SEQUENCE DETECTION ASSAY

摘要

The invention concerns a method for detecting a target sequence within a variable target region of a nucleic acid molecule; said method comprising: a)providing a sample containing or suspected of containing said nucleic acid molecule; b)contacting said sample with an oligonucleotide probe complementary to the target region that includes said target sequence, in the presence of a label that emits a signal which is different when the probe is bound to said target region as compared to when the probe is in an unbound state, and further said probe comprises at least one abasic nucleotide substitution which is positioned within said probe so that it aligns with a mutagenic/polymorphic site within the target region outside said target sequence when said probe binds to said target region, but wherein the temperature at which the probe hybridizes to the target region or dissociates from the target region is substantially unaffected by the nature of the nucleotide at said mutagenic/polymorphic site in the target region; c)monitoring said signal whilst changing the temperature of the sample and determining the temperature at which the signal changes as a result of either binding of the probe to the target region or destabilization of a duplex formed by the binding of a probe to the target region; and d)relating the temperature determined to the presence or absence of said target sequence within said target region. The invention also concerns probes and kits for use in the method.