Design and optimization of the test-system component composition for laboratory diagnosing of lumpy skin disease by PCR

  • М.D. Almezhanova; К.A. Shorayeva; A.A. A.A. Tlepov; N.T. Tumenbayeva; N.N. Mukhami; K.D. Zakarya; 1 - Research Institute for Biological Safety Problems, Kazakhstan, Gvardeyskiy; 2 - Taraz State University named after M.Kh. Dulati, Kazakhstan, Taraz

Abstract

The actual direction of veterinary diagnostics is the development of diagnostic test systems for thedetection of infectious agents, which will help maintain a favorable ecological environment. The interaction of human society and nature has become one of the most important problems of our time. Onthe way of solving environmental problems, the world scientists are developing different methods ofcombating and preventing infectious diseases.The actual direction of veterinary diagnostics is the development of diagnostic test-systems for identifying infectious agents. The development of a domestic test-system for laboratory diagnosing lumpyskin disease of cattle (LSD) using modern molecular-genetic methods is the most important task that willallow detecting deoxyribonucleic acid (DNA) of LSD virus quickly and accurately in the disease earlystages. The aim of study is the design and optimization of the test-system component composition for laboratory diagnosing LSD by polymerase chain reaction (PCR). The given article presents the results ofthe design and optimization of the test-system component composition and working out the temperatureand time regime of the PCR.During the experiment was optimized the concentrations of the reaction mixture main components– primers, dNTPs, MgCl2, and Taq DNA polymerase. According to the obtained experimental data, 0,25μl (5 units/μl) of Taq DNA polymerase, 1 μl (20 pmol) of primers, 25 mM MgCl2 – 2 μl and dNTPs Mix(10 mM) – 1 μl are enough to produce a specific product of LSD in a volume of 25 μl for PCR with theprocessed temperature and time regime.As a result of the conducted studies, for the first time in the Republic of Kazakhstan have been processed the PCR conditions to identify the LSD virus.Based on the results obtained will be created a test-system in the veterinary practice of the Republicfor the diagnosing LSD based by PCR.Key words: lumpy skin disease, polymerase chain reaction, test-system, diagnosis, environment.
Published
2020-03-26
How to Cite
N.N. MUKHAMI; K.D. ZAKARYA;, М.D. Almezhanova; К.A. Shorayeva; A.A. A.A. Tlepov; N.T. Tumenbayeva;. Design and optimization of the test-system component composition for laboratory diagnosing of lumpy skin disease by PCR. Eurasian Journal of Ecology, [S.l.], v. 62, n. 1, p. 58-66, mar. 2020. ISSN 2617-7358. Available at: <https://bulletin-ecology.kaznu.kz/index.php/1-eco/article/view/1023>. Date accessed: 28 may 2020.
Section
ОЦЕНКА ДЕЙСТВИЯ ЗАГРЯЗНИТЕЛЕЙ ОКРУЖАЮЩЕЙ СРЕДЫ НА БИОТУ И ЗДОРОВЬЕ НАСЕЛЕНИЯ