JP5601746B2 - 同一の試料を用いた二段階核酸検査方法 - Google Patents
同一の試料を用いた二段階核酸検査方法 Download PDFInfo
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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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- 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
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/14—Heterocyclic carbon compound [i.e., O, S, N, Se, Te, as only ring hetero atom]
- Y10T436/142222—Hetero-O [e.g., ascorbic acid, etc.]
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Description
また、遺伝子組換え体の判別検査にも、上記と同様の手法も用いられている。
本発明は、このような従来技術の問題に鑑みなされたものであり、DNAを用いて、従来よりも効率の良い検査方法を提供することを目的とする。
即ち、本発明は、以下の(1)〜(12)を提供するものである。
上記(1)または(2)の方法のように、通常は、特殊設備や熟練技術者が不要で安価で短時間に検査結果が得られるような第1ステージの簡易検査をおこない、問題が発生したときに第2ステージの精密検査をおこなうという二段階での検査手法を実施することにより、効率よく信頼性の高い検査を行うことができる。
また、上記(3)の方法のように、第2ステージの精密検査として核酸の塩基配列を読み取る方法を採用する方式によれば、この方法は最も信頼度の高い核酸の一次構造の解析検査方法であるため、第1ステージの簡易検査にどのような方法を用いても、最終的に高い信頼性の検査を確保することができる。
本発明の第一の検査方法は、試料核酸中に目的の配列が含まれているかどうかを検査する方法であって、最初に試料核酸に対し簡易な検査を行い、目的の配列が含まれていると判定された場合若しくは含まれていないと判定された場合に、同一試料核酸に対し精密な検査を行うことを特徴とするものである。本発明の第二の検査方法は、試料核酸中に目的の配列が規定量以上含まれているかどうかを検査する方法であって、最初に試料核酸に対し簡易な定量的検査を行い、目的の配列が規定量以上含まれていると判定された場合に、同一試料核酸に対し精密な検査を行うことを特徴とするとするものである。このように検査を二段階に分け、問題があるときだけ精密な検査を行うようにすれば、最適なコストで信頼性の高い検査を提供することができる。この手法は、問題が発生する件数の少ない状況に適した検査方法である。
以下、本発明の検査方法の好ましい態様を図を用いて説明する。
なお、試料DNAを回収し再使用する場合においても、また、分割保存し検査ごとに使用する場合においても、第1ステージの簡易検査は、本実施の形態で説明したものに限定されるわけではない。この他、1種類または複数種類の特定のDNAの塩基配列を同定できる手法であれば、いかなるものも採用することができる。
図11に示す目的塩基配列ii31が試料DNAの塩基配列26の中に存在する相対量を定量的に検出できるかどうかを、本発明の検査方法で調べた。第1ステージの簡易検査としてリアルタイムPCR法を、第二ステージの簡易検査としてはDNA塩基配列の読み取りを、それぞれ用いた。リアルタイムPCR法は、前述のように試料DNAを破損させない検査法であるが、使用する試料DNAが極端に少量で行える検査法のため、ここでは検査後に試料DNAを回収せず、予め試料DNAを二分してその片方を第一ステージのリアルタイムPCRに用い、もう片方を第二ステージの塩基配列の読み取りに用いた。
Claims (4)
- 試料核酸中に目的の配列が含まれているかどうかを検査する方法であって、最初に試料核酸に対しマイクロチップ化したデバイス上で簡易な検査を行い、目的の配列が含まれていると判定された場合若しくは含まれていないと判定された場合に、同一試料核酸に対し精密な検査を行う核酸の検査方法であって、試料核酸を二分し、一方を簡易な検査の試料核酸とし、他方を精密な検査の試料核酸とし、それらを前記同一デバイス上に保持しておくことを特徴とし、簡易な検査がDNAハイブリダイゼーション、PCR−RFLP法、又はリアルタイムPCR法による検査であり、精密な検査がDNA塩基配列の読み取りによる検査である核酸の検査方法。
- 目的の配列が、生物種に特異的に存在する配列であることを特徴とする請求項1に記載の核酸の検査方法。
- 目的の配列が、遺伝子組み換え体に特異的に存在する配列であることを特徴とする請求項1に記載の核酸の検査方法。
- 複数種類の目的配列を1回の簡易な検査で同時に検査できることを特徴とする請求項1に記載の核酸の検査方法。
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2006072581A JP5601746B2 (ja) | 2006-03-16 | 2006-03-16 | 同一の試料を用いた二段階核酸検査方法 |
EP06840372.4A EP1995326B1 (en) | 2006-03-16 | 2006-12-26 | Two stage methods for examining nucleic acid by using one and the same sample |
PCT/JP2006/325982 WO2007108193A1 (ja) | 2006-03-16 | 2006-12-26 | 同一の試料を用いた二段階核酸検査方法 |
US11/662,597 US20090053820A1 (en) | 2006-03-16 | 2006-12-26 | Two-step nucleic acid testing method using the same sample |
CNA2006800007201A CN101528944A (zh) | 2006-03-16 | 2006-12-26 | 使用同一样本的二阶段核酸检测方法 |
Applications Claiming Priority (1)
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JP2006072581A JP5601746B2 (ja) | 2006-03-16 | 2006-03-16 | 同一の試料を用いた二段階核酸検査方法 |
Publications (2)
Publication Number | Publication Date |
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JP2007244291A JP2007244291A (ja) | 2007-09-27 |
JP5601746B2 true JP5601746B2 (ja) | 2014-10-08 |
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Country Status (5)
Country | Link |
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US (1) | US20090053820A1 (ja) |
EP (1) | EP1995326B1 (ja) |
JP (1) | JP5601746B2 (ja) |
CN (1) | CN101528944A (ja) |
WO (1) | WO2007108193A1 (ja) |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
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CA2002076A1 (en) * | 1988-11-21 | 1990-05-21 | Brent A. Burdick | Diagnostic kit and method using a solid phase capture means for detecting nucleic acids |
JP4540846B2 (ja) * | 1998-04-30 | 2010-09-08 | マックス−プランク−ゲゼルシャフト・ツア・フェルデルング・デア・ヴィッセンシャフテン・エー・ファオ | 所望の生物学的特性を与えるクローンを発現ライブラリーから同定する新規方法 |
WO2000044935A2 (en) * | 1999-01-29 | 2000-08-03 | Bavarian Nordic Research Institute A/S | Multiplex real-time pcr |
JP3502906B2 (ja) * | 1999-09-03 | 2004-03-02 | 独立行政法人食品総合研究所 | 遺伝子組換えトウモロコシ及びこれを含む加工食品からの組換え遺伝子の検知方法 |
JP4693198B2 (ja) * | 1999-09-09 | 2011-06-01 | 横河電機株式会社 | バイオチップ |
JP3600891B2 (ja) | 2001-04-24 | 2004-12-15 | 財団法人理工学振興会 | Sine法による動物種判別方法 |
JP2002345498A (ja) | 2001-05-28 | 2002-12-03 | Fisheries Research Agency | 日本産サバ属の種判別方法 |
WO2003100382A1 (en) * | 2002-05-20 | 2003-12-04 | Northrop Grumman Corporation | Automatic point source biological agent detection system |
JP4497846B2 (ja) * | 2002-06-07 | 2010-07-07 | 三菱化学メディエンス株式会社 | ヒトアデノウイルスの検出法 |
EP1581650A4 (en) * | 2002-10-11 | 2006-08-23 | Univ Arizona State | MOLECULAR SIGNATURES AND ASSAY FOR FLUOROQUINOLINE RESISTANCE IN BACILLUS OF CHARCOAL |
US7745115B2 (en) * | 2003-04-02 | 2010-06-29 | New York University | Method for the surveillance for biological, chemical and radiological agents |
US20040229211A1 (en) * | 2003-05-13 | 2004-11-18 | Yeung Wah Hin Alex | Sensitive diagnostic testing methodology using multiplex real time PCR with one dye (MOD) and its use as in severe acute respiratory syndrome (SARS) |
EP1582599A1 (en) | 2004-03-31 | 2005-10-05 | Takara Bio Inc. | Method for purifying microbeads |
JP2006072581A (ja) | 2004-08-31 | 2006-03-16 | Sharp Corp | 共同購入システム、並びに、それに用いる共同購入サービスサーバおよび端末 |
-
2006
- 2006-03-16 JP JP2006072581A patent/JP5601746B2/ja active Active
- 2006-12-26 CN CNA2006800007201A patent/CN101528944A/zh active Pending
- 2006-12-26 US US11/662,597 patent/US20090053820A1/en not_active Abandoned
- 2006-12-26 EP EP06840372.4A patent/EP1995326B1/en not_active Not-in-force
- 2006-12-26 WO PCT/JP2006/325982 patent/WO2007108193A1/ja active Application Filing
Also Published As
Publication number | Publication date |
---|---|
EP1995326A4 (en) | 2010-05-05 |
EP1995326B1 (en) | 2013-10-16 |
US20090053820A1 (en) | 2009-02-26 |
CN101528944A (zh) | 2009-09-09 |
JP2007244291A (ja) | 2007-09-27 |
EP1995326A1 (en) | 2008-11-26 |
WO2007108193A1 (ja) | 2007-09-27 |
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