Two-step hybridization and capture of a polynucleotide

Details Two-step hybridization and capture of a polynucleotide Two-step hybridization and capture of a polynucleotide

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 21/00 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2287570

A method for capturing a target polynucleotide in a sample onto a solid support with an attached immobilized probe by using a capture probe and two different hybridization conditions, which preferably differ in temperature only, is disclosed. The two hybridization conditions control the order of hybridization, where the first hybridization condition allows the capture probe and the target polynucleotide to hybridize, and the second hybridization condition allows the capture probe and the immobilized probe to hybridize, to produce a complex of immobilized probe, capture probe and target polynucleotide. A method for determining the presence of a target polynucleotide in a sample by capturing a target polynucleotide, amplifying the captured target polynucleotide and detecting amplified sequences is also disclosed.

L'invention concerne un procédé permettant de capturer un polynucléotide cible dans un échantillon posé sur un support solide pourvu d'une sonde fixe, à l'aide d'une sonde de capture et de deux conditions d'hybridation différentes présentant, de préférence, uniquement une différence de température. Les deux conditions d'hybridation commandent l'ordre d'hybridation, la première condition d'hybridation permettant l'hybridation de la sonde de capture et du polynucléotide cible et la seconde condition d'hybridation permettant l'hybridation de la sonde de capture et de la sonde immobilisée, afin de produire un complexe contenant une sonde immobilisée, une sonde de capture et un polynucléotide cible. L'invention concerne également un procédé permettant de déterminer la présence d'un polynucléotide cible dans un échantillon par capture d'un polynucléotide cible, par amplification du polynucléotide cible capturé et par détection de séquences amplifiées.

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