State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei, 230026, China.
Anal Chem. 2023 Dec 5;95(48):17808-17817. doi: 10.1021/acs.analchem.3c03877. Epub 2023 Nov 16.
The timely detection of diseases and the accurate identification of pathogens require the development of efficient and reliable diagnostic methods. In this study, we have developed a novel specific multivariate probe termed MRTFP (multivariate real-time fluorescent probe) by assembling strand exchange three-way-junction (3WJ) structures. The 3WJ structures were incorporated into a four-angle probe (FP) and a hexagonal probe (HP), to target the multivariate genes of . The FP and HP enable single-step and multiplexed detection in RT-LAMP (real-time loop-mediated isothermal amplification) with exceptional sensitivity and specificity. Encouragingly, real food samples contaminated with ( and ) can be readily identified and distinguished with a minimum detectable concentration (MDC) of 10 CFU/mL without the need for further culture. The introduction of MRTFP allows for simultaneous detection of dual or three targets in a single tube for LAMP, thereby improving detection efficiency. The MRTFP simplifies the design of robust multivariate probes, exhibits excellent stability, and avoids interference from multiple probe units, offering significant potential for the development of specific probes for efficient and accurate disease detection and pathogen identification.
疾病的及时检测和病原体的准确识别需要开发高效可靠的诊断方法。在本研究中，我们通过组装链交换三链结（3WJ）结构开发了一种称为 MRTFP（多变量实时荧光探针）的新型特异多变量探针。3WJ 结构被整合到四角探针（FP）和六角形探针（HP）中，以靶向. 的多变量基因。FP 和 HP 能够在 RT-LAMP（实时环介导等温扩增）中进行单步和多重检测，具有出色的灵敏度和特异性。令人鼓舞的是，受污染的实际食物样本（和）可以被快速识别和区分，最低可检测浓度（MDC）为 10 CFU/mL，无需进一步培养。MRTFP 允许在单个管中同时检测 LAMP 的双或三个靶标，从而提高了检测效率。MRTFP 简化了稳健的多变量探针的设计，表现出优异的稳定性，并避免了多个探针单元的干扰，为开发用于高效准确疾病检测和病原体识别的特异性探针提供了巨大的潜力。
