Pulsed-multiline excitation for color-blind fluorescence...

G - Physics – 01 – J

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G01J 3/30 (2006.01) G01J 3/10 (2006.01) G01J 3/44 (2006.01) G01N 21/64 (2006.01) H01J 65/08 (2006.01)

Patent

CA 2484336

The present invention provides a technology called Pulse-Multiline Excitation or PME. This technology provides a novel approach to fluorescence detection with application for high throughput identification of informative SNPs, which could lead to more accurate diagnosis of inherited disease, better prognosis of risk susceptibilities, or identification of sporadic mutations. The PME technology has two main advantages that significantly increase fluorescence sensitivity : (1) optimal excitation of all fluorophores in the genomic assay and (2) "color-blind" detection, which collects considerably more light than standard wavelength resolved detection. This technology differs significantly from the current state-of-the-art DNA sequencing instrumentation, which features single source excitation and color dipersion for DNA sequence identification. Successful implementation of the PME technology will have broad application for routine usage in clinical diagnostics, forensics, and general sequencing methodologies and will have the capability, flexibility, and portability of targeted sequence variation assays for a large majority of the population.

La présente invention concerne une technique baptisée "excitation à lignes multiples par impulsions" (Pulse-Multiline Excitation, ou PME). Cette technique constitue une nouvelle approche de la détection par fluorescence appliquée pour l'identification à haut débit de SNP d'informations, ce qui permet d'effectuer un diagnostic plus précis d'une maladie héréditaire, de mieux pronostiquer les risques ou d'identifier les mutations sporadiques. La technique PME possède deux avantages qui augmentent sensiblement la sensibilité de la fluorescence: (1) l'excitation optimale de tous les fluorophores dans un dosage génomique et (2) la détection "non chromatisée" qui collecte considérablement plus de lumière que la détection de longueurs d'ondes à résolution standard. Cette technique diffère sensiblement du matériel d'avant-garde utilisé actuellement pour les séquençage d'ADN utilisant l'excitation à source unique et l'identification des séquences d'ADN par dispersion de couleurs. La mise en oeuvre réussie de la technologie de PME va avoir une large application dans l'utilisation courante en diagnostique clinique, en médecine légale et les procédés généraux de séquençage; elle pourra avoir la capacité, la souplesse et la portabilité des dosages à variation de séquences cibles pour une grande majorité de la population.

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