Email this article Rapid immunochemical method for the detection of orthopoxviruses (Orthopoxvirus, Chordopoxvirinae, Poxviridae)
- Authors: Poltavchenko A.G.1, Ersh A.V.1, Taranov O.S.1, Yakubitskiy S.N.1, Filatov P.V.1
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Affiliations:
- State Research Center of Virology and Biotechnology «Vector»
- Issue: Vol 64, No 6 (2019)
- Pages: 291-297
- Section: ORIGINAL RESEARCH
- Submitted: 03.02.2020
- Accepted: 03.02.2020
- Published: 20.12.2019
- URL: https://virusjour.crie.ru/jour/article/view/256
- DOI: https://doi.org/10.36233/0507-4088-2019-64-6-291-297
- ID: 256
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Full Text
Abstract
Introduction. The abolition of smallpox vaccination has led to the disappearance of population immunity to pox viruses. However, the threat of infection by pathogenic orthopoxviruses persists and determines the need to develop sensitive and operational methods for indicating pathogens.
Objectives and purposes. Development of a sensitive, fast and easy-to-use immunochemical test for the detection of orthopoxviruses in the «point of care» format.
Material and methods. We used preparations of cultural vaccinia virus (VV) with varying degrees of purification, polyclonal antibodies from hyperimmune rabbit serum, and equipment from a previously developed autonomous kit for dot-immunoassay on flat protein arrays.
Results and discussion. It has been established that rabbit polyclonal antibodies can be used in a single-stage dotanalysis, both as a capture agent immobilized on a substrate and as a detection reagent bound with colloidal gold particles. It is shown that the effectiveness of the detection of VV is inversely related to the degree of purification of viruses from sub-viral structures. The sensitivity of the rapid detection of viruses in a crude preparation was about 30 times higher than in pure viral material. The increase in sensitivity, presumably, occurs due to binding to the capture antibodies of subviral structures, which form large aggregates of sensitized gold particles. The test does not detect cross-reactions with heterogeneous viruses (measles, rubella and chickenpox) that cause exantematous diseases.
Conclusion. The one-stage variant of the dot-immunoassay reduces the analysis time to 40 minutes and improves the detection sensitivity of orthopoxviruses in crude viral preparations to the range of 105-104 PFU / ml. Full makeup, ease of analysis and the ability to visually accounting for results allow the test to be used outside of laboratories.
Objectives and purposes. Development of a sensitive, fast and easy-to-use immunochemical test for the detection of orthopoxviruses in the «point of care» format.
Material and methods. We used preparations of cultural vaccinia virus (VV) with varying degrees of purification, polyclonal antibodies from hyperimmune rabbit serum, and equipment from a previously developed autonomous kit for dot-immunoassay on flat protein arrays.
Results and discussion. It has been established that rabbit polyclonal antibodies can be used in a single-stage dotanalysis, both as a capture agent immobilized on a substrate and as a detection reagent bound with colloidal gold particles. It is shown that the effectiveness of the detection of VV is inversely related to the degree of purification of viruses from sub-viral structures. The sensitivity of the rapid detection of viruses in a crude preparation was about 30 times higher than in pure viral material. The increase in sensitivity, presumably, occurs due to binding to the capture antibodies of subviral structures, which form large aggregates of sensitized gold particles. The test does not detect cross-reactions with heterogeneous viruses (measles, rubella and chickenpox) that cause exantematous diseases.
Conclusion. The one-stage variant of the dot-immunoassay reduces the analysis time to 40 minutes and improves the detection sensitivity of orthopoxviruses in crude viral preparations to the range of 105-104 PFU / ml. Full makeup, ease of analysis and the ability to visually accounting for results allow the test to be used outside of laboratories.
About the authors
A. G. Poltavchenko
State Research Center of Virology and Biotechnology «Vector»
Author for correspondence.
Email: poltav@vector.nsc.ru
ORCID iD: 0000-0003-2408-5611
Dr. Sci. Biol., Lead Researcher of the department
Kol’tsovo, Novosibirsk Region, 630559
Россия
A. V. Ersh
State Research Center of Virology and Biotechnology «Vector»
Email: fake@neicon.ru
ORCID iD: 0000-0002-9220-1250
Kol’tsovo, Novosibirsk Region, 630559
Россия
O. S. Taranov
State Research Center of Virology and Biotechnology «Vector»
Email: fake@neicon.ru
ORCID iD: 0000-0002-6746-8092
Kol’tsovo, Novosibirsk Region, 630559
Россия
S. N. Yakubitskiy
State Research Center of Virology and Biotechnology «Vector»
Email: fake@neicon.ru
Kol’tsovo, Novosibirsk Region, 630559
Россия
P. V. Filatov
State Research Center of Virology and Biotechnology «Vector»
Email: fake@neicon.ru
ORCID iD: 0000-0001-7763-3808
Kol’tsovo, Novosibirsk Region, 630559
Россия
References
- Rimoin A.W., Graham B.S. Whither monkeypox vaccination. Vaccine. 2011; 29(Suppl. 4): D60-4. DOI: https://doi.org/10.1016/j.vaccine.2011.09.004
- Бондарев В.П., Терентьев А.И., Мельников С.А., Бондарева Т.А. Внедрение таблетированной оспенной вакцины «ТЭОВАК» в серийное производство для обеспечения биологической безопасности населения Российской Федерации. Проблемы особо опасных инфекций. 2010; (2): 66-8.
- Fitzgibbon J.E., Sagripanti J. Simultaneous identification of orthopoxviruses and alphaviruses by oligonucleotide macroarray with special emphasis on detection of variola and Venezuelan equine encephalitis viruses. J. Virol. Methods. 2006; 131(2): 160-7. DOI: https://doi.org/10.1016/j.jviromet.2005.08.007
- Максютов Р.А. Комплексный подход к видоспецифичной детекции вируса оспы коров. Проблемы особо опасных инфекций. 2016; (4): 60-3. DOI: https://doi.org/10.21055/0370-1069-2016-4-60-63
- Pulford D., Meyer H., Brightwell G., Damon I., Kline R., Ulaeto D. Amplification refractory mutation system PCR assays for the detection of variola and Orthopoxvirus. J. Virol. Methods. 2004; 117(1): 81-90. DOI: https://doi.org/10.1016/j.jviromet.2004.01.001
- Probst A., Besse A., Favry E., Imbert G., Tanchou V., Castell F.A., et al. Human CD4 T cell epitopes selective for Vaccinia versus Variola virus. Mol. Immunol. 2013; 53(4): 453- 9. DOI: https://doi.org/10.1016/j.molimm.2012.10.011
- Chen L., Wang H., Guo T., Xiao C., Liu L., Zhang X., et al. A rapid point-of-care test for dengue virus-1 based on a lateral flow assay with a near-infrared fluorescent dye. J. Immunol. Methods. 2018; 456: 23-7. DOI: https://doi.org/10.1016/j.jim.2018.02.005
- Ерш А.В., Полтавченко А.Г., Пьянков С.А., Агафонов А.П., Кривенчук Н.А., Буторин Д.В. Метод комплексной оценки гуморального иммунитета к детским вакциноуправляемым вирусным инфекциям. Вопросы вирусологии. 2015; 60(1): 45-9.
- Poltavchenko A.G., Nechitaylo O.V., Filatov P.V., Ersh A.V., Gureyev V.N. Multiplex method for initial complex testing of antibodies to blood transmitted diseases agents. J. Virol. Methods. 2016; 236: 231-6. DOI: https://doi.org/10.1016/j.jviromet.2016.08.003
- Скоупс Р. Методы очистки белков. Пер с англ. М.: Мир; 1985: 66-72.
- Poltavchenko A.G., Zaytzev B.N., Ersh A.V., Korneev D.V., Taranov O.S., Filatov P.V., et al. The selection and optimization of the detection system for self-contained multiplexed dot-immunoassay. J. Immunoassay Immunochem. 2016; 37(5): 540-54. DOI: https://doi.org/10.1080/15321819.2016.1174134
- Poltavchenko A.G., Zaitsev B.N., Ersh A.V., Taranov O.S., Korneev D.V., Nikonov A.M. Selection of Substrate Material for Protein Matrices. Prot. Met. Phys. Chem+. 2016; 52(2): 301-7. DOI: https://doi.org/10.1134/S2070205116020234
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