FIELD: virology; veterinary science.
SUBSTANCE: invention relates to veterinary virology, to means of molecular diagnostics, in particular, to differentiation of the vaccine strain "Archie" of the causative agent of canine parvoviral enteritis from other strains and field isolates of PVE using a highly sensitive HRM-analysis using a tandem fluorescent dye SN v428. Main advantage of the present invention is the possibility of performing differential analysis in a short period of time – not more than 2 hours. Developed method was used for the first time to solve this problem. In the target DNA region of the VP1 gene of the causative agent of parvoviral enteritis, there are two nucleotide mutations unique to the vaccine strain "Archie", which made it possible to develop highly specific original oligonucleotide primers CPV-F2524 and CPV-R2676, calculated for amplification of the target DNA region of the VP1 gene of the causative agent of parvoviral enteritis of size 176 bp and analysis of maximum melting points of PCR products using tandem fluorescent dye SN v428.
EFFECT: developed method is characterized by 100% analytical specificity; in 95% confidence interval diagnostic sensitivity for this method is 99.05-100%, diagnostic specificity – 98.62-100%.
2 cl, 4 dwg, 6 tbl, 3 ex
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