Solid phase recombinase polymerase amplification for human pathogen identification
AssigneesNational Technology & Engineering Solutions of Sandia, LLC, Resonantia Diagnostics, Inc.
InventorsJoshua Podlevsky, Darren W. Branch, Adam Bolotsky, Matthew Jones
Life of this patent
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Abstract
The present invention is directed to methods and devices for identification of pathogens, and more particularly to solid phase recombinase polymerase amplification (SP-RPA) methods and devices for massively multiplexed solid-phase isothermal identification of human pathogens from patient samples. The principal application for multiplexed solid-phase isothermal identification of human pathogens is point-of-care diagnostics.
Claims20
A sample analysis device, comprising:
- (a) a sample and reagent input module;
- (b) a multiplex reaction module in communication with the input module, the multiplex reaction module comprising a plurality of sample handling chambers for performing recombinase polymerase amplification assay, wherein the recombinase polymerase amplification assay comprises at least one forward primer immobilized on a solid support and at least one reverse primer comprising a detectable label;
- wherein the forward primer and the reverse primer are complementary to a target nucleic acid;
- wherein the at least one forward primer is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, and SEQ ID NO:11;
- and
- wherein the at least one reverse primer is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, and SEQ ID NO:12:
- (c) a data analysis module in communication with the multiplex reaction module, wherein the data analysis module analyzes for the presence of the detectable label;
- and
- (d) a waste output module for receiving waste from the data analysis module.
A method of detecting a target nucleic acid in a patient sample, the method comprising the steps of:
- (a) obtaining a sample from a patient, the sample containing nucleic acid;
- (b) amplifying the nucleic acid on a multiplex array using a recombinase polymerase amplification (RPA) assay with at least one forward primer and at least one reverse primer, wherein the forward primer is immobilized on a solid support and the reverse primer contains a detectable label;
- and wherein the forward primer and the reverse primer are complementary to the target nucleic acid;
- wherein the at least one forward primer is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, and SEQ ID NO:11;
- and
- wherein the at least one reverse primer is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, and SEQ ID NO:12;
- and
- (c) detecting for the presence of the detectable label on the target nucleic acid.
Dependencies are inferred from the claim text.