摘要
本研究根据杀鲑气单胞菌(Aeromonas salmonicida)的铁载受体(fstA)基因的保守序列设计特异性引物和探针,建立了一种基于重组酶聚合酶扩增(recombinase polymerase amplification,RPA)与侧向流试纸条(lateral flow strips,LFS)相结合的可视化快速检测杀鲑气单胞菌的方法。结果显示:本研究建立的基于RPA-LFS的杀鲑气单胞菌检测方法在37℃孵育25 min即可完成,特异性良好,与其他致病菌无交叉反应;灵敏度实验结果显示,该菌的纯培养物和人工污染的鲤鱼组织中的最低检测限均为单次反应1 CFU(colony forming unit)。本研究建立的基于RPA-LFS的杀鲑气单胞菌检测方法具有特异、灵敏、快速等优点,检测结果可通过肉眼直接观察,操作简单,不依赖昂贵的仪器设备,适用于养殖业中杀鲑气单胞菌的现场检测。
This study established a visualized rapid on-site detection method for Aeromonas salmonicida based on recombinase polymerase amplification(RPA)combined with lateral flow strips(LFS).Specific primers and probes were designed targeting the conserved sequence of fstA gene of A.salmonicida.The results showed that the RPA-LFS method established in this study could finish the detection of A.salmonicida by incubation at 37℃for 25 min with reasonable specificity.No cross-reaction to other pathogenic bacteria was observed.The sensitivity test results showed that the limits of detection with pure culture and artificially spiked carp tissues were both 1 colony.The limitations of detection with pure culture and artificially sharp carp tissues were one colony-forming unit(CFU)per reaction.The RPA-LFS method established in this study had the advantages of specificity,sensitivity,and rapidity.The detection results could be observed with the naked eye directly.The procedure was simple without dependence on sophisticated equipment.The method is suitable for the on-site detection of A.salmonicida in the aquaculture industry.
作者
谷庆花
李铭远
司鑫鑫
高嵩
GU Qing-hua;LI Ming-yuan;SI Xin-xin;GAO Song(School of Pharmacy,Jiangsu Ocean University,Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening,Lianyungang 222005,Jiangsu,China)
出处
《淡水渔业》
CSCD
北大核心
2022年第1期52-57,共6页
Freshwater Fisheries
基金
江苏省研究生科研与实践创新计划(SJCX20-1240)。
