Life Technologies Corporation

Oxoid Limited

Olga Petrauskene

Craig Cummings

Lily Wong

Paolo Vatta

Robert Tebbs

Pius Brzoska

Manohar R Furtado

Priya Balachandran

Jen-Kuei Liu

Patrick Zoder

Melissa Barker

Sueh-Ning Liew

Somaya Bit

Arlene Nunez

Payman Fatemi

Allison De Los Reyes

Michael Brewer

Manohar Furtado

Catherine O'CONNELL

Michael Schumaker

Sharon Matheny

Craig A Cummings

Disclosed is a multi-primer amplification assay, method and kits for detectingspecies and closely related species utilizing a plurality of oligonucleotide primers in contact with a sample in a single vessel and detecting the amplification product, wherein the presence of an amplification product indicatesin the sample.

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The present teachings generally relate to methods and kits incorporating a discriminating positive control for determining whether a particular microorganism or group of microorganisms are present in a sample, for example but not limited to a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample. According to certain methods, at least part of a starting material, for example but not limited to, a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample can be combined with a culture medium and incubated under conditions suitable for microbial growth followed by extracting microorganism and added control nucleic acids for analysis. The extracted nucleic acids are amplified and the amplified nucleic acids are detected, directly or indirectly, and the fidelity of the methods and the presence or absence of the corresponding microorganism is determined because the discriminating positive control provides both confirmatory results for the methods used but eliminates false positive results.

Discriminatory positive/extraction control DNA

Embodiments of the disclosure relate to isolated nucleic acid sequences, methods of use thereof, and workflows for detection of severalspecies in a sample, particularly in a food or environmental sample. Embodiments of the disclosure may also be used to detect one or more species or strains offrom each other, for examplemay be detected independently of otherspp. Some embodiments also describe a duplexed assay that can detect(formerly-),, and. Kits for detection ofare also described. In some embodiments, methods and kits of the disclosure may comprise a TAQMAN&#xae; assay. In some embodiments, 0.2-2 cfu ofspp. are detected using the compositions, methods and kits after a 24-28 hour enrichment period.

Embodiments of the disclosure relate to compositions of isolated nucleic acid sequences, methods, workflows and kits of use thereof for detection of, subspecies, serovar() in a sample, particularly in a food (egg, poultry) or environmental sample (including poultry-related environmental samples). Embodiments of the disclosure may also be used to differentially detectwithout cross-reacting withC or other closely relatedspecies. In some embodiments, methods and kits of the disclosure may comprise a TaqMan&#xae; assay. Following sample enrichment, methods of detection may be completed in approximately 3 hours.

Described are compositions, methods and kits for detection and/or differential detection of serovars ofsubsp.serovar such as S. Heidelberg and S. Hadar in samples. Some embodiments relate to multiplex amplification based molecular assays.

Disclosed are methods and kits for the specific detection ofO157:H7 and notO55:H7 from samples such as: complex food matrices, water, beverages, fermentation broths, forensic & biological samples, and environmental samples including food processing and manufacturing surfaces. In some embodiments, a method of the disclosure comprises: hybridizing at least a first pair of polynucleotide primers to at least a first target polynucleotide sequence, hybridizing at least a second pair of polynucleotide primers to at least a second target polynucleotide sequence, amplifying the at least first and at least second target polynucleotide sequences, and detecting the first and second amplified target polynucleotide sequence products, wherein the detection of both the first amplified target polynucleotide sequence product and the second amplified target polynucleotide sequence product is indicative of the presence ofO157:H7 in a sample and notO55:H7.

Embodiments of the disclosure relate to isolated nucleic acid sequences, methods of use thereof, and workflows for detection of severalspecies in a sample, particularly in a food or environmental sample. Embodiments of the disclosure may also be used to detect one or more species or strains offrom each other, for examplemay be detected independently of otherspp. Some embodiments also describe a duplexed assay that can detect(formerly incertae-sedis),, and. Kits for detection ofare also described. In some embodiments, methods and kits of the disclosure may comprise a TAQMAN&#xae; assay. In some embodiments, 0.2-2 cfu ofspp. are detected using the compositions, methods and kits after a 24-28 hour enrichment period.

Disclosed are primer and probe compositions, PCR assays, methods and kits for the specific detection ofO157:H7 and notO55:H7 from a variety of samples. In some embodiments, methods for specifically detectingO157:H7 comprise: hybridizing at least a first pair of polynucleotide primers to at least a first target polynucleotide sequence, hybridizing at least a second pair of polynucleotide primers to at least a second target polynucleotide sequence, amplifying said at least first and said at least second target polynucleotide sequences, and detecting said at least first and said at least second amplified target polynucleotide sequence products, wherein the detection of both the first amplified target polynucleotide sequence product and the second amplified target polynucleotide sequence product is indicative of the presence ofO157:H7 in a sample and notO55:H7.

Growing community of inventors

San Carlos, CA, United States of America

Olga Petrauskene