首页|Development of a multiplex recombinase polymerase amplification coupled with lateral flow dipsticks for the simultaneous rapid detection of Salmonella spp.,Salmonella Typhimurium and Salmonella Enteritidis
Development of a multiplex recombinase polymerase amplification coupled with lateral flow dipsticks for the simultaneous rapid detection of Salmonella spp.,Salmonella Typhimurium and Salmonella Enteritidis
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Objectives:Salmonella spp.is a world-leading foodborne pathogen and its rapid detection is essential for ensuring food safety.Conventional methods require expensive instruments,considerable operational skills and cannot provide fast mobile on-site systems to detect Salmonella in food.Materials and Methods:A visual method was established based on multiple recombinase polymerase amplification(RPA)coupled with lateral flow dipsticks(LFD)for the simultaneous detection of Salmonella spp.,Salmonella Enteritidis and SalmonellaTyphimurium in vitro and food.Results:The optimal volume and temperature for the multiplex RPA-LFD method were determined to be 25 μL and 38 ℃,respectively.The re-action process was completed within 25 min and the results were observed visually.The limits of detection(LODs)were 2.8×102,5.9×102,and 7.6×102 CFU/mL for Salmonella spp.,S.Enteritidis and S.Typhimurium,respectively.Meanwhile,the results of the established method showed no cross-reactivity between the Salmonella cells and other common foodborne bacteria,which was highly specific for Salmonella.More import-antly,the developed method exhibited good performance in artificially contaminated chicken samples with the LODs of 2.8×103,5.9×103,and 7.6×103 CFU/mL for Salmonella spp.,S.Enteritidis,and S.Typhimurium,respectively.Finally,the application of the multiple RPA-LFD methods in retailed food samples displayed that this method was effective and practical for the detection of Salmonella spp.in food.Conclusion:The developed multiplex RPA-LFD method provides a new sensitive and rapid alternative for the specific detection of Salmonella spp.and its important serovars in food.
Department of Food Science&Technology,MOST-USDA Joint Research Center for Food Safety and NMPA Key Laboratory for Testing Technology of Pharmaceutical Microbiology,School of Agriculture&Biology,and State Key Lab of Microbial Metabolism,Shanghai Jiao Tong University,Shanghai,China
Department of Human Nutrition,Food and Animal Sciences,University of Hawaii at Manoa,Honolulu,HI,USA
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