College of Chemistry and Chemical Engineering, State Key Laboratory of Chemical Safety, China University of Petroleum (East China), Qingdao 266580, China.
School of Chemistry, The University of New South Wales, Sydney, New South Wales 2052, Australia.
Anal Chem. 2024 Apr 16;96(15):6065-6071. doi: 10.1021/acs.analchem.4c00648. Epub 2024 Apr 3.
The conventional lateral flow immunoassay (LFIA) method using colloidal gold nanoparticles (Au NPs) as labeling agents faces two inherent limitations, including restricted sensitivity and poor quantitative capability, which impede early viral infection detection. Herein, we designed and synthesized CsPbBr perovskite quantum dot-based composite nanoparticles, CsPbBr@SiO@FeO (CSF), which integrated fluorescence detection and magnetic enrichment properties into LFIA technology and achieved rapid, sensitive, and convenient quantitative detection of the SARS-CoV-2 virus N protein. In this study, CsPbBr served as a high-quantum-yield fluorescent signaling probe, while SiO significantly enhanced the stability and biomodifiability of CsPbBr. Importantly, the SiO shell shows relatively low absorption or scattering toward fluorescence, maintaining a quantum yield of up to 74.4% in CsPbBr@SiO. Assembly of FeO nanoparticles mediated by PEI further enhanced the method's sensitivity and reduced matrix interference through magnetic enrichment. Consequently, the method achieved a fluorescent detection range of 1 × 10 to 5 × 10 pg·mL after magnetic enrichment, with a limit of detection (LOD) of 58.8 pg·mL, representing a 13.3-fold improvement compared to nonenriched samples (7.58 × 10 pg·mL) and a 2-orders-of-magnitude improvement over commercial colloidal gold kits. Furthermore, the method exhibited 80% positive and 100% negative detection rates in clinical samples. This approach holds promise for on-site diagnosis, home-based quantitative tests, and disease procession evaluation.
传统的胶体金纳米粒子(Au NPs)标记侧向流动免疫分析(LFIA)方法存在两个固有局限性,包括限制的灵敏度和较差的定量能力,这阻碍了早期病毒感染的检测。在此,我们设计并合成了基于 CsPbBr 钙钛矿量子点的复合纳米粒子 CsPbBr@SiO@FeO(CSF),它将荧光检测和磁性富集特性集成到 LFIA 技术中,实现了 SARS-CoV-2 病毒 N 蛋白的快速、灵敏、方便的定量检测。在这项研究中,CsPbBr 用作高量子产率荧光信号探针,而 SiO 显著提高了 CsPbBr 的稳定性和生物修饰性。重要的是,SiO 壳层对荧光的吸收或散射相对较低,在 CsPbBr@SiO 中保持高达 74.4%的量子产率。PEI 介导的 FeO 纳米粒子的组装通过磁性富集进一步提高了方法的灵敏度并减少了基质干扰。因此,该方法在磁性富集后实现了荧光检测范围为 1×10 到 5×10 pg·mL-1,检测限(LOD)为 58.8 pg·mL-1,与未富集样品(7.58×10 pg·mL-1)相比提高了 13.3 倍,与商业胶体金试剂盒相比提高了 2 个数量级。此外,该方法在临床样本中的阳性检出率为 80%,阴性检出率为 100%。这种方法有望用于现场诊断、家庭定量检测和疾病进程评估。