METHOD FOR DIFFERENTIATING STRAINS OF PSEUDOMONAS AERUGINOSA USING MOLECULAR GENETIC TYPING Russian patent published in 2024 - IPC C12Q1/68 C12N15/10 

FIELD: medicine; microbiology.

SUBSTANCE: invention refers to medical microbiology, namely to methods of molecular-genetic typing of Pseudomonas aeruginosa strains, which can be used in epidemiological diagnostics of nosocomial purulent-septic, nosocomial infections on the basis of intraspecific typing of agent. Method for differentiating Pseudomonas aeruginosa strains using INDEL-typing involves DNA extraction from the analysed P. aeruginosa strain, performing PCR with specific primers and recording the reaction using electrophoresis. From the DNA of the analysed strain, four common INDEL genes having deletions of a certain size are detected, namely IND-PA0168, PA3324, PA0659 and PA1451, with their subsequent amplification using designed specific primers, detecting two alternative alleles: to gene IND-PA0168 — primers AGGACATGGACTGCGTGG and CCTGCTGCAGGCACTTGG with a length of the amplified fragment of 100 bp or 114 bp; to gene IND-PA3324 — primers GAGTCCCGGATGTGGTAGTC and GATCACGCTCCACAGGTTGA with a length of the amplified fragment of 99 bp or 113 bp; to gene IND-PA0659 — primers GCGGCATCATCATCGAGAAG and CTGGGCGGTGTGTAGGGA with a length of the amplified fragment of 106 bp or 112 bp; to gene IND-PA1451 — primers CAATTTCAGCGCCGGCGG and CGAAGACGAACTGGACGAAC with a length of the amplified fragment of 100 bp or 109 bp. Differentiation results are recorded visually after electrophoresis in 8% polyacrylamide gel in the presence of a DNA molecular weight marker. Common or different origin of the analysed strains of P. aeruginosa is established by comparing the identified INDEL-genotypes with each other and with known genotypes of the identification table.

EFFECT: method enables reliable, fast and low cost intraspecific differentiation of Pseudomonas aeruginosa strains.

3 cl, 3 tbl, 2 ex