BRANDMEIER, Julian, Natalia JURGA, Tomasz GRZYB, Antonín HLAVÁČEK, Radka OBOŘILOVÁ, Petr SKLÁDAL, Zdeněk FARKA and Hans-Heiner GORRIS. Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay. Analytical Chemistry. Washington, DC: American Chemical Society, 2023, vol. 95, No 10, p. 4753-4759. ISSN 0003-2700. Available from: https://dx.doi.org/10.1021/acs.analchem.2c05670.
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Basic information
Original name Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay
Authors BRANDMEIER, Julian (276 Germany, belonging to the institution), Natalia JURGA, Tomasz GRZYB (616 Poland), Antonín HLAVÁČEK (203 Czech Republic), Radka OBOŘILOVÁ (203 Czech Republic, belonging to the institution), Petr SKLÁDAL (203 Czech Republic, belonging to the institution), Zdeněk FARKA (203 Czech Republic, belonging to the institution) and Hans-Heiner GORRIS (276 Germany, guarantor, belonging to the institution).
Edition Analytical Chemistry, Washington, DC, American Chemical Society, 2023, 0003-2700.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 7.400 in 2022
RIV identification code RIV/00216224:14310/23:00130445
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1021/acs.analchem.2c05670
UT WoS 000939526700001
Keywords in English Covid-19; SARS-CoV-2; immunoassay; photon-upconversion nanoparticle; ULISA; digital detection
Tags CF NANO, rivok
Tags International impact, Reviewed
Changed by Changed by: Mgr. Eva Dubská, učo 77638. Changed: 1/3/2024 20:54.
Abstract
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 SARS-CoV-2, the digital ULISA additionally improved the LOD by a factor of 10 compared to the analog readout.
Links
GA21-03156S, research and development projectName: Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů
Investor: Czech Science Foundation
LM2018127, research and development projectName: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
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