Method for the qualitative and quantitative detection of dna...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12Q 1/18 (2006.01)

Patent

CA 2215493

A method for the qualitative and quantitative detection of damages to DNA, characterised in that it includes the steps of binding the target DNA to a sensitised substrate, subjecting the DNA to the action of a test composition comprising at least one damage-generating product, subjecting the DNA to the action of a composition containing at least one cell extract active in repairing the DNA and containing markers, and detecting directly or indirectly the markers optionally incorporated into the DNA, if the repair has been effective, wherein a washing step is included between said steps and the binding and damaging steps can be reversed. The invention also provides a method for capturing the DNA directly from the cells with a lysis solution and a method for determining the repair modulation effect of some molecules. Said method is suitable for detecting DNA damage and the repair inhibition modulating activity of one or more molecules.

L'objet de l'invention est un procédé de détection qualitative et quantitative de lésions sur de l'ADN, qui se caractérise en ce qu'il comprend les différentes étapes suivantes: fixation de l'ADN cible sur un support sensibilisé, action d'une composition à tester comprenant au moins un produit lésant, action d'une composition comprenant au moins un extrait cellulaire possédant une activité réparatrice sur cet ADN, ledit extrait contenant des marqueurs, révélation directe ou indirecte des marqueurs éventuellement incorporés dans l'ADN, en cas de réparation; lesdites étapes étant séparées par au moins une étape de lavage, les étapes de fixation et de lésion pouvant être inversées. L'invention a également pour objet le procédé de capture de l'ADN directement à partir de cellules avec une solution de lyse, et un procédé de détermination de l'effet modulateur de la réparation de certaines molécules. Application à la détection de lésions de l'ADN et de l'action modulatrice d'inhibition de réparation d'au moins une molécule.

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