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  

作　　者：Zeqiang Zhan Shoukui He Yan Cui Jinzeng Yang Xianming Shi 詹泽强;何守魁;崔妍;杨金增;史贤明(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)

机构地区：[1]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 [2]Department of Human Nutrition,Food and Animal Sciences,University of Hawaii at Manoa,Honolulu,HI,USA

出　　处：《Food Quality and Safety》2024年第2期302-311,共10页食品品质与安全研究（英文版）

基　　金：supported by the National Key R&D Program of China(No.2019YFE0119700);the National Natural Science Foundation of China(No.32172316).

摘　　要：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 Salmonella Typhimurium in vitro and food.Results:The optimal volume and temperature for the multiplex RPA-LFD method were determined to be 25μL and 38°C,respectively.The reaction process was completed within 25 min and the results were observed visually.The limits of detection(LODs)were 2.8×10^(2),5.9×10^(2),and 7.6×10^(2) 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 importantly,the developed method exhibited good performance in artificially contaminated chicken samples with the LODs of 2.8×10^(3),5.9×10^(3),and 7.6×10^(3) 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.

关 键 词：SALMONELLA multiplex detection lateral flow dipstick recombinase polymerase amplification meat product 

分 类 号：Q523[生物学—生物化学]