METHOD FOR IDENTIFICATION OF DNA TISSUE OF COMMON QUAIL (COTURNIX COTURNIX) IN DRY FODDER AND SEMI-FINISHED MEAT PRODUCTS

FIELD: veterinary microbiology.SUBSTANCE: invention relates to methods for determining species identity of meat by polymerase chain reaction. DNA is recovered by sorption method from common quail (Coturnix coturnix) tissue. Polymerase chain reaction with fluorescent detection is carried out with 45 amplification cycles in real time using quail DNA genome-specific oligonucleotide primers, probes, fluorescent dyes: for a specific signal for quail-JOE/Yellow and Cy5/Red-for an internal control sample in form of suspension of bacteriophage T4 with concentration of 5×10phage particles per 1 mcl, positive control sample in the form of a mixture containing fragments of quail genomes and bacteriophage T4 with a nucleotide sequence: T4F TACATATAAATCACGCAAAGCT4R TAGTATGGCTAATCTTATTGGT4P CY5 ACATTGGCACTGACCGAGTTC, taken in ratio 1:1. Accumulation of fluorescent signals through channels of corresponding fluorescent dyes is measured. Results are interpreted based on presence or absence of intersection of the fluorescence curve with the 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. For internal control sample phage lysate of bacteriophage T4 is used, and for positive control sample-fragments of genomes of native bacteriophage T4 and common quail tissue (Coturnix coturnix) with the following nucleotide sequence: C.cot-F: 5'-CTAGGAGGCGTACTTGC-3'-forward primer C.cot-R: 5'-GTGGGCGGAATGTTATG-3'-reverse primer C.cot-Z: 5'-CAGTACTTATCCTCCTTCTAATCC-3' is a probe.EFFECT: method increases accuracy of identification of species identity, simplifies the process.1 cl, 1 ex, 5 tbl Изобретение относится к ветеринарной микробиологии, в частности к методам определения видовой принадлежности мяса с помощью полимеразной цепной реакции. Сорбционным методом из ткани перепелки обыкновенной (Coturnix coturnix) выделяют ДНК. Проводят полимеразную цепную реакцию с флуоресцентной детекцией с проведением 45 циклов амплификации в реальном времени с использованием специфичных для участка генома ДНК перепелки олигонуклеотидных праймеров, зондов, флуоресцентных красителей: для специфического сигнала для перепелки - JOE/Yellow и Cy5/Red - для внутреннего контрольного образца в виде суспензии бактериофага Т4 с концентрацией 5×10фаговых частиц на 1 мкл, положительного контрольного образца в виде смеси, содержащей