METHOD FOR WOODPECKER (PICIDAE) TISSUE DNA DETERMINATION IN DRY FEED 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. DNA is recovered from the woodpecker bird (Picidae). Polymerase chain reaction with fluorescent detection is carried out with 45 cycles of real-time amplification using bird-specific DNA of birds of the family Woodpecker (Picidae) oligonucleotide primers, probes, fluorescent dyes. For a positive control sample, a mixture containing DNA genome fragment of woodpecker family and bacteriophage T4 with the following nucleotide sequences is used: T4F TACATATAAATCACGCAAAGC – forward primer; T4R TAGTATGGCTAATCTTATTGG – reverse primer; T4P CY5 ACATTGGCACTGACCGAGTTC – probe; Pic F: 5'-GTCACAACATTTCATAGTATC-3' – forward primer; Pic R: 5'-TTCAGCATACTTGCTTCTTAC-3' – reverse primer; Pic P: FAM-5'-GAGCAGCACTAACCTCATATGCAG-3-BHQ1 – probe. Accumulating fluorescent signals through channels of corresponding fluorescent dyes is measured, results are interpreted based on presence or absence of intersection of fluorescence curve with threshold line, if fluorescence signal accumulation curves reach 35th cycle, 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 reaction result is negative.

EFFECT: method increases accuracy of identification of species identity, simplifies the preparation process and reduces its cost.

1 cl, 5 tbl