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.
Significance and impact of the study: 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.
Keywords: PCR; Salmonella; bcfD gene; detection; loop-mediated isothermal amplification.
© 2014 The Society for Applied Microbiology.