Abstract
To facilitate the detection of Salmonella and to be able to rapidly and conveniently determine the species/subspecies present, we developed and tested a generic and differential FRET-PCR targeting their tetrathionate reductase response regulator gene. The differential pan-Salmonella FRET-PCR we developed successfully detected seven plasmids that contained partial sequences of S. bongori and the six S. enterica subspecies. The detection limit varied from ∼5 copies of target gene/per PCR reaction for S. enterica enterica to ∼200 for S. bongori. Melting curve analysis demonstrated a T m of ∼68°C for S. enterica enterica, ∼62.5°C for S. enterica houtenae and S. enterica diarizonae, ∼57°C for S. enterica indica, and ∼54°C for S. bongori, S. enterica salamae and S. enterica arizonae. The differential pan-Salmonella FRET-PCR also detected and determined the subspecies of 4 reference strains and 47 Salmonella isolated from clinically ill birds or pigs. Finally, we found it could directly detect and differentiate Salmonella in feline (5/50 positive; 10%; one S. enterica salamae and 4 S. enterica enterica) and canine feces (15/114 positive; 13.2%; all S. enterica enterica). The differential pan-Salmonella FRET-PCR failed to react with 96 non-Salmonella bacterial strains. Our experiments show the differential pan-Salmonella FRET-PCR we developed is a rapid, sensitive and specific method to detect and differentiate Salmonella.
Highlights
Salmonella are important zoonotic pathogens which affect the health of people and animals worldwide
The S. enterica enterica serotypes are primarily associated with warmblooded animals while S. bongori and the remaining five S. enterica subspecies are mainly found in cold-blooded animals and the environment [3]
While most human and animal infections are associated with S. enterica enterica, the growing popularity of reptiles as pets has led to an increasing number of infections with S. bongori and S. enterica subspecies other than S. enterica enterica [2,4,5,6]
Summary
Salmonella are important zoonotic pathogens which affect the health of people and animals worldwide. The genus Salmonella consists of only 2 species, S. bongori and S. enterica, with the latter containing 6 subspecies: enterica (I), salamae (II), arizonae (IIIa), diarizonae (IIIb), houtenae (IV), and indica (VI) [1,2]. While most human and animal infections are associated with S. enterica enterica, the growing popularity of reptiles as pets has led to an increasing number of infections with S. bongori and S. enterica subspecies other than S. enterica enterica [2,4,5,6]. A retrospective analysis of over 75,000 isolates collected between 1985–2009 showed that S. enterica subspecies salamae, arizonae, diarizonae and houtenae caused invasive disease in people significantly more frequently than did S. enterica enterica [2]
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