CA2139070A1 - Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction - Google Patents

Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction

Info

Publication number
CA2139070A1
CA2139070A1 CA002139070A CA2139070A CA2139070A1 CA 2139070 A1 CA2139070 A1 CA 2139070A1 CA 002139070 A CA002139070 A CA 002139070A CA 2139070 A CA2139070 A CA 2139070A CA 2139070 A1 CA2139070 A1 CA 2139070A1
Authority
CA
Canada
Prior art keywords
rna
dna
nucleic acid
pcr
detection
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.)
Granted
Application number
CA002139070A
Other languages
French (fr)
Other versions
CA2139070C (en
Inventor
Burton W. Blais
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
567782 Bcltd
Original Assignee
Kalyx Biosciences Inc
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 Kalyx Biosciences Inc filed Critical Kalyx Biosciences Inc
Priority to CA2139070A priority Critical patent/CA2139070C/en
Publication of CA2139070A1 publication Critical patent/CA2139070A1/en
Priority to US08/718,596 priority patent/US5827661A/en
Application granted granted Critical
Publication of CA2139070C publication Critical patent/CA2139070C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

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
    • C12Q1/6804—Nucleic acid analysis using immunogens

Abstract

This invention relates to a new and novel method for significantly enhancing the sensitivity of assays aimed at detecting the presence of target deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequences in a sample. Specifically, this method applies to techniques that employ polymerase chain reactions (PCR?) or other nucleic amplificatio techniques to amplify copies of the target DNA or RNA (via reverse-transclil,tase followed by PCR, Murakawa et ah 1988 DNA 7-287) to allow for detection. The method of thepresent invention combines the following four steps for the first time: DNA amplicon synthesis and amplification in a modified PCR using a target nucleic acid sequence as a template; amplicon transcription into RNA and amplification; RNA:DNA hybrid formation;
and immunochemical detection of heteroduplex nucleic acid sequences. This method extends the standard PCR techniques to accommodate amplification of copies of the PCR product and transcription into RNA, which, following binding to DNA sequences facilitates immunochemical detection of the RNA:DNA hybrids thereby formed. This combination of steps results in a new assay with increased simplicity, speed and sensitivity, including a greatly improved signal-to-noise ratio, over standard nucleic acid detection methods. This improved method will be extremely useful for performing analyses in the foiensic, clinical, agricultural and biological fields.

Claims (6)

1. A method of nucleic acid sequence detection comprising the steps:
a) DNA amplicon synthesis utilizing polymerase chain reaction technique and priming oligonucleotides to which an appropriate RNA polymerase promoter has been appended;
b) transcription of amplicons into RNA sequences using an RNA polymerase;
c) formation of RNA:DNA hybrids; and d) immunochemical detection of RNA:DNA hybrids using procedures comprising antibodies directed to RNA:DNA hybrids.
2. A method as claimed in claim 1, wherein amplicon transcription into RNA is performed in such a manner as to significantly amplify the number of copies of the amplicons.
3. A method as claimed in claim 1 or 2, wherein the DNA amplicon synthesis contains an RNA polymerase promoter sequence.
4. A method as claimed in claim 1 or 2, wherein the DNA amplicon synthesis contains the RNA polymerase promoter T7.
5. A method as claimed in claims 1 through 4, wherein the RNA:DNA hybrid formation has one of the nucleic acid sequences attached to a solid support.
6. A method as claimed in claims 1 through 5, wherein the immunochemical detection is performed by a monoclonal or polyclonal antibody that recognizes the target RNA:DNA hybrid.
CA2139070A 1994-12-23 1994-12-23 Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction Expired - Fee Related CA2139070C (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CA2139070A CA2139070C (en) 1994-12-23 1994-12-23 Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction
US08/718,596 US5827661A (en) 1994-12-23 1996-09-23 Enhancing detection polymerase chain reaction assays by RNA transcription and immunodetection of RNA:DNA hybrids

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CA2139070A CA2139070C (en) 1994-12-23 1994-12-23 Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction

Publications (2)

Publication Number Publication Date
CA2139070A1 true CA2139070A1 (en) 1996-06-24
CA2139070C CA2139070C (en) 2010-03-30

Family

ID=4154924

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2139070A Expired - Fee Related CA2139070C (en) 1994-12-23 1994-12-23 Method for enhancing detection ability of nucleic acid assays employing polymerase chain reaction

Country Status (2)

Country Link
US (1) US5827661A (en)
CA (1) CA2139070C (en)

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US7399589B2 (en) * 1998-02-06 2008-07-15 Digene Corporation Immunological detection of RNA:DNA hybrids on microarrays
US6686151B1 (en) 1998-02-06 2004-02-03 Digene Corporation Immunological detection of RNA:DNA hybrids on microarrays
US6087112A (en) * 1998-12-30 2000-07-11 Oligos Etc. Inc. Arrays with modified oligonucleotide and polynucleotide compositions
US20030180789A1 (en) * 1998-12-30 2003-09-25 Dale Roderic M.K. Arrays with modified oligonucleotide and polynucleotide compositions
US6232068B1 (en) * 1999-01-22 2001-05-15 Rosetta Inpharmatics, Inc. Monitoring of gene expression by detecting hybridization to nucleic acid arrays using anti-heteronucleic acid antibodies
JP2002535015A (en) * 1999-01-29 2002-10-22 エイブイアイ バイオファーマ， インコーポレイテッド Non-invasive method for detecting target RNA
US6271002B1 (en) 1999-10-04 2001-08-07 Rosetta Inpharmatics, Inc. RNA amplification method
US7439016B1 (en) 2000-06-15 2008-10-21 Digene Corporation Detection of nucleic acids by type-specific hybrid capture method
US7601497B2 (en) 2000-06-15 2009-10-13 Qiagen Gaithersburg, Inc. Detection of nucleic acids by target-specific hybrid capture method
NZ530343A (en) * 2001-06-22 2007-01-26 Marshfield Clinic Methods and oligonucleotides for the detection of E. coli 0157:H7
US7015317B2 (en) * 2002-05-02 2006-03-21 Abbott Laboratories Polynucleotides for the detection and quantification of hepatitis B virus nucleic acids
CA2559401A1 (en) * 2003-12-24 2005-07-07 Advanomics Corp. Direct identification and mapping of rna transcripts
US20060240442A1 (en) * 2005-04-20 2006-10-26 Vevea Dirk N Methods and oligonucleotides for the detection of Salmonella SP., E coli 0157:H7, and Listeria monocytogenes
US20060246463A1 (en) * 2005-04-20 2006-11-02 Vevea Dirk N Methods and oligonucleotides for the detection of Salmonella SP., E coli 0157:H7, and Listeria monocytogenes
AU2006247597B2 (en) * 2005-05-13 2010-02-11 Bio-Rad Laboratories, Inc. Identifying statistically linear data
CA2613442C (en) * 2005-06-30 2016-08-23 Archemix Corp. Materials and methods for the generation of fully 2'-modified nucleic acid transcripts
US8101385B2 (en) 2005-06-30 2012-01-24 Archemix Corp. Materials and methods for the generation of transcripts comprising modified nucleotides
EP2759307B1 (en) 2006-03-29 2016-11-09 Merial Limited Vaccine against Streptococci
US20080076121A1 (en) * 2006-09-22 2008-03-27 Paul Kenneth Wolber Microarray nuclease protection assay
CA2673029C (en) * 2006-12-22 2017-03-28 Archemix Corp. Materials and methods for the generation of transcripts comprising modified nucleotides
US8288520B2 (en) * 2008-10-27 2012-10-16 Qiagen Gaithersburg, Inc. Fast results hybrid capture assay and system
ES2644516T3 (en) * 2009-01-28 2017-11-29 Qiagen Gaithersburg, Inc. Method and test for sample preparation of large sequence specific volume
US9797000B2 (en) 2009-05-01 2017-10-24 Qiagen Gaithersburg Inc. Non-target amplification method for detection of RNA splice-forms in a sample
EP2478087B1 (en) 2009-09-14 2017-01-18 QIAGEN Gaithersburg, Inc. Compositions and methods for recovery of nucleic acids or proteins from tissue samples fixed in cytology media
CN102822189B (en) * 2010-01-29 2016-06-29 奇亚根盖瑟斯堡股份有限公司 The method and composition of (multiplex analysis) is analyzed for the sequence-specific purification of nucleic acid and MULTIPLE COMPOSITE body
JP2013517803A (en) 2010-01-29 2013-05-20 キアジェン ゲイサーズバーグ インコーポレイテッド Method for determining and confirming the presence of HPV in a sample
JP2013528049A (en) 2010-05-19 2013-07-08 キアゲン ガイサーズバーグ アイエヌシー． Methods and compositions for sequence-specific purification and multiplex analysis of nucleic acids
US9885092B2 (en) 2011-02-24 2018-02-06 Qiagen Gaithersburg Inc. Materials and methods for detection of HPV nucleic acids
CN102676506A (en) * 2012-05-30 2012-09-19 曹际娟 Mononuclear cell proliferation listeria monocytogenes nucleic acid standard sample and establishing method and application thereof

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ZA853756B (en) * 1984-06-01 1986-01-29 Miles Lab Nucleic acid hybridization assay employing immobilized rna probes
US4683202A (en) * 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4683105A (en) * 1985-10-31 1987-07-28 Westinghouse Electric Corp. Testable, fault-tolerant power interface circuit for normally de-energized loads
CA1340807C (en) * 1988-02-24 1999-11-02 Lawrence T. Malek Nucleic acid amplification process
US5130238A (en) * 1988-06-24 1992-07-14 Cangene Corporation Enhanced nucleic acid amplification process
CA2020958C (en) * 1989-07-11 2005-01-11 Daniel L. Kacian Nucleic acid sequence amplification methods
US5215899A (en) * 1989-11-09 1993-06-01 Miles Inc. Nucleic acid amplification employing ligatable hairpin probe and transcription
US5166051A (en) * 1990-08-08 1992-11-24 Genesis Labs, Inc. Membranes, membrane overlays for exclusion of erythrocytes, and method for immunoassay of whole blood analytes

Also Published As

Publication number Publication date
US5827661A (en) 1998-10-27
CA2139070C (en) 2010-03-30

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