Graduate Institute of Toxicology, College of Medicine, National Taiwan University, No.1, Sec. 1, Jen Ai Rd, Taipei, 100, Taiwan.
School of Medicine, Chung Shan Medical University, No.110, Sec. 1, Chien Kuo N. Rd, Taichung, 402, Taiwan.
J Nanobiotechnology. 2024 Jan 3;22(1):5. doi: 10.1186/s12951-023-02191-9.
We have successfully generated oligonucleotide aptamers (Apts) and monoclonal antibodies (mAbs) targeting the recombinant nucleocapsid (N) protein of SARS-CoV-2. Apts were obtained through seven rounds of systematic evolution of ligands by exponential enrichment (SELEX), while mAbs were derived from the 6F6E11 hybridoma cell line. Leveraging these Apts and mAbs, we have successfully devised two innovative and remarkably sensitive detection techniques for the rapid identification of SARS-CoV-2 N protein in nasopharyngeal samples: the enzyme-linked aptamer-antibody sandwich assay (ELAAA) and the hybrid lateral flow strip (hybrid-LFS). ELAAA exhibited an impressive detection limit of 0.1 ng/mL, while hybrid-LFS offered a detection range of 0.1 - 0.5 ng/mL. In the evaluation using ten nasopharyngeal samples spiked with known N protein concentrations, ELAAA demonstrated an average recovery rate of 92%. Additionally, during the assessment of five nasopharyngeal samples from infected individuals and ten samples from healthy volunteers, hybrid-LFS displayed excellent sensitivity and specificity. Our study introduces a novel and efficient on-site approach for SARS-CoV-2 detection in nasopharyngeal samples. The reliable hybrid Apt-mAb strategy not only advances virus diagnostic methods but also holds promise in combating the spread of related diseases.
我们成功地生成了针对 SARS-CoV-2 重组核衣壳 (N) 蛋白的寡核苷酸适体 (Apts) 和单克隆抗体 (mAbs)。Apts 通过指数富集的配体系统进化 (SELEX) 进行了七轮筛选获得，而 mAbs 则源自 6F6E11 杂交瘤细胞系。利用这些 Apts 和 mAbs，我们成功设计了两种创新且非常灵敏的检测技术，用于快速鉴定鼻咽样本中的 SARS-CoV-2 N 蛋白：酶联适体-抗体夹心检测 (ELAAA) 和杂交侧向流条带 (hybrid-LFS)。ELAAA 展现出令人印象深刻的检测限为 0.1ng/mL，而 hybrid-LFS 的检测范围为 0.1-0.5ng/mL。在使用含有已知 N 蛋白浓度的十个鼻咽样本进行评估时，ELAAA 平均回收率为 92%。此外，在对五名感染个体的鼻咽样本和十名健康志愿者的样本进行评估时，hybrid-LFS 显示出优异的灵敏度和特异性。本研究引入了一种新颖且高效的鼻咽样本中 SARS-CoV-2 检测的现场方法。可靠的混合 Apt-mAb 策略不仅推进了病毒诊断方法，还有望用于对抗相关疾病的传播。
