The rapid ELISA method for detection of orthopoxviruses

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Abstract

Introduction. Following the successful eradication of smallpox, mass vaccination against this disease was discontinued in 1980. The unvaccinated population continues to be at risk of infection due to military use of variola virus or exposure to monkeypox virus in Africa and non-endemic areas. In cases of these diseases, rapid diagnosis is of great importance, since the promptness and effectiveness of therapeutic and quarantine measures depend on it.

The aim of work is to develop a kit of reagents for enzyme-linked immunosorbent assay (ELISA) for fast and highly sensitive detection of orthopoxviruses (OPV) in clinical samples.

Materials and methods. The efficiency of virus detection was evaluated by single-stage ELISA in the cryolisate of CV-1 cell culture samples infected with vaccinia, cowpox, rabbitpox, and ectromelia viruses, as well as in clinical samples of infected rabbits and mice.

Results. The method of rapid ELISA was shown to allow the detection of OPV in crude viral samples in the range of 5.0 × 102–5.0 × 103 PFU/ml, and in clinical samples with a viral load exceeding 5 × 103 PFU/ml.

Conclusions. The assay involves a minimum number of operations and can be performed within 45 minutes, which makes it possible to use it in conditions of a high level of biosecurity. Rapid ELISA method was developed using polyclonal antibodies, which significantly simplifies and reduces the cost of manufacturing a diagnostic system.

About the authors

Nikita D. Ushkalenko

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: ushkalenko_nd@vector.nsc.ru
ORCID iD: 0000-0002-2171-7444

PhD student

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Anna V. Ersh

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: ersh_av@vector.nsc.ru
ORCID iD: 0000-0002-9220-1250

Candidate of Sciences in Biology, Research Officer

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Pavel V. Filatov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Author for correspondence.
Email: filatov_pv@vector.nsc.ru
ORCID iD: 0000-0001-7763-3808

Candidate of Sciences in Biology, Research Officer

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Alexander G. Poltavchenko

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: poltav@vector.nsc.ru
ORCID iD: 0000-0003-2408-5611

Doctor of Sciences in Biology, Head Scientist Researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

References

  1. Shchelkunov S.N., Marennikova S.S., Moyer R.W. Orthopoxviruses pathogenic for humans. In: Orthopoxviruses Pathogenic for Humans. Springer Science & Business Media; 2005. https://doi.org/10.1007/b107126 https://elibrary.ru/ueqvtt
  2. Richter J. Poxviruses. In: Tropical Dermatology. Elsevier; 2017: 152–65. https://doi.org/10.1016/B978-0-323-29634-2.00013-4
  3. Buller R.M.L. 170 – Poxviruses. In: Cohen J., Powderly W.G., Opal S.M., eds. Infectious Diseases. Elsevier; 2017: 1452–7.e1. https://doi.org/10.1016/B978-0-7020-6285-8.00170-2
  4. Sklenovská N. Monkeypox virus. In: Malik Y.S., Singh R.K., Dhama K., eds. Animal-Origin Viral Zoonoses. Singapore: Springer; 2020: 39–68. https://doi.org/10.1007/978-981-15-2651-0_2
  5. Supotnitskiy M.V. Natural smallpox, monkeypox. In: Supotnitskiy M.V. Biological Warfare. Introduction to the Epidemiology of Artificial Epidemic Processes and Biological Diseases [Biologicheskaya voyna. Vvedenie v epidemiologiyu iskusstvennykh epidemicheskikh protsessov i biologicheskikh porazheniy]. Moscow: Kafedra, Russkaya panorama; 2013: 834–86. (in Russian)
  6. Whitley R.J. Smallpox: a potential agent of bioterrorism. Antiviral Res. 2003; 57(1-2): 7–12. https://doi.org/10.1016/S0166-3542(02)00195-X
  7. Rimoin A.W., Mulembakani P.M., Johnston S.C., Lloyd Smith J.O., Kisalu N.K., Kinkela T.L., et al. Major increase in human monkeypox incidence 30 years after smallpox vaccination campaigns cease in the Democratic Republic of Congo. Proc. Natl Acad. Sci. USA. 2010; 107(37): 16262–7. https://doi.org/10.1073/pnas.1005769107
  8. Shchelkunov S.N. An increasing danger of zoonotic orthopoxvirus infections. PLoS Pathog. 2013; 9(12): e1003756. https://doi.org/10.1371/journal.ppat.1003756
  9. Townsend M.B., MacNeil A., Reynolds M.G., Hughes C.M., Olson V.A., Damon I.K., et al. Evaluation of the Tetracore Orthopox BioThreat® antigen detection assay using laboratory grown orthopoxviruses and rash illness clinical specimens. J. Virol. Methods. 2013; 187(1): 37–42. https://doi.org/10.1016/j.jviromet.2012.08.023
  10. Gavrilova E.V., Shcherbakov D.N., Maksyutov R.A., Shchelkunov S.N. Development of real-time PCR assay for specific detection of cowpox virus. J. Clin. Virol. 2010; 49(1): 37–40. https://doi.org/10.1016/j.jcv.2010.06.003 https://elibrary.ru/mxekaf
  11. Shchelkunov S.N., Shcherbakov D.N., Maksyutov R.A., Gavrilova E.V. Species-specific identification of variola, monkeypox, cowpox, and vaccinia viruses by multiplex real-time PCR assay. J. Virol. Methods. 2011; 175(2): 163–9. https://doi.org/10.1016/j.jviromet.2011.05.002
  12. Maksyutov R.A. Complex approach to species-specific detection of cowpox virus. Problemy osobo opasnykh infektsiy. 2016; (4): 60–3. https://doi.org/10.21055/0370-1069-2016-4-60-63 https://elibrary.ru/xgsxtz (in Russian)
  13. Stern D., Olson V.A., Smith S.K., Pietraszczyk M., Miller L., Miethe P., et al. Rapid and sensitive point-of-care detection of Orthopoxviruses by ABICAP immunofiltration. Virol. J. 2016; 13(1): 207. https://doi.org/10.1186/s12985-016-0665-5
  14. Rimoin A.W., Graham B.S. Whither monkeypox vaccination. Vaccine. 2011; 29(Suppl. 4): D60–4. https://doi.org/10.1016/j.vaccine.2011.09.004
  15. Stern D., Pauly D., Zydek M., Miller L., Piesker J., Laue M., et al. Development of a genus-specific antigen capture ELISA for orthopoxviruses – target selection and optimized screening. PLoS One. 2016; 11(3): e0150110. https://doi.org/10.1371/journal.pone.0150110
  16. Czerny C.P., Meyer H., Mahnel H. Establishment of an ELISA for the detection of orthopox viruses based on neutralizing monoclonal and polyclonal antibodies. Zentralbl. Veterinarmed. B. 1989; 36(7): 537–46. https://doi.org/10.1111/j.1439-0450.1989.tb00641.x
  17. Poltavchenko A.G., Ersh A.V., Filatov P.V., Ushkalenko N.D., Yakubitskiy S.N., Sergeev A.A., et al. Rapid detection of orthopoxviruses. Problemy osobo opasnykh infektsiy. 2021; (3): 106–13. https://doi.org/10.21055/0370-1069-2021-3-106-113 https://elibrary.ru/ywbpsy (in Russian)
  18. Nakane P.K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J. Histochem. Cytochem. 1974; 22(12): 1084–91. https://doi.org/10.1177/22.12.1084
  19. Ushkalenko N., Ersh A., Sergeev A., Filatov P., Poltavchenko A. Evaluation of rapid dot-immunoassay for detection orthopoxviruses using laboratory-grown viruses and animal’s clinical specimens. Viruses. 2022; 14(11): 2580. https://doi.org/10.3390/v14112580
  20. Poltavchenko A.G., Ersh A.V., Taranov O.S., Yakubitskiy S.N., Filatov P.V. Rapid immunochemical method for the detection of orthopoxviruses (Orthopoxvirus, Chordopoxvirinae, Poxviridae). Voprosy virusologii. 2019; 64(6): 291–7. https://doi.org/10.36233/0507-4088-2019-64-6-291-297 https://elibrary.ru/tjsbcf (in Russian)
  21. Sergeev A.A., Bulychev L.E., P’yankov O.V., Sergeev A.A., Bodnev S.A., Kabanov A.S., et al. Sensitivity of different animal species to monkeypox virus. Problemy osobo opasnykh infektsiy. 2012; (1): 88–91. https://doi.org/10.21055/0370-1069-2012-1(111)-88-91 https://elibrary.ru/orihiz (in Russian)
  22. Poltavchenko A., Ersh A., Filatov P., Yakubitskiy S. Rapid protocol of dot-immunnoassay for orthopoxviruses detection. J. Virol. Methods. 2020; 279: 113859. https://doi.org/10.1016/j.jviromet.2020.113859 https://elibrary.ru/wxwshr
  23. Gilchuk I., Gilchuk P., Sapparapu G., Lampley R., Singh V., Kose N., et al. Cross-neutralizing and protective human antibody specificities to poxvirus infections. Cell. 2016; 167(3): 684–94.e9. https://doi.org/10.1016/j.cell.2016.09.049

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2. Fig. 1. Calibration plot for vaccinia virus

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3. Fig. 2. Calibration plot for rabbitpox virus

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Copyright (c) 2023 Ushkalenko N.D., Ersh A.V., Filatov P.V., Poltavchenko A.G.

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