METHOD OF DETECTING PHYLOGENETIC SUBLINES OF BEIJING MYCOBACTERIUM TUBERCULOSIS GENOTYPE IN REAL TIME FORMAT Russian patent published in 2021 - IPC C12Q1/686 C12Q1/689 G01N33/569 C12R1/32 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, specifically to a method of determining phylogenetic sublines of Beijing M. tuberculosis genotype. Simultaneous analysis of codon 58 in mutT2 gene and codon 48 in mutT4 gene is carried out by PCR in real-time format. Oligonucleotide primers are used: 5'-CGGTTGCACGGTCTTGGT, 5'-GCAATATCGTCGCCCACAC, 5'-ACAAGCCAAATCACGTCGAC, 5'-AACCCTCCAGCCGATGTTT, and fluorescent-labelled probes: FAM-5'-CTGGGACTCGAGGTCGC-RTQ1, HEX-5'-ACTGCGACTCGAGGTCG-BHQ2, ROX-5'-AGCGGTCCCGGTCC-BHQ2, Cy5-5'-AGCGGTCCGGGTCC-BHQ2. PCR results are evaluated between 15–35 cycles. When recording the exponential growth of the fluorescence signal on the HEX channel at wavelength of 555 nm and the Cy5 channel at wavelength of 670 nm, the strain is attributed to the modern subline of Beijing genotype M. tuberculosis, FAM-channel with wavelength of 510 nm and Cy5-channel with wave length of 670 nm indicates membership of strain to ancient subline, according to FAM-channel with wavelength of 510 nm and ROX-channel with wavelength of 600 nm, it is determined whether strain belongs to early ancient subline.

EFFECT: method enables fast and reliable detection of subline genotype Beijing Mycobacterium tuberculosis, analysis of large collections of Mycobacterium tuberculosis strains.

1 cl, 3 dwg, 3 ex