title: Detection of Salmonella spp. using a generic and differential FRET-PCR.
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.
authors.affiliation: Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, P. R. China ; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, P. R. China.
authors.forename: Jilei
authors.initials: J
authors.lastname: Zhang
authors.forename: Lanjing
authors.initials: L
authors.lastname: Wei
authors.forename: Patrick
authors.initials: P
authors.lastname: Kelly
authors.forename: Mark
authors.initials: M
authors.lastname: Freeman
authors.forename: Kirsten
authors.initials: K
authors.lastname: Jaegerson
authors.forename: Jiansen
authors.initials: J
authors.lastname: Gong
authors.forename: Bu
authors.initials: B
authors.lastname: Xu
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Pan
authors.forename: Chuanling
authors.initials: C
authors.lastname: Xu
authors.forename: Chengming
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000592:standards*; D002415:Cats; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004285:Dogs; D005243:Feces / Q000382:microbiology; D031541:Fluorescence Resonance Energy Transfer; D057230:Limit of Detection; D009691:Nucleic Acid Denaturation; D010088:Oxidoreductases / Q000145:classification / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry / Q000382:microbiology; D012475:Salmonella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012481:Salmonella Infections, Animal / Q000382:microbiology; D013552:Swine / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010088:Oxidoreductases; C022727:tetrathionate reductase