US 2025/0043335 A1 · METHOD FOR DETECTING MUTATION IN TARGET BASE SEQUENCE OF NUCLEIC ACID, METHOD FOR SELECTIVELY INHIBITING AMPLIFICATION OF NUCLEIC ACID, AND KITS FOR PERFORMING SAME

Abstract

One purpose of the present invention is to provide a simple method by which a mutant gene can be detected at a lower mutant gene content than the mutant gene detection limit by the ordinary clamp-PCR method, and a kit for performing the method. Another purpose of the present invention is to provide a simple method by which, in a nucleic acid amplification reaction, the amplification of a target base sequence can be selectively inhibited, and a kit for performing the method. In a nucleic acid amplification reaction, a diallylamine/sulfur dioxide copolymer or an acid addition salt thereof comprising a constituent unit (A) having a structure represented by general formula (I-a) or (I-b): wherein R1 represents an alkyl group having 1-10 carbon atoms that may have a hydroxyl group, a cycloalkyl group having 5-10 carbon atoms or an aralkyl group having 7-10 carbon atoms, and a constituent unit (B) having a structure represented by general formula (I-c): is used as an agent of enhancing nucleic acid amplification inhibition by a clamp nucleic acid.

Claims22

Dependent claims

A method for detecting mutation in a target base sequence of a nucleic acid in a sample relative to a standard base sequence, comprising the steps of:

subjecting the nucleic acid in the sample to a nucleic acid amplification reaction as a template with
a clamp nucleic acid having a base sequence complementary to the standard base sequence and containing at least one artificial nucleotide residue, and
a diallylamine/sulfur dioxide copolymer that comprises
a constituent unit (A) having a structure represented by general formula (I-a) or general formula (I-b), or a structure of an acid addition salt thereof:
wherein R1 represents an alkyl group having 1 to 10 carbon atoms, and
a constituent unit (B) having a structure represented by general formula (I-c):
and
determining the presence of the mutation based on the total amount of nucleic acid amplification products obtained by the nucleic acid amplification reaction, the number of amplification cycles of the nucleic acid amplification reaction required for the total amount of the nucleic acid amplification products to reach a threshold value, the amount of nucleic acids with the mutation in the nucleic acid amplification products or the number of amplification cycles of the nucleic acid amplification reaction required for the amount of nucleic acids with the mutation in the nucleic acid amplification products to reach a threshold value.

5 more elements

Dependencies are inferred from the claim text.

Method for detecting mutation in target base sequence of nucleic acid, method for selectively inhibiting amplification of nucleic acid, and kits for performing same

Life of this patent

Method for detecting mutation in target base sequence of nucleic acid, method for selectively inhibiting amplification of nucleic acid, and kits for performing same

Life of this patent

Abstract

Claims22