FIELD: biotechnology.
SUBSTANCE: method for indirectly determining the titer of infectious activity of canine mastadenovirus in raw materials for vaccines using real-time PCR, which includes the following steps: isolation of canine mastadenovirus DNA using a 3M guanidine thiocyanate solution, 75% ethyl alcohol solution and 80% isopropyl alcohol solution; carrying out real-time PCR with detection of the fluorescent signal of logistic curves for the studied samples using developed original oligonucleotides:
CAdV-T-F17638 primer with design 5'-GAAGCTACTATTATGAGACCA-3',
CAdV-T-R17702 primer with design 5'-TCTGTGTTTCTGTCTTGCAA-3',
CAdV-T-P17667-probe with design 5' -FAM-CGGGTCAATCATCTCAAC-RTQ1-3',
determination of the quantitative assessment cycle based on real-time PCR data and calculation of the titer of infectious activity of canine mastadenovirus in raw materials for the vaccine based on the developed logarithmic regression function.
EFFECT: possibility to quickly and with a high degree of reliability calculate the titer of infectious activity of CAdV in raw materials for vaccines.
2 cl, 5 ex, 3 dwg, 6 tbl
