Method for concurrent amplification and real time detection...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2396301

The present invention provides a method of detecting a genetic polymorphism in an individual or between individuals. The method comprises the following steps, (1) obtaining a sample containing nucleic acid from an individual; (2) contacting the sample, under conditions which permit primer-initiated nucleic acid amplification and nucleic acid cleavage, with (i) a primer suitable for initiating amplification, (ii) an indicator system which provides a signal proportional to the amount of amplification product, and (iii) a sequence specific nucleic acid cleavage agent; and (3) measuring the signal produced by the indicator system against time.Cleavage of the amplification product by the cleavage agent results in an inhibition of the rate of accumulation of amplification product comprising the sequence recognised by the cleavage agent relative to the rate of accumulation of amplification product not comprising the sequence recognised by the cleavage agent.

La présente invention concerne un procédé de détection d'un polymorphisme génétique dans un individu ou entre plusieurs individus. Ledit procédé consiste (1) à prélever un échantillon contenant des acides nucléiques sur un individu, (2) à mettre l'échantillon, dans des conditions permettant l'amplification d'acides nucléiques initiée par amorceur et le clivage d'acides nucléiques, en contact avec (I) un amorceur pouvant initier l'amplification, (II) un système indicateur produisant un signal proportionnel à la quantité de produit d'amplification, et (III) un agent de clivage d'acides nucléiques spécifique à la séquence, et (3) à mesurer le signal produit par le système indicateur par rapport au temps. Le clivage du produit d'amplification par l'agent de clivage entraîne une inhibition de la vitesse d'accumulation du produit d'amplification contenant la séquence reconnue par l'agent de clivage par rapport à la vitesse d'accumulation du produit d'amplification ne contenant pas la séquence reconnue par l'agent de clivage.

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