FIELD: biotechnology; molecular biology; medical microbiology.
SUBSTANCE: set of reagents for the detection of the plague microbe is proposed. The kit includes magF19 immunomagnetic particles — conjugates of monoclonal antibodies to Y. pestis F19 surface antigens with nanomagnetic particles of 200 nm in size — for magnetically controlled separation of bacteria of the Yersinia pestis species in samples, and a set of oligonucleotide primers for the detection of bacteria of the Yersinia pestis species by loop isothermal amplification, containing oligonucleotides with the sequences of SEQ ID 2, SEQ ID 3, SEQ ID 4, SEQ ID 5, SEQ ID 6. The following is proposed: a method of the detection of a plague microbe, in which, at the first stage, magnetically controlled separation of bacteria of the Yersinia pestis species in samples using a set of immunomagnetic particles magF19 is carried out, at the second stage bacteria of the Yersinia pestis species are detected using loop isothermal amplification with the help of the set of reagents for the detection of the plague microbe. Synthesis and accumulation of target gene fragments is carried out at 63°C for a maximum of 40 min. Determination of the target gene is carried out by staining the products of the amplification reaction with the intercalating dye SYT082.
EFFECT: claimed invention allows for 1–2 hours with high specificity to detect the presence of Yersinia pestis plague pathogen in samples of pure cultures at concentrations from 10 to 1,000 mc. in ml depending on strains.
2 cl, 3 dwg, 3 tbl, 5 ex
