Analytical method and kit
AssigneeThe Secretary of State for Defence
InventorsDavid James Squirrell, Martin Alan Lee
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Abstract
Analytical methods using RNA probes for the detection or analysis of nucleic acid sequences is 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.
Claims16
A method for detecting the presence or amount of a target nucleic acid within a sample, said method comprising conducting an amplification reaction in the presence of (a) an RNA probe which is specific for at least a portion of said target nucleic acid;
- (b) an enzyme which hydrolyses RNA when in double stranded form and (c) one or more enzymes and/or reagents necessary to convert adenosine monophosphate produced to adenosine triphosphate;
- adding to the sample bioluminescent reagents which react to the presence of ATP, detecting a signal from said biolumineseent reagents and relating that to the presence or amount of the target nucleic acid sequence.
A method according to any one of the preceding claims wherein bioluminescent reagents comprise luciferin and luciferase.
A method according to any one of the preceding claims wherein the bioluminescent reagents are added throughout the amplification reaction.
A kit for use in method according to any one of the preceding claims analysis comprising at least one RNA probe which is specific for a target sequence, an enzyme which can hydrolyse RNA when in double stranded form, and an amplification primer.
Dependencies are inferred from the claim text.