THE SELECTION AND OPTIMIZATION OF PRIMERS FOR TYPE ASSIGNMENT OF FOOT-AND-MOUTH DISEASE VIRUS OF TYPES A, O
The existing methods of foot-and-mouth disease (FMD) diagnosis and the most common virus types detected in the Republic are briefly described. The methods of selection and optimization of species-specific primers are presented, and the results of the experiment on the type assignment of FMD virus using reverse transcription PCR are discussed. To run polymerase chain reaction (PCR) by the developed primers, the temperature regime of primer annealing and denaturation time of the site was set. The analysis of amplification products was performed by using 2.5% agarose gel. To confirm the authenticity of the obtained results, sequencing of PCR products obtained by the developed primers was performed. The sequencing revealed that the amplification products contained the RNA of FMD virus of types A and O. Diagnostic reactions to other types of the virus were negative. Consequently, the selected species-specific primers are characterized by high specificity and sensitivity and ensured valid results.