title: Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene.
abstract: In this study, we developed and validated a loop-mediated isothermal amplification (LAMP) assay for Salmonella detection targeting bcfD gene, a conserved fimbrial operon gene existing in Salmonella. The Salmonella LAMP assay we developed successfully amplified 44 Salmonella strains (14 standard strains and 30 clinical isolates), but none of 9 non-Salmonella standard strains (Proteus mirabilis, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Shigella flexneri, Shigella sonnei, Klebsiella pneumoniae, Campylobacter jejuni and Vibrio parahemolyticus). The detection limit was 5 CFU of Salmonella pure culture or 200 CFU of artificially spiked faeces per reaction system (equivalent to 5000 CFU g(-1) of faeces), and this method could directly detect Salmonella in chicken faeces free of pre-enrichment in a reaction time of 25 min. Our experiments show that the LAMP method we developed is a rapid, sensitive, specific and practical method for Salmonella detection. The Salmonella LAMP assay can potentially serve as new on-site diagnostics in the food and agricultural industries.
  
  
  CONCLUSIONS
  A loop-mediated isothermal amplification (LAMP) assay was established to detect Salmonella targeting bcfD gene, a conserved fimbrial operon gene. The detection limit was 5 CFU of Salmonella pure culture or 200 CFU of artificially spiked faeces per reaction system (equivalent to 5000 CFU g(-1) of faeces), and this method could directly detect Salmonella in chicken faeces free of pre-enrichment in a reaction time of 25 min. The Salmonella LAMP assay is a rapid, sensitive, specific and practical method for Salmonella detection and can potentially serve as new on-site diagnostics in the food and agricultural industries.
authors.affiliation: Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, China.
authors.forename: L
authors.initials: L
authors.lastname: Zhuang
authors.forename: J
authors.initials: J
authors.lastname: Gong
authors.forename: Q
authors.initials: Q
authors.lastname: Li
authors.forename: C
authors.initials: C
authors.lastname: Zhu
authors.forename: Y
authors.initials: Y
authors.lastname: Yu
authors.forename: X
authors.initials: X
authors.lastname: Dou
authors.forename: X
authors.initials: X
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Xu
authors.forename: C
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D002645:Chickens / Q000382:microbiology*; D005243:Feces / Q000382:microbiology; D010861:Fimbriae, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009876:Operon; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000382:microbiology*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D012481:Salmonella Infections, Animal / Q000382:microbiology*; D012477:Salmonella enteritidis / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
keywords: PCR; Salmonella; bcfD gene; detection; loop-mediated isothermal amplification