Русский
Development of a dot-assay for detecting antibodies to tick-borne encephalitis virus using recombinant EDIII-TBEV antigen and IgY antibodies
DOI: https://dx.doi.org/10.18565/epidem.2024.14.2.63-9
Poltavchenko A.G., Filatov P.V., Ersh A.V., Ushkalenko N.D., Shanshin D.V., Kolosova E.A., Shcherbakov D.N.
State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia
Materials and methods. Probes based on colloidal gold and secondary immunoreagents (Au-SpA, Au-SpG, Au-IgG-a/Hum and Au-IgY-a/Hum) were manufactured and studied in a dot analysis of human immunoglobulins of A, M and G classes. Experimental kits for dot-immunoassay of IgG to TBEV were created based on the probes and EDIII-TBEV.
Results. Experimental assessment of the characteristics of the kits using panels of blood serum samples containing and not containing IgG to TBEV, as well as panels of samples containing antibodies to pathogens of other flavivirus infections (dengue fever and yellow fever) showed 100% sensitivity and specificity of the dot analysis, which indicates the high selectivity of the recombinant EDIII-TBEV antigen. Probes based on polyclonal IgY-a/Hum are not inferior in sensitivity and provide a 10-20% more intense optical signal compared to probes based on mouse monoclonal IgG-a/Hum.
Conclusion. Probes based on IgY-a/Hum and recombinant EDIII-TBEV antigen can be considered as promising reagents for antibody analysis, in particular, for dot analysis on flat substrates. Kits created on their basis can be used to diagnose tick-borne encephalitis in remote areas with poorly developed laboratory infrastructure.
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About the Autors
Alexander G. Poltavchenko, ВD, Leading Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; poltav@vector.nsc.ru; https://orcid.org/0000-0003-2408-5611
Pavel V. Filatov, Cand. Biol. Sci., Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; filatov_pv@vector.nsc.ru; https://orcid.org/0000-0001-7763-3808
Anna V. Ersh, Cand. Biol. Sci., Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; ersh_av@vector.nsc.ru; https://orcid.org/0000-0002-9220-1250
Nikita D. Ushkalenko, Postgraduate Student, Junior Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; ushkalenko_nd@vector.nsc.ru; https://orcid.org/0000-0002-2171-7444
Daniil V. Shanshin, Junior Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; shanshin_dv@vector.nsc.ru; https://orcid.org/0000-0001-9985-1582
Evgeniуa A. Kolosova, Junior Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; kurchanovaea@gmail.com; https://orcid.org/0000-0002-8967-4719
Dmitry N. Shcherbakov, Cand. Biol. Sci., Leading Researcher, State Scientific Center for Virology and Biotechnology «Vector», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Koltsovo, Novosibirsk Region, Russia; scherbakov_dn@vector.nsc.ru; https://orcid.org/0000-0001-8023-4453
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