Patent ID: 6248526
Filing Date: 2001-06-19
Classification: C12Q,Y10S

Abstract:
A process for the detection of a target nucleic acid in a sample comprising single-stranded DNA, said process comprising:(a) contacting said sample with a forward and/or reverse primer, wherein at least one primer carries a label or part of a label system in a 3' terminal portion of said primer which is deliberately mismatched relative to the target nucleic acid, said mismatched portion amounting to at least one nucleotide, under conditions sufficient to allow hybridization of the complementary portions of said forward and/or reverse primer and said target nucleic acid;(b) subsequently exposing said sample to nucleic acid polymerase having a 3' to 5' proofreading or functionally equivalent nuclease activity, or a mixture of enzymes having such proofreading activity, under conditions sufficient to permit said 3' to 5' proofreading activity to cleave said forward and/or reverse primer in said 3' mismatched portion, thereby releasing said label or part of a label system;(c) detecting and/or measuring the release of said label or part of a label system.