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Revista de Salud Animal

Print version ISSN 0253-570XOn-line version ISSN 2224-4700


MARTINEZ-GARCIA, Yuneilys Aleli et al. Evaluation of a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk. Rev Salud Anim. [online]. 2018, vol.40, n.2, e03. ISSN 0253-570X.

Infectious and toxigenic pathogens transmitted through food have been recognized as the cause of Foodborne Diseases (FBD). The main pathogens of concern are Salmonella spp. and Staphylococcus aureus. These bacteria are associated with acute gastrointestinal outbreaks caused by contaminated food. One of the causes of this contamination is the poor hygiene during the processing and handling of food. Foodborne outbreaks caused by milk and dairy products contaminated reach up to 6 %. The objective of the present work was to develop and standardize a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk. The principle of the method is the simultaneous amplification of the genes yfiR for Salmonella spp. and ARNr 23S for Staphylococcus aureus. The thermodynamic properties of each primer were determined and the main critical parameters of the dPCR assay were optimized. The duplex PCR demonstrated its specificity for the target microorganisms and the detection limit of the assay was up to 1 pg/μL of DNA and 102 cfu/mL in unpasteurized milk. In the milk samples analyzed, 46.7 % were contaminated with S. aureus and none of the samples were positive to Salmonella spp. The results suggest that the developed duplex PCR is sensible and specific for the simultaneous detection of both pathogens from raw milk.

Keywords : Salmonella spp.; Staphylococcus aureus; duplex PCR; raw milk.

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