Engineering Research Center of Bio-process, Ministry of Education, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, P. R. China.
Anhui Province Institute of Product Quality Supervision & Inspection, Hefei 230051, P. R. China.
Anal Chem. 2022 Jan 18;94(2):1357-1364. doi: 10.1021/acs.analchem.1c04638. Epub 2021 Dec 28.
Detection of pathogenic bacteria is of vital significance for combating and preventing infectious diseases. In this work, we developed a multivalent aptamer probe (Multi-VAP)-based trigging isothermal circular amplification (TICA) for rapidly and ultrasensitively detecting . In this sensing system, the fluorescence of Multi-VAP was strongly quenched via the dual effect of FRET. Introduction of to the system forced the configuration change of Multi-VAP, leading to the occurrence of a TICA responsible for tuning all of the fluorescence-quenched Multi-VAP into a complete restoration state. This prominent feature allows the reasonable combination of a strong background restraint and great target signal amplification into one sensing system, which in turn benefits the improvement of the signal-to-noise ratio to ensure that the system has an ultrahigh sensitivity. Combined with the employment of an aptamer to ensure that it has excellent specificity, the can be quantitatively and qualitatively analyzed even from human serum. The total processing merely requires sample addition and incubation. The turnaround time of the complete analysis from "sample-to-result" was within 30 min. With the method to decrease the time to detect and simplify the process to operate, the assay was successfully used as a sensing platform for specific detection of as few as 9 CFU/mL .
检测病原菌对于防治传染病至关重要。在这项工作中，我们开发了一种基于多价适体探针（Multi-VAP）的触发等温循环扩增（TICA）方法，用于快速、高灵敏地检测。在这个传感系统中，Multi-VAP 的荧光通过 FRET 的双重作用被强烈猝灭。当 引入到该系统中时，Multi-VAP 的构象发生变化，导致负责将所有荧光猝灭的 Multi-VAP 调制成完整恢复状态的 TICA 发生。这一显著特征使得一个传感系统能够合理地结合强背景抑制和大目标信号放大，从而提高信噪比，确保系统具有超高的灵敏度。结合使用适体以确保其具有优异的特异性，即使来自人血清的也可以进行定量和定性分析。整个处理过程仅需要添加样品并孵育。从“样品到结果”的完整分析的周转时间在 30 分钟内。通过减少检测时间和简化操作过程的方法，该检测方法成功地用作检测低至 9 CFU/mL 的特定检测的传感平台。
