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Lactococcus lactis  subsp.  cremoris  belongs to lactic acid bacteria that play a crucial role in cheese production and it is known to be beneficial to human health. The aim of the study was to establish a rapid and accurate quantitative real-time polymerase chain reaction (qPCR) method to detect and enumerate  L. lactis  subsp.  cremoris  in artisanal raw cow’s milk cheese. Artisanal raw cow’s milk cheese samples were used to check for presence and number of  L. lactis  subsp.  cremoris  strains. The method applies a set of target-specific PCR (polymerase chain reaction) primers and a fluorogenic probe, and amplifies a part of the LACR_RS01280 gene that encodes the aminoacetone oxidase family flavin adenine dinucleotide (FAD) binding enzyme. All 5  L. lactis  subsp.  cremoris  strains examined were found to be qPCR positive. There was no signal recorded for 8 strains which belong to closely related species. The limit of detection amounted to ten copies per reaction and the assay indicated a linear dynamic range of seven logs. This method may be applied in detection and enumeration of  L. lactis  subsp.  cremoris  in cheese during its ripening. Moreover, it may be applied to examine the distribution of  L. lactis  subsp.  cremoris  during the cheese production and ripening.

Quantitative assessment of Lactococcus lactis subsp. cremoris present in artisanal raw cow’s milk cheese