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US 2016/0097090 A1Published application
Methods for Multiplexing Recombinase Polymerase Amplification
AssigneeAlere San Diego, Inc.
InventorsOlaf Piepenburg, Colin H. Williams, Niall A. Armes
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Abstract
This disclosure provides for methods and reagents for rapid multiplex RPA reactions and improved methods for detection of multiplex RPA reaction products. In addition, the disclosure provides new methods for eliminating carryover contamination between RPA processes.
Claims33
-78. (canceled)
A device, comprising,
- a reaction chamber containing dried reagent composition for a preforming a recombinase polymerase amplification (RPA) comprising
- (i) at least one recombinase,
- (ii) at least one polymerase;
- (iii) a first and second nucleic acid primer;
- (iv) a third extension blocked primer which comprises an elongation blocking group at the 3′ end, an internal tetrahydrofuran (THF) residue;
- and
- (v) at least one nuclease;
- and
- a lateral flow strip configured to detect the amplified product.
6 more elements
A method, comprising:
- obtaining a liquid sample from a human being, the liquid sample comprising a target nucleic acid;
- combining the sample with a reagent composition for preforming a recombinase polymerase amplification (RPA) to form a mixture, wherein the reagent composition comprises
- (i) at least one recombinase,
- (ii) at least one polymerase;
- (iii) a first and second nucleic acid primer;
- (iv) a third extension blocked primer which comprises an elongation blocking group at the 3′ end, an internal tetrahydrofuran (THF) residue;
- and
- (v) at least one nuclease;
- performing RPA on the mixture to produce amplicons indicative of the presence of the target nucleic acid;
- and
- detecting the presence of the amplicons produced during the performing step and determining the presence of the target nucleic acid based on the detected amplicons, wherein the presence of the amplicons is detected using a lateral flow strip.
8 more elements
Dependencies are inferred from the claim text.