Efficiency of definition of Mycoplasma bovis in milk cows with mastitis using a polymerase chain reaction in the real time on a microchip with liophilized systems

The aim of our study was to analyze cows' milk for mastitis, caused by Mycoplasma bovis, using modern molecular genetic methods - real-time polymerase chain reaction (PCR Real-time) on a microchip with lyophilized test systems. Mastitis of farm animals cause significant economic damage to livestock farmers. PCR real-time is a more rapid, effective, sensitive and chip method than the standard cultural technique, and can be used as an alternative method. In the study were done molecular-genetic study of milk samples of cows with hidden and clinically manifested mastitis. For Polymerase chain reaction for the detection and identification of Mycoplasma bovis from mastitis milk were used samples from cows of black-and-white breed from husbandries of the North-West region. DNA extraction was performed from clinical specimens taken from farms. For amplification microarray used RT-PCR cycler "AriaDNA" and microchip with lyophilized reagents developed SC "Lumex". In the samples were identified microorganisms Mycoplasma bovis. Ruminant mycoplasmoses are important diseases worldwide, that results in sugnificant economic loses. The method of molecular genetic diagnosis in the microchip format allows to reduce the preparing time for the study, simplify and speed up the analysis procedure, and also makes it possible to screen a large number of samples in a short time.

ПЦР в режиме реального времени, Mycoplasma bovis, микрочипы, микоплазмозы, диагностика, коровы, PCR real-time, Mycoplasma bovis, microarray, mycoplasmosis, diagnostics, cows

1. Испытания универсального лабораторного метода диагностики мико-плазмозов животных / А. А. Сухинин, С. А. Макавчик, М. В. Виноходова, О. В. Прасолова // Вопр. нормативноправового регулирования в ветеринарии. - 2015. - № 1. - С. 40-46.

2. Применение полимеразной цепной реакции в молекулярной диагностике инфекционных болезней животных : учеб. пособие / А. А. Сухинин, С. А. Макавчик, О. В. Прасолова, М. В. Виноходова.- Санкт-Петербург : Изд-во ФГБОУ ВО СПБГАВМ, 2017. - 96 с.

3. Сухинин, А. А. Бактериологический и молекулярно-генетический метод для выделения и идентификации Mycoplasma bovis у крупного рогатого скота / А. А. Сухинин, Л. И. Смирнова, С. А. Макавчик // Вопр. нормативноправового регулирования в ветеринарии. - 2016. - № 4. - C. 80-84.

4. Fox, L. K. Mycoplasma mastitis: a review of transmission and control / L. K. Fox, J. H. Kirk, A. Britten // J. of veterinary medicine. B, Infectious diseases and veterinary public health. - 2005. - Vol. 52, iss. 4. - P. 153-60.

5. Incidence of clinical mastitis in dairy herds grouped in three categories by bulk milk somatic cell counts / H. W. Barkema [et al.] // J. Dairy Science. -1998. - Vol. 81, № 2. - P. 411-419.

6. Makavchik, S. A. Identification Bovis bacteria by polymerase chain reaction and sequencing / S. A. Makavchik, A. A. Sukhinin, S. R. Abgaryan, I. V. Belkina // Research j. of pharmaceutical, biological and chemical sciences. - 2019. - Vol. 10, iss. 1. - P. 2004-2012.

7. Pfutzner, H. Mycoplasma bovis as an agent of mastitis, pneumonia, arthritis and genital disorders in cattle / H. Pfutzner, K. Sachse // Rev. Scientifiqueet techn. (off. Intern. Epizootics). - 1996. -Vol. 15, № 4. - P. 1477-1494.

8. Shivakumar, V. Biofilms community behavior by bacteria / V. Shivakumar, D. Chakravortty // Resonance. - 2014. -Vol. 19, Iss. 11. - P. 1005-1016.

9. Simmons, W. L. The Vsa proteins modulate susceptibility of Mycoplasma pulmonis to complement killing, heamadsorption, and adherence to polystyrene / W. L. Simmons, K. Dybvig // Infections and Immunology -2003. - Vol. 71, № 10. - P. 5733-5738.

10. Trends In Therapeutic and Prevention Strategies for Management of Bovine Mastitis: An Overview / Tiwari J. G. [et al.] // J. of Vaccines and Vaccination. - 2013. - Vol. 4, iss. 2. - P. 1-11.