College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao, 266580, People's Republic of China.
Anal Bioanal Chem. 2023 Feb;415(4):545-554. doi: 10.1007/s00216-022-04437-1. Epub 2022 Nov 22.
Serological antibody tests are useful complements of nuclei acid detection for SARS-CoV-2 diagnosis, which can significantly improve diagnostic accuracy. However, antibody detection in serum or plasma remains challenging to do with high sensitivity. In this study, Ag nanoparticles with ultra-thin Au shells embedded with 4-mercaptobenzoic acid (MBA) (Ag@Au) were manufactured and then assembled onto FeO surface by electrostatic interaction to construct the FeO-Ag@Au nanoparticles (NPs) with magnetic-Raman-colorimetric properties. Based on the composite nanoparticles, a colorimetric and Raman dual-mode lateral flow immunoassay (LFIA) for ultrasensitive identification of SARS-CoV-2 nucleocapsid (N) protein antibody was constructed. The magnetic nanoparticles (FeO NPs) were acted as the core and coated a layer of Ag@Au particles on the surface by electrostatic interaction to prepare FeO-Ag@Au NPs, which can amplify the SERS signal due to multiple Ag@Au particles concentrated on a single magnetic nanoparticle. Moreover, the FeO-Ag@Au NPs facilitated pre-purifying sample using magnetic separation, and complex matrix interference would be greatly decreased in the detection. The FeO-Ag@Au NPs modified with N protein recognized and bound with N protein antibodies, which were trapped on the T-line, forming color band for observing detection. Under optimal conditions, the N protein antibodies could be qualitatively detected in colorimetric mode with the visual limit of 10 mg/mL and quantitatively detected by SERS signals between 10 and 10 mg /mL with 0.08 pg/mL detection limit. The coefficients variations (CV) of intra-assay was 8.0%, whereas of inter-assay was 11.7%, confirming of good reproducibility. Finally, this approach was able to discriminate between positive, negative, and weakly positive samples when detecting 107 clinical serum samples. The process enables highly sensitive quantitative assays that are valuable for evaluating disease processes and guiding treatment. Colorimetric and Raman dual-mode LFIA detection of SARS-CoV-2 N protein antibody based on FeO-Ag@Au nanoparticles.
基于 FeO-Ag@Au 纳米粒子的 SARS-CoV-2 N 蛋白抗体的比色和拉曼双模式侧向流免疫分析检测
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