METHOD FOR DETECTING DNA OF CHLOMYDIA IN FARM ANIMALS AND BIRDS Russian patent published in 2019 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention represents a method for detecting DNA of chlamydia in farm animals and birds, involving chlamydia DNA extraction by Chlamydia spp. from biological material from infected animals by sorption method, polymerase chain reaction – with addition of internal positive control and 45 amplification cycles with fluorescent detection in real time using chlamydia specific for Chlamydia spp. oligonucleotide primers, fluorescent-labeled probe, positive and negative control samples, 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 additionally taken from the infected birds, wherein for the internal control sample a bacteriophage T4 suspension 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 DNA genome of chlamydia DNA 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 specific chlamydia DNA signal Chlamydia spp; FAM/Green for internal control sample signal, if fluorescence signal accumulation curves are up to 35 cycles, 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: invention provides more accurate diagnosis of the presence of chlamydia DNA exciter and broader functional capabilities.

1 cl, 3 dwg, 4 tbl