AR077841A1 - Metodos kits y composiciones de deteccion de acido nucleico a multiples temperaturas, con sonda unica - Google Patents
Metodos kits y composiciones de deteccion de acido nucleico a multiples temperaturas, con sonda unicaInfo
- Publication number
- AR077841A1 AR077841A1 ARP100102930A ARP100102930A AR077841A1 AR 077841 A1 AR077841 A1 AR 077841A1 AR P100102930 A ARP100102930 A AR P100102930A AR P100102930 A ARP100102930 A AR P100102930A AR 077841 A1 AR077841 A1 AR 077841A1
- Authority
- AR
- Argentina
- Prior art keywords
- temperature
- variant
- probe
- temperature signal
- sample
- Prior art date
Links
- 239000000523 sample Substances 0.000 title abstract 17
- 238000000034 method Methods 0.000 title abstract 6
- 239000000203 mixture Substances 0.000 title abstract 4
- 238000001514 detection method Methods 0.000 title abstract 3
- 150000007523 nucleic acids Chemical class 0.000 title 1
- 102000039446 nucleic acids Human genes 0.000 title 1
- 108020004707 nucleic acids Proteins 0.000 title 1
- 108091093088 Amplicon Proteins 0.000 abstract 6
- 238000009396 hybridization Methods 0.000 abstract 3
- 230000003321 amplification Effects 0.000 abstract 2
- 230000001419 dependent effect Effects 0.000 abstract 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract 2
- 241001465754 Metazoa Species 0.000 abstract 1
- 238000012408 PCR amplification Methods 0.000 abstract 1
- 241000191940 Staphylococcus Species 0.000 abstract 1
- 238000007846 asymmetric PCR Methods 0.000 abstract 1
- 230000000295 complement effect Effects 0.000 abstract 1
- 238000002405 diagnostic procedure Methods 0.000 abstract 1
- 244000052769 pathogen Species 0.000 abstract 1
- 230000001717 pathogenic effect Effects 0.000 abstract 1
Classifications
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- 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
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- 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
-
- 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
- C12Q1/6858—Allele-specific amplification
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- 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
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- 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
- C12Q2600/156—Polymorphic or mutational markers
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- 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
- C12Q2600/16—Primer sets for multiplex assays
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Se proveen en la presente métodos, kits y composiciones que incluyen la discriminacion de secuencias diana por unica sonda donde diferentes amplicones diana pueden tener secuencias de hibridacion de sonda idénticas, empleando deteccion de senales de punto final de sonda a multiples temperaturas. También se proveen en la presente métodos, kits y composiciones para distinguir entre dos o más amplicones diana utilizando deteccion de punto final de sonda a multiples temperaturas. En ciertas realizaciones, se emplean métodos de amplificacion por PCR asimétrica (por ej., amplificacion por LATE-PCR). Estos métodos, kits y composiciones están relacionados con ensayos diagnosticos de Staphylococcus aureaus (patogeno virulento de animales y de humanos). Reivindicacion 1: Un método para identiticar la presencia de variantes de secuencia diana polimortica en una muestra, que comprende: a) proveer: i) una muestra que se sospecha contiene: una primera o segunda variante de una secuencia diana polimorfica, ii) una sonda marcada, iii) una relacion de senales temperatura/temperatura de la primera variante, iv) una relacion de senales temperatura/temperatura de la segunda variante, y v) un cebador directo, y vi) un cebador inverso; b) combinar dicha muestra, dicha sonda marcada, dicho cebador directo, y dicho cebador inverso para generar una muestra combinada y tratar dicha muestra combinada bajo condiciones de amplificacion de modo que: se genera un primer amplicon de simple hebra si dicha primera variante está presente, y se genera un segundo amplicon de simple hebra si dicha segunda variante está presente, en donde dichos amplicones primero y segundo de simple hebra cada uno comprende las siguientes secuencias idénticas: i) una secuencia de hibridacion de sonda, ii) un extremo 5' correspondiente a la secuencia de dicho cebador inverso, y iii) un extremo 3 complementario a dicho cebador directo; y en donde dichos amplicones primero y segundo de simple hebra no tienen identidad de secuencia completa; c) exponer dicha muestra combinada a multiples temperaturas que permiten que dicha sonda marcada se hibridice a dicha secuencia de hibridacion de sonda y produzca senales dependientes de la temperatura; d) detectar dichas senales dependientes de la temperatura a al menos dos temperaturas; e) generar una relacion de senales temperatura/temperatura experimental; y f) comparar dicha relacion de senales temperatura/temperatura experimental con dicha relacion de senales temperatura/temperatura de la primera y segunda variante, en donde una coincidencia entre dicha relacion de senales temperatura/temperatura experimental y dicha relacion de senales temperatura/temperatura de la primera y segunda variante identifica la presencia de dicha primera o segunda variante en dicha muestra.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US23299909P | 2009-08-11 | 2009-08-11 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AR077841A1 true AR077841A1 (es) | 2011-09-28 |
Family
ID=43586459
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ARP100102930A AR077841A1 (es) | 2009-08-11 | 2010-08-10 | Metodos kits y composiciones de deteccion de acido nucleico a multiples temperaturas, con sonda unica |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US9493835B2 (es) |
| EP (1) | EP2464737A4 (es) |
| AR (1) | AR077841A1 (es) |
| WO (1) | WO2011019837A1 (es) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AR077840A1 (es) | 2009-08-11 | 2011-09-28 | Univ Brandeis | Ensayos de deteccion e identificacion de especies y tipos de staphylococcus |
| WO2012095748A2 (en) * | 2011-01-14 | 2012-07-19 | Smiths Detection - Watford Limited | Single tube quantitative polymerase chain reaction (pcr) |
| US20130096845A1 (en) | 2011-10-14 | 2013-04-18 | Ze'ev Russak | Cumulative differential chemical assay identification |
| EP2617836A1 (en) * | 2012-01-23 | 2013-07-24 | Steffen Mergemeier | Method for detecting a methicillin resistant coagulase positive Staphylococcus aureus strain |
| WO2015147370A1 (en) | 2014-03-28 | 2015-10-01 | Seegene, Inc. | Detection of target nucleic acid sequences using different detection temperatures |
| US10876155B2 (en) | 2014-12-09 | 2020-12-29 | Seegene, Inc. | Differentiation of signals for target nucleic acid sequences |
| CN107208134B (zh) * | 2014-12-09 | 2021-05-11 | Seegene株式会社 | 使用不同检测温度及基准值的靶核酸序列检测 |
| ES2786974T3 (es) * | 2016-04-07 | 2020-10-14 | Illumina Inc | Métodos y sistemas para la construcción de bibliotecas de ácidos nucleicos normalizadas |
| EP3781708B1 (en) * | 2018-04-20 | 2025-01-15 | Seegene, Inc. | Method for detecting a plurality of target nucleic acid sequences in sample |
| JP7221491B2 (ja) * | 2018-09-18 | 2023-02-14 | 株式会社ミズホメディー | 複数の標的核酸を検出するキットを用いる検出方法 |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5066584A (en) | 1988-09-23 | 1991-11-19 | Cetus Corporation | Methods for generating single stranded dna by the polymerase chain reaction |
| CA2348042A1 (en) * | 2001-06-04 | 2002-12-04 | Ann Huletsky | Sequences for detection and identification of methicillin-resistant staphylococcus aureus |
| US7198897B2 (en) | 2001-12-19 | 2007-04-03 | Brandeis University | Late-PCR |
| WO2006044994A2 (en) | 2004-10-18 | 2006-04-27 | Brandeis University | Primers, probes and methods for nucleic acid amplification |
| US7838221B2 (en) | 2005-10-11 | 2010-11-23 | Geneohm Sciences, Inc. | Sequences for detection and identification of methicillin-resistant Staphylococcus aureus (MRSA) |
| JP5286261B2 (ja) | 2006-07-17 | 2013-09-11 | ブランディーズ・ユニバーシティ | 特殊化オリゴヌクレオチドならびに核酸の増幅および検出でのその使用 |
-
2010
- 2010-08-10 AR ARP100102930A patent/AR077841A1/es unknown
- 2010-08-11 EP EP10808708.1A patent/EP2464737A4/en not_active Withdrawn
- 2010-08-11 WO PCT/US2010/045199 patent/WO2011019837A1/en not_active Ceased
- 2010-08-11 US US13/389,921 patent/US9493835B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| WO2011019837A1 (en) | 2011-02-17 |
| EP2464737A1 (en) | 2012-06-20 |
| EP2464737A4 (en) | 2013-04-17 |
| US9493835B2 (en) | 2016-11-15 |
| US20120202203A1 (en) | 2012-08-09 |
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