Alere San Diego, Inc.

Abbott Diagnostics Scarborough, Inc.

Twistdx, Inc.

Asm Scientific, Inc.

Olaf Piepenburg

Niall A Armes

Colin H Williams

Derek L Stemple

Catherine Jean Greenwood

Oliver Nentwich

Mathew James David Parker

Justin Davidson

Karthikeyan Kumaravadivelu

Frank Ray Bowler

Matthew James David Parker

Alexander Schenk Zu Schweinsberg

Grzegorz Artur Orlowski

Hazel Lucy Greetham

Maulik Vinod Patel

Cheng Zhou

Austin Derfus

Austin Matthew Derfus

Murray John Whyte

Niall Antony Armes

Mathew James Parker

This invention relates to sequence specific electrochemically-labeled oligonucleotide probes for the detection of nucleic acids and methods associated therewith.

Redox labeled oligonucleotide probes and their use

The present invention features novel, diverse, hybrid and engineered recombinase enzymes, and the utility of such proteins with associated recombination factors for carrying out DNA amplification assays. The present invention also features different recombinase 'systems' having distinct biochemical activities in DNA amplification assays, and differing requirements for loading factors, single-stranded DNA binding proteins (SSBs), and the quantity of crowding agent employed.

Recombinase polymerase amplification

A flu assay system including a sample module, a microfluidic nucleic acid amplification device, and an analyzer to facilitate fully automated nested recombinase polymerase amplification (RPA) on a sample delivered to the nucleic acid amplification device via the sample module. The assay includes providing a sample to a microfluidic device, and amplifying a target polynucleotide sequence in the sample. Amplifying the target polynucleotide sequence includes performing a first round of amplification on the sample to yield a first amplification product, and performing a second round of amplification on the first amplification product to yield a second amplification product. The second amplification product includes a smaller sequence completely contained within the first amplification product produced during the first round of amplification.

Automated nested recombinase polymerase amplification

A microfluidic device includes an inlet port configured to receive a sample, a first reaction chamber fluidically coupled to the inlet port, a first pump fluidically coupled to the inlet port, a second pump fluidically coupled to a mixing chamber, a metering channel fluidically coupled to the first reaction chamber and to the mixing chamber, and one or more second reaction chambers fluidically coupled to the mixing chamber. The first pump is configured to move fluid from the inlet port to the first reaction chamber and from the first pump to the inlet port. The second pump is configured to move fluid from the second pump to the mixing chamber, from the first reaction chamber to the mixing chamber, and from the mixing chamber to the one or more second reaction chambers.

Microfluidic devices and related methods

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.

Methods for multiplexing recombinase polymerase amplification

This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third 'specificity' probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.

This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes. Further, the improved processivity of the disclosed methods may allow amplification of DNA up to hundres of megabases in length.

Growing community of inventors

Barnet, United Kingdom

Olaf Piepenburg