Detection and quantification of biomolecules using mass spectrometry
AssigneeSequenom, Inc.
InventorsDirk Johannes VAN DEN BOOM, Paul Andrew Oeth, Payam Mahboubi
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Abstract
The present invention is directed in part to a method for detecting a target nucleic acid using detector oligonucleotides detectable by mass spectrometry. This method takes advantage of the 5′ to 3′ nuclease activity of a nucleic acid polymerase to cleave annealed oligonucleotide probes from hybridized duplexes and releases labels for detection by mass spectrometry. This process is easily incorporated into a polymerase chain reaction (PCR) amplification assay. The method also includes embodiments directed to quantitative analysis of target nucleic acids.
Claims30
A method of detecting the presence or absence of a target nucleic acid, comprising:
- (a) annealing an oligonucleotide primer and a detector nucleotide to the target nucleic acid;
- (b) introducing an enzyme that extends the oligonucleotide primer in the direction of the detector oligonucleotide, wherein the enzyme cleaves and thereby releases at least a portion of the annealed detector oligonucleotide;
- and
- (c) detecting the presence or absence of the released detector oligonucleotide or portion thereof, thereby detecting the presence or absence of the target nucleic acid.
The method of 1, wherein more than one detector oligonucleotide is used to detect more than one target nucleic acid in a multiplexed reaction.
The method of 1, wherein the target nucleic acid comprises a mixture of nucleic acid.
Dependencies are inferred from the claim text.