Method of detecting nucleic acids containing a genetic variation

摘要

Provided are a composition or a kit for detecting a nucleic acid with genetic variation including a first amplification blocking nucleic acid and a second amplification blocking nucleic acid, and a method of detecting a nucleic acid with genetic variation by using the same. Based on the above, a nucleic acid with genetic variation can be detected with high sensitivity and accuracy.

主权项

1. A method of detecting a nucleic acid with genetic variation, the method comprising:
(a) incubating a sample comprising a target nucleic acid, wherein the target nucleic acid can be a nucleic acid with genetic variation or a wild-type nucleic acid differing from the nucleic acid with genetic variation, with a primer pair; a first amplification blocking nucleic acid comprising a sequence complementary to a sense strand of the wild-type nucleic acid, specific to the wild-type nucleic acid, and having a length in a range of 9 nucleotides to 13 nucleotides; and a second amplification blocking nucleic acid comprising a sequence complementary to an antisense strand of the wild-type nucleic acid, complementary to at least two consecutive nucleotides of the first amplification blocking nucleic acid, specific to the wild-type nucleic acid, and having a length of 11 nucleotides; wherein the first amplification blocking nucleic acid and the second amplification blocking nucleic acid each comprises a Peptide Nucleic Acid (PNA), Locked Nucleic Acid (LNA), Zip Nucleic Acid (ZNA), Bridged Nucleic Acid (BNA, nucleotide analogue, or combination thereof; (b) amplifying the target nucleic acid with a polymerase, thereby producing an amplification product, wherein amplification of the target nucleic acid is inhibited by the first amplification blocking nucleic acid and/or second amplification blocking nucleic acid if the target nucleic acid is the wild-type nucleic acid, but amplification of the target nucleic acid is not inhibited if the target nucleic acid is the nucleic acid with genetic variation; and (c) detecting the amplification product to detect the nucleic acid with genetic variation.