WO2012069484A3 - Method for qualitatively and quantitatively detecting specific nucleic acid sequences in real time - Google Patents

Method for qualitatively and quantitatively detecting specific nucleic acid sequences in real time Download PDF

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Publication number
WO2012069484A3
WO2012069484A3 PCT/EP2011/070699 EP2011070699W WO2012069484A3 WO 2012069484 A3 WO2012069484 A3 WO 2012069484A3 EP 2011070699 W EP2011070699 W EP 2011070699W WO 2012069484 A3 WO2012069484 A3 WO 2012069484A3
Authority
WO
WIPO (PCT)
Prior art keywords
fluorophor
marked
qualitatively
nucleic acid
real time
Prior art date
Application number
PCT/EP2011/070699
Other languages
German (de)
French (fr)
Other versions
WO2012069484A2 (en
Inventor
Elmara Graser
Timo Hillebrand
Original Assignee
Aj Innuscreen Gmbh
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Aj Innuscreen Gmbh filed Critical Aj Innuscreen Gmbh
Publication of WO2012069484A2 publication Critical patent/WO2012069484A2/en
Publication of WO2012069484A3 publication Critical patent/WO2012069484A3/en

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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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6851—Quantitative amplification

Abstract

The method and test kit are designed to qualitatively and quantitatively detect nucleic acid sequences in real time. The invention relates to the use of a DNA probe marked only with a fluorochrome (fluorophor), wherein the base marked with the fluorophor is always present in the vicinity of a guanine base and thus quenches the fluorophor without the use of a quenching dye. A mixture of duplexes is generated under hybridization conditions by means of primers, wherein the duplexes comprise the target nucleic acids deposited on the marked oligonucleotide (probe). By adding a polymerase having exonuclease activity, cutting the deposited marked oligonucleotide (probe) between the fluorophor marking and the guanine base, and thus terminating the quenching, a measurable fluorescence signal is generated.
PCT/EP2011/070699 2010-11-22 2011-11-22 Method for qualitatively and quantitatively detecting specific nucleic acid sequences in real time WO2012069484A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102010052524.3 2010-11-22
DE201010052524 DE102010052524A1 (en) 2010-11-22 2010-11-22 Method for the qualitative and quantitative detection of specific nucleic acid sequences in real time

Publications (2)

Publication Number Publication Date
WO2012069484A2 WO2012069484A2 (en) 2012-05-31
WO2012069484A3 true WO2012069484A3 (en) 2012-09-27

Family

ID=45406667

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2011/070699 WO2012069484A2 (en) 2010-11-22 2011-11-22 Method for qualitatively and quantitatively detecting specific nucleic acid sequences in real time

Country Status (2)

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DE (1) DE102010052524A1 (en)
WO (1) WO2012069484A2 (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015023677A1 (en) * 2013-08-12 2015-02-19 Bio-Rad Laboratories, Inc. Amplification reporter with base-pairing oligomers
DE102018213026A1 (en) 2018-08-03 2020-02-06 Robert Bosch Gmbh Procedure for performing real-time PCR
CN112852927A (en) * 2021-02-02 2021-05-28 中国科学院合肥物质科学研究院 Isothermal amplification system and method based on fluorescence self-inhibition probe

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008110622A1 (en) * 2007-03-14 2008-09-18 Aj Innuscreen Gmbh Method and test kit for the rapid detection of specific nucleic acid sequences, especially for detecting mutations or snps

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5210015A (en) 1990-08-06 1993-05-11 Hoffman-La Roche Inc. Homogeneous assay system using the nuclease activity of a nucleic acid polymerase
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

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008110622A1 (en) * 2007-03-14 2008-09-18 Aj Innuscreen Gmbh Method and test kit for the rapid detection of specific nucleic acid sequences, especially for detecting mutations or snps

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
SHI M M ET AL: "High throughput genotyping for the detection of a single nucleotide polymorphism in NAD(P)H quinone oxidoreductase (DT diaphorase) using TaqMan probes", MP. MOLECULAR PATHOLOGY, BMJ PUBLISHING GROUP, LONDON, GB, vol. 52, no. 5, 1 October 1999 (1999-10-01), pages 295 - 299, XP008094615, ISSN: 1366-8714, DOI: 10.1136/MP.52.5.295 *
TANAKA R ET AL: "Fully automated and super-rapid system for the detection of JAK2V617F mutation", LEUKEMIA RESEARCH, NEW YORK,NY, US, vol. 32, no. 9, 1 September 2008 (2008-09-01), pages 1462 - 1467, XP022659315, ISSN: 0145-2126, [retrieved on 20080306], DOI: 10.1016/J.LEUKRES.2007.12.019 *

Also Published As

Publication number Publication date
WO2012069484A2 (en) 2012-05-31
DE102010052524A1 (en) 2012-05-24

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