DEVELOPMENT OF A REAL-TIME PCR TEST SYSTEM FOR THE DIAGNOSTICS OF BOVINE LUMPY SKIN DISEASE VIRUS

Lumpy skin disease virus (LSDV) is currently one of the most harmful and economically significant diseases of cattle in many countries, including Kazakhstan. Due to the unfavorable situation in the country for this type of disease, a quick diagnosis of this virus is required for a timely response and taking the necessary measures. To solve these problems, a test system based on the real-time polymerase chain reaction (RT-PCR) method has been developed. When developing the test system, modern molecular biological methods were used, with the help of which the most conservative and specific regions of the GPCR gene of the LSDV were determined. Using the selected regions, species-specific pairs of primers and probes were designed and synthesized. For setting RT-PCR, the temperature regime and the ratio of the components of the reaction mixture are optimized. The developed test system was tested in terms of specificity and sensitivity using DNA from LSDV, sheep pox, goat pox, camel pox viruses and plasmids with inserts of LSDV gene fragments. The results of laboratory tests МОЛЕКУЛЯРНАЯ БИОЛОГИЯ И ГЕННАЯ ИНЖЕНЕРИЯ 33 №8 2021 Ғылыми журнал Научный журнал The scientific journal БИОҚАУІПСІЗДІК ЖӘНЕ БИОТЕХНОЛОГИЯ БИОБЕЗОПАСНОСТЬ И БИОТЕХНОЛОГИЯ BIOSAFETY AND BIOTECHNOLOGY showed that the test system works specifically, and the sensitivity is 20 attograms of plasmid DNA in one reaction, which corresponds to 5 copies of the DNA molecules of the LSDV

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