ES2398984T3 - RT-PCR rápida en una etapa - Google Patents
RT-PCR rápida en una etapa Download PDFInfo
- Publication number
- ES2398984T3 ES2398984T3 ES08006778T ES08006778T ES2398984T3 ES 2398984 T3 ES2398984 T3 ES 2398984T3 ES 08006778 T ES08006778 T ES 08006778T ES 08006778 T ES08006778 T ES 08006778T ES 2398984 T3 ES2398984 T3 ES 2398984T3
- Authority
- ES
- Spain
- Prior art keywords
- pcr
- temperature
- sample
- stage
- seconds
- 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.)
- Active
Links
- 238000003757 reverse transcription PCR Methods 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 claims abstract description 28
- 239000002299 complementary DNA Substances 0.000 claims abstract description 17
- 230000003321 amplification Effects 0.000 claims abstract description 16
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 16
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 8
- 239000011541 reaction mixture Substances 0.000 claims abstract description 6
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 claims description 43
- 102100034343 Integrase Human genes 0.000 claims description 42
- 238000006243 chemical reaction Methods 0.000 claims description 38
- 230000000694 effects Effects 0.000 claims description 20
- 230000001419 dependent effect Effects 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 241000589499 Thermus thermophilus Species 0.000 claims description 3
- 239000000523 sample Substances 0.000 description 37
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 30
- 238000011533 pre-incubation Methods 0.000 description 25
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 19
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 19
- 238000009396 hybridization Methods 0.000 description 16
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 108020004414 DNA Proteins 0.000 description 10
- 238000010839 reverse transcription Methods 0.000 description 10
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 108020004999 messenger RNA Proteins 0.000 description 7
- 238000004925 denaturation Methods 0.000 description 6
- 230000036425 denaturation Effects 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 150000007523 nucleic acids Chemical group 0.000 description 5
- 238000003753 real-time PCR Methods 0.000 description 5
- 241000620137 Carboxydothermus hydrogenoformans Species 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 108010006785 Taq Polymerase Proteins 0.000 description 4
- 108010085671 Thermus thermophilus DNA polymerase Proteins 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000014509 gene expression Effects 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 102100028161 ATP-binding cassette sub-family C member 2 Human genes 0.000 description 3
- 108010066419 Multidrug Resistance-Associated Protein 2 Proteins 0.000 description 3
- 241000589596 Thermus Species 0.000 description 3
- 108010020713 Tth polymerase Proteins 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 241000713869 Moloney murine leukemia virus Species 0.000 description 2
- 108010010677 Phosphodiesterase I Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000000833 heterodimer Substances 0.000 description 2
- 229910001437 manganese ion Inorganic materials 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000002797 proteolythic effect Effects 0.000 description 2
- 108010068698 spleen exonuclease Proteins 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 208000030507 AIDS Diseases 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 241000713838 Avian myeloblastosis virus Species 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 1
- 241000204666 Thermotoga maritima Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000002820 assay format Methods 0.000 description 1
- 208000005266 avian sarcoma Diseases 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000001036 exonucleolytic effect Effects 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000009830 intercalation Methods 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 238000012423 maintenance Methods 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
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 239000002751 oligonucleotide probe Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 1
- 238000005382 thermal cycling Methods 0.000 description 1
- 230000010512 thermal transition Effects 0.000 description 1
- 229920001187 thermosetting polymer Polymers 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000005758 transcription activity Effects 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000011144 upstream manufacturing 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6846—Common amplification features
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (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)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP07007173 | 2007-04-05 | ||
| EP07007173 | 2007-04-05 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2398984T3 true ES2398984T3 (es) | 2013-03-25 |
Family
ID=38453943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES08006778T Active ES2398984T3 (es) | 2007-04-05 | 2008-04-03 | RT-PCR rápida en una etapa |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US8119353B2 (enExample) |
| EP (1) | EP1978109B1 (enExample) |
| JP (1) | JP5191041B2 (enExample) |
| ES (1) | ES2398984T3 (enExample) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9175332B2 (en) * | 2010-07-29 | 2015-11-03 | Roche Molecular Systems, Inc. | Generic PCR |
| WO2012045668A1 (en) | 2010-10-04 | 2012-04-12 | Roche Diagnostics Gmbh | Method for cell lysis in a rt-pcr reaction buffer |
| WO2013101290A1 (en) * | 2011-12-29 | 2013-07-04 | Qiagen | Methods of using telomeres as markers for aging |
| DK2855692T3 (da) | 2012-05-24 | 2019-10-14 | Univ Utah Res Found | Ekstrem pcr |
| ES2916153T3 (es) * | 2015-11-05 | 2022-06-28 | Univ Utah Res Found | PCR extrema para transcripción inversa |
| JP2018023360A (ja) * | 2016-07-29 | 2018-02-15 | セイコーエプソン株式会社 | 核酸増幅反応方法、試薬、および試薬の使用方法 |
| GB201812192D0 (en) | 2018-07-26 | 2018-09-12 | Ttp Plc | Variable temperature reactor, heater and control circuit for the same |
| CA3126398A1 (en) * | 2019-01-17 | 2020-07-23 | Northwestern University | Rapid reverse transcription quantitative polymerase chain reaction |
| EP4269592A4 (en) | 2020-12-25 | 2025-01-01 | Nippon Gene Co., Ltd | MODIFIED DNA POLYMERASE |
Family Cites Families (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5105599A (en) * | 1989-02-24 | 1992-04-21 | Highland Supply Corporation | Means for securing a decorative cover about a flower pot |
| US5561058A (en) | 1986-08-22 | 1996-10-01 | Hoffmann-La Roche Inc. | Methods for coupled high temperatures reverse transcription and polymerase chain reactions |
| US5455170A (en) * | 1986-08-22 | 1995-10-03 | Hoffmann-La Roche Inc. | Mutated thermostable nucleic acid polymerase enzyme from Thermus species Z05 |
| WO1991009944A2 (en) | 1989-12-22 | 1991-07-11 | F.Hoffmann-La Roche Ag | High temperature reverse transcriptases |
| US5374553A (en) * | 1986-08-22 | 1994-12-20 | Hoffmann-La Roche Inc. | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
| US5118801A (en) | 1988-09-30 | 1992-06-02 | The Public Health Research Institute | Nucleic acid process containing improved molecular switch |
| US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
| EP0620859A1 (en) | 1992-09-29 | 1994-10-26 | Cedars-Sinai Medical Center | One step rna and combined one step rna and dna polymerase chain reaction for detection of rare rna or rna and dna |
| US5538848A (en) | 1994-11-16 | 1996-07-23 | Applied Biosystems Division, Perkin-Elmer Corp. | Method for detecting nucleic acid amplification using self-quenching fluorescence probe |
| US5876924A (en) * | 1994-06-22 | 1999-03-02 | Mount Sinai School Of Medicine | Nucleic acid amplification method hybridization signal amplification method (HSAM) |
| AU726501B2 (en) * | 1996-06-04 | 2000-11-09 | University Of Utah Research Foundation | Monitoring hybridization during PCR |
| ATE390491T1 (de) | 1996-06-04 | 2008-04-15 | Univ Utah Res Found | Behälter zum durchführen und beobachten biologischer prozesse |
| US7179590B2 (en) | 2000-04-18 | 2007-02-20 | Roche Molecular Systems, Inc | High temperature reverse transcription using mutant DNA polymerases |
| CA2439402A1 (en) * | 2001-03-02 | 2002-09-12 | University Of Pittsburgh Of The Commonwealth System Of Higher Education | Pcr method |
| US20030108898A1 (en) * | 2001-07-03 | 2003-06-12 | Paul Choppa | Method for detection, quantitation and breakpoint cluster region determination of fusion transcripts |
| CN101245376A (zh) * | 2003-01-17 | 2008-08-20 | 香港中文大学 | 作为妊娠相关病症的诊断标志物的循环mRNA |
-
2008
- 2008-04-01 JP JP2008094907A patent/JP5191041B2/ja active Active
- 2008-04-03 EP EP08006778A patent/EP1978109B1/en active Active
- 2008-04-03 ES ES08006778T patent/ES2398984T3/es active Active
- 2008-04-03 US US12/061,700 patent/US8119353B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| EP1978109A1 (en) | 2008-10-08 |
| JP2008253263A (ja) | 2008-10-23 |
| JP5191041B2 (ja) | 2013-04-24 |
| EP1978109B1 (en) | 2012-12-05 |
| US8119353B2 (en) | 2012-02-21 |
| US20080248535A1 (en) | 2008-10-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2398984T3 (es) | RT-PCR rápida en una etapa | |
| Kiddle et al. | GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use | |
| ES2759872T3 (es) | Reacción de amplificación de extensión y mellado para la amplificación exponencial de los ácidos nucleicos | |
| ES2523140T3 (es) | Aumento de la confianza en las identificaciones de alelos con el recuento molecular | |
| BR112020000809A2 (pt) | uso da proteína cas, método para detecção de molécula de ácido nucleico, e kit | |
| CA2730761C (en) | Improved lysis and reverse transcription for mrna quantification | |
| ES2874143T3 (es) | Métodos y composiciones para reducir códigos de barras, moleculares redundantes creados en reacciones de prolongación de cebadores | |
| CN107446919B (zh) | 一种恒温条件下合成核酸的方法及试剂盒 | |
| KR20120084320A (ko) | Ｔｈｄ 프라이머 타겟 검출 | |
| CN114507716B (zh) | 检测样品中靶核酸的方法 | |
| KR102560137B1 (ko) | 극소량의 희귀 단일 염기 변이체 검출용 프라이머 및 이를 이용하여 극소량의 희귀 단일 염기 변이체를 특이적이고 민감하게 검출하는 방법 | |
| ES2685710T3 (es) | Kits que comprenden controles para amplificación de ácido nucleico | |
| CN101680040A (zh) | 改良的诺如病毒rna检测方法 | |
| CN107922969A (zh) | 包含核酸组装体的结构切换、核酸消化和扩增的串联反应的生物传感器 | |
| US7919244B2 (en) | Nucleic acid detection method involving the direct generation of a measurable signal | |
| US20210102237A1 (en) | Means and methods for nucleic acid target detection | |
| ES2350525T3 (es) | Nuevo formato de detección para las reacciones en cadena de la polimerasa en tiempo real de inicio en caliente. | |
| WO2019051732A1 (zh) | 一种恒温条件下合成核酸的方法及试剂盒 | |
| JP2020530434A (ja) | Ｒｎａおよびｄｎａからの核酸ライブラリーの作製 | |
| ES2533732T3 (es) | Método para lisis celular en un tampón de reacción de RT-PCR | |
| JP5887078B2 (ja) | 合成ｓｉＲＮＡ検出方法 | |
| JP7160142B2 (ja) | 核酸増幅用試薬及び核酸増幅法 | |
| ES3015628T3 (en) | Method for amplifying nucleotide sequence, and sequence determination method | |
| BR102014029398A2 (pt) | composição para reação de transcrição reversa hot-start, métodos de preparação, e kit para reação de transcrição reversa hot-start | |
| Marx et al. | A mutated thermostable Thermus aquaticus DNA polymerase with reverse transcriptase activity for one step RNA pathogen detection |