Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, 836 Ohya Suruga-ku, Shizuoka, 422-8529, Japan.
Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya Suruga-ku, Shizuoka, 422-8529, Japan.
Anal Chim Acta. 2022 May 15;1207:339817. doi: 10.1016/j.aca.2022.339817. Epub 2022 Apr 9.
Signal amplification have been centralized in developing the highly reliable biosensor for analyte detection with a narrow detection window. We proposed an aptasensor to provide a highly sensitive early-stage diagnostic platform of dengue virus NS1 protein (DENV-NS1) by dual-approach - colorimetric and electrochemical detection. This work utilized two different aptamers specific to DENV-NS1: One conjugated to gold nanoparticles (AuNPs), forming AuNPs-Apt and its complementary sequence aptamer, forming AuNPs-Apt. The unbound Apt of AuNPs-Apt by DENV-NS1 were to hybridize to AuNPs-Apt and induced a 3D-nanoassembled formation, resulting in DENV-NS1 concentration-dependent plasmonic color change. Occurrence of the hybridization of Apt and Apt, the 3D-assembled hybridized aptamers of AuNPs was incubated with methylene blue (MB) solution, which intercalated a high number of MB molecules within the duplex structure of aptamers, and the complex was captured on the Apt-conjugated disposable gold electrode (DGE). The developed aptamer-based biosensor showed high sensitivity with colorimetric response down to 1.28 pg/mL and electrochemical approach down to 30 fg/mL of DENV-NS1 with good selectivity. This work showcases an advanced utilization of aptamer and its complementary anti-sense aptamer in signal amplification and nanocarrier for biosensing.
信号放大已集中在开发用于分析物检测的高可靠性生物传感器上，检测窗口较窄。我们提出了一种适体传感器，通过比色和电化学检测的双重方法为登革热病毒 NS1 蛋白（DENV-NS1）提供高度敏感的早期诊断平台。这项工作利用了两种针对 DENV-NS1 的不同适体：一种与金纳米粒子（AuNPs）结合，形成 AuNPs-Apt 及其互补序列适体，形成 AuNPs-Apt。未与 DENV-NS1 结合的 AuNPs-Apt 被用于与 AuNPs-Apt 杂交，并诱导 3D 纳米组装形成，导致 DENV-NS1 浓度依赖性等离子体颜色变化。发生 Apt 和 Apt 的杂交后，AuNPs 的 3D 组装杂交适体与亚甲蓝（MB）溶液孵育，MB 分子大量插入适体的双链结构中，该复合物被捕获在 Apt 偶联的一次性金电极（DGE）上。基于适体的生物传感器具有高灵敏度，比色响应低至 1.28 pg/mL，电化学方法低至 30 fg/mL 的 DENV-NS1，具有良好的选择性。这项工作展示了适体及其互补反义适体在信号放大和生物传感中的纳米载体的先进应用。
