Patent ID: 8323929

Claim:
A method for detecting the presence or absence of at least one single nucleotide variation in at least one of target nucleic acids in a sample, comprising: a) obtaining a sample comprising target nucleic acids, wherein at least one of the target nucleic acids is suspected to contain at least one single nucleotide variation; b) selecting at least one signal primer comprising a 5′ portion and a 3′ portion, wherein said 3′ portion of the at least one signal primer comprises at least one target binding sequence that specifically hybridizes to the at least one single nucleotide variation of 3′ portion of said at least one target nucleic acid; c) selecting hybridization conditions such that the at least one signal primer hybridizes to its corresponding complementary sequence in the at least one of target nucleic acids that is suspected to contain the at least one single nucleotide variation, wherein said 5′ portion of the at least one signal primer does not hybridize to a sequence in the at least one of target nucleic acids and a complement of at least part of said 5′ portion of the at least one signal primer hybridizes to a 3′ end of a reporter probe; d) mixing said at least one signal primer with the target nucleic acids and at least one amplification primer under the selected hybridization conditions so that said at least one amplification primer hybridizes to a region of said at least one of target nucleic acids that is upstream of a region of said at least one target nucleic acid hybridized to said 3′ portion of said at least one signal primer, and wherein said at least one amplification primer and said at least one signal primer are extended under the selected hybridization conditions if the at least one signal primer does not form a mismatch with the 3′ portion of said at least one of target nucleic acids; e) hybridizing a nucleic acid sequence to an extended portion of said at least one signal primer and synthesizing a complement of said at least one signal primer; f) hybridizing said complement of said at least one signal primer to at least one reporter probe comprising a label, wherein said at least one reporter probe does not hybridize to the at least one of the target nucleic acids but hybridizes to at least a portion of 5′ portion of said complement of said at least one signal primer, and g) detecting a hybridized complex formed by said complement of said at least one signal primer and said at least one reporter probe, wherein the presence of the label of said at least one reporter probe on said hybridized complex indicates that the at least one single nucleotide variation is present in the at least one of target nucleic acids in said sample.