Universal nucleic acid probe set and method for utilization thereof

A probe set for detecting a target nucleic acid, comprising two probes:

• (1) a fluorescent probe (A) and a binding probe (B),
• wherein
• the fluorescent probe (A) has a sequence (a) which contains, in a 3′-terminal nucleotide unit thereof, a fluorescent substance (d),
• the binding probe (B) comprises, on a 3′ end portion thereof, a sequence (b1), and on a 5′ end portion thereof, a sequence (b2),
• the sequence (b1) hybridizes to the sequence (a),
• the sequence (b2) hybridizes to a target nucleic acid sequence (C),
• the fluorescent substance (d) is a fluorescent substance that changes in a fluorescent character upon interacting with guanine, and
• at least one of the nucleotide units of the sequence (a) is an artificial nucleotide unit that is configured to raise a dissociation temperature between the sequence (a) and the sequence (b1) as compared to that of where the sequence (a) does not include the artificial nucleotide unit;
• or
• (2) a fluorescent probe (A′) and a binding probe (B′),
• wherein
• the fluorescent probe (A′) has a sequence (a′) which contains, in a 5′-terminal nucleotide unit thereof, a fluorescent substance (d′),
• the binding probe (B′) comprises, on a 5′ end portion thereof, a sequence (b1′), and on a 3′ end portion thereof, a sequence (b2′),
• the sequence (b1′) hybridizes to the sequence (a′),
• the sequence (b2′) hybridizes to a target nucleic acid sequence (C′),
• the fluorescent substance (d′) is a fluorescent substance that changes in a fluorescent character upon interacting with guanine, and
• at least one of the nucleotide units of the sequence (a′) is an artificial nucleotide unit(s) that is configured to raise a dissociation temperature between the sequence (a′) and the sequence (b1′) as compared to that of where the sequence (a′) does not include the artificial nucleotide unit.

A probe set for detecting a target nucleic acid sequence, comprising three probes:

• a fluorescent probe (A1), a fluorescent probe (A2), and a binding probe (B),
• wherein
• the fluorescent probe (A1) has a sequence (a1) which contains, in a 5′-terminal nucleotide unit thereof, a fluorescent substance (d1),
• the fluorescent probe (A2) has a sequence (a2) which contains, in a 3′-terminal nucleotide unit thereof, a fluorescent substance (d2),
• the binding probe (B) comprises, on a 5′ end portion thereof, a sequence (b1-1), on a 3′ end portion thereof, a sequence (b1-2), and between the sequence (b1-1) and the sequence (b1-2), a sequence (b2),
• the sequence (b1-1) hybridizes to the sequence (a1),
• the sequence (b1-2) hybridizes to the sequence (a2),
• the sequence (b2) hybridizes to a target nucleic acid sequence (C),
• each of the fluorescent substances (d1) and (d2) is a fluorescent substance that changes in a fluorescent character upon interacting with guanine,
• at least one of the nucleotide units of the sequence (a1) is an artificial nucleotide unit that is configured to raise a dissociation temperature between the sequence (a1) and the sequence (b1-1) as compared to that of where the sequence (a1) does not include the artificial nucleotide unit, and
• at least one of the nucleotide units of the sequence (a2) is an artificial nucleotide unit that is configured to raise a dissociation temperature between the sequence (a2) and the sequence (b1-2) as compared to that of where the sequence (a2) does not include the artificial nucleotide unit.