Method for detection of single nucleotide variant

A method for detecting single nucleotide variants in a gene, comprising:

• (a) providing a test sample containing a target gene sequence to be detected, wherein the target gene sequence is a wild-type nucleotide sequence, a single nucleotide variant sequence, or a combination thereof;
• (b) adding a specific primer pair to perform a gene amplification reaction to produce an amplified product, wherein the amplified product includes the target gene sequence, with one end of the amplified product has a sequence capable of synthesizing copper nanoclusters and the other end has a biotin modification;
• (c) digesting the amplified product with a restriction enzyme to produce a digested product and an undigested amplified product;
• wherein the restriction enzyme can only digest one of the wild-type nucleotide sequence or the single nucleotide variant sequence;
• wherein the digested product includes a digested fragment with the sequence capable of synthesizing copper nanoclusters and a digested fragment with a biotin modification;
• (d) adding streptavidin-coated magnetic beads to precipitate the undigested amplified product and the biotin-modified digested fragment with a magnet to obtain a supernatant;
• (e) conducting a copper nanocluster synthesis reaction in the supernatant, wherein more double-stranded DNA template copper nanoclusters are produced when the supernatant contains more digested fragments with the sequence capable of synthesizing copper nanoclusters;
• and
• (f) interpreting the results by determining the condition of the target gene sequence based on the amount of double-stranded DNA template copper nanoclusters.