Probe set for identification of nucleotide mutation, and method for identification of nucleotide mutation

摘要

Patent No. 596528 Disclosed is a probe set to be used for identifying by fluorescent real-time PCR the type of the nucleotide at a mutation site of NS3 protease of HCV, said probe set comprising: a detection probe composed of an oligonucleotide having: a nucleotide sequence comprising said mutation site in said target nucleic acid, said mutation site comprising a nucleotide of interest; or a nucleotide sequence complementary to said nucleotide sequence; said oligonucleotide having a fluorescent substance attached to the 5-end and a quencher attached to the 3-end; and a counter probe composed of an oligonucleotide having: a nucleotide sequence comprising said mutation site, said mutation site comprising a nucleotide different from said nucleotide of interest; or a nucleotide sequence complementary to said nucleotide sequence, and wherein said detection probe has a modification introduced such that the melting temperature thereof is not less than 3°C higher than the melting temperature of the counter probe, and wherein the detection probe and counter probe are selected from the group consisting of gcatcttccgggctgc, ggcatcttccgggttg, ggcatcttccggtttgc, ggcatcttccggactgc, ggcatcttccggtctgc, tccgggctgctgtat, tcggggctgctgtatg, tcctggctgctgtgtg, tcaaggctgctgtatgca, tccgggttgctatatgca, tccggactgctatatgca, cgtgtgttggactgtct, gtgtgttgggctgtct, cgtgtgttggtctgtct, ccaggcctgtctcctac, ccaggcctatctcctac, ccaggccctctcctac, tccgggttgctgtgt, tccggtttgctgtatgca, tccggactgctgtgtg, tccggtatgctgtatgca