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ČSN EN ISO 6579-4 This document specifies a horizontal in vitro method for the molecular identification and differentiation of the monophasic variant of Salmonella enterica subsp. enterica serovar Typhimurium (1,4,[5],12:i:-) lacking the second H phase H:1,2, starting from isolates. The method detects specific DNA sequences of an intergenic region of the first H phase flagellin cluster for identification of Salmonella enterica subsp. enterica serovar Typhimurium (further called Salmonella Typhimurium) and specific DNA sequences of genes associated with second H phase flagellar antigen expression. The method is applicable for: - differentiation of the isolate under analysis between monophasic Salmonella Typhimurium and the monophasic variant of another Salmonella on-Typhimurium serovar that has the same antigenic formula; - identification of the isolate under analysis being either monophasic Salmonella Typhimurium or (biphasic) Salmonella Typhimurium. This document is applicable for the analysis of a pure culture belonging to the genus Salmonella, isolated from: - products intended for human consumption; - products intended for animal feeding; - environmental samples in the area of food and feed production and handling; - samples from the primary production stage. This document can also be applied in other domains for identification of monophasic Salmonella Typhimurium (e.g. environmental, human health, animal health).