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Construction of specific primers for identification of Leuconostoc mesenteroides subspecies

Bacteria p. Leuconostoc is a technologically important group of lactic acid bacteria that is part of starter cultures for production of various dairy products. Two species are most important in the dairy industry: Leuconostoc lactis and Leuconostoc mesenteroides, which includes three subspecies: dextranicum, mesenteroides and cremoris. The main problem of identifying representatives of the p. Leuconostoc that these microorganisms can often be misidentified as enterococci or lactobacilli. In comparison with traditional methods of species detection, the establishment of species identity using PCR is characterized by universality, a deeper level of species differentiation, high reproducibility and reliability. The article presents the results of designing specific primers for Leuconostoc mesenteroides ssp. mesenteroides and Leuconostoc mesenteroides ssp. dextranicum. The specificity of developed primers was confirmed by in silico testing using available Leuconostoc mesenteroides genomic sequences, and experimentally using DNA samples of Leuconostoc mesenteroides clear cultures. The taxonomic affiliation of 5 isolates of leuconostocci isolated from natural samples was established using the developed primers. Methodological Instructions have been developed that regulate the procedure for determining the taxonomic position of bacteria of genus Leuconostoc to a subspecies. Methodological guidelines for identification of leuconostocs will be used in collections of industrial microorganisms for the accurate identification of deposited strains.