FIELD: veterinary science; virology.
SUBSTANCE: invention relates to veterinary virology, to molecular diagnostics. Disclosed is a method for genotyping feline calicivirus isolates and strains by phylogenetic analysis using original oligonucleotide primers for a highly variable region of ORF2-ORF3 genes. Method involves using the developed original oligonucleotide primers with the following design: ORF2-F7250-primer - 5'- TTTCTGCCTCCTACATGGG-3' and ORF3-R7573-primer -5 - TATGAGTAAGGGTCRACCCTA-3' and includes the following stages of analysis and use of the following reaction components: – extraction of RNA from positive and negative controls and analysed samples on glass fibre filters GF/F using 5M guanidine thiocyate solution and mixture of 80 % propanol-2 and 70 % ethanol, taken in ratio 4:1, evaluating the degree of purity of the eluates and controlling the concentration of RNA in the sample; performing reverse transcription and real-time amplification reaction Q with developed original oligonucleotide primers and annealing temperature equal to 55 °C; detection of PCR products using gel electrophoresis; purification of amplicons, determination of degree of purity of PCR products and determination of concentration of amplified fragments of cDNA; sequencing of samples and carrying out phylogenetic analysis based on data on nucleotide sequences of analysed samples of feline calicivirus strains. Developed method is specific with respect to the feline calicivirus genome.
EFFECT: method enables differentiation of calicivirus isolates and strains for creating effective vaccine preparations with respect to the causative agent of one or another genotype and diagnostic homologous test systems.
3 cl, 5 dwg, 7 tbl, 7 ex
