@article{Zhou2026, 
author = {Shu Zhou and Yuxi Xu and Huan Liao and Hailong Ou and Dan Qi and Yatao Wu and Yunyi Liu and Juan Li and Jiaxuan Li and Bi Shi and Fei Zhu and Siran Zhang and Jason H. Huang and Erxi Wu and Xiaoxiao Hu},
title = {Dual-mode aptamer-driven biosensing platform for ultrasensitive and mutation-resilient detection of the SARS-CoV-2 nucleocapsid protein},
year = {2026},
journal = {Genes & Diseases},
volume = {13},
number = {3},
keywords = {Aptamer, Aptamer-antibody sandwich assay, Dynamic light scattering, Mutation, SARS-CoV-2 N protein},
url = {https://www.sciopen.com/article/10.1016/j.gendis.2025.101943},
doi = {10.1016/j.gendis.2025.101943},
abstract = {Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a significant global health threat because of its rapid evolution and high mutation rate, which limits the performance of existing molecular diagnostics. This study presents a dual-mode, aptamer-based detection platform that combines high sensitivity with mutation resilience. Using a computer-assisted X-aptamer Systematic Evolution of Ligands by EXponential enrichment (SELEX) approach, we identified NP14, a high-affinity, dual-target DNA aptamer that specifically binds to the SARS-CoV-2 nucleocapsid (N) protein at its N-terminal domain. Analyses via molecular docking, aptamer truncation, and targeted mutagenesis revealed that NP14 interacted with both SARS-CoV-2 and SARS-CoV N proteins and identified key nucleotides C24 and G27 of the P1 region and structural determinants critical for its high-affinity binding. Building on this discovery, we engineered a dual-mode biosensing system by integrating NP14 into a multicolor dynamic light scattering-enhanced enzyme-linked aptamer-antibody assay (MD ELAAA). MD ELAAA synergistically combines two complementary detection strategies: ⅰ) non-aggregative plasmonic colorimetry for visual signal detection and ⅱ) dynamic light scattering for ultrasensitive quantitative analysis, in which Au/Ag nanomaterials are used to amplify optical and scattering signals. This system achieves a sensitivity of 0.43 TCID50/mL, representing a 47-fold improvement over standard methods. By integrating high sensitivity, specificity, variant recognition, and dual-mode signal output, the MD ELAAA platform enables reliable detection of low-abundance SARS-CoV-2 antigens. Its robust performance supports early-stage diagnostics and high-throughput variant monitoring, establishing MD ELAAA as a robust platform for next-generation viral detection and surveillance.}
}