METHOD FOR DETECTING DNA OF AVNITHOSIS AGENT (CHLAMYDOPHILA PSITTACI) IN BIRDS Russian patent published in 2019 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention is a method for detecting DNA of an ornithosis agent (Chlamydophila psittaci) in birds, involving recovering DNA from a biological material by a sorption method, polymerase chain reaction – with addition of internal positive control and successively carrying out no more than 40 amplification cycles with fluorescent detection in real time using oligonucleotide primers, fluorescent-labeled probe, positive and negative control samples of DNA-specific DNA orbital pathogen (Chlamydophila psittaci) specific DNA segment , measuring accumulation of a fluorescent signal through channels of corresponding fluorescent dyes for a specific signal and an internal control signal and interpreting the results based on the presence/absence of crossing the fluorescence curve with the threshold line, according to the invention, the biological material is collected from infected birds, 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 of recombinant plasmid DNA, containing a fragment of the genome of the DNA exciter ornithosis (Chlamydophila psittaci) and a fragment of the bacteriophage T4 genome, taken in ratio of 1:1, wherein accumulation of fluorescent signal is measured in channels: JOE(HEX)/Yellow for the specific DNA signal of the ornithosis agent (Chlamydophila psittaci) and FAM/Green for the internal control signal, if the accumulation curves of the fluorescent signal are up to 35 cycles, the DNA of the ornithosis agent (Chlamydophila psittaci) is present and the result of the reaction is considered positive, and if the curves do not cross the threshold line or cross it after 35 cycles, then the DNA of the ornithosis agent (Chlamydophilapsittaci) is absent and the result of the reaction – negative.

EFFECT: invention enables authentically diagnosing an ornithosis DNA exciter (Chlamydophila psittaci) in birds.

1 cl, 3 dwg, 4 tbl