A method for determining the integrity of nucleic acid

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2198896

The subject invention is directed at a method for determining the integrity of nucleic acid isolated from a specimen of eukaryotic origin comprising subjecting the specimen to nucleic acid isolation followed by amplification using a pair of oligonucleotide primers in a manner known per se, said primers being capable of amplifying nucleic acid specific for RNA encoding a product of a housekeeping gene, said RNA being low abundance RNA, said gene being present in all cell types of a eukaryotic species, and said method further comprises determining whether amplification of the nucleic acid specific for the RNA has occurred. In particular, the method comprises amplifying mRNA. Optionally, the method can also comprise quantifying the amplification product. As this method is of particular relevance for diagnosis of diseases in humans, the eukaryotic species is preferably a human being.

L'invention concerne un procédé permettant de déterminer l'intégrité d'un acide nucléique isolé d'un échantillon d'origine eucaryote, et consistant à soumettre l'échantillon à un processus d'isolement de l'acide nucléique suivi d'une amplification à l'aide d'une paire d'amorces oligonucléotidiques d'une manière connue en soi, lesdites sondes pouvant amplifier l'acide nucléique spécifique à un ARN codant un produit domestique, cet ARN étant peu abondant, ledit gène étant présent dans tous les types de cellules d'une espèce eucaryote, le procédé consistant en outre à déterminer si l'amplification de l'acide nucléique spécifique à l'ARN s'est effectivement produite. En particulier, le procédé comprend l'amplification de l'ARNm. Eventuellement, le procédé peut également consister à quantifier le produit d'amplification. Ce procédé étant particulièrement utile au diagnostic de maladies chez les humains, l'espèce eucaryote est de préférence d'origine humaine.

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