SET OF OLIGONUCLEOTIDE PRIMERS AND FLUORESCENT-LABELLED PROBES AND METHOD FOR DETECTING DNA OF AGENTS OF GLANDERS AND MELIOIDOSIS BY PCR METHOD WITH PRODUCT DETECTION IN REAL TIME Russian patent published in 2020 - IPC C12N1/20 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. Disclosed is a set of oligonucleotide primers and fluorescent-labeled probes for PCR-PB, which enables simultaneous detection of bacterial specific B. mallei DNA fragments of the target gene, having the nucleotide sequence SEQ ID No: 1, and bacteria specific B. pseudomallei DNA fragments of the target gene having the nucleotide sequence SEQ ID No: 2. For amplification of specific B. mallei PCR-PB DNA fragments, a set of specific oligonucleotides includes: a direct primer - "BmF" having a nucleotide sequence SEQ ID No: 3, a reverse primer - "BmR", having nucleotide sequence SEQ ID No: 4, fluorescent-labeled probe - "Bm_FAM" having nucleotide sequence SEQ ID No: 7. For amplification of specific DNA fragments of B. pseudomallei PCR-RV set of oligonucleotides includes: direct primer - "BpF", having nucleotide sequence SEQ ID No: 5, reverse primer - "BpR" having nucleotide sequence SEQ ID No: 6, fluorescent-labeled probe - "Bp_Cu5", having nucleotide sequence SEQ Sh№: 8. To detect B. mallei and B. pseudomallei DNA, a reaction is carried out using two pairs of oligonucleotide primers, homologous to fragments of SEQ ID NO: 1 V. mallei and SEQ ID No: 2 B. pseudomallei, as well as two corresponding oligonucleotide TaqMan probes with fluorescent dyes differing by the absorption and emission spectrum, which enables to detect signals from different targets.

EFFECT: present invention enables simultaneous detection of a Burkholderia mallei excipient and a Buriholderia pseudomallei exciter in high-specificity for 2 hours in pure culture samples in concentration of 100 microbial cells/ml.

2 cl, 5 dwg, 2 tbl, 3 ex