FIELD: biotechnology.
SUBSTANCE: method of indirect determining of the titer of infectious activity of canine alpha-coronavirus production strain named RICH in raw materials for a vaccine using real-time RT-PCR is described. The method includes the stages of compiling a calibration panel of positive control samples of canine alpha-coronavirus production strain RICH, extracting RNA from virus-containing raw materials for the vaccine and controls, compiling a reaction mixture for reverse transcription and real-time PCR with oligonucleotides, and performing reverse transcription. The next stages are taking into account the fluorescent signal of each sample using a fluorimeter, determining the value of the threshold amplification cycle, establishing the relationship between the threshold amplification cycle and the titer of infectious activity of the strain in the raw materials for the vaccine, as well as determining the titer of the infectious activity of the strain.
EFFECT: development of a highly sensitive, highly specific, express method that does not involve the use of cell lines for indirectly determining the titer of infectious activity of canine alpha-coronavirus production strain named RICH in raw materials for a cultured vaccine using the RT-RT-PCR method.
5 cl, 10 dwg, 7 tbl, 6 ex
