Life Technologies Corporation

Applied Biosystems, LLC

Applera Corporation

Pe Corporation

Paolo Vatta

Shaheer Khan

Craig Cummings

Olga Petrauskene

Lily Wong

Peter Virgil Fisher

Pius Brzoska

Manohar R Furtado

Robert Tebbs

Priya Balachandran

Patrick Zoder

Melissa Barker

Craig A Cummings

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.

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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.

The invention provides novel dye-labeled ribonucleotide analogs and methods for synthesizing those analogs. The compounds of the invention are especially useful for DNA sequencing by the polymerase chain reaction.

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San Mateo, CA, United States of America

Paolo Vatta