eifcЭпизоотология Иммунобиология Фармакология СанитарияEpizootology Immunobiology Pharmacology Sanitation2224-168XИнститут экспериментальной ветеринарии им. С.Н. Вышелесскогоeifc-193Research ArticleИММУНОБИОЛОГИЯIMMUNOBIOLOGYРазработка ИФА-диагностикума для выявления специфических антител к вирусу африканской чумы свинейDevelopment of ELISA-diagnosticum for detection of specific antibodies to African swine fever virusБорисовецД. С.BorisovetsD. S.

Борисовец Д.С., кандидат ветеринарных наук, доцент

г. Минск

Родригез-МаллонА. А.Rodriguez-MallonA. A.

Родригез-Маллон А.А., руководитель проекта

г. Гавана

ТяпшаЮ. И.TyapshaYu. I.

Тяпша Ю.И., кандидат ветеринарных наук, доцент

г. Минск

ЗгировскаяА. А.ZgirovskayaA. A.

Згировская А.А., кандидат биологических наук

г. Минск

ГерасименкоВ. И.GerasimenkoV. I.

Герасименко В.И., младший научный сотрудник

г. Минск

РУП «Институт экспериментальной ветеринарии им. С.Н. Вышелесского»BelarusЦентр генетической инженерии и биотехнологийCuba202529122025022935Copyright © Борисовец Д.С., Родригез-Маллон А.А., Тяпша Ю.И., Згировская А.А., Герасименко В.И., 20252025Борисовец Д.С., Родригез-Маллон А.А., Тяпша Ю.И., Згировская А.А., Герасименко В.И.Borisovets D.S., Rodriguez-Mallon A.A., Tyapsha Y.I., Zgirovskaya A.A., Gerasimenko V.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://eifc.belal.by/jour/article/view/193

Разработаны рекомбинантные антигены вируса африканской чумы свиней (АЧС) путем клонирования и экспрессии белков, кодируемых двумя генами данного вируса – p72 (B646L) и p30 (CP204L). Отработаны параметры адсорбции рекомбинантных белков вируса африканской чумы свиней и определены оптимальные концентрации компонентов ИФА-диагностикума для выявления антител к антигенам вируса АЧС. Разработан ИФА-диагностикум для выявления специфических антител к вирусу африканской чумы свиней. Проведена иммунизация кроликов и получены специфические сыворотки к вирусу АЧС. Проведен подбор оптимальных концентраций компонентов диагностикума для выявления антител к антигену вируса АЧС. Проведена проверка разработанного ИФА-диагностикума по показателю специфичности и чувствительности.

Recombinant antigens of the African swine fever virus (ASF) have been developed by cloning and expressing proteins encoded by two genes of this virus: p72 (B646L) and p30 (CP204L). The parameters of adsorption of recombinant proteins of the African swine fever virus have been worked out and optimal concentrations of ELISA diagnostic components have been determined for the detection of antibodies to antigens of the African swine fever virus. An ELISA diagnosticum has been developed to detect specific antibodies to the African swine fever virus. Rabbits were immunized and specific serums for the African swine fever virus were obtained. The selection of optimal concentrations of diagnostic components for the detection of antibodies to the ASF virus antigen was carried out. The developed ELISA diagnostic was tested in terms of specificity and sensitivity.

африканская чума свинейрекомбинантные антигеныИФА-диагностикумспецифичностьчувствительностьAfrican swine fever virusRecombinant antigensELISA diagnosticspecificitysensitivityReferencesКашкин, К. П. Методические разработки по клинической иммунологии / К. П. Кашкин, Е. Н. Степанова, Л. М. Скуинь. – М. : Медицина, 1997. – 518 с.Кашкин, К. П. Методические разработки по клинической иммунологии / К. П. Кашкин, Е. Н. Степанова, Л. М. Скуинь. – М. : Медицина, 1997. – 518 с.Gladue, D. P. Recombinant ASF live attenuated virus strains as experimental vaccine candidates / D. P. Gladue, M. V. Borca // Viruses. – 2022. – Vol. 14. – № 5. – P. 878.Gladue, D. P. Recombinant ASF live attenuated virus strains as experimental vaccine candidates / D. P. Gladue, M. V. Borca // Viruses. – 2022. – Vol. 14. – № 5. – P. 878.Viral vector vaccines against ASF: Problems and prospectives / R. K. Ravilov [et al.] // Frontiers in Veterinary Science. – 2022. – Vol. 9. – P. 830244.Viral vector vaccines against ASF: Problems and prospectives / R. K. Ravilov [et al.] // Frontiers in Veterinary Science. – 2022. – Vol. 9. – P. 830244.Vaccination with a gamma irradiation-inactivated African swine fever virus is safe but does not protect against a challenge / J. Pikalo [et al.] // Frontiers in Immunology. – 2022. – Vol. 13. – P. 832264.Vaccination with a gamma irradiation-inactivated African swine fever virus is safe but does not protect against a challenge / J. Pikalo [et al.] // Frontiers in Immunology. – 2022. – Vol. 13. – P. 832264.Adenovirus-vectored African Swine Fever Virus antigen cocktails are immunogenic but not protective against intranasal challenge with Georgia 2007/1 isolate / S. Lokhandwala [et al.] // Veterinary microbiology. – 2019. – Vol. 235. – P. 10–20.Adenovirus-vectored African Swine Fever Virus antigen cocktails are immunogenic but not protective against intranasal challenge with Georgia 2007/1 isolate / S. Lokhandwala [et al.] // Veterinary microbiology. – 2019. – Vol. 235. – P. 10–20.A solid-phase enzyme linked immunosorbent assay using monoclonal antibodies, for the detection of african swine fever virus antigens and antibodies / M. I. Vidal, M. Stiene, J. Henkel [et al.] // J. Virol. Methods. – 1997. – 66 (2). – Р. 211–218.A solid-phase enzyme linked immunosorbent assay using monoclonal antibodies, for the detection of african swine fever virus antigens and antibodies / M. I. Vidal, M. Stiene, J. Henkel [et al.] // J. Virol. Methods. – 1997. – 66 (2). – Р. 211–218.World Organization of Animal Health (OIE). African swine fever. Manual of Standards for diagnostic test and vaccines. – 2008.World Organization of Animal Health (OIE). African swine fever. Manual of Standards for diagnostic test and vaccines. – 2008.High doses of inactivated African swine fever virus are safe, but do not confer protection against a virulent challenge / E. Cadenas-Fernández [et al.] // Vaccines. – 2021. – Vol. 9. – № 3. – P. 242.High doses of inactivated African swine fever virus are safe, but do not confer protection against a virulent challenge / E. Cadenas-Fernández [et al.] // Vaccines. – 2021. – Vol. 9. – № 3. – P. 242.Immunization of pigs by DNA prime and recombinant vaccinia virus boost to identify and rank African swine fever virus immunogenic and protective proteins / J. K. Jancovich [et al.] // Journal of virology. – 2018. – Vol. 92. – №. 8. – P. 10.1128/jvi.02219-17.Immunization of pigs by DNA prime and recombinant vaccinia virus boost to identify and rank African swine fever virus immunogenic and protective proteins / J. K. Jancovich [et al.] // Journal of virology. – 2018. – Vol. 92. – №. 8. – P. 10.1128/jvi.02219-17.DNA-protein vaccination strategy does not protect from challenge with African swine fever virus Armenia 2007 strain / S. Y. Sunwoo [et al.] // Vaccines. – 2019. – Vol. 7, № 1. – P. 12.DNA-protein vaccination strategy does not protect from challenge with African swine fever virus Armenia 2007 strain / S. Y. Sunwoo [et al.] // Vaccines. – 2019. – Vol. 7, № 1. – P. 12.Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein-dye binding / M.M. Bradford // Anal. Biochem. – 1976. – Vol.72, № 1-2. – P. 248–254.Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein-dye binding / M.M. Bradford // Anal. Biochem. – 1976. – Vol.72, № 1-2. – P. 248–254.

The authors declare that there are no conflicts of interest present.