Faculty of Chemistry & Environmental Science, Guangdong Ocean University, Zhanjiang, 524088, China; Research Center for Coastal Environmental Protection and Ecological Resilience, Guangdong Ocean University, Zhanjiang, 524088, China.
Faculty of Chemistry & Environmental Science, Guangdong Ocean University, Zhanjiang, 524088, China.
Anal Chim Acta. 2024 Apr 22;1299:342406. doi: 10.1016/j.aca.2024.342406. Epub 2024 Feb 27.
The hybridization chain reaction (HCR), as one of the nucleic acid amplification technologies, is combined with fluorescence signal output with excellent sensitivity, simplicity, and stability. However, current HCR-based fluorescence sensing methods still have some defects such as the blocking effect of the HCR combination with fluorophores and the aggregation-caused quenching (ACQ) phenomenon of traditional fluorophores. Herein, a triplex DNA-based aggregation-induced emission probe (AIE-P) was designed as the fluorescent signal transduction, which is able to provide a new platform for HCR-based sensing assay. The AIE-P was synthesized by attaching the AIE fluorophores to terminus of the oligonucleotide through amido bond, and captured the products of HCR to form triplex DNA. In this case, the AIE fluorophores were located in close proximity to generate fluorescence. This assay provided turn-on fluorescence efficiency with a high signal-to-noise ratio and excellent amplification capability to solve the shortcoming of HCR-based fluorescence sensing methods. It enabled sensitive detection of Vibrio parahaemolyticus in the range of 10-10 CFU mL, and with a low limit of detection down to 39 CFU mL. In addition, this assay expressed good specificity and practicability. The triplex DNA-based AIE probe forms a universal molecular tool for developing HCR-based fluorescence sensing methods.
杂交链式反应 (HCR) 作为一种核酸扩增技术，与荧光信号输出相结合，具有出色的灵敏度、简单性和稳定性。然而，目前基于 HCR 的荧光传感方法仍然存在一些缺陷，例如 HCR 与荧光团的结合的阻断效应和传统荧光团的聚集猝灭 (ACQ) 现象。在此，设计了一种基于三链 DNA 的聚集诱导发射探针 (AIE-P) 作为荧光信号转导，为基于 HCR 的传感分析提供了一个新平台。AIE-P 通过酰胺键将 AIE 荧光团连接到寡核苷酸的末端合成，捕获 HCR 的产物以形成三链 DNA。在这种情况下，AIE 荧光团靠近以产生荧光。该测定法提供了高信噪比和出色的扩增能力的开启荧光效率，以解决基于 HCR 的荧光传感方法的缺点。它能够在 10-10 CFU mL 的范围内灵敏地检测副溶血性弧菌，检测限低至 39 CFU mL。此外，该测定法表现出良好的特异性和实用性。基于三链 DNA 的 AIE 探针形成了开发基于 HCR 的荧光传感方法的通用分子工具。
