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

Abstract

The actual direction of veterinary diagnostics is the development of diagnostic test systems for the
detection 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. On
the way of solving environmental problems, the world scientists are developing different methods of
combating 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 lumpy
skin disease of cattle (LSD) using modern molecular-genetic methods is the most important task that will
allow detecting deoxyribonucleic acid (DNA) of LSD virus quickly and accurately in the disease early
stages. 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 of
the design and optimization of the test-system component composition and working out the temperature
and 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, 0,25
μl (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 the
processed 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 Republic
for the diagnosing LSD based by PCR.
Key words: lumpy skin disease, polymerase chain reaction, test-system, diagnosis, environment.