Nanobiosensors Lab, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, P.O. Box 14399-56191, Tehran, Iran.
Department of Chemistry, College of Science, Sultan Qaboos University, Box 36, Al-Khod 123, Sultan Qaboos, Oman.
Mikrochim Acta. 2023 Aug 24;190(9):368. doi: 10.1007/s00604-023-05947-0.
A dual-targeting nanobiosensor has been developed for the simultaneous detection of AMELX and AMELY genes based on the different fluorescence signals emitted from gold and silver nanoclusters, AuNCs and AgNCs respectively. In our design, both catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) have been used as isothermal, enzyme-free and simple methods for signal's amplification. The working principle is based on the initiation of a cascade of CHA-HCR reactions when AMELX is present, in which AuNCs, synthesized on the third hairpin, are aggregated on the surface of the dsDNA product, performing the phenomenon of aggregation induced emission (AIE) and enhancing their fluorescence signal. On the other hand, the presence of the second target, AMELY, is responsible for the enhancement of the fluorescence signal corresponding to AgNCs by the same phenomenon, via hybridizing to the free end of the dsDNA formed and at the same time to the probe of silver nanoclusters fixing it closer to the surface of the dsDNA product. Such a unique design has the merits of being simple, inexpensive, specific and stable and presents rapid results. The detection limits of this assay for AMELX and AMELY are as low as 3.16 fM and 23.6 fM respectively. Moreover, this platform showed great performance in real samples. The design has great promise for the application of dual-targeting nanobiosensors to other biomarkers.
已经开发出一种双靶向纳米生物传感器，用于基于金和银纳米簇分别发出的不同荧光信号同时检测 AMELX 和 AMELY 基因。在我们的设计中，催化发夹组装（CHA）和杂交链式反应（HCR）都被用作等温、无酶和简单的信号放大方法。工作原理基于当存在 AMELX 时 CHA-HCR 反应的级联起始，其中在第三发夹上合成的 AuNCs 在 dsDNA 产物的表面上聚集，表现出聚集诱导发射（AIE）现象并增强其荧光信号。另一方面，第二个靶标 AMELY 的存在通过相同的现象导致对应于 AgNCs 的荧光信号增强，通过与形成的 dsDNA 的游离端杂交，同时与固定在 dsDNA 产物表面附近的银纳米簇的探针杂交。这种独特的设计具有简单、廉价、特异性和稳定性的优点，并呈现出快速的结果。该测定法对 AMELX 和 AMELY 的检测限分别低至 3.16 fM 和 23.6 fM。此外，该平台在实际样品中表现出出色的性能。该设计有望将双靶向纳米生物传感器应用于其他生物标志物。
