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.
Department of Chemical and Biological Engineering, University at Buffalo, State University of New York , Buffalo, New York 14260, United States.
Anal Chem. 2017 Apr 4;89(7):4085-4090. doi: 10.1021/acs.analchem.6b04958. Epub 2017 Mar 27.
The effective monitoring, identification, and quantification of pathogenic bacteria is essential for addressing serious public health issues. In this study, we present a universal and facile one-step strategy for sensitive and selective detection of pathogenic bacteria using a dual-molecular affinity-based Förster (fluorescence) resonance energy transfer (FRET) platform based on the recognition of bacterial cell walls by antibiotic and aptamer molecules, respectively. As a proof of concept, Vancomycin (Van) and a nucleic acid aptamer were employed in a model dual-recognition scheme for detecting Staphylococcus aureus (Staph. aureus). Within 30 min, by using Van-functionalized gold nanoclusters and aptamer-modified gold nanoparticles as the energy donor and acceptor, respectively, the FRET signal shows a linear variation with the concentration of Staph. aureus in the range from 20 to 10 cfu/mL with a detection limit of 10 cfu/mL. Other nontarget bacteria showed negative results, demonstrating the good specificity of the approach. When employed to assay Staph. aureus in real samples, the dual-recognition FRET strategy showed recoveries from 99.00% to the 109.75% with relative standard derivations (RSDs) less than 4%. This establishes a universal detection platform for sensitive, specific, and simple pathogenic bacteria detection, which could have great impact in the fields of food/public safety monitoring and infectious disease diagnosis.
有效监测、识别和量化致病菌对于解决严重的公共卫生问题至关重要。在本研究中,我们提出了一种通用且简便的一步法策略,用于使用基于抗生素和适体分子分别识别细菌细胞壁的双分子亲和性Förster(荧光)共振能量转移(FRET)平台,对致病菌进行灵敏和选择性检测。作为概念验证,万古霉素(Van)和核酸适体被用于检测金黄色葡萄球菌(Staph. aureus)的模型双识别方案中。在 30 分钟内,通过分别将 Van 功能化的金纳米簇和适体修饰的金纳米颗粒用作能量供体和受体,FRET 信号显示出与 Staph. aureus 浓度在 20 到 10 cfu/mL 范围内的线性变化,检测限为 10 cfu/mL。其他非靶标细菌显示阴性结果,表明该方法具有良好的特异性。当用于检测实际样品中的金黄色葡萄球菌时,双识别 FRET 策略的回收率在 99.00%到 109.75%之间,相对标准偏差(RSD)小于 4%。这为敏感、特异和简单的致病菌检测建立了一个通用的检测平台,在食品/公共安全监测和传染病诊断等领域具有重要影响。