摘要
目的建立食品中沙门氏菌、单核细胞增生李斯特菌和蜡样芽胞杆菌多重重组酶聚合酶扩增(recombinase polymerase amplification,RPA)快速检测方法。方法选择沙门氏菌invA基因、单增李斯特菌hlyA基因和蜡样芽孢杆菌16S RNA序列为目标基因进行扩增,建立并优化多重RPA扩增体系和扩增条件;评价反应体系的特异性和灵敏度,并对人工污染食品样品和实际样品进行检测。结果多重RPA反应体系能够在20min完成3种目标基因的扩增,特异性强;对沙门氏菌、单增李斯特菌和蜡样芽孢杆菌的灵敏度分别为2.70×10^(5)、1.30×10^(5)、1.44×10^(4)CFU/mL,能够用于人工污染样品和实际样品的检测。结论本研究建立的多重RPA等温扩增方法特异性强,操作快速、简单,可为食源性致病菌的快速检测提供新方向。
Objective To establish a rapid detection method for Salmonella,Listeria monocytogenes and Bacillus cereus by multiple recombinase polymerase amplification(RPA).Methods Selecting Salmonella invA gene,Listeria monocytogenes hlyA gene and Bacillus cereus 16 S RNA sequence as target genes for amplification,and establishing and optimizing a multiple RPA amplification system and amplification conditions;the specificities and sensitivities of the reaction systems were evaluated and artificially contaminated food samples and actual samples were tested.Results The multiple RPA reaction system was able to complete the amplification of the 3 kinds of target genes in 20 min,with strong specificity;the sensitivities to Salmonella,Listeria monocytogenes and Bacillus cereus were 2.70×10^(5),1.30×10^(5),1.44×10^(4) CFU/mL,respectively,and could be used for detecting artificially contaminated samples and actual samples.Conclusion The multiple RPA isothermal amplification method established in this study is specific,rapid and simple,and can provide a new direction for rapid detection of foodborne pathogenic bacteria.
作者
王凤娇
秦超
廖倩
林佳艺
阮佳
WANG Feng-Jiao;QIN Chao;LIAO Qian;LIN Jia-Yi;RUAN Jia(Department of Public Health,Chengdu Medical College,Chengdu 610500,China;Chengdu Xindu District Center for Disease Control and Prevention,Chengdu 610500,China;Chengdu Wuhou District Center for Disease Control and Prevention,Chengdu 610041,China)
出处
《食品安全质量检测学报》
CAS
北大核心
2021年第20期8121-8127,共7页
Journal of Food Safety and Quality
基金
成都医学院科研基金自科项目(CYZ17-19)
四川省科技创新苗子工程资助项目(2019047)。
