Abstract: A method of detecting the presence or absence of variant nucleotides within target nucleic acid sequences, said method comprising the use of at least one pair of target-specific amplification primers capable of hybridizing to target nucleic acid sequences, and the use of at least one set of diagnostic primers, each set consisting of at least two types of diagnostic primers capable of hybridizing to corresponding target nucleic acid sequences 3?-relative to said amplification primers, such that each set of diagnostic primers is semi-nested relative to the corresponding pair of amplification primers. Additionally, the use of said method for genotyping, for diagnostic screening, for assaying the cytosine methylation status and for quantification of nucleic acids is disclosed.

Abstract: A method of detecting the presence or absence of variant nucleotides within target nucleic acid sequences, said method comprising the use of at least one pair of target-specific amplification primers capable of hybridizing to target nucleic acid sequences, and the use of at least one set of diagnostic primers, each set consisting of at least two types of diagnostic primers capable of hybridizing to corresponding target nucleic acid sequences 3?-relative to said amplification primers, such that each set of diagnostic primers is semi-nested relative to the corresponding pair of amplification primers. Additionally, the use of said method for genotyping, for diagnostic screening, for assaying the cytosine methylation status and for quantification of nucleic acids is disclosed.