veterinaryВетеринария сегодняVeterinary Science Today2304-196X2658-6959"Veinard"veterinary-205Research ArticleБОЛЕЗНИ СВИНЕЙPORCINE DISEASESИспользование рекомбинантных белков рЗО, pK205R и pB602L в серодиагностике африканской чумы свинейUse of p30,pK205R and pB602L recombinant proteins for African swine fever serodiagnosisЯкуповМ. Р.YakupovM. R.mapat.89@mail.ruЯковлеваА. С.YakovlevaA. S.yakovleva_as@arriah.ruЩербаковА. В.ScherbakovA. V.ascherbakov@arriah.ruФГБУ «ВНИИЗЖ», г. ВладимирRussian Federation201502042018033747Copyright © Якупов М.Р., Яковлева А.С., Щербаков А.В., 20182018Якупов М.Р., Яковлева А.С., Щербаков А.В.Yakupov M.R., Yakovleva A.S., Scherbakov A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://veterinary.arriah.ru/jour/article/view/205

Экспрессией в £ coli получены рекомбинантные белки рЗО, pK205R и pB602L вируса африканской чумы свиней. На основе рекомбинантных антигенов разработан непрямой вариант иммуноферментного анализа для обнаружения антител к вирусу африканской чумы свиней. Диагностическая специфичность тест-систем pK205R-и рВ6021_-ИФА составляла 99,8%, рЗО-ИФА - 99,5%. На референтной панели положительных сывороток диагностическая чувствительность составляла 100% для рЗО-ИФА и pK205R-H®A и 94,12% - для рВ6021_-ИФА. При исследовании сыворотокот экспериментально и естественно инфицированных животных рЗО-ИФА и pK205R-H®A превосходили по чувствительности коммерческие тест-системы и были сравнимы с иммуноферментным анализом, который Всемирная организация здравоохранения животных рекомендует для серодиагностики африканской чумы свиней. Международные сличительные испытания по диагностике африканской чумы свиней подтвердили высокую специфичность и чувствительностьтест-систем рЗО-ИФА и pK205R-H®A.

P30, pK205R and pB602L recombinant proteins of African swine fever (ASF) virus were generated by expression in £ coli. Indirect ELISA for detection of antibodies to ASF virus was developed on the base of the said recombinant proteins. Diagnostic specificity of pK205R-and pB602L-ELISA tests systems was 99,8%, рЗО-ELISA - 99,5%. Diagnostic sensitivity p30-ELISA/pK205R-ELISA and pB602L-ELISA was 100% and 94,12%, respectively, when the reference panel of positive sera was used. The sensitivity рЗО-ELIS A and pK205R-ELIS A used for testing sera from experimentally and naturally infected animals was found to be higher than that one of commercial test systems and comparable to the ELISA recommended by the World Animal Health Organization for ASF serological diagnosis. High рЗО-ELISA and pK205R-ELISA specificity and sensitivity were confirmed by results of international proficiency testings for ASF diagnosis.

африканская чума свинейиммуноферментный анализрекомбинантные белкиантителаAfrican swine fever (ASF)enzyme-linked immunosorbent assay (ELISA)recombinant proteinsantibodiesReferencesМаниатис Т., Фрич Э., Сэмбрук Д. Методы генетической инженерии. Молекулярное клонирование. - М.: Мир, 1984, - 480 с.Маниатис Т., Фрич Э., Сэмбрук Д. Методы генетической инженерии. Молекулярное клонирование. - М.: Мир, 1984, - 480 с.Тимина А.М.,Якупов М.Р., Щербаков A.B. Результаты участия в международных сличительных испытаниях по диагностике африканской чумы свиней//Ветеринария сегодня. - 2015. - №1 (12). - С. 9-14.Тимина А.М.,Якупов М.Р., Щербаков A.B. Результаты участия в международных сличительных испытаниях по диагностике африканской чумы свиней//Ветеринария сегодня. - 2015. - №1 (12). - С. 9-14.African swine fever //Manual of DiagnosticTestsand Vaccines forTerrestrial Animals (Mammals, Birds and Bees) / OIE. - 7th ed. - Paris, France, 2012. - Vol. 2, Chapter 2.8.1. - P. 1067-1079.African swine fever //Manual of DiagnosticTestsand Vaccines forTerrestrial Animals (Mammals, Birds and Bees) / OIE. - 7th ed. - Paris, France, 2012. - Vol. 2, Chapter 2.8.1. - P. 1067-1079.African swine fever virus serodiagnosis: a general review with a focus on the analyses of African serum samples / C. Cubillos, S. Gómez-Sebastian, N. Moreno [et al.] //Virus Res. - 2013. - Vol. 173, № 1. - P. 159-167.African swine fever virus serodiagnosis: a general review with a focus on the analyses of African serum samples / C. Cubillos, S. Gómez-Sebastian, N. Moreno [et al.] //Virus Res. - 2013. - Vol. 173, № 1. - P. 159-167.Antigenic properties and diagnostic potential of African swine fever virus protein pp62 expressed in insect cells/С. Gallardo, E. Blanco, J.M. Rodriguez [et al.] //J. Clin. Microbiol. - 2006. - Vol. 44, № 3. - P. 950-956.Antigenic properties and diagnostic potential of African swine fever virus protein pp62 expressed in insect cells/С. Gallardo, E. Blanco, J.M. Rodriguez [et al.] //J. Clin. Microbiol. - 2006. - Vol. 44, № 3. - P. 950-956.Escribano J.M., Pastor M.J., Sânchez-Vizcaino J.M. Antibodies to bovine serum albumin in swine sera: implications for false-positive reactions in the serodiagnosis of African swine fever // Am. J. Vet. Res. - 1989. - Vol. 50, № 7. - P. 1118-1122.Escribano J.M., Pastor M.J., Sânchez-Vizcaino J.M. Antibodies to bovine serum albumin in swine sera: implications for false-positive reactions in the serodiagnosis of African swine fever // Am. J. Vet. Res. - 1989. - Vol. 50, № 7. - P. 1118-1122.High level expression ofthe major antigenic African swine fever virus proteins p54 and рЗО in baculovirus and their potential use as diagnostic reagents / J.M. Oviedo, F. Rodriguez, P. Gômez-Puertas [et al.] //J. Virol. Methods. - 1997. - Vol. 64, № 1. - P. 27-35.High level expression ofthe major antigenic African swine fever virus proteins p54 and рЗО in baculovirus and their potential use as diagnostic reagents / J.M. Oviedo, F. Rodriguez, P. Gômez-Puertas [et al.] //J. Virol. Methods. - 1997. - Vol. 64, № 1. - P. 27-35.Highly specific confirmatory western blot test of African swine fever virus antibody detection using the recombinant virus protein p54 / C.l. Alcaraz, F. Rodriguez, J.M. Oviedo [et al.] // J. Virol. Methods. - 1995.- Vol. 52. - P. 111-119.Highly specific confirmatory western blot test of African swine fever virus antibody detection using the recombinant virus protein p54 / C.l. Alcaraz, F. Rodriguez, J.M. Oviedo [et al.] // J. Virol. Methods. - 1995.- Vol. 52. - P. 111-119.Optimization and validation of recombinant serological tests for African swine fever diagnosis based on detection ofthe рЗО protein produced in Trichoplusia ni larvae / D.M. Pérez-Filgueira, F. Gonzâlez-Camacho, C. Gallardo [et al.] // J. Clin. Microbiol. - 2006. - Vol. 44, № 9. - P. 3114-3121.Optimization and validation of recombinant serological tests for African swine fever diagnosis based on detection ofthe рЗО protein produced in Trichoplusia ni larvae / D.M. Pérez-Filgueira, F. Gonzâlez-Camacho, C. Gallardo [et al.] // J. Clin. Microbiol. - 2006. - Vol. 44, № 9. - P. 3114-3121.Recombinant antigen targets for serodiagnosis of African swine fever / C. Gallardo, A.L. Reis, G. Kalema-Zikusoka [et al.] // Clin, and Vaccine Immunol. - 2009. - Vol. 16, -P. 1012-1020.Recombinant antigen targets for serodiagnosis of African swine fever / C. Gallardo, A.L. Reis, G. Kalema-Zikusoka [et al.] // Clin, and Vaccine Immunol. - 2009. - Vol. 16, -P. 1012-1020.Sancyez-Vizcaino J.M. African swine fever// Dis. of Swine / ed. B.E. Straw [et al.]. - 9th ed. - Ames, Iowa, 2006. - P. 291-298.Sancyez-Vizcaino J.M. African swine fever// Dis. of Swine / ed. B.E. Straw [et al.]. - 9th ed. - Ames, Iowa, 2006. - P. 291-298.Serodiagnosis of African swine fever using the recombinant protein рЗО expressed in insect larvae / M.G. Barderas, A. Wigdorovitz, F. Merelo [et al.] // J. Virol. Methods. - 2000. - Vol. 89, № 1 -2. - P. 129-136.Serodiagnosis of African swine fever using the recombinant protein рЗО expressed in insect larvae / M.G. Barderas, A. Wigdorovitz, F. Merelo [et al.] // J. Virol. Methods. - 2000. - Vol. 89, № 1 -2. - P. 129-136.Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus/A.L. Reis, R.M.E. Parkhouse, A.R. Penedos [et al.] //J. Gen. Virol. - 2007. - Vol. 88. - P. 2426-2434.Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus/A.L. Reis, R.M.E. Parkhouse, A.R. Penedos [et al.] //J. Gen. Virol. - 2007. - Vol. 88. - P. 2426-2434.

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