期刊
ANALYTICAL CHEMISTRY
卷 95, 期 10, 页码 4753-4759出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c05670
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This article investigates a novel approach using upconversion nanoparticles for the detection of the COVID-19 virus and its SARS-CoV-2 variant. The upconversion-linked immunosorbent assay (ULISA) demonstrated a higher sensitivity than conventional assays, with a limit of detection of 1.3 pg/mL. Furthermore, the digital ULISA improved the detection limit by a factor of 10 compared to analog readout for SARS-CoV-2.
The COVID-19 crisis requires fast and highly sensitive tests for the early stage detection of the SARS-CoV-2 virus. For detecting the nucleocapsid protein (N protein), the most abundant viral antigen, we have employed upconversion nanoparticles that emit short-wavelength light under near-infrared excitation (976 nm). The anti-Stokes emission avoids autofluorescence and light scattering and thus enables measurements without optical background interference. The sandwich upconversion-linked immunosorbent assay (ULISA) can be operated both in a conventional analog mode and in a digital mode based on counting individual immune complexes. We have investigated how different antibody combinations affect the detection of the wildtype N protein and the detection of SARS-CoV-2 (alpha variant) in lysed culture fluid via the N protein. The ULISA yielded a limit of detection (LOD) of 1.3 pg/mL (27 fM) for N protein detection independent of the analog or digital readout, which is approximately 3 orders of magnitude more sensitive than conventional enzyme-linked immunosorbent assays or commercial lateral flow assays for home testing. In the case of SARSCoV-2, the digital ULISA additionally improved the LOD by a factor of 10 compared to the analog readout.
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