Method for the detection of cytosine methylations in dna...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2540378

The invention relates to a method for analyzing cytosine methylations in DNA sequences, according to which non-methylated cytosines are first converted into uracil while 5-methylcytosine remains unmodified. The DNA is then amplified by means of a polymerase and at least one primer whose 5 end is connected to a probe via a linker. The probe is intramolecularly hybridized onto the amplified products in accordance with the methylation state of the DNA, hybridization being detectable via different detection systems. The inventive method is particularly suitable for diagnosing and predicting cancer diseases and other diseases associated with a modification of the methylation state as well as for predicting undesired effects of medicaments.

Procédé de détection de méthylation de cytosine dans des séquences d'ADN, selon lequel des cytosines non méthylées sont d'abord converties en uracile, tandis que la 5-méthylcytosine reste inchangée. Ensuite, l'ADN est amplifié à l'aide d'une polymérase et d'au moins une amorce dont la terminaison 5' est reliée à une sonde via une séquence de liaison. Un hybridation intramoléculaire de la sonde se produit sur les produits amplifiés en fonction de l'état de méthylation de l'ADN. L'hybridation peut être prouvée à l'aide de différents systèmes de détection. Le procédé selon la présente invention est particulièrement adapté pour le diagnostic et le pronostic de maladies cancéreuses et d'autres maladies associées à une modification de l'état de méthylation, ainsi que pour prédire des effets non désirés de médicaments.

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