Additives for use in cycling probe reactions

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2295206

A method for detecting a target nucleic acid molecule is provided, comprising the steps of (a) reacting a mixture comprising (i) a target nucleic acid molecule; (ii) a single-stranded nucleic acid probe containing a scissile linkage; (iii) an enzyme capable of cleaving the probe portion of a double- stranded target-probe complex at the scissile linkage; and (iv) ribosomal protein and/or spermine, under conditions and for a time sufficient to allow the target nucleic acid and probe to hybridize to each other and form a double- stranded target-probe complex, followed by cleavage of the probe and cycling of the target to a new uncleaved probe, such that one or more portions of the cleaved nucleic acid probe are released from the target-probe complex; and (b) determining whether cleaved portions of the nucleic acid probe are produced, and thereby detecting the presence of the target nucleic acid.

L'invention concerne un procédé pour détecter une molécule d'acide nucléique cible. Ce procédé consiste à (a) faire réagir un mélange comprenant (i) une molécule d'acide nucléique cible; (ii) une sonde d'acide nucléique à simple brin contenant une liaison pouvant être scindée; (iii) une enzyme assurant le clivage de la partie de sonde d'un complexe cible-sonde à doubles brins au niveau de la liaison pouvant être scindée; et (iv) une protéine ribosomique et/ou diaminopropyl putressine, dans des conditions et pendant un laps de temps suffisants por que l'acide nucléique cible et la sonde s'hybrident l'un à l'autre et forment un complexe cible-sonde à double brins, avant le clivage de la sonde et la formation d'un cycle de la cible jusqu'à une nouvelle sonde non clivée, de telle sorte que une ou plusieurs parties de la sonde d'acide nucléique clivée soient libérées du complexe cible-sonde, et (b) déterminer la production de parties clivées de la sonde d'acide nucléique, ce qui correspond à la présence d'un acide nucléique cible.

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