METHOD FOR DETECTION OF DNA OF NODULAR DERMATITIS VIRUS (LSDV) IN ANIMAL BIOLOGICAL MATERIAL BY MEANS OF POLYMERASE CHAIN REACTION IN REAL TIME Russian patent published in 2020 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention is a method for detecting DNA of nodular dermatitis virus (LSDV) in biological material of animals using a real-time polymerase chain reaction involving DNA recovery of an infectious animal pathogen by a sorption method, staged one-stage polymerase chain reaction with fluorescent detection with 40 amplification cycles in real time using oligonucleotide primers, probes, fluorescent dyes specific for the DNA genome region of the causative agent of the infectious animal disease: for a specific animal signal and FAM/Green for an internal control sample in the form of a bacteriophage T4 suspension with concentration of 5×10³ phage particles per 1 mcl, positive control sample in the form of a mixture containing in the same volume ratio fragments of animal genomes and bacteriophage T4 with a nucleotide sequence: T4F: 5'-TACATATAAATCACGCAAAGC-3' – direct primer; T4R: 5'- TAGTATGGCTAATCTTATTGG-3' – reverse primer; T4P: CY5-5'-ACATTGGCACTGACCGAGTTC-3'-BHQ1 – probe, measurement of accumulation of fluorescent signals via channels of corresponding fluorescent dyes, interpretation of results based on the presence or absence of the intersection of the fluorescence curve with the threshold line, if the fluorescence signal accumulation curves are up to 35 cycles, 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 result of the reaction is negative, according to the invention, nodular dermatitis virus (LSDV) DNA is recovered from the animal's biological material, and the bacteriophage T4 phage lysate is used for the internal control sample, and for a positive control sample – fragments of genomes of native bacteriophage T4 and fragment of genome of nodular dermatitis virus (LSDV) with the following nucleotide sequence: LSDV-F 5-TTTAGGAGATAAGATTGTCAC-3' direct primer; LSDV-R 5'-GGAGATTTTATTATGAGTGGC-3' reverse primer; LSDV-P ROX-5'-GGTTAATTTTTTACCACAAATAGG-3'-BHQ2 probe, wherein the DNA of the nodular dermatitis virus (LSDV) uses a ROX / Orange fluorescent dye.

EFFECT: invention widens functional capabilities, increases sensitivity and simplifies the process of preparing an internal control sample.

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