 前記PCR溶液を含む容器は、PCRチューブまたは重合マイクロチャンバーである、請求項1または2に記載の方法。
- 前記誘電特性は、インピーダンス、誘電損失、誘電率またはアドミタンスである、請求項1〜3のいずれか1項に記載の方法。
- 前記(c)段階において、前記電場を発生させる段階は、周波数1Hz〜100MHzの交流を使用する、請求項1〜4のいずれか1項に記載の方法。
- 前記(c)段階において、前記電場を発生させる段階は、1mV〜10Vの平均交流電圧[平均AC電圧(Vrms)]を使用する、請求項1〜5のいずれか1項に記載の方法。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020020070141A KR100858080B1 (ko) | 2002-11-12 | 2002-11-12 | 전기적 신호를 측정하는 pcr 증폭 산물을 검출하는 방법 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2004159657A true JP2004159657A (ja) | 2004-06-10 |
JP4094530B2 JP4094530B2 (ja) | 2008-06-04 |
Family
ID=32171627
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003380668A Expired - Fee Related JP4094530B2 (ja) | 2002-11-12 | 2003-11-11 | 電気的信号を用いてpcr産物を検出する方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US7135294B2 (ja) |
EP (1) | EP1420070A1 (ja) |
JP (1) | JP4094530B2 (ja) |
KR (1) | KR100858080B1 (ja) |
CN (1) | CN1499194A (ja) |
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US7157232B2 (en) * | 2000-12-13 | 2007-01-02 | The Regents Of The University Of California | Method to detect the end-point for PCR DNA amplification using an ionically labeled probe and measuring impedance change |
US7250775B1 (en) * | 2003-11-12 | 2007-07-31 | The Regents Of The University Of California | Microfluidic devices and methods based on measurements of electrical admittance |
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US7799557B2 (en) * | 2003-12-10 | 2010-09-21 | Samsung Electronics Co., Ltd. | Polymerase chain reaction (PCR) module and multiple PCR system using the same |
US8697433B2 (en) * | 2003-12-10 | 2014-04-15 | Samsung Electronics Co., Ltd. | Polymerase chain reaction (PCR) module and multiple PCR system using the same |
US7553625B2 (en) * | 2003-12-22 | 2009-06-30 | John Wayne Cancer Institute | Method and apparatus for in vivo collection of circulating biological components |
US20050153309A1 (en) * | 2003-12-22 | 2005-07-14 | David Hoon | Method and apparatus for in vivo surveillance of circulating biological components |
KR100590569B1 (ko) * | 2004-02-14 | 2006-06-15 | 삼성전자주식회사 | 실시간 중합효소연쇄반응 검사 방법 |
CN101506380A (zh) * | 2006-08-11 | 2009-08-12 | 香港科技大学 | 用电导或电化学活跃标签实时定量分析与监察核酸扩增的方法与系统 |
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KR100969671B1 (ko) * | 2008-03-28 | 2010-07-14 | 디지탈 지노믹스(주) | 고감도 바이오 센서 및 이를 포함하는 바이오 칩 그리고이를 제조하는 방법 |
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KR20120045909A (ko) * | 2010-11-01 | 2012-05-09 | 엘지전자 주식회사 | 복수 종의 표적 핵산을 실시간으로 검출하는 장치 및 방법 |
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CN102604827B (zh) * | 2012-03-31 | 2014-01-29 | 中国科学院上海微系统与信息技术研究所 | 非接触式电导法实现pcr反应过程的实时检测系统和方法 |
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KR101950210B1 (ko) * | 2012-07-24 | 2019-02-22 | 주식회사 미코바이오메드 | 히터 유닛이 반복 배치된 열 블록을 포함하는 전기화학적 신호를 검출하기 위한 실시간 pcr 장치, 및 이를 이용한 실시간 pcr 방법 |
KR101946339B1 (ko) * | 2012-08-29 | 2019-04-25 | 주식회사 미코바이오메드 | 히터 유닛이 반복 배치된 열 블록을 포함하는 전기화학적 신호를 검출하기 위한 실시간 pcr 장치, 및 이를 이용한 실시간 pcr 방법 |
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KR101983580B1 (ko) * | 2012-08-29 | 2019-05-29 | 주식회사 미코바이오메드 | 히터 유닛이 반복 배치된 열 블록을 포함하는 전기화학적 신호를 검출하기 위한 pcr 칩, 이를 포함하는 pcr 장치, 및 이를 이용한 실시간 pcr 방법 |
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- 2003-09-09 CN CNA031584195A patent/CN1499194A/zh active Pending
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Also Published As
Publication number | Publication date |
---|---|
US7135294B2 (en) | 2006-11-14 |
KR100858080B1 (ko) | 2008-09-10 |
KR20040042021A (ko) | 2004-05-20 |
JP4094530B2 (ja) | 2008-06-04 |
US20040100284A1 (en) | 2004-05-27 |
CN1499194A (zh) | 2004-05-26 |
EP1420070A1 (en) | 2004-05-19 |
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