Patent ID: 8329405
Filing Date: 2012-12-11
Classification: C12Q

Abstract:
1. A method for detecting a presence or an absence of a mutant DNA sequence in a sample, the method comprising the steps of: a) contacting the sample with an oligonucleotide system under hybridization conditions so as to form a reaction mixture, said oligonucleotide system including a reverse primer and a mutagenic primer, wherein i) said reverse primer comprises: ii) said mutagenic primer comprises: b) adding with appropriate substrates and cofactors, at a suitable ionic and pH environment, both a temperature resistant DNA polymerase and said high temperature resistant restriction endonuclease to said reaction mixture under predetermined reaction conditions, such that, if the mutant DNA sequence is present in the sample, said reverse primer and said mutagenic primer hybridize to the same and prime the DNA polymerase reaction to obtain a first specific amplified product; c) cycling the hybridization of said oligonucleotide system so that a second specific amplified product is extended comprising the 5′ end region of the reverse primer forming a double stranded recognition site for said restriction endonuclease so that the high temperature resistant restriction endonuclease cuts specifically at the recognition sequence and induces the generation of a signal by the coupled detection system; and d) detecting the generated signal.