To solve one of the most important tasks of food hygiene - ensuring foodstuff biosafety, the leading place is given to the use of nucleic acid sequencing methods that allow to identify and describe the nucleotide sequences of genomes both in individual microorganisms and metagenomic characteristics of microbial communities in environmental objects, in humans and animals. The purpose of the review was to analyze the main areas of sequencing application in food microbiology, biotechnology and epidemiology. Material and methods. The collection and analysis of scientific and informational materials published in domestic and foreign publications from Scopus, Web of Science, PubMed, RSCI databases, official reports of the World Health Organization (WHO), the European Food Safety Agency (EFSA) and other published sources has been carried out. Results. The review presents generalized characteristics of the most well-known sequencing formats based on different principles of reading and processing genetic information: capillary sequencing by Sanger, pyrosequencing, sequencing by ligation, sequencing by DNA synthesis, semiconductor high-performance sequencing, sequencing of individual DNA molecules. It is shown that multilocus analysis (MLST, MLVA) increases the specificity of genotyping and intraspecific identification of food-borne pathogens. It is used to assess the pathogenic potential and study the mechanisms of evolutionary variability of bacterial pathogens, leading to polymorphism of clonal lines and the appearance of strain-specific differences, to monitor the different genotypes of pathogens in production and other objects of the food chain. Nanopore sequencing is promising for studying bacterial plasmids and localization of resistance genes of MDR bacteria. Genome-wide analysis (WGS) makes it possible to identify detailed characteristics of pathogens of infectious diseases, including uncultivated and previously unknown microorganisms, as well as to study the features of the organization of individual pro- and eukaryotic genomes, gene regulation systems, pathogenicity factors and protein expression. The role of WGS methods for determining the degree of genetic kinship of infectious agents isolated from various sources in the investigation of outbreaks and sporadic food poisoning and infections is shown. The possibilities of metagenomic sequencing in the study of species composition, genetic diversity and metabolic potential of microbial communities are demonstrated. The main advantage of metagenomics is the possibility of direct sequencing of the genomes of microorganisms living in the human body, animals, water, soil and other objects. Metagenomic analysis plays a leading role in the detection of unexplored taxa, uncultivated and difficult-to-cultivate forms of a wide range of microorganisms. The most important direction of metagenomics is the study of the human or animal microbiome, the analysis of the relationship between the intestinal microbiota and metabolism, as well as the formation of the nutritional status of a person. Theoretical and practical aspects of the implementation of high-performance sequencing methods in the system of food biosafety assessment and control are reviewed: safety assessment of genetically modified strains of food producers, development of fundamentally new algorithms for investigating outbreaks of infectious diseases, creation of network systems for early diagnosis and notification of food infections of bacterial and viral etiology. Conclusion. Sequencing should become a standard methodology in the field of food safety for the identification and characterization of food pathogens, including those with resistance to antibiotics and adverse technological effects. The introduction of WGS into the food safety assessment and control system will make it possible to assess real food contamination with new and newly returning pathogens, create electronic databases and scientifically substantiate the most adequate measures for the prevention of food infections and ensuring the health of the population.