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1. WO1996005323 - METHOD FOR THE AMPLIFICATION AND/OR DETECTION OF A NUCLEIC ACID SEQUENCE, DETECTION REAGENT AND USES THEREOF

Publication Number WO/1996/005323
Publication Date 22.02.1996
International Application No. PCT/FR1995/001047
International Filing Date 03.08.1995
Chapter 2 Demand Filed 08.03.1996
IPC
C12Q 1/68 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
CPC
C12Q 1/6865
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6844Nucleic acid amplification reactions
6865Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
Applicants
  • CIS BIO INTERNATIONAL [FR/FR]; Route nationale 306 F-91400 Saclay, FR (AllExceptUS)
  • SAUVAIGO, Sylvie [FR/FR]; FR (UsOnly)
  • BAZIN, Hervé [FR/FR]; FR (UsOnly)
  • LIVACHE, Thierry [FR/FR]; FR (UsOnly)
Inventors
  • SAUVAIGO, Sylvie; FR
  • BAZIN, Hervé; FR
  • LIVACHE, Thierry; FR
Agents
  • CABINET ORES; 6, avenue de Messine F-75008 Paris, FR
Priority Data
94/0989610.08.1994FR
Publication Language French (FR)
Filing Language French (FR)
Designated States
Title
(EN) METHOD FOR THE AMPLIFICATION AND/OR DETECTION OF A NUCLEIC ACID SEQUENCE, DETECTION REAGENT AND USES THEREOF
(FR) METHODE D'AMPLIFICATION ET/OU DE DETECTION D'UNE SEQUENCE D'ACIDE NUCLEIQUE, REACTIF DE DETECTION ET LEURS APPLICATIONS
Abstract
(EN)
Method for the amplification of at least one nucleic acid sequence characterized in that it comprises the steps of (1) solution hybridizing at least one target sequence with a polynucleotidic construct comprising at least one single-strand domain (A) corresponding to a complementary nucleotide sequence of the target sequence and a double strand domain (8, B') forming a promoter, (2) separating the nucleic acids; (3) digesting using a nuclease specific to single strand nucleic acids of polynucleotide constructs, namely those not having produced hybrids with the target sequence, and (4) transcripting, in a solution and in the presence of a DNA dependent RNA-polymerase, the target sequence hybridized in step (1), in the form of multiple RNA copies of the target sequence. According to the method of the invention, the amplified sequence can also be detected or quantified. The invention also concerns the polynucleotide constructs for carrying out the above-mentioned method.
(FR)
L'invention est relative à un procédé d'amplification d'au moins une séquence d'acide nucléique, caractérisé en ce qu'il comprend: (1) l'hybridation en solution d'au moins une séquence cible avec une construction polynucléotidique comprenant au moins un domaine (A) simple-brin correspondant à une séquence nucléotidique complémentaire de la séquence cible et un domaine (B, B') double-brin constituant un promoteur, (2) la séparation des acides nucléiques, (3) la digestion par une nucléase spécifique des acides nucléiques simple-brin des fragments simple-brin des constructions polynucléotidiques, c'est-à-dire notamment celles n'ayant pas formé d'hydrides avec la séquence cible, et (4) la transcription en solution et en présence d'une ARN-polymérase ADN-dépendante, de la séquence cible ayant hybridé à l'étape (1), sous la forme de copies multiples d'ARN de ladite séquence cible, lequel procédé peut comprendre également la détection et/ou la quantification de la séquence amplifiée. L'invention englobe également des constructions polynucléotidiques permettant la mise en ÷uvre du procédé ci-dessus.
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