Nucleic acid detection method using nucleotide probes...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2228712

A nucleic acid detection method wherein (a) a large nucleic acid probe, in particular one having more than 100 nucleotides, is hybridised in a solution, said probe being capable of hybridising with a predetermined RNA or DNA in a nucleic acid-containing sample, and bearing detection elements and first capture elements on a solid medium; (b) the unhybridised nucleotide sequences in the sample are decomposed using an enzyme suitable for decomposing unhybridised nucleic acids; (c) the hybrids produced in (b) are captured by contacting them with a solid medium coated with second capture elements interacting with said first capture elements of said probe; and (d) the hybrids or probes are detected after the hybrids have been denatured using said detection elements.

La présente invention a pour objet un procédé de détection d'acides nucléiques caractérisé en ce qu'il comporte les étapes suivantes: a) on réalise l'hybridation en solution d'une sonde d'acide nucléique de grande taille, notamment de plus de 100 nucléotides, capable de s'hybrider avec un ARN ou un ADN donné dans un échantillon contenant des acides nucléiques, ladite sonde portant des éléments de détection et des premiers éléments de capture sur support solide; b) on réalise la dégradation des séquences nucléotidiques non hybridées dans l'échantillon par un enzyme dégradant les acides nucléiques non hybridées; c) on effectue la capture des hybrides obtenus à l'étape b) en les mettant en présence d'un support solide revêtu des deuxièmes éléments de capture interagissant avec lesdits premiers éléments de capture de ladite sonde et d) on effectue la détection des hybrides ou des sondes après dénaturation des hybrides par l'intermédiaire desdits éléments de détection.

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