Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University (TMDU) , 2-3-10 Kanda-Surugadai , Chiyoda , Tokyo 101-0062 , Japan.
Department of Molecular Virology, Graduate School of Medical and Dental Sciences , Tokyo Medical and Dental University (TMDU) , 1-5-45 Yushima , Bunkyo , Tokyo 113-8510 , Japan.
Langmuir. 2019 Feb 5;35(5):1798-1806. doi: 10.1021/acs.langmuir.8b01586. Epub 2018 Aug 31.
Resistive pulse sensing (RPS) is an analytical technique for detecting particles with nano- to micrometer diameters, such as proteins, viruses, and bacteria. RPS is a promising tool for diagnosis as it can analyze the characteristics of target particles individually from ion current blockades as pulse waveforms. However, it is difficult to discriminate analog targets because RPS merely provides physical information such as size, shape, concentration, and charge density of the analyte. Influenza A virus, which is 80-120 nm in diameter, has various subtypes, demonstrating the diversity of virus characteristics. For example, highly pathogenic avian influenza infections in humans are recognized as an emerging infectious disease with high mortality rates compared with human influenza viruses. Distinguishing human from avian influenza using their differing biological characteristics would be challenging using RPS. To develop a highly selective diagnostic system for infectious diseases, we combined RPS with molecular recognition. Gold nanoparticles (GNPs) that have human influenza A (H1N1 subtype) virus-specific sialic acid receptors on the surface were prepared as a virus label for RPS analysis. A sulfobetaine and sialic acid (ligand) hybrid surface was formed on the GNPs for the suppression of nonspecific interaction. The results show a size change of viruses derived from specific interactions with GNPs. In contrast, no size shift was observed when nonspecific sialic acid receptor-immobilized GNPs were used. Detection of viruses by individual particle counting could be a new facet of diagnosis.
电阻脉冲感应（RPS）是一种用于检测纳米到微米直径颗粒的分析技术，例如蛋白质、病毒和细菌。RPS 是一种很有前途的诊断工具，因为它可以根据脉冲波形中离子电流阻塞来分析目标颗粒的个体特征。然而，由于 RPS 仅提供分析物的大小、形状、浓度和电荷密度等物理信息，因此很难区分模拟目标。直径为 80-120nm 的甲型流感病毒有多种亚型，表现出病毒特征的多样性。例如，与人类流感病毒相比，高致病性禽流感感染在人类中被认为是一种具有高死亡率的新发传染病。使用 RPS 区分人类和禽流感可能具有挑战性，因为它们具有不同的生物学特征。为了开发一种高度选择性的传染病诊断系统，我们将 RPS 与分子识别相结合。在表面上制备了具有人类甲型流感（H1N1 亚型）病毒特异性唾液酸受体的金纳米颗粒（GNPs），作为 RPS 分析的病毒标记物。在 GNPs 上形成了磺基甜菜碱和唾液酸（配体）混合表面，以抑制非特异性相互作用。结果表明，病毒的大小变化源于与 GNPs 的特异性相互作用。相比之下，当使用非特异性唾液酸受体固定化的 GNPs 时，未观察到大小移动。通过逐个颗粒计数检测病毒可能是诊断的一个新方面。
