Cheng Dan, Yu Mengqun, Fu Fei, Han Weiye, Li Gan, Xie Jianping, Song Yang, Swihart Mark T, Song Erqun
Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University , Chongqing 400715, People's Republic of China.
College of Life Sciences, Southwest University , Chongqing 400715, People's Republic of China.
Anal Chem. 2016 Jan 5;88(1):820-5. doi: 10.1021/acs.analchem.5b03320. Epub 2015 Dec 19.
Food poisoning and infectious diseases caused by pathogenic bacteria such as Staphylococcus aureus (SA) are serious public health concerns. A method of specific, sensitive, and rapid detection of such bacteria is essential and important. This study presents a strategy that combines aptamer and antibiotic-based dual recognition units with magnetic enrichment and fluorescent detection to achieve specific and sensitive quantification of SA in authentic specimens and in the presence of much higher concentrations of other bacteria. Aptamer-coated magnetic beads (Apt-MB) were employed for specific capture of SA. Vancomycin-stabilized fluorescent gold nanoclusters (AuNCs@Van) were prepared by a simple one-step process and used for sensitive quantification of SA in the range of 32-10(8) cfu/mL with the detection limit of 16 cfu/mL via a fluorescence intensity measurement. And using this strategy, about 70 cfu/mL of SA in complex samples (containing 3 × 10(8) cfu/mL of other different contaminated bacteria) could be successfully detected. In comparison to prior studies, the developed strategy here not only simplifies the preparation procedure of the fluorescent probes (AuNCs@Van) to a great extent but also could sensitively quantify SA in the presence of much higher concentrations of other bacteria directly with good accuracy. Moreover, the aptamer and antibiotic used in this strategy are much less expensive and widely available compared to common-used antibodies, making it cost-effective. This general aptamer- and antibiotic-based dual recognition strategy, combined with magnetic enrichment and fluorescent detection of trace bacteria, shows great potential application in monitoring bacterial food contamination and infectious diseases.
由金黄色葡萄球菌(SA)等致病细菌引起的食物中毒和传染病是严重的公共卫生问题。一种特异性、灵敏且快速检测此类细菌的方法至关重要。本研究提出了一种策略,该策略将适体和基于抗生素的双识别单元与磁性富集和荧光检测相结合,以实现对真实样本中以及存在高得多浓度其他细菌情况下的SA进行特异性和灵敏的定量分析。用适体包被的磁珠(Apt-MB)特异性捕获SA。通过简单的一步法制备了万古霉素稳定的荧光金纳米簇(AuNCs@Van),并通过荧光强度测量用于在32 - 10⁸ cfu/mL范围内对SA进行灵敏定量分析,检测限为16 cfu/mL。使用该策略,可以成功检测复杂样本(含有3×10⁸ cfu/mL其他不同污染细菌)中约70 cfu/mL的SA。与先前的研究相比,此处开发的策略不仅在很大程度上简化了荧光探针(AuNCs@Van)的制备过程,而且能够在存在高得多浓度其他细菌的情况下直接灵敏地对SA进行定量分析,且准确性良好。此外,与常用抗体相比,该策略中使用的适体和抗生素成本低得多且易于获得,具有成本效益。这种基于适体和抗生素的通用双识别策略,结合微量细菌的磁性富集和荧光检测,在监测细菌性食品污染和传染病方面显示出巨大的潜在应用价值。