WO2000077253A1 - Appareil et procede d'examen des genes - Google Patents
Appareil et procede d'examen des genes Download PDFInfo
- 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
Links
- 238000000034 method Methods 0.000 title claims description 47
- 108090000623 proteins and genes Proteins 0.000 title claims description 47
- 238000002360 preparation method Methods 0.000 claims abstract description 367
- 238000012408 PCR amplification Methods 0.000 claims abstract description 258
- 108091033319 polynucleotide Proteins 0.000 claims abstract description 88
- 102000040430 polynucleotide Human genes 0.000 claims abstract description 88
- 239000002157 polynucleotide Substances 0.000 claims abstract description 88
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 50
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 40
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 32
- 238000001514 detection method Methods 0.000 claims abstract description 24
- 239000002699 waste material Substances 0.000 claims abstract description 10
- 239000000872 buffer Substances 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 208
- 239000000758 substrate Substances 0.000 claims description 64
- 239000007850 fluorescent dye Substances 0.000 claims description 48
- 230000001678 irradiating effect Effects 0.000 claims description 38
- 238000002372 labelling Methods 0.000 claims description 38
- 239000007853 buffer solution Substances 0.000 claims description 31
- 230000003321 amplification Effects 0.000 claims description 30
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 30
- 238000012360 testing method Methods 0.000 claims description 22
- 230000002068 genetic effect Effects 0.000 claims description 21
- 239000003398 denaturant Substances 0.000 claims description 20
- 239000002773 nucleotide Substances 0.000 claims description 19
- 125000003729 nucleotide group Chemical group 0.000 claims description 19
- 238000007689 inspection Methods 0.000 claims description 17
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 16
- 229910052710 silicon Inorganic materials 0.000 claims description 16
- 239000010703 silicon Substances 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 14
- 238000010998 test method Methods 0.000 claims description 9
- 239000011148 porous material Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 3
- 239000006285 cell suspension Substances 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 238000005464 sample preparation method Methods 0.000 claims 1
- 238000011109 contamination Methods 0.000 abstract description 5
- 239000011541 reaction mixture Substances 0.000 abstract 2
- 239000000523 sample Substances 0.000 description 274
- 210000004027 cell Anatomy 0.000 description 76
- 239000000047 product Substances 0.000 description 58
- 210000004369 blood Anatomy 0.000 description 36
- 239000008280 blood Substances 0.000 description 36
- 238000003745 diagnosis Methods 0.000 description 13
- 239000013307 optical fiber Substances 0.000 description 13
- 238000012545 processing Methods 0.000 description 12
- 108020004414 DNA Proteins 0.000 description 11
- 238000001917 fluorescence detection Methods 0.000 description 11
- 102000039446 nucleic acids Human genes 0.000 description 9
- 108020004707 nucleic acids Proteins 0.000 description 9
- 150000007523 nucleic acids Chemical class 0.000 description 9
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 8
- 210000000265 leukocyte Anatomy 0.000 description 8
- 241000700605 Viruses Species 0.000 description 7
- 238000000746 purification Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 239000000975 dye Substances 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 230000002550 fecal effect Effects 0.000 description 5
- 238000009396 hybridization Methods 0.000 description 5
- 230000000717 retained effect Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 206010027476 Metastases Diseases 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
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- 238000011897 real-time detection Methods 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000012780 transparent material Substances 0.000 description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 3
- 101710130181 Protochlorophyllide reductase A, chloroplastic Proteins 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000005530 etching Methods 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 201000005202 lung cancer Diseases 0.000 description 3
- 208000020816 lung neoplasm Diseases 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 206010016952 Food poisoning Diseases 0.000 description 2
- 208000019331 Foodborne disease Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- CGNLCCVKSWNSDG-UHFFFAOYSA-N SYBR Green I Chemical compound CN(C)CCCN(CCC)C1=CC(C=C2N(C3=CC=CC=C3S2)C)=C2C=CC=CC2=[N+]1C1=CC=CC=C1 CGNLCCVKSWNSDG-UHFFFAOYSA-N 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 230000005661 hydrophobic surface Effects 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 101100191561 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) PRP3 gene Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- -1 for example Chemical compound 0.000 description 1
- 230000005861 gene abnormality Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 238000000206 photolithography Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
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.
Landscapes
- 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
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 |
Family
ID=14235988
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/JP1999/003209 WO2000077253A1 (fr) | 1999-06-16 | 1999-06-16 | Appareil et procede d'examen des genes |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2000077253A1 (fr) |
Cited By (6)
| 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)
| 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 |
-
1999
- 1999-06-16 WO PCT/JP1999/003209 patent/WO2000077253A1/fr active Application Filing
Patent Citations (7)
| 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)
| 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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