Method for the detection of specific nucleic acid sequences...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12N 15/00 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2354682

A method for rapid and efficient detection of a target DNA or RNA sequence is provided. A primer having a 3'-hydroxyl group at one end and having a sequence of nucleotides sufficiently homologous with an identifying sequence of nucleotides in the target DNA is selected. The primer is hybridized to the identifying sequence of nucleotides on the DNA or RNA sequence and a reporter molecule is synthesized on the target sequence by progressively binding complementary nucleotides to the primer, where the complementary nucleotides include nucleotides labeled with a fluorophore. Fluorescence emitted by fluorophores on single reporter molecules is detected to identify the target DNA or RNA sequence.

Cette invention a trait à une technique permettant une détection rapide et efficace d'une séquence cible d'ADN ou d'ARN. On sélectionne, dans le cadre de cette invention, une amorce possédant un groupe 3'-hydroxy sur l'une de ses extrémités et pourvue d'une séquence nucléotidique suffisamment homologue avec une séquence d'identification de nucléotides dans l'ADN ciblé. On hybride l'amorce à la séquence d'identification de nucléotides sur la séquence d'ADN ou d'ARN et on synthétise une molécule reporter sur la séquence cible par fixation progressive de nucléotides complémentaires à l'amorce, ces nucléotides complémentaires comportant des nucléotides marqués à l'aide d'un fluorophore. La fluorescence émise par les fluorophores sur les molécules du reporter est détectée aux fins de l'identification de la séquence d'ADN ou d'ARN cible.

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