THEILERIA ANNULATA-НЫҢ РЕКОМБИНАНТТЫ ПРОТЕИНІН АЛУ ЖӘНЕ ОНЫҢ ДИАГНОСТИКАЛЫҚ ҚҰНДЫЛЫҒЫН АНЫҚТАУ
DOI:
https://doi.org/10.56339/2305-9397-2023-2-1-46-56Ключевые слова:
theileriosis, Theileria annulata surface protein, TaSP, recombinant protein, ELISA.Аннотация
Для эффективной борьбы с тейлериозом крупного рогатого скота необходимы мероприятия по предотвращению распространения инфекции на благополучные регионы. В этой ситуации важным моментом является проведение систематических и достаточно масштабных серологических исследований восприимчивого поголовья, для чего необходимо наличие у ветеринарных специалистов высокочувствительных и специфичных диагностикумов.
В рамках данных исследований была создана экспрессионная векторная система и отработан протокол очистки и рефолдинга рекомбинантного поверхностного протеина Theileria annulata (surface protein of T.annulata, TaSP). SDS-PAGE анализ очищенных проб показал достаточно высокий выход чистых препаратов рекомбинантного белка TaSP T. annulata с молекулярной массой 32 кДа. Полученный рекомбинантный протеин был использован в качестве антигена при оптимизации протокола иммуноферментного анализа (ИФА) для определения специфических антител в пробах плазмы крови в которых методом полимеразной цепной реакции (ПЦР), было определено наличие ДНК T.annulata. Результаты тестирования выборки условно положительных и отрицательных проб, определили сероактивность и достаточно высокий уровень специфичности полученного рекомбинантного антигена. Предварительные результаты показали перспективность использованного протеина для дополнительных исследований диагностической ценности и определения возможности его использования в качестве компонента иммунологических тест-систем.
Библиографические ссылки
ӘДЕБИЕТТЕР ТІЗІМІ
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Saaid, A.A. The protection afforded to cattle immunized with Theileria annulata infected cell line is enhanced by subunit vaccine candidate TaSP [Text] / A.A. Saaid [and etc.] // Transboundary and Emerging Diseases. - 2020. - Vol. 67. - P. 26-34.
Gubbels, M. J. Development of an indirect Tams1 enzyme-linked immunosorbent assay for diagnosis of Theileria annulata infection in cattle [Text] / M. J. Gubbels [and etc.] // Clinical Diagnostic Laboratory Immunology. - 2000. - Vol. 7. - № 3. - P. 404-411.
Jaramillo Ortiz, J.M. Development of an indirect ELISA based on a recombinant chimeric protein for the detection of antibodies against bovine babesiosis [Text] / J.M. Jaramillo Ortiz [and etc.] // Veterinary Sciences. - 2018. - Vol. 5. - № 1. - P. 13.
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REFERENCES
Zhao, S. Evaluating an indirect rMPSP enzyme-linked immunosorbent assay for the detection of bovine Theileria infection in China [Text] / S. Zhao [and etc.] // Parasitology Research. - 2017. - Vol. 116. - P. 667-676.
Dikov, G. Spravochnik po parazitozam sel’skohozyajstvennyh zhivotnyh v Respublike Kazakhstan [Text] / G. Dikov [and etc.] // Almaty. - 1994. - V. 1. - № 994. - P. 144-173.
Turganbayeva, G. Study of ixodid ticks on existence of blood parasites [Text] /
G. Turganbayeva [and etc.] // Journal of International Scientific Publications, Agruculture and Food. - 2016. - Vol. 2. - P. 229-239.
Sang, C. Tick distribution and detection of Babesia and Theileria species in Eastern and Southern Kazakhstan [Text] / C. Sang [and etc.] // Ticks and Tick-Borne Diseases. - 2021. - Vol. 12. - № 6. - P. 101817.
Guo, H. Molecular survey and characterization of Theileria annulata and Ehrlichia ruminantium in cattle from Northwest China [Text] / H. Guo [and etc.] // Parasitology international. - 2018. - Vol. 67. - № 6. - P. 679-683.
Yu, Z. Tick-borne pathogens and the vector potential of ticks in China [Text] / Z.Yu [and etc.] // Parasites & vectors. - 2015. - Vol. 8. - № 1. - P. 1-8.
Perfilyeva, Y. V. Tick-borne pathogens and their vectors in Kazakhstan–A review [Text] / Y. V. Perfilyeva [and etc.] // Ticks and tick-borne diseases. - 2020. - Vol. 11. - № 5. - P. 101498.
Seitzer, U. From molecule to diagnostic tool: Theileria annulata surface protein TaSP [Text] / U. Seitzer [and etc.] // Parasitology research. - 2007. - Vol. 101. - P. 217-223.
Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 [Text] // Nature. - 1970. - Vol. 227. - № 5259. - P. 680-685.
Kuibagarov, M. Theileria and Babesia infection in cattle – First molecular survey in Kazakhstan [Text] / M. Kuibagarov [and etc.] // Ticks and Tick-borne Diseases. - 2023. - Vol. 14.
- № 1. - P. 102078.
Manuja, A. Comparison of cellular schizont, soluble schizont and soluble piroplasm antigens in ELISA for detecting antibodies against Theileria annulata [Text] / A. Manuja [and etc.] // Veterinary parasitology. - 2000. - Vol. 87. - № 2-3. - P. 93-101.
Rajendran, C. Diagnosis of tropical bovine theileriosis by ELISA with recombinant merozoite surface protein of Theileria annulata (Tams1) [Text] / C. Rajendran [and etc.] // Journal of parasitic diseases. - 2014. - Vol. 38. - P. 41-45.
Prabhakaran, H.S. Evaluation of sporozoite and macroschizont antigen (Spm2) of Theileria annulata for its diagnostic potential [Text] / H.S. Prabhakaran [and etc.] // Ticks and Tick-Borne Diseases. - 2021. - Vol. 12. - № 4. - P. 101691.
Tian, Z. Development of an indirect ELISA based on the recombinant Spm2 protein for detection of tropical theileriosis [Text] / Z. Tian [and etc.] // Acta tropica. - 2018. - Vol. 182.
- P. 232-236.
Mohmad, A. Development of a recombinant TaSP-based Dot-ELISA for detection of Theileria annulata infection in cattle [Text] / A. Mohmad [and etc.] // Ticks and tick-borne diseases. - 2018. - Vol. 9. - № 6. - P. 1416-1420.
Elati, K. Sequence polymorphisms in a Theileria annulata surface protein (TaSP) known to augment the immunity induced by live attenuated cell line vaccine [Text] / K. Elati [and etc.] // Transboundary and Emerging Diseases. - 2022.
Saaid, A.A. The protection afforded to cattle immunized with Theileria annulata infected cell line is enhanced by subunit vaccine candidate TaSP [Text] / A.A. Saaid [and etc.] // Transboundary and Emerging Diseases. - 2020. - Vol. 67. - P. 26-34.
Gubbels, M. J. Development of an indirect Tams1 enzyme-linked immunosorbent assay for diagnosis of Theileria annulata infection in cattle [Text] / M. J. Gubbels [and etc.] // Clinical Diagnostic Laboratory Immunology. - 2000. - Vol. 7. - № 3. - P. 404-411.
Jaramillo Ortiz, J.M. Development of an indirect ELISA based on a recombinant chimeric protein for the detection of antibodies against bovine babesiosis [Text] / J.M. Jaramillo Ortiz [and etc.] // Veterinary Sciences. - 2018. - Vol. 5. - № 1. - P. 13.
Lira-Amaya, J.J. Comparative Study of Indirect Fluorescent Antibody, ELISA, and Immunochromatography Tests for Serological Diagnosis of Bovine Babesiosis Caused by Babesia bovis [Text] / J.J. Lira-Amaya [and etc.] // Animals. - 2021. - Vol. 11. - № 12. - P. 3358.