Kits, solid supports and methods for point-of-care...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) B01J 19/00 (2006.01)

Patent

CA 2715890

Compositions and methods for amplifying and detecting solution-state polynucleotide targets in a single device are described. In one aspect, a method for a coupled isothermal amplification and detection process utilizes a coated solid support, including a solid substrate, a cationic layer, and a plurality of target- specific probes attached to the coated solid support. Polynucleotide targets in the sample are amplified by an isothermal amplification process involving in situ hybridization onto the coated solid support. The entire process can be carried out with a high degree of specificity under low salt conditions in less than one hour. Further aspects of the present invention include methods for coupled hybridization/detection of polynucleotide targets, coat-ed silicon biosensors optimized for use with the coupled detection systems to provide visual detection of polynucleotide targets under visible light conditions, and kits for practicing the above described methods.

La présente invention concerne des compositions et des procédés d'amplification et de détection de polynucléotides ciblés à l'état de solution dans un seul dispositif. Un aspect de l'invention concerne un procédé d'amplification isotherme couplée à une détection utilisant un support enduit solide constitué d'un substrat solide, d'une couche cationique, et d'une pluralité de sondes spécifiques de la cible, fixées au support solide enduit. Les polynucléotides ciblés de l'échantillon subissent une amplification isotherme avec hybridation in situ sur le support solide enduit. La totalité du processus peut s'exécuter en moins d'une heure, avec un degré élevé de spécificité, et dans des conditions de faible salinité. D'autres aspects de la présente invention concernent des procédés couplant l'hybridation et la détection des polynucléotides ciblés, des capteurs biologiques en silicium enduit destinés aux systèmes de détection couplés, permettant une détection visuelle en lumière visible des polynucléotides ciblés, et des nécessaires permettant de mettre en uvre les procédés de l'invention.

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