Patent ID: 9528157
Filing Date: 2016-12-27
CPC Classification: C12Q

Claim Text:
1. A method for determining the presence or absence of a variant nucleotide(s) in a diagnostic region of a target nucleic acid sequence in a sample, comprising: (a) providing a variant specific first primer and a second primer which are capable of amplifying a product comprising a sequence covering that of the diagnostic region of the target nucleic sequence via an amplification process, wherein the variant specific first primer is identical or substantially identical to a first region, the first region overlaps the 5′ part of the diagnostic region of the target nucleic acid sequence, and the 3′ end of the first region overlaps the variant nucleotide(s), and in which the variant specific first primer comprises a 3′ terminus nucleotide complimentary to the variant nucleotide(s), and the second primer comprises a sequence based on that of a second region located downstream of the diagnostic region of the target nucleic acid sequence, wherein and/or (b) carrying out an amplification reaction on a reaction mixture comprising the target nucleic acid using a nucleic acid polymerase, the blocking oligonucleotide probe, optionally a detector probe and the pair of the first and second primers under conditions which are permissive for the amplification process; and (c) detecting the amplified products by a melting profile analysis to determine the presence of the variant nucleotide(s) in the diagnostic region of the target nucleic acid sequence where the variant specific first primer comprises a label or by a change in a detectable signal where the reaction comprises the detector probe.