Chimeric primers with hairpin conformations and methods of using same

Patent metadata

Field	Value
Publication number	US-12473586-B2
Application number	US-202318394469-A
Country	US
Kind code	B2
Filing date	Dec 22, 2023
Priority date	Jun 29, 2009
Publication date	Nov 18, 2025
Grant date	Nov 18, 2025

Abstract

Methods and compositions for nucleic acid amplification, detection, and genotyping techniques are disclosed. In one embodiment, a nucleic acid molecule having a target-specific primer sequence; an anti-tag sequence 5′ of the target-specific primer sequence; a tag sequence 5′ of the anti-tag sequence; and a blocker between the anti-tag sequence and the tag sequence is disclosed. Compositions containing such a nucleic acid molecule and methods of using such a nucleic acid molecule are also disclosed.

First claim

The invention claimed is:

1 . A method for amplifying a target nucleic acid comprising:
(a) providing a first primer pair comprising:
(i) a first primer comprising:
a first target-specific primer sequence;
a universal anti-tag sequence 5′ of the target-specific primer sequence;
a universal tag sequence 5′ of the universal anti-tag sequence;
a blocker between the universal anti-tag sequence and the universal tag sequence; and
a chromophore attached to the universal tag sequence; and

(ii) a second primer comprising:
a second target-specific primer sequence;
a universal anti-tag sequence 5′ of the target-specific primer sequence;
a universal tag sequence 5′ of the universal anti-tag sequence;
a blocker between the universal anti-tag sequence and the universal tag sequence; and
a chromophore attached to the universal tag sequence; and

(b) a label nucleic acid molecule comprising:
(i) a universal anti-tag sequence complementary to the universal tag sequences of the first primer pair; and
(ii) a chromophore capable of Forster Resonance Energy Transfer with the chromophores of the first primer pair; and
(d) amplifying the target nucleic by combining the first primer pair, the label nucleic acid molecule, and a sample comprising the target nucleic acid under conditions suitable for amplification of the target nucleic acid.

2 . The method of claim 1 , wherein the amplification of the target nucleic acid is catalyzed by a polymerase that has strand displacement activity but does not have exonuclease activity.

3 . The method of claim 1 , further comprising detecting the amplified nucleic acid.

4 . The method of claim 3 , wherein the detection is in real-time.

5 . The method of claim 1 , wherein the sample comprises at least a second target nucleic acid.

6 . The method of claim 1 , wherein the target nucleic acid is an amplicon.

Assignees

• Luminex Corp

Inventors

• Whitman Douglas F
• Zhang Hongwei

Classifications

• C12Q1/6813

  Hybridisation assays · CPC title

• C12Q1/68

  involving nucleic acids · CPC title

• C12Q1/6834

  Enzymatic or biochemical coupling of nucleic acids to a solid phase · CPC title

• C12Q1/6823

  Release of bound markers · CPC title

• C12Q1/6816

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title