Analytical method and kit

Abstract

RNA-containing probes and kits comprising RNA-containing probes for the detection and analysis of nucleic acid sequences are described. These probes are contacted with a sample suspected of containing the nucleic acid sequence and if they form duplexes, they are hydrolysed. This may be done, for example during an amplification reaction. AMP generated as a result of the hydrolysis is converted to ATP. The ATP may then be detected using bioluminescent reagents. Inclusion of modified adenosine in at least one probe means that the signal arising from one probe will give rise to a different and distinguishable bioluminescent signal thus enabling the use of for example an internal control in bioluminescently-reported nucleic acid tests.

1 . A kit for use in analysis, the kit comprising at least two RNA-containing probes, each of which is specific for a target sequence, wherein one of said probes comprises at least one modified adenosine. 
     
     
         2 . The kit of  claim 1 , which further comprises an enzyme or enzyme mixture which can hydrolyse an RNA-containing probe when in double stranded form. 
     
     
         3 . The kit of  claim 1 , which further comprises means for converting AMP to ATP or modified AMP to modified ATP. 
     
     
         4 . The kit  claim 1 , which further comprises a bioluminescent reagent. 
     
     
         5 . The kit of  claim 4 , wherein the bioluminescent reagent comprises luciferase and/or luciferin. 
     
     
         6 . The kit of  claim 5 , wherein the luciferase is a recombinant thermostable luciferase. 
     
     
         7 . The kit of  claim 1 , which further comprises at least two amplification primers. 
     
     
         8 . The kit of  claim 1 , wherein one of the RNA-containing probes comprises an oligonucleotide comprising a DNA polymer able to function as a primer in a nucleic acid amplification reaction, linked to an RNA-containing probe region by way of a blocking group which prevents an extension reaction amplifying the probe region of the oligonucleotide. 
     
     
         9 . An RNA-containing probe which includes a modified form of adenosine. 
     
     
         10 . An RNA-containing probe which comprises an RNA/DNA heteropolymer which contains at least one adenosine or a modified form of adenosine. 
     
     
         11 . The RNA-containing probe of  claim 10 , which comprises a DNA polymer wherein one or more deoxy-adenosine nucleotides are replaced by adenosine nucleotides or modified adenosine nucleotides. 
     
     
         12 . An oligonucleotide comprising a DNA polymer able to function as a primer in a nucleic acid amplification reaction, linked to an RNA-containing probe region by way of a blocking group which prevents an extension reaction amplifying the probe region of the oligonucleotide.