ES2188357A1 - Method for the amplification by reverse transcription coupled to a nested or semi-nested reaction PCR in a single step, of a target RNA, and its applications in the identification of enterovirus and the discrimination between poliovirus and non-polio enterovirus. - Google Patents

Method for the amplification by reverse transcription coupled to a nested or semi-nested reaction PCR in a single step, of a target RNA, and its applications in the identification of enterovirus and the discrimination between poliovirus and non-polio enterovirus.

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Publication number
ES2188357A1
ES2188357A1 ES200100567A ES200100567A ES2188357A1 ES 2188357 A1 ES2188357 A1 ES 2188357A1 ES 200100567 A ES200100567 A ES 200100567A ES 200100567 A ES200100567 A ES 200100567A ES 2188357 A1 ES2188357 A1 ES 2188357A1
Authority
ES
Grant status
Application
Patent type
Prior art keywords
rna
enterovirus
nested
method
target
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
ES200100567A
Other languages
Spanish (es)
Other versions
ES2188357B1 (en )
Inventor
Seiz Antonio Madejon
Lopez Gemma Limones
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.)
Biotools Biotechnologial & Medical Laboratories Sa
Biotools Biotechnologial & Med
Original Assignee
BIOTOOLS BIOTECHNOLOGIAL & MEDICAL LABORATORIES, S.A.
BIOTOOLS BIOTECHNOLOGIAL & MED
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

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Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/26Preparation of nitrogen-containing carbohydrates
    • C12P19/28N-glycosides
    • C12P19/30Nucleotides
    • C12P19/34Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids

Abstract

Method for the amplification by reverse transcription coupled to a nested or semi-nested reaction PCR in a single step, of a target RNA, and its applications in the identification of enterovirus and the discrimination between poliovirus and non-polio enterovirus. The method consists in performing the reverse transcription of a target RNA contained in a sample by employing an enzyme with the appropriate activity and applying a heat treatment for denaturalising the target RNA and a low temperature heat cycle. From the DNAc obtained, a fragment can be amplified using nested or semi-nested PCR. The method is useful in the amplification of RNA and in the detection and identification of macromolecular entities that include RNA, for example, in the identification of enterovirus and in the discrimination of poliovirus and non-polio enterovirus.
ES200100567A 2001-03-12 2001-03-12 Method for amplification by reverse transcription coupled to a reaction of nested or semi-nested PCR in a single step, of a target RNA, and their applications in the identification of enterovirus and poliovirus discrimination between non-polio enteroviruses and. Expired - Fee Related ES2188357B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
ES200100567A ES2188357B1 (en) 2001-03-12 2001-03-12 Method for amplification by reverse transcription coupled to a reaction of nested or semi-nested PCR in a single step, of a target RNA, and their applications in the identification of enterovirus and poliovirus discrimination between non-polio enteroviruses and.

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES200100567A ES2188357B1 (en) 2001-03-12 2001-03-12 Method for amplification by reverse transcription coupled to a reaction of nested or semi-nested PCR in a single step, of a target RNA, and their applications in the identification of enterovirus and poliovirus discrimination between non-polio enteroviruses and.

Publications (2)

Publication Number Publication Date
ES2188357A1 true true ES2188357A1 (en) 2003-06-16
ES2188357B1 ES2188357B1 (en) 2005-01-01

Family

ID=8497045

Family Applications (1)

Application Number Title Priority Date Filing Date
ES200100567A Expired - Fee Related ES2188357B1 (en) 2001-03-12 2001-03-12 Method for amplification by reverse transcription coupled to a reaction of nested or semi-nested PCR in a single step, of a target RNA, and their applications in the identification of enterovirus and poliovirus discrimination between non-polio enteroviruses and.

Country Status (1)

Country Link
ES (1) ES2188357B1 (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5641864A (en) * 1986-08-22 1997-06-24 Hoffman-La Roche Inc. Kits for high temperature reverse transcription of RNA
US6013488A (en) * 1996-07-25 2000-01-11 The Institute Of Physical And Chemical Research Method for reverse transcription
US6406891B1 (en) * 1998-09-28 2002-06-18 Board Of Regents, The University Of Texas System Dual RT procedure for cDNA synthesis

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5641864A (en) * 1986-08-22 1997-06-24 Hoffman-La Roche Inc. Kits for high temperature reverse transcription of RNA
US6013488A (en) * 1996-07-25 2000-01-11 The Institute Of Physical And Chemical Research Method for reverse transcription
US6406891B1 (en) * 1998-09-28 2002-06-18 Board Of Regents, The University Of Texas System Dual RT procedure for cDNA synthesis

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CARNINCI et al. Thermostabilization and thermoactivation of thermolabile enzymes by trehalose and its application for the synthesis of full length cDNA. Enero 1998. Proc. Natl. Acad. Sci. Vol. 95. Pßginas 520-524. Figura 2; pßgina 522. *
PERRIN YOLANDE et al. Original reverse transcription polymerase chain reaction method to obtain the full-length cDNA of rice tungro spherical virus. Enero 1999. Journal of Virological Methods. Vol. 79. Pßginas 161-168. *

Also Published As

Publication number Publication date Type
ES2188357B1 (en) 2005-01-01 grant

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