METHOD FOR IDENTIFICATION OF SPECIES IDENTITY OF MUTTON AND BEEF IN FOOD RAW MATERIALS, FODDER AND FOOD PRODUCTS Russian patent published in 2019 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention is a method for identification of species identity of mutton and beef in food raw materials, fodder and food products, including extraction of DNA from mutton (Ovis) and beef (Bos) by a sorption method, setting of a single-stage polymerase chain reaction, carrying out 45 amplification cycles with real-time detection using mutant DNA-specific mutton (Ovis) and beef (Bos) oligonucleotide primers, probes, dyes, internal control sample in form of bacteriophage suspension and positive control samples – containing DNA genome fragments of mutton (Ovis) and beef (Bos), measuring specific signals and control signal and interpreting the results, wherein further investigating food raw materials and food products containing mutton DNA of mutton (Ovis) and beef (Bos), polymerase chain reaction is carried out with fluorescent detection using a thermal cycler of Rotor-Gene Q type and accumulation of fluorescent signals is measured through channels of corresponding fluorescent dyes: JOE / Yellow for specific signal for mutton (Ovis); ROX / Orange – for beef (Bos) and Cy5 / Red – for an internal control sample, the interpretation of results is carried out based on the presence or absence of the intersection of the fluorescence curve with the threshold line, if the curves of accumulation of the fluorescent signal reach 35 cycles, the result of the reaction 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, wherein the internal reference sample is a suspension of bacteriophage T4 with concentration of 5×10³ copies of nucleotide sequences per 1 mcl, and for a positive control sample – a mixture containing fragments of mutton genomes (Ovis), beef (Bos) and bacteriophage T4 taken in ratio 1:1:1 with the following nucleotide sequences: Ovis F GCCTCATCTCCCTCCAACAG forward primer; Ovis R CGGAAGCCTGTAATTACAGCTC reverse primer; Ovis P R6G-CTCATGTCTGTCCTTTGGTGTTATGAATGC-BHQ1 probe; Bos F AACAGCATCATTCTACCCACTT forward primer; Bos R ACCTAAATTCCTATTCTAACACTG reverse primer; Bos P ROX-ACGACTTACATACTCCACTGCACTCACG-BHQ2 probe; T4F TACATATAAATCACGCAAAGC; T4R TAGTATGGCTAATCTTATTGG; T4P CY5 ACATTGGCACTGACCGAGTTC.

EFFECT: invention makes it possible to increase accuracy of beef or mutton species identity identification.

1 cl, 5 dwg, 5 tbl