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US 2019/0078147 A1Published application
Detection of recombinase polymerase amplification using dual-hapten probe
AssigneeAlere San Diego, Inc.
InventorsMichael L. Powell, Frank Ray Bowler, Catherine Jean Greenwood, Olaf Piepenburg, Niall A. Armes
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Abstract
This disclosure relates to methods and compositions for detecting a target nucleic acid sequence using a dual-hapten probe. More specifically, the present disclosure relates to using recombinase polymerase amplification (RPA) and a dual-hapten probe to detect a target nucleic acid sequence. In some cases, the detection is on lateral flow strips.
Claims20
A recombinase polymerase amplification composition comprising;
- a crowding agent;
- an oligonucleotide probe with a dual-hapten leaving group;
- and
- a nuclease enzyme.
1 more element
A device, comprising:
- a lateral flow strip, comprising
- a sample application zone;
- a reagent zone downstream of the sample application zone and in fluid communication therewith, the reagent zone comprising dried RPA reagent composition for amplifying a target nucleic acid, a binding agent specific for an amplified target nucleic acid product and a detection molecule;
- at least one test zone downstream of the reagent zone and in fluid communication therewith, the test zone comprising an immobilized capture molecule specific for the amplified target nucleic acid product;
- and
- a control zone downstream of the test zone.
3 more elements
A method of detecting amplification products, comprising:
- contacting a sample suspected of containing a target nucleic acid of interest with RPA reagents for amplifying the target nucleic acid, an oligonucleotide probe comprising a nucleic acid sequence complementary to the target nucleic acid and a covalently linked dual-hapten leaving group, and a nuclease;
- amplifying the target nucleic acid to produce a target nucleic acid product;
- and
- detecting the target nucleic acid product by detecting free dual-hapten moieties cleaved from the oligonucleotide probe hybridized to the target nucleic acid.
1 more element
Dependencies are inferred from the claim text.