Multiparameter analysis of comprehensive nucleic acids and...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/00 (2006.01) C07H 21/04 (2006.01) C07K 16/00 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01) G01N 33/543 (2006.01) G01N 33/574 (2006.01)

Patent

CA 2465288

A highly sensitive assay is disclosed which utilizes a method for gene specific primed amplification of mRNA libraries from rare cells and rare transcripts found in blood.. The assay allows detection of rare, mRNA (10 copies/cell) found in 1 to 10 cells isolated through immunomagnetic enrichment. The assay is an improvement over multiplex PCR and allows efficient detection of rare coding sequences for circulating carcinoma cells in the blood. The methods are useful in profiling of cells isolated from tissues or body fluids and serves as an adjunct to clinical diagnosis of diverse carcinomas including early stage detection and classification of circulating tumor cells. Monitoring of nucleic acid and protein profiles of cells either in conventional or microarray formats, facilitates management of therapeutic intervention including staging, monitoring response to therapy, confirmation of remission and detection of regression.

L'invention concerne un dosage biologique hautement sensible, qui utilise un procédé pour l'amplification, à amorçage spécifique de gènes, de bibliothèques d'ARNm provenant de cellules rares et de transcrits rares trouvés dans le sang. Le dosage biologique selon l'invention permet de détecter de l'ARNm rare (10 copies/cellule) trouvé dans 1 à 10 cellules isolées par enrichissement immunomagnétique. Le dosage biologique selon l'invention constitue une amélioration par rapport aux PCR multiplex et permet la détection efficace de séquences de codage rares pour des cellules carcinomateuses circulantes dans le sang. L'invention concerne également des procédés permettant le profilage de cellules isolées dans des tissus ou des fluides corporels et pouvant servir de compléments au diagnostic clinique de divers carcinomes, incluant la détection précoce et la classification des cellules tumorales circulantes. La surveillance de profils d'acides nucléiques et de protéines de cellules, dans des formats conventionnels ou bien dans des jeux ordonnés de micro-échantillons, facilite la gestion de l'intervention thérapeutique, incluant la mise en place de la thérapie, la surveillance de la réponse à la thérapie, la confirmation de la rémission et la détection d'une régression.

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