食品与机械2025，Vol.41Issue(6)：44-50,7.DOI:10.13652/j.spjx.1003.5788.2025.80343
基于RPA辅助增强荧光偏振的沙门氏菌检测
Salmonella detection based on fluorescence polarization enhanced by recombinase polymerase amplification
摘要
Abstract
[Objective]To develop a novel fluorescence polarization(FP)detection strategy and achieve highly sensitive Salmonella detection by incorporating recombinase polymerase amplification(RPA)technology.[Methods]The detection mechanism relies on homologous sequence recognition between target DNA and specific primers:When Salmonella targets exist,primer and target DNA binding triggers strand displacement and initiates DNA amplification.To enhance sensitivity,two key design features are implemented:5'-end modification of primers with 6-carboxyfluorescein(6-FAM)as a fluorescent reporter,and selective digestion of unbound primers by exonuclease Ⅰ(Exo Ⅰ).[Results]During RPA amplification,the substantially increased molecular weight of amplicons restricts the rotational freedom of 6-FAM-labeled DNA compounds,resulting in significantly enhanced FP signals.After optimization,this method achieves a detection limit of 11 CFU/mL for Salmonella with excellent specificity.[Conclusion]This method provides an efficient Salmonella detection.Its modular design principle can also be readily adapted for rapid diagnosis of other foodborne pathogens.关键词
沙门氏菌/重组酶聚合酶扩增(RPA)/引物修饰/荧光偏振/定量检测Key words
Salmonella/recombinase polymerase amplification(RPA)/primer modification/fluorescence polarization/quantitative detection引用本文复制引用
薛鹏鹏,陈伟,徐建国,涂佳,屈玮..基于RPA辅助增强荧光偏振的沙门氏菌检测[J].食品与机械,2025,41(6):44-50,7.基金项目
国家重点研发项目(编号:2024YFF0618100) （编号:2024YFF0618100）
安徽省自然科学基金项目(编号:2308085MB57) （编号:2308085MB57）
上海市科委项目(编号:22N31900500) （编号:22N31900500）
