METHOD FOR DETECTING DNA OF RHINOTRACHEITIS VIRUS (BOVINE HERPES VIRUS 1, BOHV-1) IN CATTLE Russian patent published in 2019 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention is a method for detecting DNA of rhinotracheitis virus (bovine herpes virus 1, BoHV-1) in cattle, involving DNA recovery by sorption method, staged one-stage polymerase chain reaction with simultaneous amplification cycles with real-time detection using oligonucleotide primers, probes, dyes and control samples specific for genome segment of rhinotracheitis virus (bovine herpes virus I, BoHV-1) in the form of internal and positive, measurement of specific signal and control signal via channels of corresponding dyes and interpretation of results, according to the invention, fluorescent detection is carried out, measuring the JOE(HEX)/Yellow accumulation of the fluorescent signal for a specific DNA signal of rhinotracheitis virus (bovine herpes virus 1, BoHV-1), and FAM/Green channel signal internal control sample and interpretation of results is carried out based on presence/absence of intersection of fluorescence curve with threshold line, if the accumulation curves of the fluorescent signal reach 35th cycle, the result of the reaction is considered to be positive, and if the curves do not cross the threshold line or cross it after 35 cycles, the reaction result is negative, wherein for internal control sample suspension of bacteriophage T4 with concentration of 5×10³ copies of nucleotide sequences at 1 mcl, and for a positive control sample there used is a mixture of recombinant plasmid DNA containing a fragment of genome of DNA of rhinotracheitis virus (bovine herpes virus 1, BoHV-1) and a fragment of genome of bacteriophage T4, taken in ratio 1:1.

EFFECT: invention enables authentically diagnosing a causative agent of cattle rhinotracheitis virus DNA.

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