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Post a Comment A fluorescent microspheres-based microfluidic test system for the detection of immunoglobulin G to SARS-CoV-2
- Authors: Shakurov R.I.1, Shansky Y.D.1, Prusakov K.A.1, Sizova S.V.1,2, Dudik S.P.1, Plotnikova L.V.3, Manuvera V.A.1, Klinov D.V.1, Lazarev V.N.1, Bespyatykh J.A.1,3, Basmanov D.V.1
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Affiliations:
- Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
- Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry
- Mendeleev University of Chemical Technology of Russia
- Issue: Vol 14, No 1 (2023)
- Pages: 44-53
- Section: Original Study Articles
- URL: https://journals.rcsi.science/clinpractice/article/view/142803
- DOI: https://doi.org/10.17816/clinpract278280
- ID: 142803
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Abstract
Background: The pandemic of the new coronavirus infection, COVID-19, is currently ongoing in the world. Over the years, the pathogen, SARS-CoV-2, has undergone a series of mutational genome changes, which has led to the spread of various genetic variants of the virus. Meanwhile, the methods used to diagnose SARS-CoV-2, to establish the disease stage and to assess the immunity, are nonspecific to SARS-CoV-2 variants and time-consumable. Thus, the development of new methods for diagnosing COVID-19, as well as their implementation in practice, is currently an important direction. In particular, application of systems based on chemically modified fluorescent microspheres (with a multiplex assay for target protein molecules) opens great opportunities.
Aim: development of a microfluidic diagnostic test system based on fluorescent microspheres for the specific detection of immunoglobulins G (IgG) to SARS-CoV-2.
Methods: A collection of human serum samples was characterized using enzyme-linked immunosorbent assay (ELISA) and commercially available reagent kits. IgG to SARS-CoV-2 in the human serum were detected by the developed immunofluorescent method using microspheres containing the chemically immobilized RBD fragment of the SARS-CoV-2 (“Kappa” variant) viral S-protein.
Results: The level of IgG in the blood serum of recovered volunteers was 9-300 times higher than that in apparently healthy volunteers, according to ELISA (p<0.001). Conjugates of fluorescent microspheres with the RBD-fragment of the S-protein, capable of specifically binding IgG from the blood serum, have been obtained. The immune complexes formation was confirmed by the fluorescence microscopy data; the fluorescence intensity of secondary antibodies in the immune complexes formed on the surface of microspheres was proportional to the content of IgG (r 0.963). The test system had a good predictive value (AUC 70.3%).
Conclusion: A test system has been developed, based on fluorescent microspheres containing the immobilized RBD fragment of the SARS-CoV-2 S-protein, for the immunofluorescent detection of IgG in the human blood serum. When testing the system on samples with different levels of IgG to SARS-CoV-2, its prognostic value was shown. The obtained results allow us to present the test system as a method to assess the level of immunoglobulins to SARS-CoV-2 in the human blood serum for the implementation in clinical practice. The test system can also be integrated into various microfluidic systems to create chips and devices for the point-of-care diagnostics.
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##article.viewOnOriginalSite##About the authors
Ruslan I. Shakurov
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Author for correspondence.
Email: ruslan.shakurov@rcpcm.org
ORCID iD: 0000-0002-5986-0676
SPIN-code: 9576-8093
Junior Research Associate
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowYaroslav D. Shansky
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: yar.shansky@rcpcm.org
ORCID iD: 0000-0003-4672-2474
SPIN-code: 7640-5940
PhD, Research Associate
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowKirill A. Prusakov
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: k.prusakov@rcpcm.org
ORCID iD: 0000-0002-7244-5741
SPIN-code: 9244-6581
Research Associate
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowSvetlana V. Sizova
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine; Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry
Email: sv.sizova@gmail.com
ORCID iD: 0000-0003-0846-4670
SPIN-code: 4322-1945
PhD, Research Associate
Russian Federation, 1a Malaya Pirogovskaya street, 119435 Moscow; MoscowStepan P. Dudik
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: stepan.dudik@rcpcm.org
ORCID iD: 0000-0002-3157-5902
SPIN-code: 8007-1870
Russian Federation, 1a Malaya Pirogovskaya street, 119435 Moscow
Lyudmila V. Plotnikova
Mendeleev University of Chemical Technology of Russia
Email: ntmdfs@gmail.com
Russian Federation, Moscow
Valentin A. Manuvera
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: vmanuvera@yandex.ru
ORCID iD: 0000-0002-2471-0563
SPIN-code: 9010-4521
PhD, Senior Research Assistant
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowDmitry V. Klinov
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: klinov.dmitry@mail.ru
ORCID iD: 0000-0001-8288-2198
SPIN-code: 9830-8561
PhD
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowVassili N. Lazarev
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: lazar0@mail.ru
ORCID iD: 0000-0003-0042-966X
SPIN-code: 1578-8932
PhD
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowJulia A. Bespyatykh
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine; Mendeleev University of Chemical Technology of Russia
Email: JuliaBes@rcpcm.org
ORCID iD: 0000-0002-4408-503X
SPIN-code: 6003-9246
PhD
Russian Federation, 1a Malaya Pirogovskaya street, 119435 Moscow; MoscowDmitriy V. Basmanov
Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine
Email: dmitry.basmanov@rcpcm.org
ORCID iD: 0000-0001-6620-7360
SPIN-code: 1801-6408
Research Associate
Russian Federation, 1a Malaya Pirogovskaya street, 119435 MoscowReferences
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