Method of detecting gene methylation and method of examining...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/09 (2006.01)

Patent

CA 2592993

It is intended to provide a method of conveniently and economically detecting the methylation of a gene contained in a biological sample. It is also intended to provide an accurate and sensitive examination method to be used in the case of examining a neoplasm by using the method of detecting gene methylation. In examining the methylation of a gene, a polysaccharide is added to a biological sample before treating it with a bisulfite. Thus, the subsequent bisulfite treatment can be conducted while requiring neither the addition of a protein-denaturing agent nor the purification of DNA. Thus methylated cytosine without uracil-modification can be detected. Moreover, a neoplasm can be examined at an elevated accuracy by detecting the methylation of genes of multiple species contained in a biological sample and calculating the sum thereof.

L'invention a pour objet un procédé consistant à détecter commodément et économiquement la méthylation d'un gène contenu dans un échantillon biologique. L'invention a également pour objet un procédé de recherche précise et sensible à utiliser dans le cas où l'on recherche un néoplasme en utilisant le procédé consistant à détecter la méthylation d'un gène. Lorsqu'on recherche la méthylation d'un gène, on ajoute un polysaccharide à un échantillon biologique avant de le traiter avec un bisulfite. Ainsi, on peut effectuer le traitement au bisulfite subséquent sans avoir recours ni à l'ajout d'un agent dénaturant les protéines, ni à la purification de l'ADN. Ainsi on peut détecter la cytosine méthylée sans modification en uracile. De plus, on peut rechercher un néoplasme avec une précision élevée en détectant la méthylation de gènes de multiples espèces contenus dans un échantillon biologique et en calculant la somme de celles-ci.

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