Antisense oligonucleotide conjugated gold nanoconstructs-based electrochemical biosensor for detection of SARS-CoV-2

IF 7.5 Q1 CHEMISTRY, PHYSICAL
Tejaswini P. Patil , Vishakha S. Parkhe , Somnath S. Kundale , Rajanish K. Kamat , Tukaram D. Dongale , Rajendra S Patil , Arpita P. Tiwari
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引用次数: 0

Abstract

Nanoconstructs of gold nanoparticles (AuNPs) conjugated with SARS-CoV-2 specific antisense oligonucleotides (ASO) have been utilized to develop sensitive electrochemical nucleic acid biosensor for the detection of SARS-CoV-2 RNA. AuNPs were prepared through a one-pot synthesis method by utilizing Poly-L-Lysine (PLL) biopolymer and as synthesised AuNP were characterized by various analytical techniques such as UV–Vis spectroscopy, X-ray Diffraction (XRD) analysis, Fourier Transform Infra-Red spectroscopy (FT-IR), zeta potential, and Transmission Electron Microscopy (TEM). Poly-L-Lysine functionalized AuNPs (PLL-AuNPs) nanoconstructs platform was employed for immobilization of SARS-CoV-2 specific antisense oligonucleotides (ASO-conjugated PLL-AuNPs) via electrostatic interactions. The PLL-AuNPs were drop casted on glassy carbon electrode (GCE) following immobilization of ASO for fabrication of electrochemical biosensor. The ASO-conjugated PLL-AuNPs nanoconstructs were characterized by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) techniques. The responsiveness of ASO-conjugated PLL-AuNPs nanoconstructs in presence SARS-CoV-2 RNA was monitored using the DPV, SWV and EIS technique, where methylene blue was employed as an electrochemical indicator for DNA-RNA hybridization detection. The biosensor exhibits a detection range for SARS-CoV-2 RNA infection ranging from 0 to 100 nM, with a limit of detection at 30.2 nM. The electrode, modified with ASO-conjugated PLL-AuNPs, was employed for the detection of SARS-CoV-2 RNA from clinical samples collected from COVID-19-positive individuals.

Abstract Image

基于反义寡核苷酸共轭金纳米结构的电化学生物传感器用于检测 SARS-CoV-2
利用与 SARS-CoV-2 特异性反义寡核苷酸(ASO)共轭的金纳米粒子(AuNPs)纳米结构开发了灵敏的电化学核酸生物传感器,用于检测 SARS-CoV-2 RNA。利用聚-L-赖氨酸(PLL)生物聚合物通过一锅合成法制备了 AuNPs,并通过紫外可见光谱、X 射线衍射(XRD)分析、傅立叶变换红外光谱(FT-IR)、ZETA 电位和透射电子显微镜(TEM)等各种分析技术对合成的 AuNPs 进行了表征。利用聚赖氨酸功能化 AuNPs(PLL-AuNPs)纳米结构平台,通过静电相互作用固定 SARS-CoV-2 特异性反义寡核苷酸(ASO 共轭 PLL-AuNPs)。在固定 ASO 后,将 PLL-AuNPs 滴铸在玻璃碳电极(GCE)上,用于制造电化学生物传感器。通过循环伏安法(CV)、差分脉冲伏安法(DPV)和电化学阻抗谱(EIS)技术对 ASO 共轭的 PLL-AuNPs 纳米结构进行了表征。利用 DPV、SWV 和 EIS 技术监测了 ASO 共轭 PLL-AuNPs 纳米结构在 SARS-CoV-2 RNA 存在下的响应性,其中亚甲基蓝被用作 DNA-RNA 杂交检测的电化学指示剂。该生物传感器对 SARS-CoV-2 RNA 感染的检测范围为 0 至 100 nM,检测限为 30.2 nM。该电极由 ASO 共轭 PLL-AuNPs 修饰,用于检测从 COVID-19 阳性个体采集的临床样本中的 SARS-CoV-2 RNA。
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来源期刊
CiteScore
8.10
自引率
1.60%
发文量
128
审稿时长
66 days
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