Laboratory of Biotechnology, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Biosens Bioelectron. 2018 Dec 30;122:16-24. doi: 10.1016/j.bios.2018.09.024. Epub 2018 Sep 7.
A new method of label free sensing approach with superior selectivity and sensitivity towards virlabel-freeon is presented here, employing the localized surface plasmon resonance (LSPR) behavior of gold nanoparticles (AuNPs) and fluorescent CdSeTeS quantum dots (QDs). Inorganic quaternary alloyed CdSeTeS QDs were capped with -cysteine via a ligand exchange reaction. Alternatively, citrate stabilized AuNPs were functionalized with 11-mercaptoundecanoic acid to generate carboxylic group on the gold surface. The carboxylic group on the AuNPs was subjected to bind covalently with the amine group of -cysteine capped CdSeTeS QDs to form CdSeTeS QDs/AuNPs nanocomposites. The fluorescence of CdSeTeS QDs/AuNPs nanocomposite shows quenched spectrum of CdSeTeS QDs at 640 nm due to the close interaction with AuNPs. However, after successive addition of norovirus-like particles (NoV-LPs), steric hindrance-induced LSPR signal from the adjacent AuNPs triggered the fluorescence enhancement of QDs in proportion to the concentration of the target NoV-LPs. A linear range of 10 to 10 g mL NoV-LPs with a detection limit of 12.1 × 10 g mL was obtained. This method was further applied on clinically isolated norovirus detection, in the range of 10-10 copies mL with a detection limit of 95.0 copies mL, which is 100-fold higher than commercial ELISA kit. The superiority of the proposed sensor over other conventional sensors is found in its ultrasensitive detectability at low virus concentration even in clinically isolated samples. This proposed detection method can pave an avenue for the development of high performance and robust sensing probes for detection of virus in biomedical applications.
本文提出了一种新的无标记传感方法,该方法对病毒具有优异的选择性和灵敏度,利用了金纳米粒子(AuNPs)的局域表面等离子体共振(LSPR)行为和荧光 CdSeTeS 量子点(QDs)。通过配体交换反应,将无机四元合金 CdSeTeS QDs 用半胱氨酸进行了封端。或者,用 11-巯基十一酸对柠檬酸稳定的 AuNPs 进行功能化,在金表面生成羧基。AuNPs 上的羧基与半胱氨酸封端的 CdSeTeS QDs 上的氨基发生共价结合,形成 CdSeTeS QDs/AuNPs 纳米复合材料。由于与 AuNPs 的紧密相互作用,CdSeTeS QDs/AuNPs 纳米复合材料的荧光在 640nm 处显示出 CdSeTeS QDs 的猝灭光谱。然而,在连续加入诺如病毒样颗粒(NoV-LPs)后,由于相邻 AuNPs 的空间位阻诱导的 LSPR 信号,导致 QDs 的荧光增强,与靶标 NoV-LPs 的浓度成正比。得到了 10 至 10 g/mL 的 NoV-LPs 的线性范围,检测限为 12.1×10 g/mL。该方法进一步应用于临床分离的诺如病毒检测,检测限为 95.0 拷贝/mL,检测范围为 10-10 拷贝/mL,比商业 ELISA 试剂盒高 100 倍。与其他传统传感器相比,该传感器的优势在于即使在临床分离的样本中,也能在低病毒浓度下实现超灵敏检测。该检测方法为开发用于生物医学应用中病毒检测的高性能和稳健感测探针铺平了道路。