WO2000077253A1 - Appareil et procede d'examen des genes - Google Patents

Appareil et procede d'examen des genes Download PDF

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Publication number
WO2000077253A1
WO2000077253A1 PCT/JP1999/003209 JP9903209W WO0077253A1 WO 2000077253 A1 WO2000077253 A1 WO 2000077253A1 JP 9903209 W JP9903209 W JP 9903209W WO 0077253 A1 WO0077253 A1 WO 0077253A1
Authority
WO
WIPO (PCT)
Prior art keywords
filter
sample preparation
pcr amplification
amplification reaction
opening
Prior art date
Application number
PCT/JP1999/003209
Other languages
English (en)
Japanese (ja)
Inventor
Hiroko Matsunaga
Katsuji Murakawa
Kazunori Okano
Yuji Miyahara
Original Assignee
Hitachi, Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hitachi, Ltd. filed Critical Hitachi, Ltd.
Priority to PCT/JP1999/003209 priority Critical patent/WO2000077253A1/fr
Publication of WO2000077253A1 publication Critical patent/WO2000077253A1/fr

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Classifications

    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

Definitions

  • the present invention relates to a sample preparation apparatus for preparing a sample for performing a gene diagnosis using whole blood or a cell culture solution, a gene test apparatus for easily performing a process from sample preparation to detection of a target arrested child, and a gene.
  • a sample preparation apparatus for preparing a sample for performing a gene diagnosis using whole blood or a cell culture solution
  • a gene test apparatus for easily performing a process from sample preparation to detection of a target arrested child
  • a gene Related to inspection method. Background art
  • JP-A-7-98314 discloses a fecal occult blood measuring device.
  • the stool sample lysate is first removed of solid components by a separation filter. Thereafter, the stool sample solution from which the solid components have been removed is dropped onto the measurement filter.
  • the measurement filter is colored.
  • the measurement filter is uncolored. Therefore, the presence or absence of fecal occult blood can be easily determined by visually judging the presence or absence of coloring of the measurement filter. Disclosure of the invention
  • a second member in which a plurality of second regions having a second filter holding a CR amplification reaction solution is formed at a lower portion, and a second member in which a plurality of third regions having a hydrophobic third filter are formed.
  • a genetic test method using a sample preparation device in which a flow path is formed (1) A buffer solution in which cells are suspended is injected into the first opening, and the cells are captured by a first filter. (2) injecting a denaturing agent into the first opening to capture the polynucleotide eluted from the cells in the second filter; and (3) fluorescently labeling the first opening.
  • the first member, the second member, and the third member are arranged in this order from above, and the first region, the second region, and the third member are
  • C 20 a flow path connecting an upper opening into which a buffer solution in which cells are suspended is injected, and a lower opening through which waste liquid flows out, wherein the flow path extends from the upper opening to the lower opening.
  • FIG. 10 is a cross-sectional view showing an example of the configuration of an arrested child detecting device for detecting the amplification process of a target gene in real time in the second embodiment of the present invention.
  • Fig. 11 shows the configuration of an abductor detection device that irradiates a laser to a plurality of sample preparation units from the lateral direction and detects the amplification process of the target gene in real time in the second embodiment of the present invention. It is a perspective view showing an example.
  • FIG. 16 is a perspective view showing a configuration example of a sample preparation device having a sample preparation unit for simultaneously processing a plurality of samples in the fifth embodiment of the present invention.
  • FIG. 27 is a sectional view showing a configuration example of a sample preparation device according to an eighth embodiment of the present invention.
  • Blood collection volume is 1 mL
  • the solution 300 passes through the first filter 110 and flows down into the space below the first filter 110, but most of the solution 300 does not pass through the first filter 110 by gravity alone. Then, a pressure is applied from above the sample preparation unit 100 to pass the solution 300 through the first filter I10. At this time, only the white blood cells 302 in the solution 300 are captured by the first filter 110.
  • the solution that has passed through the first filter 110 passes through the space below the first filter 110, the second filter 130, and the third filter 140, and flows out to the waste liquid tank. Instead of applying pressure for passing the solution through the first, second, and third filters, centrifugal force generation, suction from the lower opening, etc. can also be used. Subsequently, 3 mL of the denaturing agent 3I0 is poured into the upper space of the first filter 110 (step-2).
  • FIG. 5 is a cross-sectional view showing a configuration example of an arrested child detecting device that detects an amplification process of a target gene in real time using an optical fiber in the first embodiment of the present invention.
  • the optical fiber 131 is inserted from the upper part of the sample preparation unit 100, pressure is applied to the center of the first filter 110, the filter is pierced, a hole is made, and the fiber 31 is passed through the upper part of the second filter 130. PCR amplification products retained in
  • the pretreatment of the sample blood is performed in the same manner as in the first embodiment.
  • a third filter force is formed in the upper part, and lower openings 611-1-3, 61-2-3, 613-3, 614-3 are formed in the lower part of each third filter.
  • the fourth substrate 640 has through-holes 6 1 1—4, 6 1 2, and 6 1 1—3, 6 1 2—3, 6 13—3, 6 6 1 3-4, 6 14 —4 forces, 'formed.
  • the fourth substrate includes the inside and the inside of each second filter.
  • a Peltier element for controlling the temperature in the PCR amplification by raising and lowering the temperature of the solution held in the upper space of each second filter and the second filter is embedded.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un appareil (50) de préparation d'échantillons comprenant une unité (100) de préparation des échantillons pourvue d'une ouverture supérieure, dans laquelle on verse un tampon renfermant des cellules en suspension, d'une ouverture inférieure, qui permet d'évacuer une liqueur résiduelle, et d'un canal reliant ces deux ouvertures. Ledit canal est équipé d'un premier filtre (110) destiné à piéger les cellules, d'un deuxième filtre (130) destiné à piéger les polynucléotides élués des cellules au moyen de l'agent dénaturant que l'on a versé dans l'ouverture supérieure et à retenir un mélange de réaction de polymérisation en chaîne (PCR) renfermant une amorce marquée par fluorescence destinée à la réaction PCR et un réactif de réaction PCR servant à amplifier la copie d'une séquence de base partielle voulue du polynucléotide, et un troisième filtre (140), les trois filtres étant disposés dans cet ordre entre l'ouverture supérieure et l'ouverture inférieure. L'appareil selon l'invention comprend également un élément (200) de maintien de l'unité de préparation des échantillons; et un élément (210) permettant de contrôler la température du mélange de réaction PCR. L'utilisation de cet appareil permet de procéder en continu à une série d'étapes, à savoir le prétraiteme
PCT/JP1999/003209 1999-06-16 1999-06-16 Appareil et procede d'examen des genes WO2000077253A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/JP1999/003209 WO2000077253A1 (fr) 1999-06-16 1999-06-16 Appareil et procede d'examen des genes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP1999/003209 WO2000077253A1 (fr) 1999-06-16 1999-06-16 Appareil et procede d'examen des genes

Publications (1)

Publication Number Publication Date
WO2000077253A1 true WO2000077253A1 (fr) 2000-12-21

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003031972A1 (fr) * 2001-10-05 2003-04-17 Bml, Inc. Plaque de detection
EP1504121A2 (fr) * 2002-04-24 2005-02-09 Hitachi Chemical Research Center, Inc. Dispositif et methode de quantification a haut debit d'arn messagers a partir du sang total
JP2005295942A (ja) * 2004-04-15 2005-10-27 Kyocera Corp 遺伝子反応管およびこれを用いた遺伝子検査装置
JP2007529225A (ja) * 2004-03-16 2007-10-25 アンビオン インコーポレーティッド 分画された血液白血球からのｒｎａの抽出のための方法および試薬
JP2009510398A (ja) * 2005-09-26 2009-03-12 キアゲン ゲゼルシャフト ミット ベシュレンクテル ハフツング 流体処理方法及び流体処理装置
US7745180B2 (en) 2002-04-24 2010-06-29 Hitachi Chemical Co., Ltd. Device and method for high-throughput quantification of mRNA from whole blood

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07107999A (ja) * 1993-10-12 1995-04-25 Hitachi Ltd 遺伝子解析方法及び装置
JPH08266267A (ja) * 1995-03-24 1996-10-15 Becton Dickinson & Co 複数の液体標本を反応させるための自動化された装置
JPH08275800A (ja) * 1995-03-24 1996-10-22 Becton Dickinson & Co 核酸増幅方法及び装置
WO1997003348A1 (fr) * 1995-07-13 1997-01-30 Immunological Associates Of Denver Appareil autonome integrant l'extraction, l'amplification et la detection de l'acide nucleique
JPH10505497A (ja) * 1994-09-09 1998-06-02 ナノゲン・インコーポレイテッド 自動化された分子生物学的診断システム
JPH10239300A (ja) * 1997-02-28 1998-09-11 Kagaku Gijutsu Shinko Jigyodan 全自動遺伝子解析システム
JPH10257887A (ja) * 1996-09-30 1998-09-29 Dainippon Printing Co Ltd 遺伝子解析装置および方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07107999A (ja) * 1993-10-12 1995-04-25 Hitachi Ltd 遺伝子解析方法及び装置
JPH10505497A (ja) * 1994-09-09 1998-06-02 ナノゲン・インコーポレイテッド 自動化された分子生物学的診断システム
JPH08266267A (ja) * 1995-03-24 1996-10-15 Becton Dickinson & Co 複数の液体標本を反応させるための自動化された装置
JPH08275800A (ja) * 1995-03-24 1996-10-22 Becton Dickinson & Co 核酸増幅方法及び装置
WO1997003348A1 (fr) * 1995-07-13 1997-01-30 Immunological Associates Of Denver Appareil autonome integrant l'extraction, l'amplification et la detection de l'acide nucleique
JPH10257887A (ja) * 1996-09-30 1998-09-29 Dainippon Printing Co Ltd 遺伝子解析装置および方法
JPH10239300A (ja) * 1997-02-28 1998-09-11 Kagaku Gijutsu Shinko Jigyodan 全自動遺伝子解析システム

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003031972A1 (fr) * 2001-10-05 2003-04-17 Bml, Inc. Plaque de detection
US7981608B2 (en) 2002-04-24 2011-07-19 Hitachi Chemical Co., Ltd. Device and method for high-throughput quantification of MRNA from whole blood
EP1504121A4 (fr) * 2002-04-24 2005-06-29 Hitachi Chemical Res Ct Inc Dispositif et methode de quantification a haut debit d'arn messagers a partir du sang total
US7745180B2 (en) 2002-04-24 2010-06-29 Hitachi Chemical Co., Ltd. Device and method for high-throughput quantification of mRNA from whole blood
US7939300B2 (en) 2002-04-24 2011-05-10 Hitachi Chemical Co., Ltd Device and method for high-throughput quantification of mRNA from whole blood
US7968288B1 (en) 2002-04-24 2011-06-28 Hitachi Chemical Co., Ltd. Device and method for high-throughput quantification of mRNA from whole blood
EP1504121A2 (fr) * 2002-04-24 2005-02-09 Hitachi Chemical Research Center, Inc. Dispositif et methode de quantification a haut debit d'arn messagers a partir du sang total
US8076105B2 (en) 2002-04-24 2011-12-13 Hitachi Chemical Research Center, Inc. Device and method for high-throughput quantification of MRNA from whole blood
US8101344B2 (en) 2002-04-24 2012-01-24 Hitachi Chemical Research Center, Inc. Device and method for high-throughput quantification of mRNA from whole blood
JP2007529225A (ja) * 2004-03-16 2007-10-25 アンビオン インコーポレーティッド 分画された血液白血球からのｒｎａの抽出のための方法および試薬
JP2005295942A (ja) * 2004-04-15 2005-10-27 Kyocera Corp 遺伝子反応管およびこれを用いた遺伝子検査装置
JP2009510398A (ja) * 2005-09-26 2009-03-12 キアゲン ゲゼルシャフト ミット ベシュレンクテル ハフツング 流体処理方法及び流体処理装置
JP4779104B2 (ja) * 2005-09-26 2011-09-28 キアゲン ゲゼルシャフト ミット ベシュレンクテル ハフツング 流体処理方法及び流体処理装置

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