Department of Biochemistry and Molecular Biology, College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang, 330045, People's Republic of China.
Institute of Functional Materials and Agricultural Applied Chemistry, Jiangxi Agricultural University, Nanchang, 330045, People's Republic of China.
Mikrochim Acta. 2023 Dec 26;191(1):52. doi: 10.1007/s00604-023-06130-1.
Coconut cadang-cadang viroid (CCCVd) is an infectious single-stranded RNA (ssRNA) pathogen, which leads directly to the death of a large number of coconut palm trees and heavy economic loss to coconut farmers. Herein, a novel electrochemical impedance RNA genosensor is presented based on highly stable gold nanoparticles (AuNPs) decorated phosphorene (BP) nanohybrid with graphene (Gr) for highly sensitive, low-cost, and label-free detection of CCCVd. BP-AuNPs are environmentally friendly prepared by ultrasonic-assisted liquid-phase exfoliation of black phosphorus, accompanying direct reduction of chloroauric acid. Gr/BP-AuNPs are facilely prepared by the in situ growth of AuNPs onto the BP surface and its nanohybrid with Gr to improve environmental stability of BP. Gr/BP-AuNP-based RNA genosensor is fabricated by immobilizing the thiol-functionalized single-stranded DNA (ssDNA) oligonucleotide probe onto the surface of Gr/BP-AuNP-modified glassy carbon electrode via gold-thiol interactions, which served as an electrochemical genosensing platform for the label-free impedance detection of CCCVd by hybridization between the functionalized ssDNA probe and the complementary CCCVd ssRNA sequence in a wide linear range from 1.0 × 10 to 1.0 × 10 M with a low limit of detection of 2.8 × 10 M. This work supplies an experimental support and theoretical direction for the fabrication of RNA biosensors based on graphene-like materials and potential application for a specific diagnosis of plant RNA viral disease in Arecaceae planting industry.
椰心叶甲麻痹病毒（CCCVd）是一种传染性单链 RNA（ssRNA）病原体，它直接导致大量椰子树死亡，给椰子种植户造成严重的经济损失。在此，提出了一种基于高度稳定的金纳米粒子（AuNPs）修饰的黑磷（BP）纳米杂化材料与石墨烯（Gr）的新型电化学阻抗 RNA 基因传感器，用于高度灵敏、低成本和无标记检测 CCCVd。BP-AuNPs 通过黑磷的超声辅助液相剥离制备，伴随氯金酸的直接还原，环境友好。Gr/BP-AuNPs 通过 AuNPs 在 BP 表面的原位生长及其与 Gr 的纳米杂化制备，以提高 BP 的环境稳定性。Gr/BP-AuNP 基 RNA 基因传感器通过金-硫醇相互作用将巯基功能化的单链 DNA（ssDNA）寡核苷酸探针固定在 Gr/BP-AuNP 修饰的玻碳电极表面，作为电化学基因传感平台，用于无标记阻抗检测通过功能化 ssDNA 探针与互补 CCCVd ssRNA 序列在 1.0×10 到 1.0×10^M 的宽线性范围内杂交，检测限低至 2.8×10^M。这项工作为基于类石墨烯材料的 RNA 生物传感器的制备提供了实验支持和理论指导，并为在 Arecaceae 种植产业中对植物 RNA 病毒病进行特定诊断提供了潜在的应用。
