20 results about "Pcr assay" patented technology

The present invention provides compositions and methods for novel digital PCR (dPCR) assay designs (droplet digital or other systems) for the detection and quantification of multiple Hepatitis B virus (HBV) gene targets. The compositions and methods can further include a non-extendible blocker oligonucleotide to reduce nonspecific inter-amplicon extension.

The present application relates to multiplex polymerase chain reaction (PCR) assays, methods and systems in which a target sequence is detected by the use of a mediating probe and a complementary single-chain fluorescent reporter complex, preferably by a combination of at least two colors (color combination methods). The method of the invention differs from prior art methods by using a universal reporter complex that produces a fluorescent signal in the PCR assay. The method provided by the invention is easy to implement, and can efficiently and accurately determine the concentration of a plurality of target sequences (generally more than 10) in a complex sample.

Described herein are primers, probes, kits, compositions, and methods related to detection of Mycoplasma genitalium (MG) and mutations in quinolone resistance determining region (QRDR) of parC and gyrA.

A cleavage-based real-time PCR assay method is provided. In general terms, the assay method includes subjecting a reaction mixture comprising a) PCR reagents for amplifying a nucleic acid target, and b) flap cleavage reagents for performing a flap cleavage assay on the amplified nucleic acid target to two sets of thermocycling conditions. No additional reagents are added to the reaction between said first and second sets of cycles and, in each cycle of the second set of cycles, cleavage of a flap probe is measured.

Method for evaluating a Multiplex Polymerase Chain Reaction (Multiplex PCR) primer and probe set, comprising the steps of: a) generating all potential primers and probes from a pool of candidate sequences; b) initiating a global optimization algorithm to generate primer and probe sets from the generated pool; c) calculating an Intramolecular Score (SI), a Free Primer / Probe Concentration Score (SF), a Dimerization Score (SD), and a Hybridization Efficiency Score (SH) for each primer and probe of the set; d) calculating a Final Oligo Score (SO) for each primer and probe of the set b); e) calculating a Total Set Score (ST) as the sum of all Final Oligo Scores (SO) of each primer and probe of the set; and f) ranking each primer and probe set according to its Total Set Score (ST). There is also describe a computer program product configured for performing said method.

The present invention relates to compositions and methods for detecting group B streptococcus (GBS, Streptococcus agalactiae) and identifying the most common genes responsible for clindamycin resistant GBS, such as ermB, ermTR, ermT, lsaC and lsaE, by multiplex real-time PCR assays.

Disclosed is a recombinant adeno-associated virus (rAAV) vector that is a universal reference standard in quantitative PCR assays used to determine the genome copy titer of rAAV-transgene particles. The rAAV-Universal Standard vector includes multiple nucleotide sequences that are targets for PCR assays to support polymerization such as fragments of commonly used promoters, enhancers, polyadenylation sequences, and reporter genes found in rAAV constructs. Those multiple targets ensure that there is a positive reaction and thus provide a standard, positive control and calibrator to assure proper performance of the quantitative PCR assays without the need to use a separate reference standard for each particular rAAV particle assayed. The rAAV-Universal Standard is produced in multiple AAV serotypes and may encode a functional reporter gene to further provide a universal positive control for analytical assays of rAAV-transgene particles.

The present invention relates to compositions and methods for detecting Acinetobacter calcoaceticus-Acinetobacter baumannii complex species and identifying the most common carbapenemases found in carbapenem-resistant Acinetobacter calcoaceticus baumannii (CRAB), such as blaOXA-23-like, blaOXA-24-like, blaOXA-58-like, and blaNDM-like, by multiplex real-time PCR assays.

A method for identifying a pathogen includes obtaining a biological specimen. The method may include dividing the specimen into at least a first sample and a second sample. The method may include identifying at least one pathogen in the first sample via genetic analysis. The method may include exposing the second sample to a panel of antimicrobial agents at known concentrations based on the at least one pathogen. The method may include performing a polymerase chain reaction (PCR) assay on the second sample after exposure of the second sample to the panel of the antimicrobial agents, wherein the PCR assay includes oligonucleotides that amplify a nucleic acid sequence for one or more of the identified pathogens. The method may include analyzing a result of the PCR assay.

The present disclosure relates primer pairs, probes, kits, and methods of use thereof to detect virulent strains of Shiga toxin-producing Escherichia coli (E. coli) (STEC).

A cleavage-based real-time PCR assay method is provided. In general terms, the assay method includes subjecting a reaction mixture comprising a) PCR reagents for amplifying a nucleic acid target, and b) flap cleavage reagents for performing a flap cleavage assay on the amplified nucleic acid target to two sets of thermocycling conditions. No additional reagents are added to the reaction between said first and second sets of cycles and, in each cycle of the second set of cycles, cleavage of a flap probe is measured.

Disclosed is a recombinant adeno-associated virus (rAAV) vector that is a universal reference standard in quantitative PCR assays used to determine the genome copy titer of rAAV-transgene particles. The rAAV-Universal Standard vector includes multiple nucleotide sequences that are targets for PCR assays to support polymerization such as fragments of commonly used promoters, enhancers, polyadenylation sequences, and reporter genes found in rAAV constructs. Those multiple targets ensure that there is a positive reaction and thus provide a standard, positive control and calibrator to assure proper performance of the quantitative PCR assays without the need to use a separate reference standard for each particular rAAV particle assayed. The rAAV-Universal Standard is produced in multiple AAV serotypes and may encode a functional reporter gene to further provide a universal positive control for analytical assays of rAAV-transgene particles.

Provided are compounds, compositions, kits, systems, devices, and methods for improving an assay such as, for example, a multiplexed PCR assay (e.g., a multiplexed immuno-PCR assay). A solid support (e.g., a bead) may be provided according to some embodiments of the present invention. The solid support may comprise an encoding agent (e.g., a dye), a nucleic acid sequence (e.g., an oligonucleotide and / or primer); and a molecular recognition element (e.g., an antibody). A detection reagent may be provided according to some embodiments of the present invention. The detection reagent may comprise a molecular recognition element (e.g., an antibody) and a nucleic acid tag. In some embodiments, at least a portion of the nucleic acid sequence of the solid support and at least a portion of the nucleic acid target tag of the detection reagent are configured to participate in a nucleic acid amplification process. A solid support and detection reagent may bind to the same target, thereby forming a reagent pair.

Provided herein are a composition, system, and kit for the real-time pentaplex amplification and detection of DNA from at least one of beta-lactamase genes Klebsiella pneumoniae carbapenemase (KPC); New Delhi metallo-beta-lactamase (NDM); cefotaximase-Munich (CTX); cephamycin AmpC beta-lactamase (CMY); and oxacillinase-48 (OXA-48).

The present invention relates to a method of certifying a test animal-derived product sample, the method comprising the steps of:(a) determining a test methylation profile of one or more pre-selected methylation sites within the genomic material obtained from the test animal derived product sample; and(b) comparing the test methylation profile obtained from (a) with a reference methylation profile obtained from a control animal of the same biological taxon of the test animal from which the product sample is derived from, where the control animal is bred under a known distinct type of animal husbandry;wherein a significant similarity in the test methylation profile of (a) compared to the reference methylation profile from the control animal, is indicative of the test animal having been bred under the same distinct type of animal husbandry as the control animal and the test animal-derived product is certified so;wherein a difference in the test methylation profile of (a) compared to the reference methylation profile of the control animal, is indicative of the test animal having been bred under another distinct type of animal husbandry as the control animal; andwherein the pre-selected methylation sites are CpG sites selected from genes or regions of genomic DNA from the control and test animal that show the highest degree of methylation variation during the training of the method; andwherein the test methylation profile has significant similarity to the reference methylation profile when the test methylation profile overlaps with the reference methylation profile that is defined by multiple training samples using Principal Component analysis and / or Multi-Dimensional Scaling; andwherein the methylation profiles are determined using at least one method selected from the group consisting of PCR, methylation-specific PCR, real-time methylation-specific PCR, PCR assay using a methylation DNA-specific binding protein, quantitative PCR, a DNA chip-based assay, pyrosequencing, bisulfate pyrosequencing, Methylated DNA immunoprecipitation-sequencing and combinations thereof; andwherein the test animal is selected from livestock or poultry.

The present invention provides compositions and methods relating to novel digital PCR (dPCR) assay designs (droplet numbers or other systems) for the detection and quantification of various hepatitis B virus (HBV) gene targets. The compositions and methods may further include a non-extensible blocker oligonucleotide for reducing extension between non-specific amplicons.

The present invention relates to compositions and methods to detect Group B Streptococcus (GBS, Streptococcus agalactiae) and to identify the most prevalent genes responsible for clindamycin resistant GBS such as ermB, ermTR, ermT, lsaC and lsaE by a multiplex real-time PCR assay.

A method for identifying a pathogen includes obtaining a biological specimen. The method may include dividing the specimen into at least a first sample and a second sample. The method may include identifying at least one pathogen in the first sample via genetic analysis. The method may include exposing the second sample to a panel of antimicrobial agents at known concentrations based on the at least one pathogen. The method may include performing a polymerase chain reaction (PCR) assay on the second sample after exposure of the second sample to the panel of the antimicrobial agents, wherein the PCR assay includes oligonucleotides that amplify a nucleic acid sequence for one or more of the identified pathogens. The method may include analyzing a result of the PCR assay.

An antibody-antigen capture ELISA-PCR test set for detection of a target protein comprises a first antibody-DNA conjugate having a first antibody which is configured to bind to a target protein; and a first DNA oligo linked to the first antibody; a second antibody-DNA conjugate having a second antibody which is configured to bind to the target protein; and a second DNA oligo linked to the second antibody; and multiple immobilized DNA fragments attached to a designated surface on their attaching ends, wherein the immobilized DNA fragments are complementary to one of the first DNA oligo and the second DNA oligo.

Provided are compounds, compositions, kits, systems, devices, and methods for improving an assay such as, for example, a multiplexed PCR assay (e.g., a multiplexed immuno-PCR assay). A solid support (e.g., a bead) may be provided according to some embodiments of the present invention. The solid support may comprise an encoding agent (e.g., a dye), a nucleic acid sequence (e.g., an oligonucleotide and / or primer); and a molecular recognition element (e.g., an antibody). A detection reagent may be provided according to some embodiments of the present invention. The detection reagent may comprise a molecular recognition element (e.g., an antibody) and a nucleic acid tag. In some embodiments, at least a portion of the nucleic acid sequence of the solid support and at least a portion of the nucleic acid tag of the detection reagent are configured to participate in a nucleic acid amplification process. A solid support and detection reagent may bind to the same target, thereby forming a reagent pair.