JP2000511776A - メチル化特異的検出 - Google Patents
メチル化特異的検出Info
- Publication number
- JP2000511776A JP2000511776A JP10500779A JP50077998A JP2000511776A JP 2000511776 A JP2000511776 A JP 2000511776A JP 10500779 A JP10500779 A JP 10500779A JP 50077998 A JP50077998 A JP 50077998A JP 2000511776 A JP2000511776 A JP 2000511776A
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- nucleic acid
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- seq
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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
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
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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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/154—Methylation markers
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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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Photoreceptors In Electrophotography (AREA)
- Investigating Or Analyzing Materials By The Use Of Fluid Adsorption Or Reactions (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.メチル化されたCpG含有核酸を検出する方法であって、核酸含有検体を、 非メチル化シトシンを修飾する試薬と接触させ、該検体中のCpG含有核酸を CpG特異的オリゴヌクレオチドプライマーにより増幅し、その際、該オリゴ ヌクレオチドプライマーは修飾された非メチル化核酸とメチル化核酸とを区別 するものであり、そしてメチル化核酸を検出することを含んでなる方法。 2.増幅工程がポリメラーゼ連鎖反応(PCR)である、請求項1に記載の方法 。 3.修飾試薬が重亜硫酸塩である、請求項1に記載の方法。 4.シトシンがウラシルに修飾される、請求項1に記載の方法。 5.CpG含有核酸がプロモーター領域に存在する、請求項1に記載の方法。 6.前記プロモーターが癌抑制遺伝子のプロモーターである、請求項5に記載の 方法。 7.癌抑制遺伝子がp16、p15、EカドヘリンおよびVHLよりなる群から 選択される、請求項6に記載の方法。 8.CpG含有核酸がアンドロゲン受容体、エストロゲン受容体、TGF−β1 、TGF−β2、p130、BRCA2、NF1、NF2、TSG101、M DGI、GST−pi、カルシトニン、HIC−1、エンドセリンB受容体、 TIMP−2、06−MGMT、MLHI、MSH2およびGFAPよりなる 群から選択されるタンパク質をコードする、請求項1に記載の方法。 9.前記検体が脳、結腸、尿生殖器、肺、腎臓、造血組織、乳房、胸腺、精巣、 卵巣および子宮組織、または血清、尿、唾液、脳脊髄液、痰、射出精液、血液 および糞便よりなる群から選択される、請求項1に記載の方法。 10.前記核酸をメチル化感受性制限エンドヌクレアーゼと接触させることをさら に含む、請求項1に記載の方法。 11.前記制限エンドヌクレアーゼがMspI、HpaII、BssHII、BstUIおよびNotIより なる群から選択される、請求項10に記載の方法。 12.前記検体におけるメチル化CpG含有核酸の存在が細胞増殖性疾患を示す、 請求項1に記載の方法。 13.前記疾患が低度星状細胞腫、未分化星状細胞腫、グリア芽細胞腫、髄芽細胞 腫、結腸癌、肺癌、腎臓癌、白血病、乳癌、前立腺癌、子宮内膜癌および神経 芽細胞腫よりなる群から選択される、請求項12に記載の方法。 14.前記プライマーが配列番号1〜103および配列番号104よりなる群から選択さ れる配列を有する標的ポリヌクレオチド配列とハイブリダイズする、請求項1 に記載の方法。 15.前記プライマーが配列番号105〜207および配列番号208よりなる群から選択 される、請求項1に記載の方法。 16.非メチル化シトシンを修飾する試薬を保持する第1の容器を含む1個以上の 容器を密着拘束状態で受容するように区画化されている担体手段を含んでなる メチル化CpG含有核酸検出用キット。 17.CpG含有核酸の増幅用プライマーを保持する第2の容器をさらに含む、請 求項16に記載のキット。 18.前記試薬が重亜硫酸塩である、請求項16に記載のキット。 19.シトシンがウラシルに修飾される、請求項16に記載のキット。 20.前記プライマーが配列番号1〜103および配列番号104よりなる群から選択さ れる配列を有する標的ポリヌクレオチド配列とハイブリダイズする、請求項17 に記載のキット。 21.前記プライマーが配列番号105〜207および配列番号208よりなる群から選択 される、請求項17に記載のキット。 22.サンプルからメチル化CpG含有核酸を検出するためのキットであって、 a)非メチル化シトシンヌクレオチドを修飾する試薬、 b)対照核酸、 c)非メチル化CpG含有核酸の増幅用プライマー、 d)メチル化CpG含有核酸の増幅用プライマー、および e)対照核酸の増幅用プライマー、 を含んでなるキット。 23.核酸増幅緩衝液をさらに含む、請求項22に記載のキット。 24.非メチル化シトシンを修飾する試薬が重亜硫酸塩である、請求項22に記載の キット。 25.前記プライマーが配列番号1〜103および配列番号104よりなる群から選択さ れる配列を有する標的ポリヌクレオチド配列とハイブリダイズする、請求項22 に記載のキット。 26.前記プライマーが配列番号105〜207および配列番号208よりなる群から選択 される、請求項22に記載のキット。 27.メチル化CpG含有核酸を検出するための単離されたオリゴヌクレオチドプ ライマーであって、配列番号1〜103および配列番号104よりなる群から選択さ れる配列を有する標的ポリヌクレオチド配列とハイブリダイズするプライマー 。 28.前記プライマーの対が配列番号順で対にした配列番号105〜207および配列番 号208である、請求項27に記載のプライマー。 29.被験者由来のTIMP−2核酸中のCpG島のメチル化と関連した細胞増殖 性疾患を検出する方法であって、 被験者由来の組織または生物学的液体のサンプル中の標的核酸を、該標的核 酸がDNAであるときはTIMP−2のメチル化を検出し、該標的核酸がRN AであるときはTIMP−2 RNAのレベルを検出するTIMP−2検出試 薬と接触させ、そして TIMP−2標的核酸を検出する、 ことを含んでなり、その際、DNAのプロモーターの高メチル化、または正常 細胞のTIMP−2 RNAレベルと比較したときのTIMP−2 RNAレ ベルの低下がTIMP−2と関連した細胞増殖性疾患を示すものである、上記 方法。 30.前記試薬が核酸プローブである、請求項29に記載の方法。 31.前記プローブが検出可能に標識されている、請求項30に記載の方法。 32.前記標識が放射性同位元素、生物発光化合物、化学発光化合物、蛍光化合物 、金属キレートおよび酵素よりなる群から選択される、請求項31に記載の方法 。 33.メチル化を検出する前記試薬が制限エンドヌクレアーゼである、請求項29に 記載の方法。 34.前記制限エンドヌクレアーゼがメチル化感受性である、請求項33に記載の方 法。 35.前記制限エンドヌクレアーゼがMspI、HpaIIおよびBssHIIよりなる群から選 択される、請求項34に記載の方法。 36.前記生物学的液体が血清、尿、唾液、脳脊髄膜、痰、射出精液、血液および 糞便よりなる群から選択される、請求項29に記載の方法。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/656,716 US5786146A (en) | 1996-06-03 | 1996-06-03 | Method of detection of methylated nucleic acid using agents which modify unmethylated cytosine and distinguishing modified methylated and non-methylated nucleic acids |
US08/656,716 | 1996-06-03 | ||
US08/835,728 US6017704A (en) | 1996-06-03 | 1997-04-11 | Method of detection of methylated nucleic acid using agents which modify unmethylated cytosine and distinguishing modified methylated and non-methylated nucleic acids |
US08/835,728 | 1997-04-11 | ||
PCT/US1997/009533 WO1997046705A1 (en) | 1996-06-03 | 1997-06-03 | Methylation specific detection |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004153956A Division JP3725535B2 (ja) | 1996-06-03 | 2004-05-24 | メチル化特異的検出 |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2000511776A true JP2000511776A (ja) | 2000-09-12 |
JP2000511776A5 JP2000511776A5 (ja) | 2004-11-04 |
JP3612080B2 JP3612080B2 (ja) | 2005-01-19 |
Family
ID=27097252
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP50077998A Expired - Lifetime JP3612080B2 (ja) | 1996-06-03 | 1997-06-03 | メチル化特異的検出 |
JP2004153956A Expired - Lifetime JP3725535B2 (ja) | 1996-06-03 | 2004-05-24 | メチル化特異的検出 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004153956A Expired - Lifetime JP3725535B2 (ja) | 1996-06-03 | 2004-05-24 | メチル化特異的検出 |
Country Status (11)
Country | Link |
---|---|
US (2) | US6017704A (ja) |
EP (2) | EP1690948A3 (ja) |
JP (2) | JP3612080B2 (ja) |
AT (1) | ATE326549T1 (ja) |
CA (1) | CA2257104C (ja) |
DE (1) | DE69735894T2 (ja) |
DK (1) | DK0954608T3 (ja) |
ES (1) | ES2264165T3 (ja) |
IL (1) | IL127342A (ja) |
PT (1) | PT954608E (ja) |
WO (1) | WO1997046705A1 (ja) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
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JP2004512050A (ja) * | 2000-10-23 | 2004-04-22 | キャンサー・リサーチ・テクノロジー・リミテッド | 核酸の増幅およびプロファイリングに関連する物質および方法 |
WO2005021743A1 (ja) * | 2003-08-29 | 2005-03-10 | Tanaka, Noriaki | 核酸増幅用プライマー及びこれを用いた大腸癌の検査方法 |
WO2006064737A1 (ja) | 2004-12-13 | 2006-06-22 | National University Corporation Okayama University | 遺伝子のメチル化検出方法及びメチル化検出による新生物の検査方法 |
JP2007502121A (ja) * | 2003-08-14 | 2007-02-08 | ケース ウエスタン リザーブ ユニバーシティ | 大腸癌を検出する方法及び組成物 |
WO2008149855A1 (ja) * | 2007-06-08 | 2008-12-11 | Bio-Dixam, Llc. | メチル化核酸又は非メチル化核酸を増幅する方法 |
JP2009531065A (ja) * | 2006-03-29 | 2009-09-03 | ジョン ウェイン キャンサー インスティチュート | エストロゲン受容体アルファのメチル化、およびその使用 |
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US7700324B1 (en) | 1998-11-03 | 2010-04-20 | The Johns Hopkins University School Of Medicine | Methylated CpG island amplification (MCA) |
US6994991B1 (en) | 1998-11-18 | 2006-02-07 | North Shore - Long Island Jewish Research Institute | Identification of differentially methylated multiple drug resistance loci |
DE19853398C1 (de) * | 1998-11-19 | 2000-03-16 | Epigenomics Gmbh | Verfahren zur Identifikation von Cytosin-Methylierungsmustern in genomischer DNA |
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AUPP844899A0 (en) * | 1999-02-01 | 1999-02-25 | University Of Western Australia, The | A method for detecting methylated nucleic acids |
US7655443B1 (en) * | 1999-05-07 | 2010-02-02 | Siemens Healthcare Diagnostics, Inc. | Nucleic acid sequencing with simultaneous quantitation |
US6331393B1 (en) * | 1999-05-14 | 2001-12-18 | University Of Southern California | Process for high-throughput DNA methylation analysis |
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US6756200B2 (en) * | 2001-01-26 | 2004-06-29 | The Johns Hopkins University School Of Medicine | Aberrantly methylated genes as markers of breast malignancy |
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JP2004512050A (ja) * | 2000-10-23 | 2004-04-22 | キャンサー・リサーチ・テクノロジー・リミテッド | 核酸の増幅およびプロファイリングに関連する物質および方法 |
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JP4781267B2 (ja) * | 2003-08-14 | 2011-09-28 | ケース ウエスタン リザーブ ユニバーシティ | 大腸癌を検出する方法及び組成物 |
WO2005021743A1 (ja) * | 2003-08-29 | 2005-03-10 | Tanaka, Noriaki | 核酸増幅用プライマー及びこれを用いた大腸癌の検査方法 |
WO2006064737A1 (ja) | 2004-12-13 | 2006-06-22 | National University Corporation Okayama University | 遺伝子のメチル化検出方法及びメチル化検出による新生物の検査方法 |
JP2009531065A (ja) * | 2006-03-29 | 2009-09-03 | ジョン ウェイン キャンサー インスティチュート | エストロゲン受容体アルファのメチル化、およびその使用 |
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JP5258760B2 (ja) * | 2007-06-08 | 2013-08-07 | 合同会社Bio−Dixam | メチル化核酸又は非メチル化核酸を増幅する方法 |
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ATE326549T1 (de) | 2006-06-15 |
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CA2257104A1 (en) | 1997-12-11 |
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JP3612080B2 (ja) | 2005-01-19 |
IL127342A0 (en) | 1999-10-28 |
CA2257104C (en) | 2008-01-29 |
ES2264165T3 (es) | 2006-12-16 |
US6265171B1 (en) | 2001-07-24 |
EP0954608A1 (en) | 1999-11-10 |
PT954608E (pt) | 2006-08-31 |
JP2004290200A (ja) | 2004-10-21 |
DE69735894D1 (de) | 2006-06-22 |
JP3725535B2 (ja) | 2005-12-14 |
DK0954608T3 (da) | 2006-08-21 |
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