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Registration Number

0223U001312, 0121U108464 , R & D reports

Title

Experimental substantiation of improvement of means of laboratory diagnostics of African swine fever

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Head

Sytiuk Mykola P.,

Registration Date

27-01-2023

Organization

Institute of Veterinary Medicine of the National Academy of Agrarian Sciences of Ukraine

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 For the detection of ASFV DNA, it was decided to use the MEB recommended primers to the target conserved region of the B646L gene of 250 nucleotide pairs (bp), which encodes the P72 protein. The table presents the nucleotide sequence and characteristics of the selected primers and fluorescent probes. Studying the effect of methods of DNA extraction from biological materials on the reproducibility and sensitivity of the PCR reaction using primers for internal control of extraction, it was established that the analytical sensitivity of the diagnostic kit at concentrations of 1.0×107 - 1.0×101 copies/cm3 was 100%. During the study of the material containing PRRSV, PTV and min. Aujeski, false results and non-specific reactions with foreign viruses were not observed, i.e. the specificity of the reaction was 100%). The obtained data also indicate a high convergence of the results of qualitative detection of ASFV DNA. As a result of the conducted research, we selected the optimal composition of the reaction mixture: 10× Taq Buffer with (NH4)2SO4, 2.5 mM MgCl2, 4 mM of each dNTP, 1 Unit of Taq DNA polymerase, 1 Unit of revertase.

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2023-01-27