Highly sensitive method for the detection of cytosine...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2435917

The invention relates to a method for detecting cytosine methylation in DNA samples. The inventive method comprises the following steps: a genomic DNA sample containing DNA to be examined and background DNA is chemically treated in such a way that all non-methylated cytosine bases are converted into uracil, the 5-methylcytosine bases remaining unchanged; the chemically treated DNA samples are amplified using at least two primer oligonucleotides and one polymerase, the DNA to be examined being preferred as a template as opposed to the background DNA; the amplificates are analysed; and the methylation status in the DNA to be examined is inferred from the presence of an amplificate and/or from the analysis of further positions.

L'invention concerne un procédé permettant la détection de la méthylation de la cytosine dans des échantillons d'ADN, caractérisé en ce qu'il comprend les étapes suivantes : on traite un échantillon d'ADN génomique renfermant l'ADN à analyser et l'ADN de base, chimiquement, de façon que toutes les bases de cytosine non méthylées soient transformées en uracile, cependant que les bases de 5-méthylcytosine demeurent inchangées ; les échantillons d'ADN traités chimiquement sont amplifiés en utilisant au moins 2 oligonucléotides primer et une polymérase, l'ADN à analyser étant préféré comme modèle par rapport à l'ADN de base, en ce que les amplificats sont analysés, et en ce que l'état de méthylation dans l'ADN à examiner est déduit à partir de la présence d'un amplificat et/ou de l'analyse d'autres positions.

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