Abstract: Compositions for detecting flavivirus nucleic acids. Particularly described are compositions for detecting West Nile virus nucleic acids in the 3? non-coding region. These compositions are preferably oligonucleotides comprising nucleotide sequences that are substantially complementary to a West Nile virus target nucleic acid. These compositions preferably comprise a detectable moiety.
Abstract: There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, “on device” assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.
Abstract: The present invention provides systems, devices, methods, kits, and compositions for nucleic acid analysis using digital PCR. In particular, methods are provided to analyze high titer samples that cannot be divided into a sufficient number of partitions containing zero nucleic acid molecules per partition to allow for Poisson analysis (digital PCR analysis).
Abstract: In some aspects, the present disclosure provides methods for identifying sequence variants in a nucleic acid sample. In some embodiments, a method comprises identifying sequence differences between sequencing reads and a reference sequence, and calling a sequence difference that occurs in at least two different circular polynucleotides, such as two circular polynucleotides having different junctions, as the sequence variant. In some aspects, the present disclosure provides compositions and systems useful in the described methods.
Abstract: The present invention provides an improved method and device for preparing, extracting, separating and/or purifying a biomolecule, for example, nucleic acid from a biological sample. The device comprises a heat-deformable material, such that an inner chamber adopts a second configuration having a chamber volume less than the chamber volume of a first configuration, thereby expelling at least a part of a processed sample through a second opening from the device, and thereby recovering the biomolecule-containing composition.
Abstract: The invention generally relates to methods for increasing the amount of DNA available for analysis when using partitioned samples and parallel processing. For example, double-stranded DNA can be dissociated into two single-stranded components, and the single strands partitioned into different droplets prior to analysis. The disclosed methods are useful for performing digital PCR analysis on samples where the target DNA is not in abundance, for example when the sample originates from a body fluid or an FFPE sample.
Abstract: A fluorescence detection apparatus for analyzing samples located in a plurality of wells in a thermal cycler and methods of use are provided. In one embodiment, the apparatus includes a support structure attachable to the thermal cycler and a detection module movably mountable on the support structure. The detection module includes one or more channels, each having an excitation light generator and an emission light detector both disposed within the detection module. When the support structure is attached to the thermal cycler and the detection module is mounted on the support structure, the detection module is movable so as to be positioned in optical communication with different ones of the plurality of wells. The detection module is removable from the support structure to allow easy replacement.
Type:
Grant
Filed:
December 28, 2018
Date of Patent:
July 28, 2020
Assignee:
Bio-Rad Laboratories, Inc.
Inventors:
Igor Kordunsky, Jeffrey A. Goldman, Michael J. Finney
Abstract: The present invention relates to probes and primers beneficial for conducting amplification assays, such as those including loop-mediated isothermal amplification reactions. Also described herein are methods for detecting targets using such probes and/or primers.
Type:
Grant
Filed:
January 27, 2016
Date of Patent:
July 28, 2020
Assignee:
National Technology & Engineering Solutions of Sandia, LLC
Inventors:
Robert Meagher, Chung-Yan Koh, Yooli Kim Light, Cameron Scott Ball
Abstract: This invention provides methods of using labeled primers or probes for nucleic acid target detection and to detect the identity or presence of a nucleotide at certain positions in nucleic acid sequences with single molecule sensitivity using nanopore detection, and sets of oligonucleotide primers for use in such methods, as well as methods of quantitative PCR coupled with nanopore detection.
Type:
Grant
Filed:
February 12, 2015
Date of Patent:
July 21, 2020
Assignee:
The Trustees of Columbia University in the City of New York
Inventors:
Chuanjuan Tao, Shiv Kumar, Minchen Chien, Jingyue Ju
Abstract: Disclosed are methods for identifying the presence or absence of a target nucleic acid from one or more organisms in a biological sample, said method comprising: (a) spinning the sample at a rotational velocity sufficient to pellet cellular debris and fluorescence inhibitors present within the sample and reduce fluorescence interference or quenching in the sample; and (b) directly amplifying and detecting the target nucleic acid in the sample.
Abstract: A process for detecting Haemophilus influenzae nucleic acid in a sample includes producing an amplification product by amplifying a Haemophilus influenzae nucleotide sequence and measuring the amplification product to detect Haemophilus influenzae in the sample. Some embodiments allow direct serotype determination in a single step assay. Also provided are reagents and methods for detecting and distinguishing Haemophilus influenzae from other infectious agents. A kit is provided for detecting and quantifying Haemophilus influenzae in a sample.
Type:
Grant
Filed:
January 31, 2017
Date of Patent:
June 16, 2020
Assignee:
THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETRARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES
Inventors:
Jennifer Thomas, Xin Wang, Cynthia Hatcher, Raydel Anderson, Mary Jordan Theodore, Leonard W. Mayer
Abstract: A fluorescence detection apparatus for analyzing samples located in a plurality of wells in a thermal cycler and methods of use are provided. In one embodiment, the apparatus includes a support structure attachable to the thermal cycler and a detection module movably mountable on the support structure. The detection module includes one or more channels, each having an excitation light generator and an emission light detector both disposed within the detection module. When the support structure is attached to the thermal cycler and the detection module is mounted on the support structure, the detection module is movable so as to be positioned in optical communication with different ones of the plurality of wells. The detection module is removable from the support structure to allow easy replacement.
Type:
Grant
Filed:
September 27, 2017
Date of Patent:
June 2, 2020
Assignee:
Bio-Rad Laboratories, Inc.
Inventors:
Igor Kordunsky, Jeffrey A. Goldman, Michael J. Finney
Abstract: A process for detecting Haemophilus influenzae nucleic acid in a sample includes producing an amplification product by amplifying a Haemophilus influenzae nucleotide sequence and measuring the amplification product to detect Haemophilus influenzae in the sample. Some embodiments allow direct serotype determination in a single step assay. Also provided are reagents and methods for detecting and distinguishing Haemophilus influenzae from other infectious agents. A kit is provided for detecting and quantifying Haemophilus influenzae in a sample.
Type:
Grant
Filed:
January 31, 2017
Date of Patent:
June 2, 2020
Assignee:
THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES
Inventors:
Jennifer Thomas, Xin Wang, Cynthia Hatcher, Raydel Anderson, Mary Jordan Theodore, Leonard W. Mayer
Abstract: Provided herein are biological assay mixtures for use in a real-time PCR and High Resolution Melt Analysis (HRMA) comprising: a universal Gram-positive forward and reverse primer pair; a universal Gram-negative forward and reverse primer pair; at least two biological probes, each one comprising a fluorophore, able to detect an amplicon prepared by the universal primer pairs; and an oligonucleotide intercalating dye for the HRMA. Methods for detecting a plurality of bacterial strains in the assay, comprise: contacting a biological sample with the biological assay mixture; preparing a plurality of amplicons for both Gram-positive and Gram-negative bacteria using one of the universal primer pairs; and detecting the amplicons using a plurality of biological probes comprising a fluorophore of one or more color emitters able to hybridize to known bacterial strains, wherein if no bacterial strain is identified, then determining whether Gram-positive and Gram-negative strains exist by using HRMA.
Abstract: The present invention relates to a kit and a method of linear amplification of a least one nucleic acid target in a sample, said method comprising: (a) contacting each target in the sample with a nucleic acid polymerase and a primer comprising a component preventing copying of the primer by the nucleic acid polymerase; and at least one nuclease blocking nucleotide; (b) generating a primer extension product; (c) preventing priming by the 3?-end of the primer extension product, and (d) repeating steps b) and c) at least once.
Abstract: Methods for the rapid detection of the presence or absence of Zika virus in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers and probes targeting Zika virus and kits are provided that are designed for the detection of Zika virus.
Type:
Grant
Filed:
March 10, 2017
Date of Patent:
May 19, 2020
Assignee:
Roche Molecular Systems, Inc.
Inventors:
Slav Dugenny, Ellen H. Fiss, Jeffery Fong, Marintha Heil, Eugene Spier, Jingtao Sun
Abstract: Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.
Type:
Grant
Filed:
December 13, 2017
Date of Patent:
May 12, 2020
Assignee:
EXACT SCIENCES DEVELOPMENT COMPANY, LLC
Inventors:
Hatim T. Allawi, William G. Weisburg, Graham P. Lidgard, Michael W. Kaiser, Abram M. Vaccaro, Gracie Shea
Abstract: The present invention discloses a method for non-invasively detecting EGFR gene mutations in subjects, comprising the following steps: designing primers according to EGFR gene exons; extracting plasma DNAs in subjects; connecting the extracted plasma DNAs with tagging linkers; PCR pre-amplifying the tagging linkers connected plasma DNAs; cyclising the pre-amplified DNAs to obtain cyclised DNAs; PCR amplifying the cyclised DNAs using the designed primers; and high throughput sequencing the PCR amplified product and analyzing the EGFR gene mutations. The present invention also discloses a corresponding kit.
Abstract: There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, “on device” assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.
Abstract: The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.
Type:
Grant
Filed:
February 1, 2018
Date of Patent:
May 5, 2020
Assignee:
Bio-Rad Laboratories, Inc.
Inventors:
Darren Roy Link, Michael Weiner, David Marran, Jonathan M. Rothberg