METHOD FOR IDENTIFICATION OF BEAR TISSUE (URSUS) IN DRY FODDER AND SEMI-FINISHED MEAT PRODUCTS Russian patent published in 2020 - IPC C12Q1/68 

FIELD: microbiology.

SUBSTANCE: invention relates to veterinary microbiology, in particular to methods for determining species of meat using polymerase chain reaction. In method of identification of bears (Ursus) tissue DNA in dry fodders and semi-finished meat products, including DNA extraction from animal tissue by sorption method, polymerase chain reaction with fluorescent detection with 45 cycles of amplification in real time using oligonucleotide primers specific for the DNA genome region, probes, fluorescent dyes: for a specific signal for an animal – JOE/Yellow and Cy5/Red – for an internal control sample in the form of a bacteriophage T4 suspension with concentration of 5×10³ copies of nucleotide sequences per 1 mcl, positive control sample in the form of a mixture containing fragments of animal genomes and bacteriophage T4 with a nucleotide sequence: T4F: 5'-TACATATAAATCACGCAAAGC-3' – forward primer; T4R: 5'-TAGTATGGCTAATCTTATTGG-3' – reverse primer; T4P: SU5-5'-ACATTGGCACTGACCGAGTTC-3'-BHQ1 – probe, taken in ratio of 1:1, and measurement of accumulation of fluorescent signals through 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, DNA is recovered from bear tissue (Ursus) and the internal control sample is bacteriophage T4 phagolysate, and for the positive control sample – fragments of native bacteriophage T4 and bear tissue (Ursus) genomes with the following nucleotide sequence: Ursus F CCCGATAGACCTCACCACCT – forward primer, Ursus R CACCCCATGGGTTACACCTT – reverse primer, Ursus P CTTGTTCCTTTTAAGGGTTTGCTG-BHQ1 – probe.

EFFECT: high accuracy of identifying species identity, simple process of preparing and reducing its cost.

1 cl, 5 tbl