适体
指数富集配体系统进化
石英晶体微天平
单核细胞增生李斯特菌
沙门氏菌
检出限
大肠杆菌
细菌
化学
微生物学
色谱法
生物
分子生物学
生物化学
核糖核酸
遗传学
基因
吸附
有机化学
作者
Xiaofan Yu,Fang Chen,Ronghui Wang,Yanbin Li
标识
DOI:10.1016/j.jbiotec.2017.12.011
摘要
The rapid detection of foodborne pathogens is critical to ensure food safety. The objective of this study is to select aptamers specifically bound to Escherichia coli O157:H7 using the whole-bacterium SELEX (Systematic Evolution of Ligands by Exponential Enrichment) and apply the selected aptamer to a QCM (quartz crystal microbalance) sensor for rapid and sensitive detection of target bacteria. A total of 19 rounds of selection against live E. coli O157:H7 and 6 rounds of counter selection against a mixture of Staphylococcus aureus, Listeria monocytogenes, and Salmonella Typhimurium, were performed. The aptamer pool from the last round was cloned and sequenced. One sequence S1 that appeared 16 times was characterized and a dissociation constant (Kd) of 10.30 nM was obtained. Subsequently, a QCM aptasensor was developed for the rapid detection of E. coli O157:H7. The limit of detection (LOD) and the detection time of the aptasensor was determined to be 1.46 × 103 CFU/ml and 50 min, respectively. This study demonstrated that the ssDNA aptamer selected by the whole-bacterium SELEX possessed higher sensitivity than previous work and the potential use of the constructed QCM aptasensor in rapid screening of foodborne pathogens.
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