Detection of <i>Salmonella enterica </i>subspecies <i>enterica </i>serovar <i>Enteritidis </i>in food and environmental samples, methods and compositions therefor

摘要

Embodiments of the disclosure relate to compositions of isolated nucleic acid sequences, methods, workflows and kits of use thereof for detection of Salmonella enterica, subspecies enterica, serovar Enteritidis (S. Enteritidis) 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 detect S. Enteritidis without cross-reacting with S. Paratyphi C or other closely related Salmonella species. In some embodiments, methods and kits of the disclosure may comprise a TaqMan® assay. Following sample enrichment, methods of detection may be completed in approximately 3 hours.

主权项

1. A method for determining the presence of S. Enteritidis in a sample comprising:
combining the sample with a culture medium for enriching Salmonella species for a time sufficient to generate a sample enriched for said species; extracting nucleic acid from at least some of the enriched sample to obtain extracted nucleic acid; contacting the extracted nucleic acid with at least one primer pair selected from a first primer pair of SEQ ID NO:1 and SEQ ID NO:3, a second primer pair of SEQ ID NO:10 and SEQ ID NO: 12, and a third primer pair of SEQ ID NO: 13 and SEQ ID NO: 15, under real-time polymerase chain reaction (PCR) amplification conditions to generate amplified nucleic acid; and detecting at least some of the amplified nucleic acid, thereby determining the presence of S. Enteritidis in the sample, wherein the method does not cross-react with S. Paratyphi C.