Real time homogeneous nucleotide assay

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2262520

A method for performing real time, homogeneous assay of a target nucleic acid comprising annealing a labeled, ribo-oligonucleotide probe to the target DNA sequence and degrading a portion of the probe with RNase H to release labelled fragments. Sequential measurements of the released fragments permit kinetic characterization of the presence of the target sequence. Preferably, the assay is integrated into a polymerase chain reaction so that target amplification can be detected in real time.

La présente invention a pour objet un dosage en temps réel et homogène d'un acide nucléique cible, s'effectuant par hybridation d'une sonde de ribo-oligonucléotide marquée avec la séquence d'ADN cible et par altération d'une partie de la sonde avec RNase H en vue de la libération de fragments marqués. Des mesures successives des fragments libérés permettent la caractérisation cinétique de la présence de la séquence cible. Le dosage est de préférence intégré dans une réaction en chaîne de la polymérase de façon à permettre la détection en temps réel de l'amplification cible.

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