Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Department of Microbiology, Shizuoka Institute of Environment and Hygiene, 232-1 Yainaba, Fujieda 426-0083, Japan.
ACS Appl Mater Interfaces. 2020 Sep 30;12(39):43522-43534. doi: 10.1021/acsami.0c14729. Epub 2020 Sep 21.
A novel biosensing system based on graphene-mediated surface-enhanced Raman scattering (G-SERS) using plasmonic/magnetic molybdenum trioxide nanocubes (mag-MoO NCs) has been designed to detect norovirus (NoV) via a dual SERS nanotag/substrate platform. A novel magnetic derivative of MoO NCs served as the SERS nanotag and the immunomagnetic separation material of the biosensor. Single-layer graphene oxide (SLGO) was adopted as the 2D SERS substrate/capture platform and acted as the signal reporter, with the ability to accommodate an additional Raman molecule as a coreporter. The developed SERS-based immunoassay achieved a signal amplification of up to ∼10-fold resulting from the combined electromagnetic and chemical mechanisms of the dual SERS nanotag/substrate system. The developed biosensor was employed for the detection of NoV in human fecal samples collected from infected patients by capturing the virus with the aid of NoV-specific antibody-functionalized magnetic MoO NCs. This approach enabled rapid signal amplification for NoV detection with this biosensing technology. The biosensor was tested and optimized using NoV-like particles within a broad linear range from 10 fg/mL to 100 ng/mL and a limit of detection (LOD) of ∼5.2 fg/mL. The practical applicability of the developed biosensor to detect clinical NoV subtypes in human fecal samples was demonstrated by effective detection with an LOD of ∼60 RNA copies/mL, which is ∼10-fold lower than that of a commercial enzyme-linked immunosorbent assay kit for NoV.
一种基于石墨烯介导的表面增强拉曼散射（G-SERS）的新型生物传感系统，使用等离子体/磁性三氧化钼纳米立方体（mag-MoO NCs），通过双 SERS 纳米标签/基底平台设计用于检测诺如病毒（NoV）。MoO NCs 的新型磁性衍生物用作 SERS 纳米标签和生物传感器的免疫磁性分离材料。单层氧化石墨烯（SLGO）被用作 2D SERS 基底/捕获平台，并充当信号报告器，具有容纳作为核心报告器的额外拉曼分子的能力。基于 SERS 的免疫测定法通过利用诺如病毒特异性抗体功能化的磁性 MoO NCs 捕获病毒，实现了高达约 10 倍的信号放大，这是由于双重 SERS 纳米标签/基底系统的电磁和化学机制的结合。该生物传感器用于检测从感染患者收集的人粪便样本中的 NoV，通过 NoV 特异性抗体功能化的磁性 MoO NCs 捕获病毒。这种方法使该生物传感技术能够快速进行 NoV 检测信号放大。该生物传感器使用 NoV 样颗粒在 10 fg/mL 至 100 ng/mL 的宽线性范围内进行了测试和优化，检测限（LOD）约为 5.2 fg/mL。通过有效检测临床 NoV 亚型在人粪便样本中的开发的生物传感器的实际适用性证明了这一点，LOD 约为 60 RNA 拷贝/mL，比商业酶联免疫吸附测定试剂盒对 NoV 的检测低约 10 倍。
