DK2802671T3 - Fremgangsmåder og kits til stuetemperatur-in situ-påvisning af en mål-nukleinsyre i en biologisk prøve - Google Patents
Fremgangsmåder og kits til stuetemperatur-in situ-påvisning af en mål-nukleinsyre i en biologisk prøve Download PDFInfo
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- DK2802671T3 DK2802671T3 DK12806823.6T DK12806823T DK2802671T3 DK 2802671 T3 DK2802671 T3 DK 2802671T3 DK 12806823 T DK12806823 T DK 12806823T DK 2802671 T3 DK2802671 T3 DK 2802671T3
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- nucleic acid
- target nucleic
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- hybridization
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- IIACRCGMVDHOTQ-UHFFFAOYSA-M sulfamate Chemical compound NS([O-])(=O)=O IIACRCGMVDHOTQ-UHFFFAOYSA-M 0.000 description 1
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- WGTODYJZXSJIAG-UHFFFAOYSA-N tetramethylrhodamine chloride Chemical compound [Cl-].C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C(O)=O WGTODYJZXSJIAG-UHFFFAOYSA-N 0.000 description 1
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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
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
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- Chemical & Material Sciences (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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- Engineering & Computer Science (AREA)
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- Physics & Mathematics (AREA)
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- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (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)
Claims (15)
1. Fremgangsmåde til påvisning afen mål-nukleinsyre i en biologisk prøve, omfattende trinnene at: a) hybridisere mindst én probe til mål-nukleinsyren i prøven ved en temperatur i området på 19 grader Celsius til 25 grader Celsius, hvor proben: 1) er til stede i en hybridiseringsbuffer omfattende 1-10 mM base og med en pH i området på 10 til 13, og 2) omfatter en nukleotidsekvens der er komplementær med en nukleotidsekvens i mål-nukleinsyren og mindst én detekterbar markør; b) vaske prøven for at fjerne prober der er ikke er hybridiseret til mål-nukleinsyren; og c) påvise den mindst ene detekterbare markør på proben efter hybridisering til mål-nukleinsyren i prøven, derved påvise mål-nukleinsyren i prøven.
2. Fremgangsmåden ifølge krav 1, hvor hybridiseringsbufferen har en pH i området på 11 til 12, og/eller hvor hybridiseringsbufferen omfatter 1-5 mM base, 20-60% formamid, og 5-40% dextransulfat.
3. Fremgangsmåden ifølge krav 1 eller 2, hvor basen er NaOH, og eventuelt hvor hybridiseringsbufferen omfatter 1,0-3,0 mM NaOH, 30-50% formamid, og 10% dextransulfat, eller hvor hybridiseringsbufferen omfatter 4,2 mM NaOH, 42% formamid, og 28% dextransulfat.
4. Fremgangsmåden ifølge et hvilket som helst af kravene 1-3, hvor varigheden af trin a) er 5 minutter til 15 minutter.
5. Fremgangsmåden ifølge et hvilket som helst af kravene 1-4, hvor proben er en enkelt-strenget oligonukleotidprobe omfattende 20 til 50 nukleotider.
6. Fremgangsmåden ifølge et hvilket som helst af kravene 1-4, hvor proben er fremstillet af et genomisk fragment.
7. Fremgangsmåden ifølge et hvilket som helst af kravene 1-6, hvor den biologiske prøve omfatter epitelceller, hudceller, knoglemarvceller, blastomerer, spermceller, oocyter, eller pollegemer, eller en kombination deraf.
8. Fremgangsmåden ifølge et hvilket som helst af kravene 1-7, hvor trin b) udføres ved en temperatur i området på 19 grader Celsius til 25 grader Celsius.
9. Fremgangsmåde til påvisning afen mål-nukleinsyre i en biologisk prøve, omfattende trinnene at: a) denaturere en biologisk prøve omfattende mål-nukleinsyren ved at bringe prøven i kontakt med en opløsning omfattende en base og 50% til 80% alkohol; b) hybridisere mindst én probe til mål-nukleinsyren i prøven, hvor proben: 1) er til stede i en hybridiseringsbuffer omfattende 1-10 mM base og med en pH i området på 10 til 13, og 2) omfatter en nukleotidsekvens der er komplementær med en nukleotidsekvens i mål-nukleinsyren og mindst én detekterbar markør; c) vaske prøven i en vaskebuffer med en pH i området på 10 til 13, hvor vaskebufferen omfatter 1-10 mM base; og d) påvise den mindst ene detekterbar markør på prober der har hybridiseret til mål-nukleinsyren i prøven, derved påvise mål-nukleinsyren i prøven, hvor hvert af trinnene a), b) og c) udføres ved en temperatur i området på 19 grader Celsius til 25 grader Celsius.
10. Kit til påvisning afen mål-nukleinsyre i en biologisk prøve, omfattende: a) en denatureringsbuffer omfattende 0,03M til 0,17M base og 50 til 80% alkohol; b) en hybridiseringsbuffer omfattende 1-10 mM base og med en pH i området 10 til 13; og c) en vaskebuffer omfattende 1-10 mM base, hvor vaskebufferen har en pH i området på 10 til 13.
11. Kittet ifølge krav 10, yderligere omfattende én eller flere proteaser til forbehandling af prøven, fx hvor den ene eller flere proteaser omfatter pepsin.
12. Kittet ifølge krav 10 or 11, yderligere omfattende mindst én pro beomfattende en nukleotidsekvens der er komplementær med en nukleotidsekvens i mål-nukleinsyren og mindst én detekterbar markør.
13. Kittet ifølge et hvilket som helst af kravene 10-12, hvor denatureringsbufferen omfatter 0,07M natriumhydroxid og 70% ethanol.
14. Kittet ifølge et hvilket som helst af kravene 10-13, hvor hybridiseringsbufferen omfatter 1-5 mM base, 20-60% formamid, og 5-40% dextransulfat.
15. Kittet ifølge et hvilket som helst af kravene 10-14, hvor vaskebufferen omfatter 1,75 til 2 mM NaOH, og eventuelt hvor vaskebufferen yderligere omfatter 2xSSC.
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| US201161569656P | 2011-12-12 | 2011-12-12 | |
| PCT/US2012/069178 WO2013090386A2 (en) | 2011-12-12 | 2012-12-12 | Methods and kits for room temperature in situ detection of a target nucleic acid in a biological sample |
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| DK (1) | DK2802671T3 (da) |
| WO (1) | WO2013090386A2 (da) |
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| US9145584B2 (en) | 2011-12-12 | 2015-09-29 | Cellay, Inc. | Methods and kits for room temperature in situ detection of a target nucleic acid in a biological sample |
| US20140161972A1 (en) * | 2012-12-09 | 2014-06-12 | National Sun Yat-Sen University | Method for forming conductive film at room temperature |
| BR112016010354B1 (pt) | 2013-11-07 | 2019-09-17 | Cellay, Inc. | Métodos e kits para detectar um carcinoma de próstata e prever prognósticos de doença para cânceres de próstata |
| US11021739B2 (en) * | 2015-02-04 | 2021-06-01 | Alma Mater Studiorum Uneverseta Di Bologna | Additive for accelerating hybridization |
| US10457981B2 (en) * | 2016-01-08 | 2019-10-29 | Abbott Molecular Inc. | Hybridization buffers |
| US10577645B2 (en) * | 2016-03-18 | 2020-03-03 | Norgen Biotek Corp. | Methods and kits for improving global gene expression analysis of human blood, plasma and/or serum derived RNA |
| CN109996875B (zh) * | 2016-08-24 | 2023-11-28 | 注入技术公司 | 使用dtbp浓缩微生物或提取核酸的方法 |
| US11287422B2 (en) | 2019-09-23 | 2022-03-29 | Element Biosciences, Inc. | Multivalent binding composition for nucleic acid analysis |
| KR20240094019A (ko) * | 2019-09-23 | 2024-06-24 | 엘리먼트 바이오사이언스, 인크. | 세포적으로 처리가능한 핵산 서열분석 방법 |
| EP4573213A1 (en) | 2022-08-15 | 2025-06-25 | Element Biosciences, Inc. | Spatially resolved surface capture of nucleic acids |
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| US5753433A (en) * | 1909-12-05 | 1998-05-19 | Boehringer Mannheim Gmbh | Method for the sensitive detection of nucleic acids |
| US5753437A (en) | 1987-10-13 | 1998-05-19 | The United States Of America As Represented By The Department Of Health & Human Services | Method of diagnosing cancer susceptibility or metastatic potential |
| US5082830A (en) | 1988-02-26 | 1992-01-21 | Enzo Biochem, Inc. | End labeled nucleotide probe |
| US5714327A (en) | 1990-07-19 | 1998-02-03 | Kreatech Diagnostics | Platinum-containing compounds, methods for their preparation and applications thereof |
| NL9001639A (nl) | 1990-07-19 | 1992-02-17 | Amc Amsterdam | Pt-houdende verbinding, werkwijze voor de bereiding ervan, alsmede toepassing van dergelijke verbindingen. |
| US5719024A (en) | 1993-08-18 | 1998-02-17 | Applied Spectral Imaging Ltd. | Method for chromosome classification by decorrelation statistical analysis and hardware therefore |
| EP1019420B1 (en) | 1995-05-09 | 2003-08-06 | Kreatech Biotechnology B.V. | Methods for the production of platinum-based linkers between labels and bio-organic molecules, for labelling bio-organic molecules, for detecting biological substances of interest |
| US6780603B1 (en) | 1995-07-21 | 2004-08-24 | Regents Of The University Of Minnesota | Analysis of alpha integrins for the diagnosis of diabetic nephropathy |
| CZ114199A3 (cs) | 1996-10-04 | 1999-09-15 | Novo Nordisk A/S | N-Substituované azaheterocyklické sloučeniny |
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| US6617148B2 (en) | 2000-06-29 | 2003-09-09 | E. I. Du Pont De Nemours And Company | Natural promoters for gene expression and metabolic monitoring in bacillus species |
| ATE334230T1 (de) | 2001-03-02 | 2006-08-15 | Stratagene California | Zusammensetzungen und verfahren mit platinverbindungen zur nukleinsäuremarkierung |
| US20040158883A1 (en) | 2002-11-05 | 2004-08-12 | Crawford Fiona C. | Organic anion transport polypeptide related protein-4 (OATPRP4) gene in Tourette syndrome and related disorders |
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| WO2009041917A1 (en) * | 2007-09-28 | 2009-04-02 | Agency For Science, Technology And Research | Method of electrically detecting a nucleic acid molecule |
| CN101363048B (zh) * | 2008-08-29 | 2011-11-23 | 芮屈生物技术（上海）有限公司 | 一种nkx2-8基因的原位杂交检测试剂盒及其检测方法和应用 |
| WO2011153354A1 (en) | 2010-06-03 | 2011-12-08 | Cellay, Inc. | Methods and kits for in situ detection of nucleotide sequences |
| US9145584B2 (en) | 2011-12-12 | 2015-09-29 | Cellay, Inc. | Methods and kits for room temperature in situ detection of a target nucleic acid in a biological sample |
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2012
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- 2012-12-12 BR BR112014014307A patent/BR112014014307A2/pt not_active IP Right Cessation
- 2012-12-12 EP EP12806823.6A patent/EP2802671B1/en not_active Not-in-force
- 2012-12-12 CN CN201280069016.7A patent/CN104145027A/zh active Pending
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- 2012-12-12 WO PCT/US2012/069178 patent/WO2013090386A2/en not_active Ceased
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2015
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Also Published As
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|---|---|
| CN105385757A (zh) | 2016-03-09 |
| US20150368701A1 (en) | 2015-12-24 |
| WO2013090386A3 (en) | 2013-08-22 |
| US9145584B2 (en) | 2015-09-29 |
| CN104145027A (zh) | 2014-11-12 |
| EP2802671A2 (en) | 2014-11-19 |
| US20130149705A1 (en) | 2013-06-13 |
| CN105385757B (zh) | 2020-04-03 |
| WO2013090386A2 (en) | 2013-06-20 |
| EP2802671B1 (en) | 2018-07-18 |
| US10400275B2 (en) | 2019-09-03 |
| BR112014014307A2 (pt) | 2019-09-24 |
| JP2015505241A (ja) | 2015-02-19 |
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