Probe including false-positive-suppressing function, method for designing the same, and method for utilizing the same

Patent metadata

Field	Value
Publication number	US-2022177966-A1
Application number	US-202217592525-A
Country	US
Kind code	A1
Filing date	Feb 4, 2022
Priority date	Feb 22, 2017
Publication date	Jun 9, 2022
Grant date	—

Abstract

An object of the present invention is to provide a means for detecting or quantitatively determining short-chain nucleic acids by simple double-strand formation with high specificity.The present invention relates to a polynucleobase probe including, in a sequence complementary to a target sequence having at least one sequence of any one of SEQ ID NOs: 1 to 10, a sequence in which at least one of bases in a portion complementary to any one sequence of SEQ ID NOs: 1 to 10 in the target sequence becomes abasic and/or is substituted; a method for designing the same; and a method for utilizing the same.

First claim

1 . A method for detecting or quantitatively determining a target nucleic acid having at least one sequence of any one of SEQ ID NOs: 1 to 10 in a test sample with high specificity,
the method comprising: preparing the test sample to detect the target nucleic acid; bringing at least one kind of polynucleobase probe in contact with the test sample, the polynucleobase probe comprising: in a sequence complementary to a target sequence having at least one sequence of any one of SEQ ID NOs: 1 to 10, a sequence in which at least one of bases in a portion complementary to any one sequence of SEQ ID NOs: 1 to 10 in the target sequence becomes abasic and/or is substituted; and/or a sequence which is cleaved to have, on an end, at least one sequence complementary to a sequence of 2 bases or less in anyone sequence of SEQ ID NOs: 1 to 10 in the target sequence; and detecting or quantitatively determining the target nucleic acid bound to the polynucleobase probe.

2 . The method according to claim 1 , wherein the polynucleobase probe is 15- to 28-mer.

3 . The method according to claim 1 , wherein a label is bound to the polynucleobase probe.

4 . The method according to claim 1 , wherein in the polynucleobase probe, in the portion complementary to any one sequence of SEQ ID NOs: 1 to 10 in the target sequence, at least one of the bases which become abasic or are substituted is located inside the portion complementary to any one sequence of SEQ ID NOs: 1 to 10 in the target sequence.

5 . The method according to claim 1 , wherein in the polynucleobase probe, in the sequence complementary to any one sequence of SEQ ID NOs: I to 10 in the target sequence, a ratio of at least one of the bases which become abasic or are substituted with respect to any one of 3 to 5 guanines and cytosines is 1.

6 . The method according to claim 1 , wherein the target nucleic acid is a miRNA.

7 . The method according to claim 1 , which the polynucleobase probe is a DNA, RNA, LNA, GNA, BNA, or PNA.

Assignees

• Yokowo Seisakusho Kk

Inventors

• Niijima Yui
• Takase Shintaro
• Suzuki Hisashi
• Ikeda Hisafumi

Classifications

• C12Q1/6876 Primary

  Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes · CPC title

• C12N15/11

  DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} · CPC title

• C12Q2600/178

  miRNA, siRNA or ncRNA · CPC title