ES2188357B1 - METHOD FOR AMPLIFICATION BY TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO. - Google Patents

METHOD FOR AMPLIFICATION BY TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO.

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Publication number
ES2188357B1
ES2188357B1 ES200100567A ES200100567A ES2188357B1 ES 2188357 B1 ES2188357 B1 ES 2188357B1 ES 200100567 A ES200100567 A ES 200100567A ES 200100567 A ES200100567 A ES 200100567A ES 2188357 B1 ES2188357 B1 ES 2188357B1
Authority
ES
Spain
Prior art keywords
nested
identification
rna
poliovirus
discrimination
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.)
Expired - Fee Related
Application number
ES200100567A
Other languages
Spanish (es)
Other versions
ES2188357A1 (en
Inventor
Antonio Madejon Seiz
Gemma Limones Lopez
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 Biotechnological and Medical Laboratories SA
Original Assignee
Biotools Biotechnological and Medical Laboratories SA
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 Biotools Biotechnological and Medical Laboratories SA filed Critical Biotools Biotechnological and Medical Laboratories SA
Priority to ES200100567A priority Critical patent/ES2188357B1/en
Publication of ES2188357A1 publication Critical patent/ES2188357A1/en
Application granted granted Critical
Publication of ES2188357B1 publication Critical patent/ES2188357B1/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • 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

Abstract

Método para la amplificación mediante transcripción en reverso acoplada a una reacción de nested o semi-nested PCR en un solo paso, de un ARN diana, y sus aplicaciones en la identificación de enterovirus y en la discriminación entre poliovirus y enterovirus no-polio. El método comprende efectuar la transcripción en reverso de un ARN diana contenido en una muestra mediante el empleo de una enzima con actividad apropiada y aplicar un tratamiento térmico para desnaturalizar el ARN diana y un ciclo térmico de baja temperatura. A partir del ADNc obtenido se puede amplificar un fragmento por nested o semi-nested PCR. El método tiene utilidad en la amplificación de ARN y en la detección e identificación de entidades macromoleculares que comprenden ARN, por ejemplo, en la identificación de enterovirus y en la discriminación de poliovirus y enterovirus no-polio.Method for amplification by reverse transcription coupled to a one-step nested or semi-nested PCR reaction of a target RNA, and its applications in the identification of enteroviruses and in the discrimination between poliovirus and non-polio enterovirus. The method comprises performing the reverse transcription of a target RNA contained in a sample by using an enzyme with appropriate activity and applying a heat treatment to denature the target RNA and a low temperature thermal cycle. From the obtained cDNA a fragment can be amplified by nested or semi-nested PCR. The method has utility in RNA amplification and in the detection and identification of macromolecular entities comprising RNA, for example, in the identification of enteroviruses and in the discrimination of poliovirus and non-polio enterovirus.

ES200100567A 2001-03-12 2001-03-12 METHOD FOR AMPLIFICATION BY TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO. 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 TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO.

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES200100567A ES2188357B1 (en) 2001-03-12 2001-03-12 METHOD FOR AMPLIFICATION BY TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO.

Publications (2)

Publication Number Publication Date
ES2188357A1 ES2188357A1 (en) 2003-06-16
ES2188357B1 true 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 TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO.

Country Status (1)

Country Link
ES (1) ES2188357B1 (en)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5310652A (en) * 1986-08-22 1994-05-10 Hoffman-La Roche Inc. Reverse transcription with thermostable DNA polymerase-high temperature reverse transcription
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. Páginas 520-524. Figura 2; página 522. \\ A 17-22 *
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. Páginas 161-168. *

Also Published As

Publication number Publication date
ES2188357A1 (en) 2003-06-16

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