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.