METHOD FOR DETECTION OF DNA OF THE BORDETLELOSIS (BORDETELLA BRONCHISEPTICA) AGENT GENOME IN FARM ANIMALS 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 genome of Bordetella bronchiseptica exciter in agricultural animals, involving DNA recovery from biological material by sorption method, staged one-stage polymerase chain reaction – with simultaneous carrying out of not more than 40 amplification cycles with real-time detection using oligonucleotide primers specific for the Bordetella bronchiseptica genome, probes, dyes and control samples in the form of internal and positive, measurement of a specific signal and a control signal through channels of corresponding dyes and interpretation of results, according to the invention, fluorescent detection is carried out, measuring the JOE(HEX)/Yellow accumulation of a fluorescent signal for a specific DNA signal of the Bordetella bronchiseptica genome, and the FAM/Green channel is a signal of internal control, interpretation of the results is carried out based on the presence/absence of the intersection of the fluorescence curve with the threshold line, if the curves of accumulation of the fluorescent signal are up to 35 cycles, the result of the reaction is considered positive, and if the curves do not cross the threshold line or cross it after 35 cycles, the result of the reaction is negative , wherein for internal control sample suspension of bacteriophage T4 with concentration of 5×10³ copies of nucleotide sequences at 1 mcl, and a positive control sample is a mixture of recombinant plasmid DNA containing a fragment of the DNA genome Bordetella bronchiseptica and a fragment of the bacteriophage T4 genome, taken in ratio 1:1.

EFFECT: invention enables reliable diagnosis of the DNA of Bordetella bronchiseptica.

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