Summary Salmonella enterica subspecies arizonae (subspecies IIIa) is most frequently associated with reptiles but is also a bacterial pathogen of poultry, primarily of young turkeys where it induces septicemia, neurologic signs, and increased mortality. Arizonosis clinical cases in broiler chickens have recently been documented in the United States, driving development of a rapid, molecular-based diagnostic for this subspecies. Subspecies arizonae is a genetically distinct subgroup of Salmonella enterica , primarily diagnosed through culture followed by serotyping or biochemical identification, which are costly in both time and laboratory resources. Real-time/quantitative PCR offers rapid and sensitive detection of Salmonella in laboratory and diagnostic samples; however, no such methodology exists to differentiate subsp. arizonae from other Salmonella . In this study, we designed a qPCR assay for subsp. arizonae. The assay is able to differentiate subsp. arizonae from other Salmonella enterica subspecies, including subsp. diarizonae (IIIb), and other non- Salmonella bacteria. Validation, including 56 different subsp. arizonae serovars, demonstrated 100% sensitivity and 100% specificity. This assay provides a rapid diagnostic option for suspect cases of arizonosis in poultry.

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