METHOD FOR DETECTING DNA OF LEPTOSPIROSIS (LEPTOSPIRA SPP) CAUSATIVE AGENT 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 of a leptospirosis agent (Leptospira spp.) in farm animals, involving recovering nucleic acid from biological material from infected animals by a sorption method, staged one-stage polymerase chain reaction with simultaneous carrying out of not more than 40 amplification cycles with fluorescent detection using specific DNA genome of leptospirosis (Leptospira spp.) causative agent of oligonucleotide primers, probes, dyes and control samples in the form of internal and positive, measurement by JOE(HEX)/Yellow channel, accumulation of a fluorescent signal for a specific signal for testing the presence of the DNA genome of the leptospirarosis (Leptospira spp.) causative agent, and on the FAM/Green channel accumulation of the signal of the internal control sample, interpretation of the results based on the presence/absence of the intersection of the fluorescence curve with the threshold line, if the fluorescent signal accumulation curves are up to 35 cycles, the leptospirosis (Leptospira spp.) DNA is present and the reaction result is considered to be positive, and if the curves do not cross the threshold line or cross it after 35 cycles, the DNA of the leptospirosis (Leptospira spp.) causative agent is absent and the result of the reaction is negative, where the internal control sample is a 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 genome of the leptospirosis (Leptospira spp.Lpts) and a fragment of the bacteriophage T4 genome, taken in ratio 1:1.

EFFECT: invention enables to simplify detection of leptospirosis causative agent.

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