CA2262012A1 - Dosage cellulaire total - Google Patents
Dosage cellulaire total Download PDFInfo
- Publication number
- CA2262012A1 CA2262012A1 CA002262012A CA2262012A CA2262012A1 CA 2262012 A1 CA2262012 A1 CA 2262012A1 CA 002262012 A CA002262012 A CA 002262012A CA 2262012 A CA2262012 A CA 2262012A CA 2262012 A1 CA2262012 A1 CA 2262012A1
- Authority
- CA
- Canada
- Prior art keywords
- assay
- cells
- rna
- specific
- primers
- Prior art date
- Legal status (The legal status 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 status listed.)
- Abandoned
Links
- 238000003556 assay Methods 0.000 title claims description 43
- 210000004027 cell Anatomy 0.000 claims abstract description 86
- 238000000034 method Methods 0.000 claims abstract description 35
- 210000001612 chondrocyte Anatomy 0.000 claims abstract description 30
- 210000002510 keratinocyte Anatomy 0.000 claims abstract description 5
- 108020004999 messenger RNA Proteins 0.000 claims description 30
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 27
- 102000016284 Aggrecans Human genes 0.000 claims description 26
- 108010067219 Aggrecans Proteins 0.000 claims description 26
- 238000003757 reverse transcription PCR Methods 0.000 claims description 18
- 239000011541 reaction mixture Substances 0.000 claims description 8
- 239000003161 ribonuclease inhibitor Substances 0.000 claims description 6
- 210000000845 cartilage Anatomy 0.000 claims description 4
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- 238000006731 degradation reaction Methods 0.000 claims description 4
- 239000003550 marker Substances 0.000 claims description 3
- 108020004513 Bacterial RNA Proteins 0.000 claims description 2
- 102000011782 Keratins Human genes 0.000 claims description 2
- 108010076876 Keratins Proteins 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 claims description 2
- 108010029485 Protein Isoforms Proteins 0.000 claims description 2
- 102000001708 Protein Isoforms Human genes 0.000 claims description 2
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- 239000012091 fetal bovine serum Substances 0.000 description 4
- 239000004033 plastic Substances 0.000 description 4
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- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 3
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 3
- 101000999998 Homo sapiens Aggrecan core protein Proteins 0.000 description 3
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- 102000007260 Deoxyribonuclease I Human genes 0.000 description 2
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 2
- 102000016911 Deoxyribonucleases Human genes 0.000 description 2
- 108010053770 Deoxyribonucleases Proteins 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 238000010240 RT-PCR analysis Methods 0.000 description 2
- 102100037968 Ribonuclease inhibitor Human genes 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
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- 239000012634 fragment Substances 0.000 description 2
- 102000047486 human GAPDH Human genes 0.000 description 2
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- 229920001778 nylon Polymers 0.000 description 2
- 108010024226 placental ribonuclease inhibitor Proteins 0.000 description 2
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- 238000010839 reverse transcription Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
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- 239000002356 single layer Substances 0.000 description 2
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- 239000012588 trypsin Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 238000009004 PCR Kit Methods 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 241000272534 Struthio camelus Species 0.000 description 1
- 239000008052 TBE buffer with ethidium bromide Substances 0.000 description 1
- 230000009830 antibody antigen interaction Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
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- 238000010804 cDNA synthesis Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
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- 238000013461 design Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000011572 manganese Substances 0.000 description 1
- 229940071125 manganese acetate Drugs 0.000 description 1
- UOGMEBQRZBEZQT-UHFFFAOYSA-L manganese(2+);diacetate Chemical compound [Mn+2].CC([O-])=O.CC([O-])=O UOGMEBQRZBEZQT-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
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- 230000004048 modification Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
- 239000012474 protein marker Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
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- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000003612 virological effect Effects 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
-
- 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
- C12Q2600/158—Expression markers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Analytical Chemistry (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (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)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Procédé de détermination de la présence de types cellulaires spécifiques dans une population cellulaire mixte. Ce procédé consiste à identifier et à détecter des marqueurs pour les types cellulaires cibles, à préparer des amorces spécifiques des marqueurs, et à utiliser la technologie de la réaction à transcriptase inverse dans le but d'amplifier et de détecter ces marqueurs. Ledit procédé est suffisamment sensible pour détecter la présence d'à peine 100 cellules cibles dans une population cellulaire. Les cellules cibles peuvent être des chondrocytes humains et des kératinocytes humains.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US2263696P | 1996-07-26 | 1996-07-26 | |
| US60/022,636 | 1996-07-26 | ||
| US3541597P | 1997-01-23 | 1997-01-23 | |
| US60/035,415 | 1997-01-23 | ||
| PCT/US1997/013131 WO1998004742A1 (fr) | 1996-07-26 | 1997-07-25 | Dosage cellulaire total |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2262012A1 true CA2262012A1 (fr) | 1998-02-05 |
Family
ID=26696163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002262012A Abandoned CA2262012A1 (fr) | 1996-07-26 | 1997-07-25 | Dosage cellulaire total |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0956364A1 (fr) |
| JP (1) | JP2000515761A (fr) |
| AU (1) | AU3895597A (fr) |
| CA (1) | CA2262012A1 (fr) |
| WO (1) | WO1998004742A1 (fr) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7598226B2 (en) | 1998-12-28 | 2009-10-06 | Corixa Corporation | Compositions and methods for the therapy and diagnosis of breast cancer |
| US6969518B2 (en) | 1998-12-28 | 2005-11-29 | Corixa Corporation | Compositions and methods for the therapy and diagnosis of breast cancer |
| AU2001253079A1 (en) * | 2000-04-03 | 2001-10-15 | Corixa Corporation | Methods, compositions and kits for the detection and monitoring of breast cancer |
| EP2381116A1 (fr) | 2000-11-16 | 2011-10-26 | California Institute of Technology | Appareil et procédés pour effectuer des analyses et criblage à haut rendement |
| WO2002081729A2 (fr) | 2001-04-06 | 2002-10-17 | California Institute Of Technology | Amplification d'acide nucleique au moyen de dispositifs microfluidiques |
| JP4355210B2 (ja) | 2001-11-30 | 2009-10-28 | フルイディグム コーポレイション | 微小流体デバイスおよび微小流体デバイスの使用方法 |
| CA2480728A1 (fr) | 2002-04-01 | 2003-10-16 | Fluidigm Corporation | Systemes d'analyse de particules microfluidiques |
| US7312085B2 (en) | 2002-04-01 | 2007-12-25 | Fluidigm Corporation | Microfluidic particle-analysis systems |
| AU2003282875A1 (en) | 2002-09-25 | 2004-04-19 | California Institute Of Technology | Microfluidic large scale integration |
| US8871446B2 (en) | 2002-10-02 | 2014-10-28 | California Institute Of Technology | Microfluidic nucleic acid analysis |
| US7604965B2 (en) | 2003-04-03 | 2009-10-20 | Fluidigm Corporation | Thermal reaction device and method for using the same |
| US7815868B1 (en) | 2006-02-28 | 2010-10-19 | Fluidigm Corporation | Microfluidic reaction apparatus for high throughput screening |
| WO2008124508A1 (fr) * | 2007-04-06 | 2008-10-16 | Genzyme Corporation | Procédés d'évaluation de cellules et de cultures de cellules |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5213961A (en) * | 1989-08-31 | 1993-05-25 | Brigham And Women's Hospital | Accurate quantitation of RNA and DNA by competetitive polymerase chain reaction |
| ES2092056T3 (es) * | 1991-09-23 | 1996-11-16 | Pfizer | Procedimiento para detectar arnm y adn especificos en celulas. |
| KR100289793B1 (ko) * | 1992-01-29 | 2001-12-01 | 테츠오 토미나가 | 폴리누클레오티드고정지지체 |
-
1997
- 1997-07-25 EP EP97936237A patent/EP0956364A1/fr not_active Withdrawn
- 1997-07-25 AU AU38955/97A patent/AU3895597A/en not_active Abandoned
- 1997-07-25 WO PCT/US1997/013131 patent/WO1998004742A1/fr not_active Ceased
- 1997-07-25 JP JP10509018A patent/JP2000515761A/ja active Pending
- 1997-07-25 CA CA002262012A patent/CA2262012A1/fr not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| AU3895597A (en) | 1998-02-20 |
| JP2000515761A (ja) | 2000-11-28 |
| EP0956364A1 (fr) | 1999-11-17 |
| WO1998004742A1 (fr) | 1998-02-05 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued |