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RNA DETECTION METHOD

  • US 20110212451A1
  • Filed: 11/13/2009
  • Published: 09/01/2011
  • Est. Priority Date: 11/13/2008
  • Status: Abandoned Application
First Claim
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1. A method for the detection of a target RNA sequence in a sample comprising the steps of:

  • (a) contacting single-stranded RNA (ssRNA) molecules present in a sample with a labelled oligonucleotide containing a sequence substantially complementary to a region of the target RNA under hybridisation conditions to provide a mixture of RNA-oligonucleotide duplexes wherein the RNA-oligonucleotide duplexes comprise the target RNA annealed to the labelled oligonucleotide;

    (b) maintaining the mixture of step (a) with an RNA-dependent RNA polymerase (RdRp) under RNA polymerisation conditions in the absence of a primer wherein said RdRp has an RNA-oligonucleotide duplex separation activity and is capable of de novo RNA synthesis such that the RdRp polymerises an RNA strand complementary to the ssRNA in the sample and releases the labelled oligonucleotide; and

    (c) detecting and/or measuring the signal generated by the released labelled oligonucleotide.

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